WorldWideScience

Sample records for human ejaculated spermatozoa

  1. STUDY OF ESTROGEN BINDING SITE ON HUMAN EJACULATED SPERMATOZOA

    Institute of Scientific and Technical Information of China (English)

    CHUJin-Shong; WANGYi-Fei

    1989-01-01

    The specific estrogen binding site for 17β-estradiol has been investigated on human spermatozoa by electron microscopec autoradiography. The results show that the binding sites were distributed over the surface of human spermatozoa: acrosomal cap, equatorial

  2. Arguments raised by the recent discovery that insulin and leptin are expressed in and secreted by human ejaculated spermatozoa.

    Science.gov (United States)

    Andò, Sebastiano; Aquila, Saveria

    2005-12-21

    The recent findings demonstrating that insulin and leptin are expressed in and secreted by human ejaculated spermatozoa raise the controversial issue related to mRNA function in male gamete. Capacitated sperm display an increased metabolism and overall energy expenditure presumably to affect the changes in sperm signaling and function during capacitation. However the relationship between the signaling events associated with capacitation and the change in sperm metabolism energy is poorly understood. It emerges from the findings here reported that both leptin and insulin may be crucial in ejaculated spermatozoa to manage their energy status. Immunoistochemical analysis revealed that in uncapacitated sperm insulin was located at the subacrosomial level, in the midpiece and through the tail while leptin was immunodetected at the equatorial segment and at the midpiece. Capacitated sperm display an overall decrease and a more uniform distribution in the signal for both hormones and this is in agreement with their enhanced release in the medium. Both hormones in ejaculated sperm somehow recapitulate the cross-talk between their signalling transductional pathways in somatic cells, resulting in the increase of phosphoinositide 3-kinase (PI3K) activity, AKT S473 and Glycogen synthase kinase 3 (GSK-3)-S9 phosphorylations. During capacitation GSK-3 phosphorylation was abolished suggesting how in capacitating sperm there is a block in glycogen synthesis. This reasonably indicates how during capacitation glycogen reserve is mobilized and this makes the glucose as energy substrate available. For instance insulin dismissed by ejaculated spermatozoa up-regulates Glucose 6-Phosphate Dehydrogenase (G6PDH), the rate-limiting enzyme in the pentose phosphate pathway (PPP), which has be shown to be crucial in the acquisition of fertilizing capability as well as to mediate gamete fusion. Insulin immunoneutralization or blockage of its release, dramatically down regulated G6PDH

  3. EFFECT OF SPERM CONCENTRATION ON EJACULATE FOR MORPHOMETRIC TRAITS OF SPERMATOZOAS OF THE PIETRAIN BREED BOARS

    Directory of Open Access Journals (Sweden)

    Dorota BANASZEWSKA

    2010-06-01

    Full Text Available An attempt to evaluate the effect of spermatozoa concentration in one ejaculate on their measurements, shape, frequency of occurrence of morphological abnormalities in spermatozoa and physical traits of boar ejaculates in Pietrain breed was made. It was concluded that there was a slight dependence between the content of spermatozoa in one ejaculate and morphometrical traits of spermatozoa. In semen with lower content of spermatozoa (I and II group, the spermatozoa had slightly longer heads (by about 0.18 μm than in semen with large spermatozoa concentration (III group. Spermatozoa in ejaculates with the lowest spermatozoa concentration were characterized by the longest flagellum and the largest total length. The total length of spermatozoa was decreasing in groups of larger concentration, which was caused by both lower length of heads and flagella. Some differences in spermatozoa shape in relation to their concentration in one ejaculate were found. Spermatozoa in ejaculates, which were classified into II group, seemed to have less prolate shape than spermatozoa in ejaculates of I and III groups. It was stated that the content of spermatozoa in one ejaculate affected the frequency of spermatozoa with morphological changes. Semen assigned to II group was distinguished by the best quality.

  4. Birth after intracytoplasmic sperm injection of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome

    Institute of Scientific and Technical Information of China (English)

    Takuya Akashi; Hideki Fuse; Yasuo Kojima; Mikiko Hayashi; Sachiko Honda

    2005-01-01

    Aim: To report a birth after intracytoplasmic sperm injection (ICSI) of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome detected by fluorescence in situ hybridization (FISH) analysis. Methods: A 35-yearold man with a normal appearance consulted our hospital because of sterility over a 5-year period. Chromosome analysis showed low-incidence mosaic Klinefelter's syndrome. Using FISH, 96% hyperploidy of the lymphocytes was found. We examined the sex chromosome of the ejaculated spermatozoa. Using FISH, we examined 200 ejaculated spermatozoa and no hyperploidy was found. Results: The 33-year-old female partner of the male patient underwent an uncomplicated controlled ovarian hyperstimulation sequence using a combined recombinant-follicle stimulating hormone (rec-FSH) + human menopausal gonadotrophin (hMG) protocol, following late luteal phase pituitary down regulation. This culminated in the retrieval of seven oocytes, six of which were fertilized with ICSI.One ICSI attempt led to clinical pregnancy with a healthy baby girl. Conclusion: We report a male patient with lowincidence mosaic Klinefelter's syndrome whose ejaculated spermatozoa were identified as being haploid by FISH before ICSI, leading to the successful pregnancy of his wife and the birth of a healthy baby girl.

  5. Effect of sperm concentration in an ejaculate on morphometric traits of spermatozoa in Duroc boars.

    Science.gov (United States)

    Kondracki, S; Wysokińska, A; Iwanina, M; Banaszewska, D; Sitarz, D

    2011-01-01

    The experimental material consisted of 75 ejaculates collected form 8 Duroc boars. The ejaculates were divided into three groups according to sperm concentration in an ejaculate. An ejaculate was obtained from each boar monthly and it was used to make microscopic preparations to examine spermatozoa morphology. In each preparation morphometric measurements were taken of fifteen randomly selected spermatozoa characterized by normal morphology. The following measurements of spermatozoa were taken: length and width of the spermatozoa head, head area, length of the flagellum, perimeter of the spermatozoon head and total spermatozoon length. The results were used to calculate indicators of spermatozoa morphology. Moreover, assessments were made of frequency of morphological defects to isolate spermatozoa with primary and secondary abnormalities following the Blom classification system. It was found that the concentration of spermatozoa in the ejaculate influenced the morphometric characteristics of spermatozoa. Ejaculates with low sperm concentrations are characterized by larger spermatozoa as compared to ejaculates with high sperm concentrations. However, sperm concentration in the ejaculate does not much influence the shape of spermatozoa.

  6. Detrimental effects of non-functional spermatozoa on the freezability of functional spermatozoa from boar ejaculate.

    Directory of Open Access Journals (Sweden)

    Maria J Martinez-Alborcia

    Full Text Available In the present study, the impact of non-functional spermatozoa on the cryopreservation success of functional boar spermatozoa was evaluated. Fifteen sperm-rich ejaculate fractions collected from five fertile boars were frozen with different proportions of induced non-functional sperm (0--native semen sample-, 25, 50 and 75% non-functional spermatozoa. After thawing, the recovery of motile and viable spermatozoa was assessed, and the functional of the spermatozoa was evaluated from plasma membrane fluidity and intracellular reactive oxygen species (ROS generation upon exposure to capacitation conditions. In addition, the lipid peroxidation of the plasma membrane was assessed by the indirect measurement of malondialdehyde (MDA generation. The normalized (with respect to a native semen sample sperm motility (assessed by CASA and viability (cytometrically assessed after staining with Hoechst 33342, propidium iodide and fluorescein-conjugated peanut agglutinin decreased (p<0.01 as the proportion of functional spermatozoa in the semen samples before freezing decreased, irrespective of the semen donor. However, the magnitude of the effect differed (p<0.01 among boars. Moreover, semen samples with the largest non-functional sperm subpopulation before freezing showed the highest (p<0.01 levels of MDA after thawing. The thawed viable spermatozoa of semen samples with a high proportion of non-functional spermatozoa before freezing were also functionally different from those of samples with a low proportion of non-functional spermatozoa. These differences consisted of higher (p<0.01 levels of intracellular ROS generation (assessed with 5-(and-6 chloromethyl-20,70-dichlorodihydrofluorescein diacetate acetyl ester; CM-H(2DCFDA and increased (p<0.01 membrane fluidity (assessed with Merocyanine 540. These findings indicate that non-functional spermatozoa in the semen samples before freezing negatively influence the freezability of functional spermatozoa.

  7. Correlation of frequency of spermatozoa morphological alterations with sperm concentration in ejaculates of Polish Landrace boars

    Directory of Open Access Journals (Sweden)

    Kondracki S.

    2013-01-01

    Full Text Available The experiments were performed on 448 ejaculates obtained from 41 Polish Landrace boars. Ejaculates collected from each boar at one-month intervals for approximately 10 months were analysed. Sperm morphometric measurements were taken from each boar and assessment of semen morphology was done on the basis of examination under a microscope of preparations made from fresh ejaculates. The ejaculates were classified based on the criterion of sperm concentration and divided into three groups. An attempt was made in the present study to assess the correlation of ejaculate parameters, morphological sperm alteration incidence and morphometric sperm parameters with the sperm concentration in ejaculates of Polish Landrace boars. It should be stated that morphometric traits of spermatozoa are related to sperm concentration. The spermatozoa in concentrated ejaculates had smaller heads than the spermatozoa in the ejaculates with lower sperm concentrations. This can mean that the high fertility of males that produce highly concentrated semen does not only result from a high sperm concentration, but also from the fact that the spermatozoa in such ejaculates have smaller heads. The highest frequency of morphologically well-formed spermatozoa was identified in ejaculates with the sperm concentration ranging from 400 to 500 thousand/mm3.

  8. A comparison of ejaculated and testicular spermatozoa aneuploidy rates in patients with high sperm DNA damage.

    Science.gov (United States)

    Moskovtsev, Sergey I; Alladin, Naazish; Lo, Kirk C; Jarvi, Keith; Mullen, J Brendan M; Librach, Clifford L

    2012-06-01

    Testicular spermatozoa are utilized to achieve pregnancy in couples with severe male factor infertility. Several studies suggest that aneuploidy rates in spermatozoa are elevated at the testicular level in infertile patients compared to ejaculates of normal controls. However, essential data regarding aneuploidy rates between ejaculated and testicular spermatozoa in the same individuals is lacking. The purpose of our study was to compare aneuploidy rates at the testicular and post-testicular level from the same patients with persistently high sperm DNA damage. Ejaculates and testicular biopsies were obtained from eight patients with persistently high DNA damage (>30%). Both ejaculated and testicular samples were analyzed for sperm DNA damage and sperm aneuploidy for chromosomes 13, 18, 21, X, and Y. In addition, semen samples from ten normozoospermic men presenting for fertility evaluation served as a control group. A strong correlation between the alteration of spermatogenesis and chromatin deterioration was observed in our study. In the same individuals, testicular samples showed a significantly lower DNA damage compared to ejaculated spermatozoa (14.9% ± 5.0 vs. 40.6% ± 14.8, P<0.05), but significantly higher aneuploidy rates for the five analyzed chromosomes (12.41% ± 3.7 vs. 5.77% ± 1.2, P<0.05). While testicular spermatozoa appear favourable for ICSI in terms of lower DNA damage, this potential advantage could be offset by the higher aneuploidy rates in testicular spermatozoa.

  9. Comparison of methods of extracting messenger Ribonucleic Acid from ejaculated Porcine (Sus Scrofa) Spermatozoa

    Science.gov (United States)

    H. D. Guthrie, G.R. Welch, and L. A. Blomberg. Comparison of Methods of Extracting Messenger Ribonucleic Acid from Ejaculated Porcine (Sus Scrofa) Spermatozoa. Biotechnology and Germplasm Laboratory, Agricultural Research Service U. S. Department of Agriculture, Beltsville, MD 20705 The purpos...

  10. Sperm treatment affects capacitation parameters and penetration ability of ejaculated and epididymal boar spermatozoa.

    Science.gov (United States)

    Matás, C; Sansegundo, M; Ruiz, S; García-Vázquez, F A; Gadea, J; Romar, R; Coy, P

    2010-11-01

    This work was designed to study how this ability is affected by different sperm treatments routinely used for in vitro fertilization (IVF) assay. In this study, boar sperm samples from epididymal or ejaculated origin were processed by three different methods: left unwashed (NW group), washed in Dulbecco's phosphate-buffered saline supplemented with 0.1% BSA (BSA group), and washed on a Percoll(®) gradient (PERCOLL group). After preparation of semen samples, changes in motility patterns were studied by CASA, calcium uptake by spectrofluorimetry, and ROS generation, spontaneous acrosome reaction, and lipid disorder by means of flow cytometry. Finally IVF assays were also performed with the different semen samples and penetrability results evaluated at 2 and 4 h post insemination (hpi). Independently of the sperm treatment, epididymal spermatozoa showed higher values of progressive motility, percentage of live cells with low lipid disorder, and penetration ability at 4 hpi than the corresponding ejaculated spermatozoa. Ejaculated spermatozoa showed higher levels of calcium uptake, ROS generation and percentage of spontaneous acrosome reaction than epididymal sperm. Regarding sperm treatments, PERCOLL group showed the highest values for some motility parameters (linearity of the curvilinear trajectory, straightness, and average path velocity/curvilinear velocity), ROS generation and penetration ability at 2 and 4 hpi; however this same group showed the lowest values for sperm curvilinear velocity and lateral head displacement. From all experimental groups, ejaculated-PERCOLL-treated spermatozoa showed the highest fertilization ability after 2 hpi. Results suggest that capacitation pathways can be regulated by suitable treatments making the ejaculated sperm able to reach capacitation and fertilize oocytes in similar levels than epididymal spermatozoa, although most of the studied capacitation-associated changes do not correlate with this ability.

  11. Comparison of intracytoplasmic sperm injection outcomes between spermatozoa retrieved from testicular biopsy and from ejaculation in cryptozoospermic men.

    Science.gov (United States)

    Amirjannati, N; Heidari-Vala, H; Akhondi, M A; Hosseini Jadda, S H; Kamali, K; Sadeghi, M R

    2012-05-01

    The infrequent presence of spermatozoa in cryptozoospermic men ejaculate is a limiting factor in the treatment of them. Sometimes, this consideration impels us to apply meticulous microscopic search in ejaculate or testicular sperm extraction (TESE) method. The aim of this study was to assess putative effectiveness of sperm origin, ejaculated or testicular, in cryptozoospermia treatment. In this context, were evaluated intracytoplasmic sperm injection (ICSI) outcomes in two parameters including fertilisation rate (2PN) and embryo quality, independently. We compared the outcome in two groups: patients who underwent ejaculate/ICSI and ones who underwent TESE/ICSI process. Nineteen ICSI cycles performed with testicular spermatozoa and the rest of cycles (n = 208) carried out with ejaculated spermatozoa. Result analysis showed similar fertilisation rate between testicular and ejaculated spermatozoa (respectively, 60% versus 68%, P ≥ 0.05). Also, on the other hand, embryo quality did not show significant differences between two groups, except grade A with low significance. With regard to almost equal performance of both methods in results and being invasive of TESE as surgical sperm retrieval method, the use of ejaculated sperm more than testicular sperm should be recommended in patients with cryptozoospermia whenever possible.

  12. Brain activation during human male ejaculation

    NARCIS (Netherlands)

    Holstege, Ger; Georgiadis, Janniko R.; Paans, Anne M.J.; Meiners, Linda C.; Graaf, Ferdinand H.C.E. van der; Reinders, A.A.T.Simone

    2003-01-01

    Brain mechanisms that control human sexual behavior in general, and ejaculation in particular, are poorly understood. We used positron emission tomography to measure increases in regional cerebral blood flow (rCBF) during ejaculation compared with sexual stimulation in heterosexual male volunteers.

  13. Cryopreservation-induced alterations in protein tyrosine phosphorylation of spermatozoa from different portions of the boar ejaculate.

    Science.gov (United States)

    Kumaresan, A; Siqueira, A P; Hossain, M S; Bergqvist, A S

    2011-12-01

    Previous studies have shown that boar sperm quality after cryopreservation differs depending on the ejaculate fraction used and that spermatozoa contained in the first 10mL (P1) of the sperm-rich fraction (SRF) show better cryosurvival than those in the SRF-P1. Since protein tyrosine phosphorylation (PTP) in spermatozoa is related with the tolerance of spermatozoa to frozen storage and cryocapacitation, we assessed the dynamics of cryopreservation-induced PTP and intracellular calcium ([Ca(2+)]i) in spermatozoa, using flow cytometry, from P1 and SRF-P1 of the boar ejaculate at different stages of cryopreservation. Sperm kinetics, assessed using a computer-assisted semen analyzer, did not differ between P1 and SRF-P1 during cryopreservation but the decrease in sperm velocity during cryopreservation was significant (Psperm PTP. The proportion of spermatozoa with PTP did not differ significantly between portions of the boar ejaculate. However at any given step during cryopreservation the percentage of spermatozoa with PTP was comparatively higher in SRF-P1 than P1. A 32kDa tyrosine phosphorylated protein, associated with capacitation, appeared after cooling suggesting that cooling induces capacitation-like changes in boar spermatozoa. In conclusion, the study has shown that the cryopreservation process induced PTP in spermatozoa and their proportions were similar between portions of SRF.

  14. Differential expression of mRNA aromatase in ejaculated spermatozoa from infertile men in relation to either asthenozoospermia or teratozoospermia.

    Science.gov (United States)

    Said, L; Saad, A; Carreau, S

    2014-03-01

    Oestrogen biosynthesis in ejaculated spermatozoa is an autonomous process, which may influence sperm functions. The purpose of this study was to evaluate the relationship between the expression of aromatase, sperm quality and seminal neutral α-glucosidase marker in semen of Tunisian infertile men: asthenozoospermia (A; n = 16), teratozoospermia (T; n = 12) and asthenoteratozoospermia (AT; n = 11) in comparison with 18 normozoospermic ones. Aromatase mRNA levels estimated by real-time PCR were reduced in groups T (52%) and AT (67%) compared to controls and inversely correlated with the percentage of normal forms. A higher coefficient of correlation was noted in presence of microcephaly or acrosome malformations (r = -0.64). The asthenozoospermic group was divided into two subgroups according to the relative amount of aromatase. The subgroup (A2) with higher aromatase transcript level was associated with an increased seminal pH, a decreased sperm viability, low sperm percentage motility and low neutral α-glucosidase semen levels. Our data highlight the involvement of aromatase in motility and morphology of spermatozoa. Thus, this enzyme could bring new insights about quality and fertilising capacity of human spermatozoa.

  15. The Enzymatic Antioxidant System of Human Spermatozoa

    Directory of Open Access Journals (Sweden)

    Cristian O’Flaherty

    2014-01-01

    Full Text Available The ejaculated spermatozoon, as an aerobic cell, must fight against toxic levels of reactive oxygen species (ROS generated by its own metabolism but also by other sources such as abnormal spermatozoa, chemicals and toxicants, or the presence of leukocytes in semen. Mammalian spermatozoa are extremely sensitive to oxidative stress, a condition occurring when there is a net increase in ROS levels within the cell. Opportunely, this specialized cell has a battery of antioxidant enzymes (superoxide dismutase, peroxiredoxins, thioredoxins, thioredoxins reductases, and glutathione s-transferases working in concert to assure normal sperm function. Any impairment of the antioxidant enzymatic activities will promote severe oxidative damage which is observed as plasma membrane lipid peroxidation, oxidation of structural proteins and enzymes, and oxidation of DNA bases that lead to abnormal sperm function. Altogether, these damages occurring in spermatozoa are associated with male infertility. The present review contains a description of the enzymatic antioxidant system of the human spermatozoon and a reevaluation of the role of its different components and highlights the necessity of sufficient supply of reducing agents (NADPH and reduced glutathione to guarantee normal sperm function.

  16. Sperm maturation in dogs: sperm profile and enzymatic antioxidant status in ejaculated and epididymal spermatozoa.

    Science.gov (United States)

    Angrimani, D S R; Lucio, C F; Veiga, G A L; Silva, L C G; Regazzi, F M; Nichi, M; Vannucchi, C I

    2014-09-01

    Spermatozoa become more susceptible to the attack of reactive oxygen species during maturation. To avoid oxidative damage, the epididymis must provide the necessary antioxidant protection. The aim of this study was to compare the canine sperm profile and the enzymatic antioxidant status of the ejaculated fractions and samples collected from the different segments of the epididymis (caput, corpus and cauda). Five adult dogs were used, and after 1-3 weeks, subsequently to bilateral orchiectomy and epididymal storage, sperm samples were collected from the different segments of the epididymis. Samples were evaluated for conventional microscopy and computer-assisted motility analysis: sperm plasma membrane permeability and the activity of the antioxidant enzymes catalase, glutathione peroxidase (GPx) and superoxide dismutase (SOD). Samples collected from the caput and corpus showed lower values for most of the motility variables evaluated, indicating different levels of immaturity. Catalase activity was observed only in ejaculated samples. Conversely, GPx activity was higher in the cauda epididymidis. Correlations were found between SOD and GPx and SOD and sperm motility in the epididymal cauda and corpus, highlighting the importance of the enzymes for the protection of spermatozoa during the transit along the epididymis.

  17. Antisperm Antibodies on the Surface of Spermatozoa before'Ejaculation from Vasectomized Men

    Institute of Scientific and Technical Information of China (English)

    文任乾; 李世勤; 王春湘; 王庆辉; 刘美意

    1997-01-01

    The antisperm antibodies (AsAbs) coated on spermatozoa of the proximal vas deferens (sperm before ejaculation, SBE) from 48 fertile men who were volunteers of vasectomy and 24 vasectomized men who asked for vasovasotomy,were determined by immunobead test (IBT) and sperm cervical mucus contact test (SCMC). The results showed that in fertile men there were no positive sam ples of SBE in IBT and SCMC. In vasectomized men positive samples of SBE were found in 79.4% for IgG, 38.2% for IgA and 35.5% for SCMC. The AsAbs on SBE could be found at the time of less than one year to more than 3 years after vasectomy. The AsAbs were still found on the semen samples at 1-3 months after vasovasotomy. Our results also indicated that the incidence of AsAbs on SBE from vasectomized men could not predict the levels of AsAbs on their ejaculated sperm after vasovasotomy. There was no significant correlation between the levels of AsAbs in serium before vasovasotomy and those on SBE from vasectomized men.

  18. Aneuploidy in human spermatozoa.

    Science.gov (United States)

    Templado, C; Vidal, F; Estop, A

    2011-01-01

    We reviewed the frequency and distribution of disomy in spermatozoa obtained by multicolor-FISH analysis on decondensed sperm nuclei in (a) healthy men, (b) fathers of aneuploid offspring of paternal origin and (c) individuals with Klinefelter syndrome and XYY males. In series of healthy men, disomy per autosome is approximately 0.1% but may range from 0.03 (chromosome 8) to 0.47 (chromosome 22). The great majority of authors find that chromosome 21 (0.18%) and the sex chromosomes (0.27%) have significantly elevated frequencies of disomy although these findings are not universal. The total disomy in FISH studies is 2.26% and the estimated aneuploidy (2× disomy) is 4.5%, more than double that seen in sperm karyotypes (1.8%). Increased disomy levels of low orders of magnitude have been reported in spermatozoa of some normal men (stable variants) and in men who have fathered children with Down, Turner and Klinefelter syndromes. These findings suggest that men with a moderately elevated aneuploidy rate may be at a higher risk of fathering paternally derived aneuploid pregnancies. Among lifestyle factors, smoking, alcohol and caffeine have been studied extensively but the compounding effects of the 3 are difficult to separate because they are common lifestyle behaviors. Increases in sex chromosome abnormalities, some autosomal disomies, and in the number of diploid spermatozoa are general features in 47,XXY and 47,XYY males. Aneuploidy of the sex chromosomes is more frequent than aneuploidy of any of the autosomes not only in normal control individuals, but also in patients with sex chromosome abnormalities and fathers of paternally derived Klinefelter, Turner and Down syndromes.

  19. Dense spermatozoa in stallion ejaculates contain lower concentrations of mRNAs encoding the sperm specific calcium channel 1, ornithine decarboxylase antizyme 3, aromatase, and estrogen receptor alpha than less dense spermatozoa.

    Science.gov (United States)

    Ing, N H; Forrest, D W; Love, C C; Varner, D D

    2014-07-15

    Stallions are unique among livestock in that, like men, they commonly receive medical treatment for subfertility. In both species, about 15% of individuals have normal semen parameters but are subfertile, indicating a need for novel analyses of spermatozoa function. One procedure for improving fertilizing capability of stallions and men is isolation of dense spermatozoa from an ejaculate for use in artificial insemination. In the current study, dense and less dense spermatozoa were purified by density gradient centrifugation from individual ejaculates from seven reproductively normal adult stallions. The RNA isolated from the spermatozoa seemed to be naturally fragmented to an average length of 250 bases, consistent with reports of spermatozoa RNA from other species. The DNAse treatment of RNA prepared from spermatozoa removed any genomic DNA contamination, as assessed by PCR with intron spanning primers for the protamine 1 (PRM1) gene. Concentrations of seven mRNAs in spermatozoa, correlated with the fertility of men and bulls, were quantified by reverse transcription polymerase chain reaction in dense and less dense spermatozoa. Concentrations of four mRNAs were two- to four-fold lower in dense spermatozoa compared with less dense spermatozoa: Encoding the spermatozoa-specific calcium channel (P 0.1). These results identify new differences in mRNA concentrations in populations of spermatozoa with dissimilar densities.

  20. Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy

    Directory of Open Access Journals (Sweden)

    Johanna L. Höög

    2015-11-01

    Full Text Available Human ejaculates contain extracellular vesicles (EVs, that to a large extent are considered to originate from the prostate gland, and are often denominated “prostasomes.” These EVs are important for human fertility, for example by promoting sperm motility and by inducing immune tolerance of the female immune system to the spermatozoa. So far, the EVs present in human ejaculate have not been studied in their native state, inside the seminal fluid without prior purification and isolation procedures. Using cryo-electron microscopy and tomography, we performed a comprehensive inventory of human ejaculate EVs. The sample was neither centrifuged, fixed, filtered or sectioned, nor were heavy metals added. Approximately 1,500 extracellular structures were imaged and categorized. The extracellular environment of human ejaculate was found to be diverse, with 5 major subcategories of EVs and 6 subcategories of extracellular membrane compartments, including lamellar bodies. Furthermore, 3 morphological features, including electron density, double membrane bilayers and coated surface, are described in all subcategories. This study reveals that the extracellular environment in human ejaculate is multifaceted. Several novel morphological EV subcategories are identified and clues to their cellular origin may be found in their morphology. This inventory is therefore important for developing future experimental approaches, and to interpret previously published data to understand the role of EVs for human male fertility.

  1. Epididymal and ejaculated cat spermatozoa are resistant to cold shock but egg yolk promotes sperm longevity during cold storage at 4 degrees C.

    Science.gov (United States)

    Hermansson, U; Axnér, E

    2007-04-15

    The aims were to evaluate the susceptibility of feline ejaculated and epididymal spermatozoa to cold shock and to evaluate the effect of egg yolk in the preservation extender. Ejaculated and epididymal spermatozoa from eight males were subjected to a slow (0.5 degrees C/min) or a fast (3 degrees C/min) cooling rate with controls kept in room temperature. Ejaculated and epididymal spermatozoa from another eight males were cooled in a plain Tris buffer (Tris) or in Tris with 20% egg yolk (EYT) and evaluated for 96 h. Subjective motility (MOT), plasma membrane integrity (PMI), and acrosome integrity (ACRI) were evaluated. Cooling did not induce sperm damage regarding PMI (P=0.6) or ACRI (P=0.19) and chilled spermatozoa had better overall MOT (P=0.046) than controls. EYT was better for MOT (P>0.05) from 48 h of cold storage than Tris. EYT was also better for overall ACRI (P0.05) for PMI or ACRI. Ejaculated spermatozoa had better overall MOT (Pegg yolk as there were no interactions (P>0.05) between source of spermatozoa and treatment (cooling or control) or between time, source and extender (P>0.05). In conclusion cat spermatozoa were tolerant to cold shock and egg yolk was beneficial for preservation of MOT and ACRI but not PMI.

  2. Differences in the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques.

    Science.gov (United States)

    Parrilla, Inma; del Olmo, David; Sijses, Laurien; Martinez-Alborcia, María J; Cuello, Cristina; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

    2012-05-01

    The present study aimed to evaluate the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques. Eighteen sperm-rich ejaculate samples from six boars (three per boar) were diluted in Beltsville Thawing Solution and split into three aliquots. The aliquots were (1) further diluted to 3×10(7) sperm/mL and stored as a liquid at 17°C for 72 h, (2) frozen-thawed (FT) at 1×10(9) sperm/mL using standard 0.5-mL straw protocols, or (3) sex-sorted with subsequent liquid storage (at 17°C for 6 h) or FT (2×10(7) sperm/mL using a standard 0.25-mL straw protocol). The sperm quality was evaluated based on total sperm motility (the CASA system), viability (plasma membrane integrity assessed using flow cytometry and the LIVE/DEAD Sperm Viability Kit), lipid peroxidation (assessed via indirect measurement of the generation of malondialdehyde (MDA) using the BIOXYTECH MDA-586 Assay Kit) and DNA fragmentation (sperm chromatin dispersion assessed using the Sperm-Sus-Halomax(®) test). Data were normalized to the values assessed for the fresh (for liquid-stored and FT samples) or the sorted semen samples (for liquid stored and the FT sorted spermatozoa). All of the four sperm-processing techniques affected sperm quality (Psperm and increased MDA generation and percentages of sperm with fragmented DNA. Significant (Pboar (effect of boars within each semen-processing technique) and intra-boar (effect of semen-processing techniques within each boar) differences were evident for all of the sperm quality parameters assessed, indicating differences in the ability of spermatozoa from individual boars to withstand the semen-processing techniques. These results are the first evidence that ejaculate spermatozoa from individual boars can respond in a boar-dependent manner to different semen-processing techniques.

  3. Direct binding of boar ejaculate and epididymal spermatozoa to porcine epididymal epithelial cells is also needed to maintain sperm survival in in vitro co-culture.

    Science.gov (United States)

    Yeste, Marc; Castillo-Martín, Míriam; Bonet, Sergi; Briz, Maria Dolors

    2012-04-01

    The aim of the present study was to compare the influence of cultured epididymal epithelial cells (EEC) from corpus, caput or cauda, oviductal epithelial cells (OEC) and non-reproductive epithelial cells (LLC-PK1) on function and survival of epididymal and ejaculated spermatozoa, in the latter case to determine whether such influence differed between morphologically normal and abnormal spermatozoa. For this purpose, either spermatozoa were directly co-cultured with EEC from caput, corpus, or cauda, OEC and LLC-PK1 cells (experiment 1) or a membrane-diffusible insert was included in these co-cultures (experiment 2). EEC cultured from the three epididymal regions did not differently affect the sperm parameters. Morphologically normal spermatozoa presented a higher ability to bind EEC, OEC, and LLC-PK1 than abnormal spermatozoa with cytoplasmic droplets or with tail/head malformations. Epididymal spermatozoa were more able to bind EEC during the first 24 h of co-culture, while ejaculated spermatozoa presented a higher capacity to bind OEC between 30 min and 3 h of co-incubation. In all cases, the ability to bind to epithelial cells was higher when they were co-cultured with EEC and OEC than with LLC-PK1. After 2 h of co-culture, the viability of epididymal spermatozoa was better maintained when they bound EEC than when they bound OEC. Conversely, the viability of ejaculated spermatozoa was better maintained when bound OEC than when bound EEC after 24 and 48 h of co-culture. Our work, apart from corroborating the involvement of morphologically normal spermatozoa in the formation of sperm reservoir, highlights the importance of direct contact spermatozoa-EEC in maintaining the sperm survival in in vitro co-culture, and also suggests that a specific binding between EEC and epididymal spermatozoa exists.

  4. In vitro inhibition of superoxide anion production and superoxide dismutase activity by zinc in human spermatozoa.

    Science.gov (United States)

    Gavella, M; Lipovac, V; Vucić, M; Sverko, V

    1999-08-01

    The in vitro effect of zinc on superoxide anion (O2-) generation and on SOD-like activity in spermatozoa of infertile men was investigated. The formation of superoxide anion was stimulated by NADPH and the level of superoxide anion was measured by the reduction of ferricytochrome c. Both Percoll-isolated (n = 14) and washed spermatozoa (n = 14) exposed to 1 mmol/L zinc (60 min, 37 degrees C), released less (p zinc-untreated spermatozoa. These results implicate a possible role for zinc as a scavenger of excessive superoxide anions produced by defective spermatozoa in semen after ejaculation. Additionally, zinc was found to dose-dependently inhibit superoxide dismutase (SOD)-like activity of spermatozoa in vitro. The inhibition of SOD-like activity by an equal concentration of zinc (1 mmol/L) was less pronounced in oligospermic (p zinc to inhibit SOD-like activity may be relevant to the physiological function of spermatozoa in fertilization. The evidence that zinc may elicit an inhibition of both superoxide anion production and SOD-like activity in human spermatozoa, indicate the existence of novel, zinc-related mechanism(s) involved in the oxidative events occurring after ejaculation, with a possible modulatory effect on germ cell function.

  5. Testicular versus ejaculated spermatozoa in ICSI cycles of normozoospermic men with high sperm DNA fragmentation and previous ART failures.

    Science.gov (United States)

    Pabuccu, E G; Caglar, G S; Tangal, S; Haliloglu, A H; Pabuccu, R

    2017-03-01

    As a part of male assessment, conventional sperm parameters including morphologic features have been dedicated as major factors influencing fertilisation and pregnancy rates in assisted reproductive technology (ART). Genomic integrity of spermatozoa has also been found to influence fertility prognosis, and hence, sperm DNA fragmentation index (DFI) has been adopted by many centres to document this entity. Despite several suggested approaches, there is lack of universal consensus on optimising fertility outcomes in males with high sperm DFI. In this context, the results from cycles using testicular spermatozoa (TESA) obtained by aspiration were compared with those of ejaculated spermatozoa (EJ) in normozoospermic subjects with high sperm DFI and previous ART failures. Clinical (41.9% versus 20%) and ongoing pregnancy rates (38.7% versus 15%) were significantly better and miscarriages were lower in TESA group when compared to EJ group. Sperm DFI should be a part of male partner's evaluation following unsuccessful ART attempts. When high DFI is detected (>30%), ICSI using testicular spermatozoa obtained by TESA seems an effective option particularly for those with repeated ART failures in terms of clinical, ongoing pregnancies and miscarriages even though conventional sperm parameters are within normal range.

  6. Apoptosis and DNA damage in human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    R John Aitken; Adam J Koppers

    2011-01-01

    DNA damage is frequently encountered in spermatozoa of subfertile males and is correlated with a range of adverse clinical outcomes including impaired fertilization, disrupted preimplantation embryonic development, increased rates of miscarriage and an enhanced risk of disease in the progeny. The etiology of DNA fragmentation in human spermatozoa is closely correlated with the appearance of oxidative base adducts and evidence of impaired spermiogenesis. We hypothesize that oxidative stress impedes spermiogenesis,resulting in the generation of spermatozoa with poorly remodelled chromatin. These defective cells have a tendency to default to an apoptotic pathway associated with motility loss, caspase activation, phosphatidylserine exteriorization and the activation of free radical generation by the mitochondria. The latter induces lipid peroxidation and oxidative DNA damage, which then leads to DNA fragmentation and cell death. The physical architecture of spermatozoa prevents any nucleases activated as a result of this apoptotic process from gaining access to the nuclear DNA and inducing its fragmentation. It is for this reason that a majority of the DNA damage encountered in human spermatozoa seems to be oxidative. Given the important role that oxidative stress seems to have in the etiology of DNA damage, there should be an important role for antioxidants in the treatment of this condition. If oxidative DNA damage in spermatozoa is providing a sensitive readout of systemic oxidative stress, the implications of these findings could stretch beyond our immediate goal of trying to minimize DNA damage in spermatozoa as a prelude to assisted conception therapy.

  7. Frozen-thawed spermatozoa from oligozoospermic ejaculates are susceptible to in situ DNA fragmentation in polyvinylpyrrolidone-based sperm-immobilization medium.

    Science.gov (United States)

    Salian, Sujit Raj; Kalthur, Guruprasad; Uppangala, Shubhashree; Kumar, Pratap; Adiga, Satish Kumar

    2012-08-01

    To elucidate the effect of sperm immobilization media that are and are not based on polyvinylpyrrolidone (PVP) on the DNA integrity of fresh and frozen-thawed spermatozoa during standard intracytoplasmic sperm injection (ICSI) conditions. Experimental prospective study. Embryology research laboratory. Forty-six ejaculates from normozoospermic and oligozoospermic men. Assessment of sperm DNA fragmentation by single-cell gel electrophoresis assay. DNA integrity of fresh and frozen-thawed spermatozoa from normozoospermic and oligozoospermic ejaculates exposed to PVP-based and non-PVP-based media. Exposure of fresh and frozen thawed spermatozoa from normozoospermic and oligozoospermic ejaculates to PVP-based medium in an ICSI dish for 30 minutes statistically significantly increased the DNA fragmentation. In contrast, the extent of DNA fragmentation in non-PVP-based medium did not statistically significantly differ from control. A PVP-based medium can induce a statistically significant amount of sperm DNA fragmentation in an ICSI dish, and frozen-thawed sperm from oligozoospermic ejaculates are more susceptible to in situ DNA fragmentation. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. Level of copper in human split ejaculate.

    Science.gov (United States)

    Skandhan, Kalanghot; Valsa, James; Sumangala, Balakrishnan; Jaya, Vasudevan

    2017-02-03

    The purpose of this study was to understand the details of splits of an ejaculate and to locate the origin of release of copper into semen. Laboratory methods routinely followed for semen analysis were carried out. Copper was estimated by employing atomic absorption spectrophotometry. First split of ejaculate showed the highest number of motile sperm, the quality of which decreased from first to third. Copper level in splits 1, 2 and 3 was 29, 23 and 22 µg%, respectively. This study concluded that copper was released from throughout the genital tract.

  9. Severe tail defects in the spermatozoa ejaculated by an English bulldog.

    Science.gov (United States)

    Rota, Ada; Manuali, Elisabetta; Caire, Simona; Appino, Simonetta

    2008-01-01

    This paper describes the case of a 2-year-old English Bulldog with severe teratozoospermia that consisted mainly of "Dag-like" defects, which is strong coiling of the tail. Although libido, semen volume and sperm concentration were normal, sperm motility was 5.0%, and 93.3% of spermatozoa exhibited morphological abnormalities affecting the tail. Transmission electron microscopic examination of the spermatozoa revealed strong folding, coiling and fracture of sperm midpieces and tails, axonemal defects and the presence of swollen and unevenly distributed mitochondria. Taking into account the dog's history and examinations and the constantly high percentage of abnormal spermatozoa over time, the defect was considered to be genetic in origin.

  10. Characteristics of spermatozoa of whole ejaculate and sperm-rich fraction of dog semen following exposure to media varying in osmolality.

    Science.gov (United States)

    Strzezek, Rafał; Fraser, Leyland

    2009-07-01

    This study aimed to analyze the effects of different osmolalities on the characteristics of spermatozoa originating from whole ejaculates (WE; including the prostatic fluid) and the sperm-rich fractions (SRF). Ejaculates, collected from four mixed-breed dogs, were exposed for 10 min at room temperature to Tris-fructose-citrate (TFC) solution with osmolality ranging from 150 to 1100 mOsm. After treatment spermatozoa were evaluated by microscopic analysis of motility and fluorescent assessments of plasma membrane integrity (carboxyfluorescein diacetate and propidium iodide, CFDA/PI) and mitochondrial function (rhodamine 123, R123). Irrespective of the sperm source, there was a complete loss of motility when spermatozoa were exposed to TFC solution with 1100 mOsm. There were no marked differences in the sperm characteristics between media with 300 and 350 mOsm, regardless of the ejaculate collection procedure. However, a marked reduction in motility of spermatozoa retrieved either from the WE or SRF was observed after exposure to different anisosmotic conditions (150, 550 and 800 mOsm). In all dogs, spermatozoa from the WE exhibited greater osmotolerance in terms of plasma membrane integrity and mitochondrial function when exposed to anisosmotic conditions (150, 550, 800 and 1100 mOsm). There were inter-dog variations in response to various osmotic conditions. The findings of this study indicated that spermatozoa from the WE tolerated exposure to a wider range of osmolality than those from the SRF. It seemed that the presence of prostatic fluid of dog semen rendered the sperm membrane structures less susceptible to osmotic stress.

  11. Comparative studies of elemental composition on ejaculated fowl, bull, rat, dog and boar spermatozoa by electron probe X-ray microanalysis.

    Science.gov (United States)

    Ashizawa, K; Ozawa, Y; Okauchi, K

    1987-01-01

    1. Comparative analyses of the concentrations of bulk and trace elements on the head, midpiece and tail regions of the ejaculated fowl, bull, rat, dog and boar spermatozoa were performed using X-ray microanalysis with scanning electron microscopy. 2. Although the pattern of distribution of elements on the surface of the three different subcellular regions was, in general, similar among all the species, there were substantial shifts in absolute concentrations. 3. Concentration of magnesium on the dog spermatozoa was significantly higher (about 2 times) than those of the other species. 4. Zinc, copper, iron and manganese concentrations were higher on fowl spermatozoa compared with those of the other species studied. 5. The Na-to-K ratios on the midpiece ranged from 1.46 (rat) to 2.26 (dog).

  12. Is prnt a Pseudogene? Identification of Ram Prt in Testis and Ejaculated Spermatozoa

    Science.gov (United States)

    Pimenta, Jorge; Domingos, Ana; Santos, Pedro; Marques, Carla C.; Cantante, Cátia; Santos, Ana; Barbas, João P.; Baptista, Maria C.; Horta, António E. M.; Viegas, Aldino; Mesquita, Patrícia; Gonçalves, João; Fontes, Carlos A.; Prates, José A. M.; Pereira, Rosa M. L. N.

    2012-01-01

    A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrPC), expressed by the prion gene (prnp), into an abnormally folded isoform (PrPSc) with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel), and prnt (that encodes the prion protein testis specific - Prt) with sprn (shadow of prion protein gene, that encodes Shadoo or Sho) genes, constitute the “prion gene complex”. Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA) immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated with its

  13. Is prnt a pseudogene? Identification of ram Prt in testis and ejaculated spermatozoa.

    Directory of Open Access Journals (Sweden)

    Jorge Pimenta

    Full Text Available A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrP(C, expressed by the prion gene (prnp, into an abnormally folded isoform (PrP(Sc with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel, and prnt (that encodes the prion protein testis specific--Prt with sprn (shadow of prion protein gene, that encodes Shadoo or Sho genes, constitute the "prion gene complex". Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated

  14. Analysis of MYO-Inositol effect on spermatozoa motility, in hyper viscous ejaculates and in patients with grades II and III varicocele

    Directory of Open Access Journals (Sweden)

    Filomena Scarselli

    2016-12-01

    Full Text Available The goal of this study is to evaluate MYOInositol effects on spermatozoa motility, in patients’ ejaculates with severe varicocele or hyper viscosity. The study included normal viscosity ejaculate from 30 patients affected by varicocele and hyper viscosity ejaculate from 33 patients without any testicular pathologies. All selected samples showed sperm concentration > 2 million/ml and progressive motility < 32%. In both groups, the pellet obtained after centrifugation in buffered medium, was divided in two aliquots, both incubated for 15 minutes at 37°C: one with MYO-Inositol and the other one, as control, only in phosphate buffered saline (PBS. Afterwards, the sperm progressive motility was assessed using Computer Assisted Sperm Analysis (CASA system. Incubation with MYO-Inositol improved sperm progressive motility in high viscosity samples compared to control group (38.9% ± 3.0 vs 24.35% ± 2.41, respectively; p ≤ 0.0001. Conversely, no statistically significant difference was observed in total sperm progressive motility in varicocele samples compared with control group (22.7% ± 2.07 vs 26.7% ± 3.31, respectively; p = 0.085. The MYO-Inositol positive effect on spermatozoa motility may depend on the type of sperm damage: heavy structural and biochemical defects which typically affects patients with varicocele are not restored by Inositol. On the contrary, MYOInositol is able to improve sperm motility in semen samples with high viscosity, since those samples show no substantial structural sperm defects.

  15. Effect of pre-freezing conditions on the progressive motility recovery rate of human frozen spermatozoa.

    Science.gov (United States)

    Zhang, X; Zhou, Y; Xia, W; Wu, H; Yao, K; Liu, H; Xiong, C

    2012-10-01

    We evaluated the effects of sperm concentration, progressive motility, sperm morphology, duration of abstinence and collection season on the progressive motility recovery rate of human frozen spermatozoa to identify characteristics that predict the progressive motility recovery rate of human frozen spermatozoa and improve the protocol for sperm collecting in sperm banks. A total of 14 190 semen samples donated at Zhejiang human sperm bank of China between September 2006 and June 2011 were collected from 1624 donors. Semen was evaluated according to WHO standard procedures for sperm concentration. Progressive motility, sperm morphology, ejaculate collection season and abstinence time were recorded. After freezing and thawing, the progressive motility was assessed. Results showed that sperm concentration, progressive motility and normal morphology were significantly associated with the progressive motility recovery rate of human frozen spermatozoa. In addition, the abstinence time and collection season also significantly affected progressive motility recovery rate. Our results indicated that sperm concentration, progressive motility and normal morphology could be valuable in predicting the progressive motility recovery rate of human frozen spermatozoa. As such, progressive motility recovery may be improved by donating semen when abstinent for 3-5 days and during seasons other than summer.

  16. A model for the importance of zinc in the dynamics of human sperm chromatin stabilization after ejaculation in relation to sperm DNA vulnerability.

    Science.gov (United States)

    Björndahl, Lars; Kvist, Ulrik

    2011-02-01

    The focus of this review is the dual functions of the sperm chromatin stabilization and how external factors can interfere with these functions. Zinc depletion after ejaculation allows for rapid and total sperm chromatin decondensation without addition of exogenous disulfide cleaving agents. Zinc depletion without concomitant repulsion of chromatin fibers induces another type of stability that requires exogenous disulfide cleaving agents to allow decondensation. It is essential to extend the present concept, that the sperm chromatin stability is based on disulfide bridges only, to include also the functions of Zn(2+). It is suggested that the chromatin stability of the ejaculated human spermatozoon is rapidly reversible due to the dual function of Zn(2+) that stabilizes the structure and prevents the formation of excess disulfide bridges by a single mechanism: the formation of zinc bridges involving protamine thiols of cysteine and potentially also imidazole groups of histidine. Extraction of zinc from the freshly ejaculated spermatozoon allows two totally different biological results: (1) immediate decondensation if chromatin fibers concomitantly are induced to repel (e.g., through phosphorylation in the ooplasm) and (2) thiols freed from Zn(2+) are available to form disulfide bridges creating a superstabilized chromatin. Spermatozoa in the zinc rich prostatic fluid (in first ejaculated fraction) represent physiology. Extraction of chromatin zinc can be caused by unphysiological exposure of spermatozoa to the zinc chelating and oxidative seminal vesicular fluid, a situation common to most assisted reproductive techniques (ART) laboratories where the entire ejaculate is collected into a single container in which spermatozoa and secretions are mixed during at least 30 min. Some men in infertile couples have low content of sperm chromatin zinc due to loss of zinc during ejaculation and liquefaction. Tests for sperm DNA integrity may give false negative results due to

  17. Seminal Plasma Characteristics and Expression of ATP-binding Cassette Transporter A1 (ABCA1) in Canine Spermatozoa from Ejaculates with Good and Bad Freezability.

    Science.gov (United States)

    Schäfer-Somi, S; Palme, N

    2016-04-01

    The composition of seminal plasma and the localization of the ATP-binding cassette transporter A1 (ABCA1) in spermatozoa from good and bad freezers were compared to frozen-thawed spermatozoa from the same dog. Ejaculates were obtained from 31 stud dogs, and the sperm-rich fraction (SRF) was kept for analysis. One aliquot was used for the analysis of concentration, progressive motility (P; CASA), viability (V; CASA) and leucocyte count, and the analysis was performed by flow cytometry (FITC-PNA/PI), SCSA and HOST. In seminal plasma, concentration of albumin, cholesterol, calcium, inorganic phosphate, sodium, potassium, zinc and copper was measured. Semen smears were prepared and evaluated for the expression of ABCA1. The remainder of each ejaculate was frozen. After thawing, the quality assessment was repeated and further smears were prepared. According to post-thaw semen quality, dogs were assigned to good freezers (n = 20) or bad freezers (n = 11), the latter were defined as 40% morphologically abnormal sperm and/or Bad freezers were older than good freezers (5.3 vs 3.4 years, p bad freezers, the percentage of sperm with ABCA1 signal in the acrosome was lower (26.3% vs 35.7%, p good freezers (p bad freezer sperm, an increase in acrosome damages coincided with an increased loss of cholesterol transporters and cell death, and a lower cholesterol concentration in seminal plasma. Follow-up studies revealed whether a relation exists between these findings.

  18. CHARACTERIZATION OF THE OLFACTORY RECEPTORS EXPRESSED IN HUMAN SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    Caroline eFlegel

    2016-01-01

    Full Text Available The detection of external cues is fundamental for human spermatozoa to locate the oocyte in the female reproductive tract. This task requires a specific chemoreceptor repertoire that is expressed on the surface of human spermatozoa, which is not fully identified to date. Olfactory receptors (ORs are candidate molecules and have been attributed to be involved in sperm chemotaxis and chemokinesis, indicating an important role in mammalian spermatozoa. An increasing importance has been suggested for spermatozoal RNA, which led us to investigate the expression of all 387 OR genes. This study provides the first comprehensive analysis of OR transcripts in human spermatozoa of several individuals by RNA-Seq. We detected 91 different transcripts in the spermatozoa samples that could be aligned to annotated OR genes. Using stranded mRNA-Seq, we detected a class of these putative OR transcripts in an antisense orientation, indicating a different function, rather than coding for a functional OR protein. Nevertheless, we were able to detect OR proteins in various compartments of human spermatozoa, indicating distinct functions in human sperm. A panel of various OR ligands induced Ca2+ signals in human spermatozoa, which could be inhibited by mibefradil. This study indicated that a variety of ORs are expressed at the mRNA and protein level in human spermatozoa and demonstrates that ORs are involved in the physiological processes.

  19. PIXE analysis of human spermatozoa isolated from seminal plasma

    Science.gov (United States)

    Maeda, K.; Sasa, Y.; Kusuyama, H.; Yoshida, K.; Uda, M.

    1990-04-01

    PIXE has been applied to the multielemental and microanalysis of human spermatozoa. This is the first attempt to determine the chemical compositions of the motile spermatozoa free from contaminations of seminal plasma without loss of component elements during washing. The spermatozoa were isolated from semen by letting them swim into a kind of physiological saline, Tyrode's solution. Relative concentrations of P, K, Ca, Ti, Fe, Zn and Br in motile spermatozoa were determined by the use of the chlorine K X-ray peak intensity for evaluating the amount of Tyrode's solution contained in the sample targets. The concentrations of calcium and iron in spermatozoa were considerably higher than in seminal plasma. The concentrations of P, K, Zn and Br in spermatozoa were not so different from those in seminal plasma.

  20. Removal of spermatozoa with externalized phosphatidylserine from sperm preparation in human assisted medical procreation: effects on viability, motility and mitochondrial membrane potential

    Directory of Open Access Journals (Sweden)

    de Agostini Ariane

    2009-01-01

    Full Text Available Abstract Background Externalization of phosphatidylserine (EPS occurs in apoptotic-like spermatozoa and could be used to remove them from sperm preparations to enhance sperm quality for assisted medical procreation. We first characterized EPS in sperms from infertile patients in terms of frequency of EPS spermatozoa as well as localization of phosphatidylserine (PS on spermatozoa. Subsequently, we determined the impact of depleting EPS spermatozoa on sperm quality. Methods EPS were visualized by fluorescently-labeled annexin V binding assay. Double staining with annexin V and Hoechst differentiates apoptotic from necrotic spermatozoa. We used magnetic-activated cell sorting using annexin V-conjugated microbeads (MACS-ANMB technique to remove EPS spermatozoa from sperm prepared by density gradient centrifugation (DGC. The impact of this technique on sperm quality was evaluated by measuring progressive motility, viability, and the integrity of the mitochondrial membrane potential (MMP by Rhodamine 123. Results Mean percentages of EPS spermatozoa were 14% in DGC sperm. Four subpopulations of spermatozoa were identified: 70% alive, 3% early apoptotic, 16% necrotic and 11% late apoptotic or necrotic. PS were localized on head and/or midpiece or on the whole spermatozoa. MACS efficiently eliminates EPS spermatozoa. MACS combined with DGC allows a mean reduction of 70% in EPS and of 60% in MMP-disrupted spermatozoa with a mean increase of 50% in sperm survival at 24 h. Conclusion Human ejaculates contain EPS spermatozoa which can mostly be eliminated by DGC plus MACS resulting in improved sperm long term viability, motility and MMP integrity. EPS may be used as an indicator of sperm quality and removal of EPS spermatozoa may enhance fertility potential in assisted medical procreation.

  1. Low Concentration of Ethylene Glycol Improved Recovery Rate of Human Spermatozoa After Vitrification (ETILEN GLIKOL KONSENTRASI RENDAH MENINGKATKAN RECOVERY RATE SPERMATOZOA MANUSIA PASCAVITRIFIKASI

    Directory of Open Access Journals (Sweden)

    Rini Widyastuti

    2016-11-01

    Full Text Available The use of cryoprotectants for the cryopreservation of human spermatozoa, oocytes, zygote, earlycleavage stage of embryos and blastocyst is an integral part of almost every human In Vitro Fertilizationprogram. Moreover, the cryopreservation of these types of cells by direct plunging into liquid nitrogen (-196°C usually requires a high concentration of cryoprotectant with a consequent of cytotoxic effect. Theaim of this study was to observe the effect of ethylene glycol concentration on the spermatozoa recoveryrate following vitrification process. Earle’s balanced salt solution + 0.25 M sukrosa + 1 % human albumineserum as basic solution supplemented with some different concentrations of etylene glycol (ie: 36.25%;18.25%; 9.12%; 4.56%; 1.14% and 0.57% were used to evaluate the motility and viability of spermatozoafollowing vitrification. Human’s spermatozoa from ejaculates with progressive motility and viability above50% were used as samples. Samples were mixed with vitrification solution and then loaded into 0.25 mLstraws, equilibrated for 10 minutes at room temperature before plunged into liquid nitrogen. Spermatozoathawing was done at 24 hours after the vitrification. The results showed that, the decrease of spermatozoamotility and viability were observed at the highest (100%, 96.70%, respectively in the samples that wereadded with vitrification medium contained 36.25% of ethylene glycol. On the other hand, the decrease ofthe spermatozoa motility and viability were found at the lowest (14.11%, 43.81 %, respectively in thesamples without ethylene glycol supplementation. It can be concluded that the highest spermatozoa recoveryrate was obtained from the vitrification using a low concentration of ethylene glycol.

  2. Immunolocalisation of ghrelin and obestatin in human testis, seminal vesicles, prostate and spermatozoa.

    Science.gov (United States)

    Moretti, E; Vindigni, C; Tripodi, S A; Mazzi, L; Nuti, R; Figura, N; Collodel, G

    2014-01-01

    The role of ghrelin and obestatin in male reproduction has not completely been clarified. We explored ghrelin and obestatin localisation in the male reproductive system. Polyclonal antibodies anti-ghrelin and anti-obestatin were used to detect the expression of these hormones in human testis, prostate and seminal vesicles by immunocytochemistry, while in ejaculated and swim up selected spermatozoa by immunofluorescence. Sertoli cells were positive for both peptides and Leydig cells for ghrelin; germ cells were negative for both hormones. Mild signals for ghrelin and obestatin were observed in rete testis; efferent ductules were the most immune reactive region for both peptides. Epididymis was moderately positive for ghrelin; vas deferens and seminal vesicles showed intense obestatin and moderate ghrelin labelling; prostate tissue expressed obestatin alone. Ejaculated and selected spermatozoa were positive for both peptides in different head and tail regions. This study confirms ghrelin localisation in Leydig and Sertoli cells; the finding that ghrelin is expressed in rete testis, epididymis, vas deferens and seminal vesicles is novel, as well as the localisation of obestatin in almost all tracts of the male reproductive system. This research could offer insights for stimulating other studies, particularly on the role of obestatin in sperm physiology, which is still obscure.

  3. Quality of boar spermatozoa from the sperm-peak portion of the ejaculate after simplified freezing in MiniFlatpacks compared to the remaining spermatozoa of the sperm-rich fraction.

    Science.gov (United States)

    Siqueira, A P; Wallgren, M; Hossain, M S; Johannisson, A; Sanz, L; Calvete, J J; Rodríguez-Martínez, H

    2011-04-15

    Boar sperm viability post-thaw differs depending on the ejaculate fraction used, with spermatozoa present in the first 10 mL of the sperm-rich fraction (SRF) (portion 1, P1, sperm-peak portion) displaying the best cryosurvival in vitro compared with that of spermatozoa from the rest of the ejaculate (portion 2 of the SRF plus the post-spermatic fraction), even when using simplified freezing routines. This viability apparently relates to the specific profile of seminal plasma in P1 (i.e., glycoprotein and bicarbonate concentrations, and pH). However, spermatozoa from P1 have not been compared with spermatozoa from the rest of the SRF (SRF-P1, usually 30-40 mL of the SRF), which is routinely used for freezing. We compared P1 with SRF-P1 in terms of sperm kinematics (using the QualiSperm™ system), while membrane integrity (SYBR-14/PI), acrosome integrity (FITC PNA/PI), and sperm membrane stability (Annexin-V) were explored using flow cytometry. As well, total protein concentration and the proteomics of the seminal plasma (SP) of both portions of the SRF were studied using two-dimensional electrophoresis (2DE), mass fingerprinting (MALDI-TOF), and collision-induced dissociation tandem mass spectrometry (CID-MS/MS) on selected peptides. The SRF portions were collected weekly from four mature boars (4-5 replicates per boar, sperm concentration: P1, 1.86 ± 0.20; SRF-P1, 1.25 ± 0.14 × 10(9) spz/mL) and processed using a quick freezing method in MiniFlatPacks. Post-thaw sperm motility reached 50%, without differences between SRF portions, but with clear inter-boar variation. Neither plasma membrane nor acrosome integrity differed (ns) between fractions. These results indicate that there are no differences in cryosurvival after quick freezing of boar spermatozoa derived from either of the two SRF portions. While P1 and SRF-P1 clearly differed in relative total protein contents, as expected, they displayed very similar protein profiles as assessed using 2DE and mass

  4. Interaction between leucocytes and human spermatozoa influencing reactive oxygen intermediates release.

    Science.gov (United States)

    Fraczek, Monika; Sanocka, Dorota; Kurpisz, Maciej

    2004-04-01

    The relationship between the presence of white blood cells (WBCs) and the fertilizing potential of human semen is still an open question. It is well known that the presence of leucocytes in human semen can be related to the production of reactive oxygen intermediates (ROI). Semen samples were obtained from 15 normozoospermic men and leucocytes were isolated from heparinized blood drawn from 15 volunteers. Lucigenin and luminol-mediated chemiluminescence assays were used to determine reactive oxygen species (ROS) generation by non-activated or activated leucocytes through 12-myristate-13-acetate or N-formyl-methionyl-leucyl-phenyalanine (FMLP) before the addition of spermatozoa isolated by swim-up or Percoll procedures. All spermatozoal fractions used in this study were characterized by defining their motility, morphology and viability. The levels of ROS formation by non-activated as well as stimulated leucocytes were significantly decreased after addition of swim-up separated spermatozoa (p levels changed depending on the type of inducing factor used for oxidative burst. Then, spermatozoa selected by swim-up procedure although with only slightly higher viability and morphology than sperm obtained from 90% Percoll fraction clearly exhibited much higher capacity to inhibit ROI secretion by receptor-stimulated leucocytes (FMLP-activation) than Percoll fractionated sperm. Such results may indicate that within normal semen may exist sperm subpopulations with different biochemical mechanisms controlling the interaction between spermatozoa and contaminating leucocytes. When ROI levels contained in normozoospermic semen are dependent on the WBCs activation, it seems that spermatozoa with preserved normal functional competence are able to defend themselves against leucocytes-derived ROI. Also for normozoospermic ejaculates, swim-up sperm may improve semen antioxidant characteristics when comparing with Percoll (90%) separated sperm. It may help for optimal sperm preparation

  5. IN VITRO EFFECT OF SILVER ENGINEERED NANOPARTICLES ON HUMAN SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    G. Terzuoli

    2012-08-01

    Full Text Available Silver (Ag NPs are among the most commercialized NPs due to their antimicrobial potential. They are highly at- tractive for possible applications in manufacture of medical device. However there is a serious lack of information concerning their impact on the human health and the environment. Moreover studies on the effects of NPs on ejaculated sperm are rather limited. For these reasons our study explored the in vitro effects of Ag NPs on human ejaculated spermatozoa. Ag NPs have been produced, characterized, and furnished by Colorobbia Industry, Sovigliana (Vinci, Florence, Italy. Aliquots of total semen were incubated at 37°C for 60 minutes (min and 120 min at the concentration of 125 μM, 250 μM, and 500 μM of engineered Ag NPs. The control was represented by specimens of semen samples treated with the same procedure without NPs. After the incubations, sperm motility was evaluated following WHO guidelines and sperm viability was eval- uated by Eosin Y test. At the end of incubation with Ag NPs the samples were processed by a Field Emission Gun-based Scanning Transmition Electron Microscope/ Energy Dispertion Spectrometry (STEM/EDS. We observed that sperm motility percentage was significantly reduced in semen samples treated with 125 μM, 250 μM and 500 μM of Ag NPs after 60 min and 120 min of incubation respect to controls (P<0.001; P<0.01, 125 μM at 60 min. Sperm viability percentage significantly decreased in a progressive manner after 125 μM (P<0.05, 250 μM (P<0.05 and 500 μM (P<0.001 Ag NPs incubation at 60 min and 120 min. We did not find any significant difference between the values assessed after 60 min of NPs incubation and those estimated after 120 min of incubation. In the control samples, the sperm motility and the sperm viability percentages significantly decreased after 120 min of incubation (P<0.001 respect to the basal values. Ag NPs were undetectable in all treated samples by STEM/EDS. These in vitro results show a

  6. Estimate of oxygen consumption and intracellular zinc concentration of human spermatozoa in relation to motility

    Institute of Scientific and Technical Information of China (English)

    Wolf-BernhardSchill; KerstinDefosse; Hans-HilhelmKoyro; NorbertWeissmann

    2003-01-01

    Aim:To investigate the human sperm oxygen/energy consumption and zinc content in relation to motility.Methods:In washed spermatozoa from 67 ejaculates,the oxygen consumption was determined.Following calculation of the total oxygen consumed by the Ideal Gas Law,the energy consumption of spermatozoa was calculated.In addition,the zinc content of the sperm was determined using an atomic absorption spectrometer.The resulting data were correlated to the vitality and motility.Results:The oxygen consumption averaged 0.24μmol/106 sperm×24h,0.28μmol/106 live sperm×24h and 0.85μmol/106 live & motile sperm×24h.Further calculations revealed that sperm motility was the most energy consuming process(164.31mJ/106 motile spermatozoa×24h),while the oxygen consumption of the total spermatozoa was 46.06mJ/106 spermatozoa ×24h.The correlation of the oxygen/energy consumption and zinc content with motility showed significant negative correlations(r=-0.759;P<0.0001 and r=-0.441;P<0.0001,respectively).However,when correlating sperm energy consumption with the zinc content,a significant positive relation(r=0.323;P=0.01)was observed.Conclusion:Poorly motile sperm are actually wasting the available energy.Moreover,our data clearly support the “Geometric Clutch Model”of the oneme function and demonstrate the importance of the outer dense fibers for the generation of sperm motility,especially progressive motility.

  7. MOLECULAR ANALYSIS OF HUMAN SPERMATOZOA: POTENTIAL FOR INFERTILITY RESEARCH

    Science.gov (United States)

    Gordon Research Conference: Mammalian Gametogenesis and Embryogenesis New London, CT, July 1-6, 2000Molecular Analysis of Human Spermatozoa: Potential for Infertility ResearchDavid Miller 1, David Dix2, Robert Reid 3, Stephen A Krawetz 3 1Reproductive ...

  8. MOLECULAR ANALYSIS OF HUMAN SPERMATOZOA: POTENTIAL FOR INFERTILITY RESEARCH

    Science.gov (United States)

    Gordon Research Conference: Mammalian Gametogenesis and Embryogenesis New London, CT, July 1-6, 2000Molecular Analysis of Human Spermatozoa: Potential for Infertility ResearchDavid Miller 1, David Dix2, Robert Reid 3, Stephen A Krawetz 3 1Reproductive ...

  9. Spermatozoa in the sperm-peak-fraction of the boar ejaculate show a lower flow of Ca(2+) under capacitation conditions post-thaw which might account for their higher membrane stability after cryopreservation.

    Science.gov (United States)

    Hossain, Md Sharoare; Johannisson, Anders; Siqueira, Amanda Pimenta; Wallgren, Margareta; Rodriguez-Martinez, Heriberto

    2011-10-01

    Boar spermatozoa collected in the ejaculate sperm peak-portion (P1, first 10 mL of the sperm-rich fraction, SRF), had shown a higher resilience to freezing and thawing compared to spermatozoa from the rest of the ejaculate (2nd portion of the SRF plus the post-sperm-rich fraction, PSRF), even when using a simplified freezing technique, as long as spermatozoa were incubated in their own seminal plasma (SP). This experiment studied the stability of P1- and SRF-P1 boar spermatozoa frozen in MiniFlatPacks (MFP), post-thaw, using flow cytometry. Since spermatozoa from either portion showed similar cryosurvival and low proportions of unstable membranes (<3%, annexin-V/propidium iodide staining), and only a tendency for SRF-P1 live spermatozoa to depict acrosome exocytosis (FITC-PNA/PI/H33342); they were explored for Ca(2+) contents using a Fluo-4 probe under in vitro capacitating conditions (mBO+ medium), as well they were tested for their ability to sustain a short Ca(2+)-ionophore (A23187) in vitro challenge. The proportions of live spermatozoa depicting high Ca(2+)-levels were initially <2% but increased over incubation time, particularly in SRF-P1(P<0.05), while proportions of live spermatozoa with low Ca(2+)-levels were basically constant over incubation time (~11-14%), for either portion. Incubation in capacitation medium did not modify the proportions of low-Ca(2+) but dramatically increased the proportions of high-Ca(2+) spermatozoa (P<0.001) already after 15 min exposure, highest for SRF-P1 spermatozoa. While the proportion of live spermatozoa with intact acrosome was significantly decreased among SRF-P1 (P<0.001), that of P1-spermatozoa remained unchanged, probably owing to the lowest relative content of cytosolic Ca(2+). The results suggest that spermatozoa in the P1-portion are more resilient to express acrosome exocytosis post-thaw compared to those bathing in the rest of the SRF-fraction when cryopreserved using a simplified technique, in MFPs.

  10. Quality of human spermatozoa: relationship between high-magnification sperm morphology and DNA integrity.

    Science.gov (United States)

    Maettner, R; Sterzik, K; Isachenko, V; Strehler, E; Rahimi, G; Alabart, J L; Sánchez, R; Mallmann, P; Isachenko, E

    2014-06-01

    The aim of this work is to establish the relationship between the morphology of Intracytoplasmic Morphologically Selected Sperm Injection (IMSI)-selected spermatozoa and their DNA integrity. The 45 ejaculates were randomly distributed into three treatment groups: normozoospermic, oligoasthenozoospermic and oligoasthenotheratozoospermic samples. The evaluation of DNA integrity was performed using the sperm chromatin dispersion test. It was established that DNA integrity of spermatozoa is strongly dependent on ejaculate quality (P 0.1). With decreased ejaculate quality, the percentage of spermatozoa with nonfragmented DNA decreased significantly (P < 0.05) independent from morphological class. Nevertheless, the rate of IMSI-selected spermatozoa with fragmented DNA within of Class 1 in normozoospermic (Group 1), in oligoasthenozoospermic (Group 2) and in oligoasthenotheratozoospermic (Group 3) samples was 21.1%, 31.8% and 54.1%, respectively. In conclusion, there is a direct relationship between morphological parameters of spermatozoa and their DNA integrity. However, the IMSI technique alone is not enough for the selection of spermatozoa with intact nuclei.

  11. ANTIOXIDANT EFFECT OF RESVERATROL IN HUMAN SPERMATOZOA AND IN RAT GERMINAL CELLS

    Directory of Open Access Journals (Sweden)

    MG. Federico

    2012-01-01

    Full Text Available Objective: To assess the antioxidant activity of Resveratrol (3,5,4’-trihydroxystilbene, RES after induction of lipid peroxidation LPO in human spermatozoa and in immature rat germinal cells. Materials and Methods: Ejaculated human spermatozoa, selected by swim up, have been incubated with tert-Butylhydroperoxide and tert-Butylhydroperoxide-RES. The localization of LPO has been performed using the probe C11-BODIPY581/591. The same assays were carried out on pachytene spermatocytes and round spermatids obtained from three Wistar rats 35 days of age. The two cellular fractions were achieved after enzymatic digestion with collagenase and subsequent fractionation on bovine serum albumin 0.5-3% gradient (STAPUT. The ultrastructure of all samples was assessed by transmission electron microscopy (TEM. Results: The midpiece of sperm tail and the whole plasma membrane of germ cells were the target of LPO. TEM analysis of sperm, quantitatively elaborated by a mathematical formula, showed a significantly lower percentage of necrosis in the samples treated with RES (P<0.01; as regards rat germinal cells, necrosis features (cytoplasmic vacuoles, disrupted chromatin and broken plasma membrane were mainly evident in the meiotic fraction without RES. Conclusions: RES, found in the skins of grape, reduces the damage induced by oxidative stress in human sperm and rat testicular germ cells; in particular spermatids appeared to be less sensitive to oxidative damage compared with spermatocytes.

  12. Effect of mitochondrial calcium uniporter blocking on human spermatozoa.

    Science.gov (United States)

    Bravo, A; Treulen, F; Uribe, P; Boguen, R; Felmer, R; Villegas, J V

    2015-08-01

    Calcium (Ca(2+) ) regulates a number of essential processes in spermatozoa. Ca(2+) is taken up by mitochondria via the mitochondrial calcium uniporter (mCU). Oxygen-bridged dinuclear ruthenium amine complex (Ru360) has been used to study mCU because it is a potent and specific inhibitor of this channel. In bovine spermatozoa, it has been demonstrated that mitochondrial calcium uptake inhibition adversely affects the capacitation process. It has been demonstrated in human spermatozoa that mCU blocking, through Ru360, prevents apoptosis; however, the contribution of the mCU to normal human sperm function has not been studied. Therefore, the aim of this study was to evaluate the effect of mCU blocking on human sperm function. Spermatozoa obtained from apparently healthy donors were incubated with 5 and 10 μm Ru360 for 4 h at 37 °C. Viability was assessed using propidium iodide staining; motility was determined by computer-aided sperm analysis, adenosine triphosphate (ATP) levels using a luminescence-based method, mitochondrial membrane potential (ΔΨm) using JC-1 staining and reactive oxygen species (ROS) production using dihydroethidium dye. Our results show that mCU blocking significantly reduced total sperm motility and ATP levels without affecting sperm viability, ΔΨm and ROS production. In conclusion, mCU contributes to the maintenance of sperm motility and ATP levels in human spermatozoa.

  13. Sperm fucosyltransferase-5 mediates spermatozoa-oviductal epithelial cell interaction to protect human spermatozoa from oxidative damage.

    Science.gov (United States)

    Huang, Venus Wenxin; Lee, Cheuk-Lun; Lee, Yin-Lau; Lam, Kevin K W; Ko, Jennifer K Y; Yeung, William S B; Ho, Pak-Chung; Chiu, Philip C N

    2015-06-01

    Oxidative damage by reactive oxygen species (ROS) is a major cause of sperm dysfunction. Excessive ROS generation reduces fertilization and enhances DNA damage of spermatozoa. Interaction between spermatozoa and oviductal epithelial cells improves the fertilizing ability of and reduces chromatin damage in spermatozoa. Our previous data showed that oviductal epithelial cell membrane proteins interact with the human spermatozoa and protect them from ROS-induced reduction in sperm motility, membrane integrity and DNA integrity. Sperm fucosyltransferase-5 (sFUT5) is a membrane carbohydrate-binding protein on human spermatozoa. In this study, we demonstrate for the first time that sFUT5 is involved in human spermatozoa-oviduct interaction and the beneficial effects of such interaction on the fertilizing ability of human spermatozoa. Anti-sFUT5 antibody-treated spermatozoa had reduced binding to oviductal membrane proteins. It is consistent with the result that affinity-purified sFUT5 is bound to the epithelial lining of human oviduct and to the immortalized human oviductal epithelial cell line, OE-E6/E7. Pretreatment of spermatozoa with anti-sFUT5 antibody and oviductal membrane proteins with sFUT5 suppressed the protective action of oviductal membrane proteins against ROS/cryopreservation-induced oxidative damage in spermatozoa. Asialofetuin, a reported sFUT5 substrate, can partly mimic the protective effect of oviductal epithelial cell membrane proteins on sperm motility, membrane and DNA integrity. The results enhance our understanding on the protective mechanism of oviduct on sperm functions.

  14. Adriamycin induces H2AX phosphorylation in human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Zhong-Xiang Li; Ting-Ting Wang; Yan-Ting Wu; Chen-Ming Xu; Min-Yue Dong; Jian-Zhong Sheng; He-Feng Huang

    2008-01-01

    Aim: To investigate whether adriamycin induces DNA damage and the formation of γH2AX (the phosphorylated form of histone H2AX) foci in mature spermatozoa. Methods: Human spermatozoa were treated with adriamycin at different concentrations. γH2AX was analyzed by immunofluorescent staining and flow cytometry and double- strand breaks (DSB) were detected by the comet assay. Results: The neutral comet assay revealed that the treatment with adriamycin at 2 μg/mL for different times (0.5, 2, 8 and 24 h), or for 8 h at different concentrations (0.4, 2 and 10 μg/mL), induced significant DSB in spermatozoa. Immunofluorent staining and flow cytometry showed that the expression of γH2AX was increased in a dose-dependent and time-dependant manner after the treatment of adriamycin. Adriamycin also induced the concurrent appearance of DNA maintenance/repair proteins RAD50 and 53BP1 with γH2AX in spermatozoa. Wortmannin, an inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, abolished the co-appearance of these two proteins with γH2AX. Conclusion: Human mature spermatozoa have the same response to DSB-induced H2AX phosphorylation and subsequent recruitment of DNA maintenance/repair proteins as somatic cells.

  15. A preliminary study on the rheological properties of human ejaculate and changes during liquefaction

    Institute of Scientific and Technical Information of China (English)

    Yong-DeShi; Lan-FengPan; Fei-KunYang; Si-QiWang

    2004-01-01

    Aim: To study the changes in rheological properties, namely the parameters of the hysteresis loops and yield stress versus time for human semen after ejaculation. Methods: Ejaculates were obtained from volunteers and immediately put into the test cup of a Brookfield Programmable DV- 11 Rheometer, by which the hysteresis loops and yield stress were determined. Results: (1) Yield stress values dropped down from more than 3000 mPa to 60 mPa inabout 5 minutes after ejaculation; (2) The shape of the hysteresis loops of shear stress versus shear rate was changed from the counter-clockwise direction, that enclosed a large area, into the clockwise direction, that enclosed a very small area. Conclusion: Human ejaculate originally possesses semi-solid or visco-elastic body behavior and in 5 minutes after liquefaction, it becomes a thixotropic fluid or shearing thinning fluid with very low viscosity. (Asian J Androl 2004 Dec; 6: 299-304)

  16. Immunomagnetic removal of cryo-damaged human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Uwe Paasch; Sonja Grunewald; Katja Wuendrich; Torsten Jope; Hans-Jurgen Glander

    2005-01-01

    Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMP, △ψm) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin Ⅴ-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin Ⅴ-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3%±2.2%. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P<0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin Ⅴ-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.

  17. Toxicity and uptake of heavy metals by human spermatozoa

    Energy Technology Data Exchange (ETDEWEB)

    Battersby, S.; Chandler, J.A.; Morton, M.S.

    1982-02-01

    The effects of metallic copper on human spermatozoa are described. Incubation with the metal caused a fall on the percentage of motile sperm, which was directly related to the surface area of copper employed and to the copper content of whole semen. These changes were accompanied by a decrease in semen zinc levels and an uptake of copper by individual sperm cells, as determined by X-ray microanalysis. Low concentrations of ionic copper caused a less marked fall in sperm motility, although the metal was generally more toxic than zinc or cadmium ions. All three metals were accumulated by spermatozoa on incubation, with cadmium causing a decrease in sperm nuclear zinc concentrations.

  18. Localization of Ureaplasma Urealyticum on Human Spermatozoa

    Institute of Scientific and Technical Information of China (English)

    徐晨; 王一飞

    1994-01-01

    Semen specimens were collected from 20 normal fertile men and 20 unexplained infertile men with U reaplasma urealyticum (U.U.) in semen. Spermatozoa of both groups were examined by immunogold technique and immunofluorescence test. A number of gold particles of the U. U. adhered to the sperm surface of infertile men were observed. Strong specific fluorescence was noticed on the sperm surface of infertile men, mostly on the midpiece and/ or postacrosomal region. A significant increase of sperm morphological abnormality, especially the swollen midpiece, the coiled tail, and head-tail angalation sperms was observed in infertile group. The sperm motility in infertile group is dramatically lower than that infertile group, It is hypothesized that teratospermia, poor motility and interference with sperm-ovum interaction might be the possible mechanisms for male infertility caused by Ureaplasma urealyticum.

  19. [The application of eosin and propidium iodide in evaluation of vitality of human spermatozoa].

    Science.gov (United States)

    Ploskonos, М В

    2014-11-01

    The article analyzes comparative assessment of vitality of spermatozoa by condition of permeability of membranes for eosin and propidium iodide and comparison of results acquired using technique of light and fluorescent microscopy. The comparison of data of light microscopy with eosin staining with data of fluorescent microscopy with propidium iodide staining demonstrated that percentage of content of spermatozoa separated from ejaculates of 28 fertile males and stained with eosin was reliably higher (34.8 ± 3.2) than percentage of content of spermatozoa with stained with propidium iodide (2.1 ± 4.0). After incubation of spermatozoa under room temperature during 24 hours percentage of unviable cells with stained eosin also was higher than in case of propidium iodide staining correspondingly (44.5 ± 3.3% and 34.7 ± 3.6%). The analysis of vitality of spermatozoa under damaging effect of oxidative stress on cell membrane developed by 4 hours incubation with 200 mkM of hydrogen peroxide (H2O2) demonstrated that under staining of spermatozoa with propidium iodide significantly higher percentage of damaged cells is detected. In such cases, eosin staining is less suitable for detection of vitality of spermatozoa (73.6 ± 5.8% against 51.7 ± 6.4%). The carried out experiment demonstrates that in case of detected effects on spermatozoa (for example, effect of oxidative stress) the light microscopy insufficiently adequate reflects degree of damage of membranes of spermatozoa. The fluorescent microscopy detects a higher percentage of spermatozoa with damaged membrane.

  20. Significance of Poisson distribution theory in analysing the interaction between human spermatozoa and zona-free hamster oocytes.

    Science.gov (United States)

    Aitken, R J; Elton, R A

    1984-11-01

    The value of Poisson distribution theory in describing and predicting the nature of sperm-egg interaction in vitro has been investigated using an interspecific in-vitro fertilization system, incorporating zona-free hamster oocytes and human spermatozoa. The frequency distribution of polyspermic oocyte penetrations in 72 experiments exhibited good agreement with the Poisson distribution at all levels of fertilization indicating that each oocyte must be of equal penetrability and that there can be no block to polyspermy in this interspecific system. Poisson distribution theory also accurately described the relationship between oocyte penetration and sperm motility in 50 out of 54 separate experiments spread across 10 serial dilution curves. For each dilution series the shape of the fitted curve was fixed but its location along the x-axis varied from donor to donor. The fixed nature of the relationship between sperm motility and egg penetration enables the results of such in-vitro fertilization experiments to be corrected for the number of motile spermatozoa in the incubation media. On the basis of these findings a protocol is described for assessing the results of the zona-free hamster oocyte penetration assay, which involves analysis of the degree of polyspermy followed by the application of Poisson distribution theory to correct the results to a standard concentration of motile spermatozoa. Changes in the penetrating ability of human spermatozoa after vasectomy and characterization of the degree of inter-ejaculate variation in penetrating potential are two clinical examples of such analyses given in the text. The statistical methods described in this paper should also be of general relevance to the study of fertilization mechanisms, in providing a rationale by which to analyse the quantitative nature of sperm-egg interaction in vitro.

  1. Relationship between ROS production, apoptosis and DNA denaturation in spermatozoa from patients examined for infertility

    National Research Council Canada - National Science Library

    Moustafa, Mohamed H; Sharma, Rakesh K; Thornton, Julie; Mascha, Edward; Abdel-Hafez, Mohammed A; Thomas, Jr, Anthony J; Agarwal, Ashok

    2004-01-01

    ...) in inducing DNA damage in ejaculated spermatozoa. We examined ejaculated spermatozoa from 31 patients examined for infertility and 19 healthy donors for apoptosis, production of ROS and DNA damage using annexin V binding, chemiluminescence assay...

  2. Effects of radiofrequency electromagnetic waves (RF-EMW) from cellular phones on human ejaculated semen: an in vitro pilot study.

    Science.gov (United States)

    Agarwal, Ashok; Desai, Nisarg R; Makker, Kartikeya; Varghese, Alex; Mouradi, Rand; Sabanegh, Edmund; Sharma, Rakesh

    2009-10-01

    To evaluate effects of cellular phone radiofrequency electromagnetic waves (RF-EMW) during talk mode on unprocessed (neat) ejaculated human semen. Prospective pilot study. Center for reproductive medicine laboratory in tertiary hospital setting. Neat semen samples from normal healthy donors (n = 23) and infertile patients (n = 9). After liquefaction, neat semen samples were divided into two aliquots. One aliquot (experimental) from each patient was exposed to cellular phone radiation (in talk mode) for 1 h, and the second aliquot (unexposed) served as the control sample under identical conditions. Evaluation of sperm parameters (motility, viability), reactive oxygen species (ROS), total antioxidant capacity (TAC) of semen, ROS-TAC score, and sperm DNA damage. Samples exposed to RF-EMW showed a significant decrease in sperm motility and viability, increase in ROS level, and decrease in ROS-TAC score. Levels of TAC and DNA damage showed no significant differences from the unexposed group. Radiofrequency electromagnetic waves emitted from cell phones may lead to oxidative stress in human semen. We speculate that keeping the cell phone in a trouser pocket in talk mode may negatively affect spermatozoa and impair male fertility.

  3. Kruger strict morphology and post-thaw progressive motility in cryopreserved human spermatozoa.

    Science.gov (United States)

    Lee, C-Y; Lee, C-T; Wu, C-H; Hsu, C-S; Hsu, M-I

    2012-05-01

    The purpose of this prospective study was to evaluate Kruger strict morphology and conventional semen analysis in predicting cryosurvival and the progressive motility recovery rate of frozen spermatozoa. Our study included 56 semen samples with >10 million spermatozoa per ejaculate. The main outcome measures were conventional semen analysis, strict morphology analysis by the Kruger method, cryosurvival rate and post-thaw sperm motility. A significant reduction in sperm motility after cryopreservation was demonstrated. The freeze-thawing process caused a 66% reduction in rapid progressive motile spermatozoa, a 45% reduction in slow progressive motile spermatozoa and a 2% reduction in nonprogressive motile spermatozoa. The cryosurvival and progressive motility recovery rates were not correlated with parameters of conventional semen analysis, such as sperm concentration, motility, WHO morphology and total motile count, but the progressive motility recovery rate was significantly correlated with the percentage of spermatozoa exhibiting Kruger normal morphology (P = 0.028). The recovery rate of rapidly progressive motility was profoundly decreased compared with slow progressive motility following the frozen-thaw procedure of semen. Kruger strict morphology assessment was a better predictor of the progressive motility recovery rate following the freezing-thaw procedure than parameters of conventional semen analysis.

  4. Bovine Circadian Locomotor Output Cycles Kaput (CLOCK) and Clusterin (CLU) mRNA Quantitation in Ejaculated Crossbred Bull Spermatozoa.

    Science.gov (United States)

    Kumar, S; Deb, R; Singh, U; Ganguly, I; Mandal, D K; Tyagi, S; Kumar, M; Sengar, G; Sharma, S; Singh, R; Singh, R

    2015-06-01

    Mammalian circadian locomotor output cycles kaput (CLOCK) gene encodes a transcription factor that affects both the persistence and the period of circadian rhythms. Earlier reports suggested that CLOCK gene might be associated with male infertility in human. Present investigation, for the first time, reports that CLOCK gene expresses differentially between good and poor quality crossbred bull semen. The relative expression of CLOCK was significantly (p < 0.05) higher among good quality bull semen than motility-impaired ones. Clusterins (CLU) are series of genes associated with a variety of physiological activities including spermatogenesis, apoptosis and degenerative disease conditions. In the present context, we also investigated that the expression of CLU gene was significantly (p < 0.05) higher among motility-impaired crossbred bull semen compared to the good quality one.

  5. Thiol oxidation by nitrosative stress: Cellular localization in human spermatozoa.

    Science.gov (United States)

    Cabrillana, María E; Uribe, Pamela; Villegas, Juana V; Álvarez, Juan; Sánchez, Raúl; Fornés, Miguel W

    2016-10-01

    Peroxynitrite is a highly reactive nitrogen species and when it is generated at high levels it causes nitrosative stress, an important cause of impaired sperm function. High levels of peroxynitrite have been shown to correlate with decreased semen quality in infertile men. Thiol groups in sperm are mainly found in enzymes, antioxidant molecules, and structural proteins in the axoneme. Peroxynitrite primarily reacts with thiol groups of cysteine-containing proteins. Although it is well known that peroxynitrite oxidizes sulfhydryl groups in sperm, the subcellular localization of this oxidation remains unknown. The main objective of this study was to establish the subcellular localization of peroxynitrite-induced nitrosative stress in thiol groups and its relation to sperm motility in human spermatozoa. For this purpose, spermatozoa from healthy donors were exposed in vitro to 3-morpholinosydnonimine (SIN-1), a compound which generates peroxynitrite. In order to detect peroxynitrite and reduced thiol groups, the fluorescent probes, dihydrorhodamine 123 and monobromobimane (mBBr), were used respectively. Sperm viability was analyzed by propidium iodide staining. Peroxynitrite generation and thiol redox state were monitored by confocal microscopy whereas sperm viability was evaluated by flow cytometry. Sperm motility was analyzed by CASA using the ISAS(®) system. The results showed that exposure of human spermatozoa to peroxynitrite results in increased thiol oxidation which is mainly localized in the sperm head and principal piece regions. Thiol oxidation was associated with motility loss. The high susceptibility of thiol groups to peroxynitrite-induced oxidation could explain, at least in part, the negative effect of reactive nitrogen species on sperm motility. DHR: dihydrorhodamine 123; mBBr: monobromobimane ONOO(-): peroxynitrite RNS: reactive nitrogen species RFI: relative fluorescence intensity SIN-1: 3-morpholinosydnonimine CASA: Computer-Aided Sperm Analysis

  6. Vitamin D is positively associated with sperm motility and increases intracellular calcium in human spermatozoa

    DEFF Research Database (Denmark)

    Blomberg Jensen, Martin; Bjerrum, Poul J; Jessen, Torben E;

    2011-01-01

    BACKGROUND The vitamin D receptor (VDR) is expressed in human spermatozoa, and VDR-knockout mice and vitamin D (VD) deficiency in rodents results in impaired fertility, low sperm counts and a low number of motile spermatozoa. We investigated the role of activated VD (1,25(OH)(2)D(3)) in human...

  7. Synergetic Effects of K, Ca, Cu and Zn in Human Semen in Relation to Parameters Indicative of Spontaneous Hyperactivation of Spermatozoa

    Science.gov (United States)

    Bolanca, Ivan; Obhodas, Jasmina; Ljiljak, Dejan; Matjacic, Lidija; Kuna, Krunoslav

    2016-01-01

    We have observed that sperm quality parameters indicative of spermatozoa hyperactivation such are lower “linearity” and “straightness”, and as showed by this research “elongation”, were more pronounced in patients with normal spermiogram compared to the group of men with reduced sperm motility who were undergoing routine in vitro fertilisation. The research encompassed 97 men diagnosed with normozoospermia (n = 20), asthenozoospermia (n = 54) and oligoasthenozoospermia (n = 23). The findings indicate that sperm quality of patients with normal spermiogram diagnosed according to WHO criteria, may be compromised by showing premature spontaneous hyperactivation which can decrease the chances of natural conception. We assessed synergistic effects of multiple chemical elements in ejaculated semen to find if premature spontaneous hyperactivation of spermatozoa can be a sign of imbalanced semen composition especially of elements K, Ca, Cu and Zn. Human semen samples showing low or high baseline status of chemical elements concentrations were found in samples from all three diagnostic groups. However, correlation of K/Ca and Cu/Zn ratios, taking into account samples from all three groups of men, were negative at statistical significance level p = 0.01. We tested if the negative correlation between K/Ca and Cu/Zn ratio works for greater number of semen samples. We found the negative correlation to be valid for 175 semen samples at statistical significance of p = 0.00002. The ratio of K/Ca and Cu/Zn, i.e. increased concentrations of K and Zn in comparison to concentrations of Ca and Cu, were associated with a decrease of “straightness” in the group of men with normal spermiogram and pronounced spontaneous hyperactivation of spermatozoa, implying that these elements act in synergy and that the balance of elements and not their absolute concentrations plays the major role in premature spermatozoa hyperactivation in ejaculated semen. PMID:27031102

  8. Synergetic Effects of K, Ca, Cu and Zn in Human Semen in Relation to Parameters Indicative of Spontaneous Hyperactivation of Spermatozoa.

    Directory of Open Access Journals (Sweden)

    Ivan Bolanca

    Full Text Available We have observed that sperm quality parameters indicative of spermatozoa hyperactivation such are lower "linearity" and "straightness", and as showed by this research "elongation", were more pronounced in patients with normal spermiogram compared to the group of men with reduced sperm motility who were undergoing routine in vitro fertilisation. The research encompassed 97 men diagnosed with normozoospermia (n = 20, asthenozoospermia (n = 54 and oligoasthenozoospermia (n = 23. The findings indicate that sperm quality of patients with normal spermiogram diagnosed according to WHO criteria, may be compromised by showing premature spontaneous hyperactivation which can decrease the chances of natural conception. We assessed synergistic effects of multiple chemical elements in ejaculated semen to find if premature spontaneous hyperactivation of spermatozoa can be a sign of imbalanced semen composition especially of elements K, Ca, Cu and Zn. Human semen samples showing low or high baseline status of chemical elements concentrations were found in samples from all three diagnostic groups. However, correlation of K/Ca and Cu/Zn ratios, taking into account samples from all three groups of men, were negative at statistical significance level p = 0.01. We tested if the negative correlation between K/Ca and Cu/Zn ratio works for greater number of semen samples. We found the negative correlation to be valid for 175 semen samples at statistical significance of p = 0.00002. The ratio of K/Ca and Cu/Zn, i.e. increased concentrations of K and Zn in comparison to concentrations of Ca and Cu, were associated with a decrease of "straightness" in the group of men with normal spermiogram and pronounced spontaneous hyperactivation of spermatozoa, implying that these elements act in synergy and that the balance of elements and not their absolute concentrations plays the major role in premature spermatozoa hyperactivation in ejaculated semen.

  9. Synergetic Effects of K, Ca, Cu and Zn in Human Semen in Relation to Parameters Indicative of Spontaneous Hyperactivation of Spermatozoa.

    Science.gov (United States)

    Bolanca, Ivan; Obhodas, Jasmina; Ljiljak, Dejan; Matjacic, Lidija; Kuna, Krunoslav

    2016-01-01

    We have observed that sperm quality parameters indicative of spermatozoa hyperactivation such are lower "linearity" and "straightness", and as showed by this research "elongation", were more pronounced in patients with normal spermiogram compared to the group of men with reduced sperm motility who were undergoing routine in vitro fertilisation. The research encompassed 97 men diagnosed with normozoospermia (n = 20), asthenozoospermia (n = 54) and oligoasthenozoospermia (n = 23). The findings indicate that sperm quality of patients with normal spermiogram diagnosed according to WHO criteria, may be compromised by showing premature spontaneous hyperactivation which can decrease the chances of natural conception. We assessed synergistic effects of multiple chemical elements in ejaculated semen to find if premature spontaneous hyperactivation of spermatozoa can be a sign of imbalanced semen composition especially of elements K, Ca, Cu and Zn. Human semen samples showing low or high baseline status of chemical elements concentrations were found in samples from all three diagnostic groups. However, correlation of K/Ca and Cu/Zn ratios, taking into account samples from all three groups of men, were negative at statistical significance level p = 0.01. We tested if the negative correlation between K/Ca and Cu/Zn ratio works for greater number of semen samples. We found the negative correlation to be valid for 175 semen samples at statistical significance of p = 0.00002. The ratio of K/Ca and Cu/Zn, i.e. increased concentrations of K and Zn in comparison to concentrations of Ca and Cu, were associated with a decrease of "straightness" in the group of men with normal spermiogram and pronounced spontaneous hyperactivation of spermatozoa, implying that these elements act in synergy and that the balance of elements and not their absolute concentrations plays the major role in premature spermatozoa hyperactivation in ejaculated semen.

  10. Physical characteristics of ejaculates produced by insemination boars depending on the interval between successive ejaculate collections

    Directory of Open Access Journals (Sweden)

    Magdalena BAJENA

    2016-06-01

    Full Text Available The ejaculate characteristics of Polish Landrace boars showed a significant correlation with the intervals between the successive ejaculate collections. The effect of insemination use intensity was however varied. Rising frequency of ejaculate collection led to a systematic and fairly even fall in ejaculate volume. Ejaculate sperm concentration remained at a relatively high level when ejaculates were collected with a frequency of 7 to 3 days but further shortening of the interval between the successive collections led to a drastic decrease in sperm concentration. An increase in ejaculate collection frequency to every four and fewer days resulted in a significant decrease in the number of spermatozoa present in the produced ejaculates and a concomitant decrease in the number of insemination doses prepared from these ejaculates, with an escalation of such changes.

  11. Binding of semenogelin I to intact human spermatozoa studied by flow cytometry and surface plasmon resonance.

    Science.gov (United States)

    Jonsson, Magnus; Frohm, Birgitta; Malm, Johan

    2010-01-01

    Approximately 1 in 10 couples is infertile. No definite cause can be found in about 25% of those cases. Studies have indicated that seminal vesicle secretion functions as an optimizer of fertilization. The Zn(2+) binding protein semenogelin I (SgI) represents a major fraction of the proteins present in seminal vesicle fluid, and it serves as a structural component of the coagulum that is formed after ejaculation. Cleavage of SgI by prostate-specific antigen results in liquefaction of the coagulum. Fragmented SgI has antibacterial effects and inhibits spermatozoa mobility. SgI has also been found complexed to eppin on spermatozoa, and this complex has been suggested to be of importance for fertility. Here, we used flow cytometry and surface plasmon resonance to study SgI regarding its association with spermatozoa and the interaction dependency on Zn(2+). The concentration of Zn(2+) in seminal plasma is approximately 100 times higher than in blood plasma, and the metal ion is known to change the structure of SgI. We found that SgI binds to spermatozoa in a concentration-dependent and saturable manner. In solution, SgI bound to spermatozoa in a non-Zn(2+)-dependent way, whereas immobilized SgI interacts with spermatozoa only in the presence of Zn(2+). It indicates that SgI must exhibit a specific structure or free flexibility to be able to interact with that ligand. Our results indicate that the association of SgI to spermatozoa is conformation dependent and specific. These findings could constitute a basis for the development of a male contraceptive.

  12. Spermatozoa-like cell invaders (nuclear vlimata) in human neoplasia.

    Science.gov (United States)

    Logothetou-Rella, H

    1993-07-01

    Spermatozoa-like cells (nuclear vlimata) have been identified in malignant cell cultures and embryonic cells, also common in the cytology and histology of all types of human neoplasia even after chemotherapy. A new mechanism of invasion of malignant cells has been described, according to which neoplastic cells behave and function as parasites using host-cells to divide, survive and eventually produce nuclear vlimata (bullets). Nuclear vlimata are the end cell products of incomplete, unequal, assymetrical division of neoplastic cells. The nuclear vlima exhibits similar morphology to spermatozoa and virus (head with, or without, tail) and invades the cytoplasm and/or nucleus of surrounding host-cells by a similar mechanism to sperm-oocyte interaction (fertilization) or viral cell infection, in the events of nuclear vlima-->tumor-->nuclear vlima-->tumor. The nuclear vlima head contains and transfers DNA, and when incorporated into the host-nucleus is indistinguishable from nucleoli and when in the cytoplasm is similar to sperm pronucleus, observed after sperm penetration of the oocyte. Function of nuclear vlimata is directly dependent on the specific extracellular matrix produced by malignant cells, consisting of glycosaminoglycans-protease-membranes. This mechanism of invasion constitutes the link of all scientific information concerning human neoplasia.

  13. Differential expression of VASA gene in ejaculated spermatozoa from normozoospermic men and patients with oligozoospermia%生殖细胞特异基因VASA在正常和少精子症患者精子中的表达研究

    Institute of Scientific and Technical Information of China (English)

    Xin Guo; Yao-Ting Gui; Ai-Fa Tang; Li-Hua Lu; Xin Gao; Zhi-Ming Cai

    2007-01-01

    Aim: To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods: Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction (RT-PCR), quantitative RT-PCR (QRT-PCR), immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results: VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion: The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.%目的:探讨生殖细胞高度特异性基因VASA在正常和少精子症患者精子中的表达,明确该基因在精子发生中的作用和相关机制.方法:正常和少精子症患者精液通过Percoll梯度分离获得精子.采用RT-PCR、realtime PCR、Western blot和免疫荧光方法分析该基因转录产物和蛋白表达.结果:VASA mRNA在射出精子中表达,正常精子VASA mRNA水平是少精子症精子的5倍;免疫荧光显示VASA蛋白主要定位在精子头部质膜和尾部,其表达强度正常组明显高于病变组,Western blot结果与免疫荧光结果相一致.结论:少

  14. Comprehensive review of the anatomy and physiology of male ejaculation: Premature ejaculation is not a disease.

    Science.gov (United States)

    Puppo, Vincenzo; Puppo, Giulia

    2016-01-01

    Human semen contains spermatozoa secreted by the testes and a mixture of components produced by the bulbo-urethral and Littre (paraurethral) glands, prostate, seminal vesicles, ampulla, and epididymis. Ejaculation is used as a synonym for the external ejection of semen, but it comprises two phases: emission and expulsion. As semen collects in the prostatic urethra, the rapid preorgasmic distension of the urethral bulb is pathognomonic of impeding orgasm, and the man experiences a sensation that ejaculation is inevitable (in women, emission is the only phase of orgasm). The semen is propelled along the penile urethra mainly by the bulbocavernosus muscle. With Kegel exercises, it is possible to train the perineal muscles. Immediately after the expulsion phase the male enters a refractory period, a recovery time during which further orgasm or ejaculation is physiologically impossible. Age affects the recovery time: as a man grows older, the refractory period increases. Sexual medicine experts consider premature ejaculation only in the case of vaginal intercourse, but vaginal orgasm has no scientific basis, so the duration of intercourse is not important for a woman's orgasm. The key to female orgasm are the female erectile organs; vaginal orgasm, G-spot, G-spot amplification, clitoral bulbs, clitoris-urethra-vaginal complex, internal clitoris and female ejaculation are terms without scientific basis. Female sexual dysfunctions are popular because they are based on something that does not exist, i.e. the vaginal orgasm. The physiology of ejaculation and orgasm is not impaired in premature ejaculation: it is not a disease, and non-coital sexual acts after male ejaculation can be used to produce orgasm in women. Teenagers and men can understand their sexual responses by masturbation and learn ejaculatory control with the stop-start method and the squeeze technique. Premature ejaculation must not be classified as a male sexual dysfunction. It has become the center of a

  15. Premature ejaculation

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/001524.htm Premature ejaculation To use the sharing features on this page, please enable JavaScript. Premature ejaculation is when a man has an orgasm ...

  16. Simple vitrification for small numbers of human spermatozoa.

    Science.gov (United States)

    Endo, Yuji; Fujii, Yoshitaka; Shintani, Kasumi; Seo, Momoyo; Motoyama, Hiroaki; Funahashi, Hiroaki

    2012-03-01

    Conventional freezing procedures and containers are not appropriate for spermatozoa from the testis because of their low number and poor in-situ motility, and various types of container have been utilized to freeze small numbers of spermatozoa. This study tried to develop a vitrification method for small numbers of spermatozoa using the Cell Sleeper, which is a closed type of cell-cryopreservation container. The container with spermatozoa were cooled in liquid nitrogen vapour and then stored in a cryotank. Sperm motility parameters improved significantly (P < 0.05) by vitrification in oil-free droplets rather than in droplets covered with oil. After vitrification of five spermatozoa per container, all spermatozoa were recovered and the viable sperm rate was significantly higher when spermatozoa were vitrified in a 3.5-ll droplet rather than in 0.5 ll (72.0% versus 38.0%; P < 0.01). Recovery, motility and viability rates of vitrified–warmed spermatozoa were similar between the Cell Sleeper and the CryoTop groups. In conclusion, the Cell Sleeper is a highly effective tool for the cryopreservation of small numbers of spermatozoa and limited cells can be vitrified quickly and simply without significant loss.

  17. Selenium in bovine spermatozoa.

    Science.gov (United States)

    Niemi, S M; Kuzan, F B; Senger, P L

    1981-05-01

    This study investigated the association of selenium with ejaculated bovine spermatozoa. Over 75% of the radioactive spermatozoa. Over 75% of the radioactive selenium-75 was released after 30 min of incubation in 2 X 10(-3) dithiothreitol. Of the selenium-75 released by dithiothreitol, 85% was associated with spermatozoal protein. Protein containing selenium-75 was found predominantly in a single band after polyacrylamide gel electrophoresis. Molecular weight was approximately 21,500 daltons.

  18. Characterization of NAADP-mediated calcium signaling in human spermatozoa

    Energy Technology Data Exchange (ETDEWEB)

    Sánchez-Tusie, A.A. [Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos (Mexico); Vasudevan, S.R.; Churchill, G.C. [Department of Pharmacology, University of Oxford, Oxford OX1 3QT, England (United Kingdom); Nishigaki, T. [Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos (Mexico); Treviño, C.L., E-mail: ctrevino@ibt.unam.mx [Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos (Mexico)

    2014-01-10

    Highlights: •Human sperm cells synthesize NAADP. •NAADP-AM mediates [Ca{sup 2+}]{sub i} increases in human sperm in the absence of [Ca{sup 2+}]{sub o}. •Human sperm have two acidic compartments located in the head and midpiece. -- Abstract: Ca{sup 2+} signaling in spermatozoa plays a crucial role during processes such as capacitation and release of the acrosome, but the underlying molecular mechanisms still remain unclear. Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca{sup 2+}-releasing second messenger in a variety of cellular processes. The presence of a NAADP synthesizing enzyme in sea urchin sperm has been previously reported, suggesting a possible role of NAADP in sperm Ca{sup 2+} signaling. In this work we used in vitro enzyme assays to show the presence of a novel NAADP synthesizing enzyme in human sperm, and to characterize its sensitivity to Ca{sup 2+} and pH. Ca{sup 2+} fluorescence imaging studies demonstrated that the permeable form of NAADP (NAADP-AM) induces intracellular [Ca{sup 2+}] increases in human sperm even in the absence of extracellular Ca{sup 2+}. Using LysoTracker®, a fluorescent probe that selectively accumulates in acidic compartments, we identified two such stores in human sperm cells. Their acidic nature was further confirmed by the reduction in staining intensity observed upon inhibition of the endo-lysosomal proton pump with Bafilomycin, or after lysosomal bursting with glycyl-L-phenylalanine-2-naphthylamide. The selective fluorescent NAADP analog, Ned-19, stained the same subcellular regions as LysoTracker®, suggesting that these stores are the targets of NAADP action.

  19. Evidence for compromised metabolic function and limited glucose uptake in spermatozoa from the teratospermic domestic cat (Felis catus) and cheetah (Acinonyx jubatus).

    Science.gov (United States)

    Terrell, Kimberly A; Wildt, David E; Anthony, Nicola M; Bavister, Barry D; Leibo, Stanley P; Penfold, Linda M; Marker, Laurie L; Crosier, Adrienne E

    2010-11-01

    Cheetahs and certain other felids consistently ejaculate high proportions (≥ 60%) of malformed spermatozoa, a condition known as teratospermia, which is prevalent in humans. Even seemingly normal spermatozoa from domestic cat teratospermic ejaculates have reduced fertilizing capacity. To understand the role of sperm metabolism in this phenomenon, we conducted a comparative study in the normospermic domestic cat versus the teratospermic cat and cheetah with the general hypothesis that sperm metabolic function is impaired in males producing predominantly pleiomorphic spermatozoa. Washed ejaculates were incubated in chemically defined medium containing glucose and pyruvate. Uptake of glucose and pyruvate and production of lactate were assessed using enzyme-linked fluorescence assays. Spermatozoa from domestic cats and cheetahs exhibited similar metabolic profiles, with minimal glucose metabolism and approximately equimolar rates of pyruvate uptake and lactate production. Compared to normospermic counterparts, pyruvate and lactate metabolism were reduced in teratospermic cat and cheetah ejaculates, even when controlling for sperm motility. Rates of pyruvate and lactate (but not glucose) metabolism were correlated positively with sperm motility, acrosomal integrity, and normal morphology. Collectively, our findings reveal that pyruvate uptake and lactate production are reliable, quantitative indicators of sperm quality in these two felid species and that metabolic function is impaired in teratospermic ejaculates. Furthermore, patterns of substrate utilization are conserved between these species, including the unexpected lack of exogenous glucose metabolism. Because glycolysis is required to support sperm motility and capacitation in certain other mammals (including dogs), the activity of this pathway in felid spermatozoa is a target for future investigation.

  20. Sperm fractions obtained following density gradient centrifugation in human ejaculates show differences in sperm DNA longevity

    Institute of Scientific and Technical Information of China (English)

    Jaime Goslvez; Stephen Johnston; Carmen Lpez-Fernndez; Altea Goslbez; Francisca Arroyo; Jose Lus Fernndez; Juan G lvarez

    2014-01-01

    Objective:To investigate the DNA longevity characteristics associated with each resultant fraction following density gradient centrifugation (DGC) in comparison to that of the original neat ejaculated sample. Methods:An aliquot of neat semen (NSS) collected from 7 patients was processed using DGC resulting in 3 fractions;Fraction 1:seminal plasma/40%gradient interface (GI);Fraction 2:40%GI/80%GI;Fraction 3:80%GI/pellet. An aliquot of each fraction and NSS was cryopreserved, thawed and incubated at 37 ℃for 24h;the increase of sperm DNA fragmentation was assessed using the Dyn-Halosperm assay following 0, 3, 6 and 24h of incubation. Results:While there was a significant reduction in the incidence of baseline sperm DNA fragmentation following DGC in Fraction 3, sperm DNA longevity was shown to be higher in the NSS than in any other sub-population following incubation. The highest levels of baseline DNA damage were found in Fractions 1 and 2;these fractions also showed the highest rate DNA fragmentation following incubation, subsequently exhibiting the lowest DNA longevity. Conclusion:1) Unnecessary incubation of spermatozoa prior to artificial insemination or in vitro fertilization, should be avoided, since sperm DNA longevity is significantly reduced after ex vivo sperm handling and 2) Although sperm selection by DCG significantly reduces the baseline levels of SDF of sperm in Fraction 3, sperm DNA longevity in this fraction was ultimately lower following 24 h incubation when compared to sperm recovered from non-centrifuged NSS.

  1. Concentration of trace elements in human semen and relation to spermatozoa quality.

    Science.gov (United States)

    Slivkova, Jana; Popelkova, Miroslava; Massanyi, Peter; Toporcerova, Silvia; Stawarz, Robert; Formicki, Grzegorz; Lukac, Norbert; Putała, Aldona; Guzik, Marek

    2009-03-01

    In this study the concentrations of trace elements such as lead, cadmium, iron, nickel, copper and zinc in the human semen (n = 47), occurrence of pathological spermatozoa, and correlations of these elements to pathological forms were investigated. For each sample of human spermatozoa at least 500 spermatozoa were evaluated. Metal contents were determined by the voltametric method and flame absorption spectrophotometry method. The concentrations of trace elements in human semen were: lead 1.49 +/- 0.40 mg x kg(- 1), cadmium 0.13 +/- 0.15 mg x kg(- 1), iron 2.59 +/- 0.21 mg x kg(- 1), nickel 0.40 +/- 0.07 mg x kg(- 1), copper 0.28 +/- 0.06 mg x kg(- 1), and zinc 153.93 +/- 67.08 mg x kg(- 1), respectively. The total percentage of pathological spermatozoa was 41.61 +/- 9.80% with predominancy of broken flagellum, flagellum torso and separated flagellum. In relation to trace elements the analysis showed correlation between copper and lead (r = -0.47), nickel and iron (r = 0.36), lead and flagellum ball (r = -0.39), cadmium and large heads (r = 0.37) and between iron and other forms of pathological spermatozoa (r = -0.32). Results of this study describe possible effects of trace elements on the spermatozoa quality in normal human sperm.

  2. DNA fragmentation of spermatozoa and assisted reproduction technology.

    Science.gov (United States)

    Henkel, Ralf; Kierspel, Eva; Hajimohammad, Marjam; Stalf, Thomas; Hoogendijk, Christiaan; Mehnert, Claas; Menkveld, Roelof; Schill, Wolf-Bernhard; Kruger, Thinus F

    2003-01-01

    Despite the ever-increasing knowledge of the fertilization process, there is still a need for better understanding of the causes of sperm DNA fragmentation and its impact on fertilization and pregnancy. For this reason, human sperm DNA fragmentation was investigated by means of the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling (TUNEL) assay and the production of reactive oxygen species (ROS) in the ejaculate and in the spermatozoa themselves. These data were correlated with fertilization and pregnancy data from IVF and intracytoplasmic sperm injection (ICSI) patients. Sperm DNA fragmentation did not correlate with fertilization rate, but there was a significantly reduced pregnancy rate in IVF patients inseminated with TUNEL-positive spermatozoa. ICSI patients exhibited the same tendency. This implies that spermatozoa with damaged DNA are able to fertilize an oocyte, but at the time the paternal genome is switched on, further development stops. The determination of ROS in the ejaculate and the percentage of ROS-producing spermatozoa revealed markedly stronger correlations between sperm functions (i.e. motility) and the percentage of ROS-producing spermatozoa. The influence of seminal leukocytes, known to produce large amounts of oxidants, on sperm DNA fragmentation should not be neglected.

  3. Cryoprotectant-free vitrification of human spermatozoa in new artificial seminal fluid.

    Science.gov (United States)

    Agha-Rahimi, A; Khalili, M A; Nottola, S A; Miglietta, S; Moradi, A

    2016-11-01

    Vitrification is a new method that has been recently introduced in Assisted Reproduction Technique programs. The aim of this study was to design a new medium similar to normal human seminal fluid (SF), formulation artificial seminal fluid (ASF), and to compare the cryoprotective potency of this medium with SF and human tubal fluid (HTF) medium. Thirty normal ejaculates were processed with the swim-up technique and sperm suspensions were divided into four aliquots: (i) fresh sample (control); (ii) vitrification in HTF medium supplemented with 5 mg/mL human serum albumin and 0.25 mol sucrose (Vit HTF); (iii) vitrification with patients' SF (Vit SF); and (iv) vitrification in ASF (Vit ASF). After warming, sperm parameters of motility, viability, and morphology were analyzed using WHO criteria. Also, sperm pellets were fixed in 2.5% glutaraldehyde and processed for scanning electron microscopy and transmission electron microscopy observations. The results showed that progressive motility (46.09 ± 10.33 vs. 36.80 ± 13.75), grade A motility (36.59 ± 11.40 vs. 16.41 ± 11.24), and normal morphology (18.74 ± 8.35 vs. 11.85 ± 5.84) and viability (68.22 ± 10.83 vs. 60.86 ± 11.72) of spermatozoa were significantly higher in Vit ASF than in Vit HTF. All parameters were better in Vit ASF than in Vit SF, but only viability was significantly different (p = 0.006). After cryopreservation, deep invagination in cytoplasm and mechanically weak point sites and folded tail were commonly observed. But, this phenomenon was more significant in Vit HTF and Vit SF than in ASF (p HTF and Vit SF than in ASF (p HTF. © 2016 American Society of Andrology and European Academy of Andrology.

  4. Effects of 1950 MHz W-CDMA-like signal on human spermatozoa.

    Science.gov (United States)

    Nakatani-Enomoto, Setsu; Okutsu, Miho; Suzuki, Satoshi; Suganuma, Ryota; Groiss, Stefan Jun; Kadowaki, Suguru; Enomoto, Hiroyuki; Fujimori, Keiya; Ugawa, Yoshikazu

    2016-09-01

    There are growing concerns about how electromagnetic waves (EMW) emitted from mobile phones affect human spermatozoa. Several experiments have suggested harmful effects of EMW on human sperm quality, motility, velocity, or the deoxyribonucleic acid (DNA) of spermatozoa. In this study, we analyzed the effects on human spermatozoa (sperm motility and kinetic variables) induced by 1 h of exposure to 1950 MHz Wideband Code Division Multiple Access (W-CDMA)-like EMW with specific absorption rates of either 2.0 or 6.0 W/kg, using a computer-assisted sperm analyzer system. We also measured the percentage of 8-hydroxy-2'-deoxyguanosine (8-OHdG) positive spermatozoa with flow cytometry to evaluate damage to DNA. No significant differences were observed between the EMW exposure and the sham exposure in sperm motility, kinetic variables, or 8-OHdG levels. We conclude that W-CDMA-like exposure for 1 h under temperature-controlled conditions has no detectable effect on normal human spermatozoa. Differences in exposure conditions, humidity, temperature control, baseline sperm characteristics, and age of donors may explain inconsistency of our results with several previous studies. Bioelectromagnetics. 37:373-381, 2016. © 2016 Wiley Periodicals, Inc.

  5. Quantitative evaluation of p53 as a new indicator of DNA damage in human spermatozoa

    Directory of Open Access Journals (Sweden)

    Salvatore Raimondo

    2014-01-01

    The aim of this study was to assess if a p53 ELISA assay could be a new indicator of DNA damage in human spermatozoa. Materials and Methods: 103 human semen samples were evaluated using both Acridine Orange test and p53 ELISA and results were compared. Results: A clear correlation between the values measured by two methods was obtained. Conclusions: If this hypothesis will be confirmed by further studies, the p53 ELISA assay could become a new and more precise indicator of DNA damage in human spermatozoa.

  6. Premature Ejaculation

    Science.gov (United States)

    ... to help delay ejaculation, including antidepressants called selective serotonin reuptake inhibitors (SSRIs). These antidepressants are available with your doctor's prescription. However, the U.S. Food and Drug Administration (FDA) has not approved the ...

  7. Acid extrusion from human spermatozoa is mediated by flagellar voltage-gated proton channel.

    Science.gov (United States)

    Lishko, Polina V; Botchkina, Inna L; Fedorenko, Andriy; Kirichok, Yuriy

    2010-02-05

    Human spermatozoa are quiescent in the male reproductive system and must undergo activation once introduced into the female reproductive tract. This process is known to require alkalinization of sperm cytoplasm, but the mechanism responsible for transmembrane proton extrusion has remained unknown because of the inability to measure membrane conductance in human sperm. Here, by successfully patch clamping human spermatozoa, we show that proton channel Hv1 is their dominant proton conductance. Hv1 is confined to the principal piece of the sperm flagellum, where it is expressed at unusually high density. Robust flagellar Hv1-dependent proton conductance is activated by membrane depolarization, an alkaline extracellular environment, endocannabinoid anandamide, and removal of extracellular zinc, a potent Hv1 blocker. Hv1 allows only outward transport of protons and is therefore dedicated to inducing intracellular alkalinization and activating spermatozoa. The importance of Hv1 for sperm activation makes it an attractive target for controlling male fertility.

  8. Resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing

    Directory of Open Access Journals (Sweden)

    Sidney Verza Jr.

    2009-10-01

    Full Text Available Objective: To study the resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing by using the fast liquid nitrogen vapor method. Material and Methods: Semen specimens were obtained from sixteen normal and oligozoospermic individuals who required disposal at the sperm bank. Five of them had testicular cancer. Specimens were thawed and an aliquot was removed for analysis. The remaining specimens were refrozen without removing the cryomedia. Repeated freeze-thaw cycles were performed until no motile sperm were observed. Sperm motility, number of motile spermatozoa and viability were determined after thawing. Resistance to cryoinjury was compared between groups and also after each refreezing cycle within groups. Results: Motile spermatozoa were recovered after five and two refreeze-thawing cycles in normozoospermic and oligozoospermic specimens, respectively. There were no significant differences in the recovery of motile spermatozoa between thaws within each group of normal and oligozoospermic specimens, but percentage motility and total number of motile spermatozoa were significantly lower in the oligozoospermic one. Specimens from men with cancer were exposed to six refreeze-thawing cycles. Although recovery of motile spermatozoa was significantly impaired after each thawing, there were no significant differences in the recovery of motile sperm between thaws in cancer and non-cancer groups. Conclusions: Human spermatozoa resist repeated cryopreservation using the fast liquid nitrogen vapor method. Normozoospermic specimens withstand refreezing for an average two cycles longer than oligozoospermic ones. Specimens from cancer patients seem to resist repeated cryoinjury similarly to non-cancer counterparts. Resistance to repeated cryoinjury was related to the initial semen quality.

  9. Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining.

    Science.gov (United States)

    Golan, R; Shochat, L; Weissenberg, R; Soffer, Y; Marcus, Z; Oschry, Y; Lewin, L M

    1997-01-01

    The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.

  10. Effect of zinc oxide nanoparticles on viability of human spermatozoa

    OpenAIRE

    Abolfazl Barkhordari; Seyedhossein Hekmatimoghaddam; Ali Jebali; Mohammad Ali Khalili; Alireza Talebi; Marzieh Noorani

    2013-01-01

    Background: The extensive use of different nanoparticles has raised great concerns about their occupational and biological safety. Objective: The aim of this study was to evaluate the cytotoxic effect of zinc oxide nanoparticles (ZnO NPs) on viability of spermatozoa. Materials and Methods: Semen samples were obtained from 15 healthy persons, and were analyzed using WHO guidelines. Each semen sample was separately incubated with different concentrations of ZnO NPs (10, 100, 500, and 1000 µg/mL...

  11. Cryoprotectant-free ultra-rapid freezing of human spermatozoa in cryogenic vials.

    Science.gov (United States)

    Zhu, J; Jin, R-T; Wu, L-M; Johansson, L; Guo, T-H; Liu, Y-S; Tong, X-H

    2014-08-01

    The objective of this study was to investigate the effect of ultra-rapid freezing (direct immersion in liquid nitrogen) on human spermatozoa in cryogenic vials (≥0.5 ml) at different concentrations of sucrose. After swim-up, the sperm suspensions (N = 58) were diluted with sperm preparation medium and divided into six aliquots: swim-up (fresh), conventional freezing group (slow freezing) and four ultra-rapid freezing groups containing sucrose at different concentrations (0.15 m, 0.20 m, 0.25 m and 0.30 m). Sperm motility, progressive motility, plasma membrane integrity, DNA stability and acrosome integrity of fresh and cooled-warmed spermatozoa were analysed. The progressive motility, plasma membrane and acrosome integrity of spermatozoa in the 0.20 m sucrose group were significantly higher than those of the slow freezing group (47.5 ± 6.8% versus 36.4 ± 8.7%, 73.2 ± 6.9% versus 63.9 ± 6.3%, 53.7 ± 10.0% versus 35.9 ± 9.7% respectively, P 0.05 respectively) between the 0.20 m sucrose and the slow freezing group. No differences were found between the ultra-rapid and slow freezing group at the other concentrations of sucrose. Our findings suggest that the method of ultra-rapid freezing of human spermatozoa in cryogenic vials with a solution containing 0.20 m sucrose results in recovery of spermatozoon of superior qualities. In contrast to slow freezing, the ultra-rapid freezing technique of human spermatozoa seems to reduce cryoinjuries and maintain important physiological characteristics of the spermatozoa after warming. © 2013 Blackwell Verlag GmbH.

  12. A radiomicroassay for cytotoxic antibody to human spermatozoa. Quantification by tritiated actinomycin d.

    Science.gov (United States)

    Sung, J S; Shizuya, H; Black, D D; Mumford, D M

    1977-03-01

    A radiomicroassay for titration of spermocytotoxic antibody is described. The assay used [3H]AACTINOMYCIN D ([3H]Act D) to label damaged spermatozoa in a fashion analogous to penetration by vital dye. Optimal conditions for and some kinetics of the assay are presented. The assay is sensitive, reliable, simple to perform and uses only small amounts of serum and spermatozoa. Applied to sperm antibody positive human postvasectomy sera, the assay compared favourably in sensitivity eith vital dye microscopic observations and with parallel titration by the Isojima's immobilization tests.

  13. Inhibition of human spermatozoa-zona pellucida binding by a combinatorially derived peptide from a synthetic target.

    Science.gov (United States)

    Pieczenik, George; Garrisi, John; Cohen, Jacques

    2006-09-01

    Intact zona-free human oocytes were screened using a combinatorial peptide library selection protocol. Pieczenik Peptide Sequence 1 (PPS1) HEHRKRG binds human spermatozoa. A complementary and unique binding sequence HNSSLSPLATPA (PPS2) was developed from the first PPS1 ligand that binds to the human zona pellucida or oolemma. Cytoplasm-free zonae from unfertilized eggs were obtained and used as an assay system to test the effects of exposure to these two ligands. Spermatozoa were inserted into evacuated zonae and their behaviour and binding activity were assessed at regular intervals. The behaviour of spermatozoa exposed to PPS1 and unlabelled spermatozoa injected into unexposed zonae was similar as far as binding was concerned (50 and 54% binding), but PPS1 exposed spermatozoa had higher motility and displacement, marked by their escape from the zona pellucida. Zonae exposed to PPS2 inhibited the interaction between injected spermatozoa and the inside of the zona when compared with controls (8.3 and 53.8% attached respectively, P movie sequence taken approximately 30 min after injection of spermatozoa into empty human zonae pellucidae shows behaviour of non-manipulated spermatozoa into zonae not exposed or exposed to ligand. This may be purchased for viewing on the Internet at www.rbmonline.com/Article/2159 (free to web subscribers).

  14. Slow cryopreservation is not superior to vitrification in human spermatozoa; an experimental controlled study

    Directory of Open Access Journals (Sweden)

    Mohamed Shehata Ali Mohamed

    2015-10-01

    Full Text Available Background: Spermatozoa cryopreservation is used for the management of infertility and some other medical conditions. The routinely applied cryopreservation technique depends on permeating cryoprotectants, whose toxic effects have raised the attention towards permeating cryoprotectants-free vitrification technique. Objective: To compare between the application of slow cryopreservation and vitrification on human spermatozoa. Materials and Methods: This was an experimental controlled study involving 33 human semen samples, where each sample was divided into three equal parts; fresh control, conventional slow freezing, and permeating cryoprotectants-free vitrification. Viability and mitochondrial membrane potential (MMP of control and post-thawing spermatozoa were assessed with the sperm viability kit and the JC-1 kit, respectively, using fluorescence-activated cell sorting analysis. Results: Significant reduction of the progressive motility, viability and MMP was observed by the procedure of freezing and thawing, while there was not any significant difference between both cryopreservation techniques. Cryopreservation resulted in 48% reduction of the percentage of viable spermatozoa and 54.5% rise in the percentage of dead spermatozoa. In addition, high MMP was reduced by 24% and low MMP was increased by 34.75% in response to freezing and thawing. Progressive motility of spermatozoa was correlated significantly positive with high MMP and significantly negative with low MMP in control as well as post-thawing specimens (r=0.8881/ -0.8412, 0.7461/ -0.7510 and 0.7603/ -0.7839 for control, slow and vitrification respectively, p=0.0001. Conclusion: Although both cryopreservation techniques have similar results, vitrification is faster, easier and associated with less toxicity and costs. Thus, vitrification is recommended for the clinical application.

  15. MOLECULAR ANALYSIS OF HUMAN SPERMATOZOA: POTENTIAL FOR INFERTILITY RESEARCH AND SCREENING

    Science.gov (United States)

    Molecular Analysis of Human Spermatozoa: Potential for Infertility Research and ScreeningDavid Miller1, David Dix2, Robert Reid3, Susan Wykes3 and Stephen Krawetz3 1Reproductive Biology Group, University of Leeds, UK2Reproductive Toxicology Division, U.S. Environmenta...

  16. RNASET2 in human spermatozoa and seminal plasma: a novel relevant indicator for asthenozoospermia.

    Science.gov (United States)

    Liu, Y; Chen, G; Lu, L; Sun, H; Guo, Q; Xue, K; Fan, Y; Ding, Z

    2013-01-01

    Adequate sperm motility is requisite for human fertilization, whereas the underlying causes or mechanisms of impaired sperm motility, asthenozoospermia, still remain largely unknown. RNASET2 (Ribonuclease T2) may be one of the effectors modulating human sperm motility. We determined if there is a correlation between RNASET2 expression levels in human semen from asthenozoospermia and fertile individuals. Thus, RNASET2 expression levels in spermatozoa and seminal plasma of healthy and asthenozoospermia individuals were evaluated using Western blot, laser scanning confocal microscope analysis, ELISA and flow cytometry. The results revealed that RNASET2 expression was identified in both human spermatozoa and seminal plasma. In spermatozoa from fertile individuals, it was localized to the acrosome, neck and the middle piece of tail regions. However, in spermatozoa from asthenozoospermia individuals (n = 67), RNASET2 staining was especially more frequent and evident in the neck and middle piece than that in fertile individuals (n = 59, p asthenozoospermia than in fertile individuals (p asthenozoospermia individuals may contribute to sperm motility impairment.

  17. Evaluación de la unión espermatozoide-ADN exógeno en espermatozoides porcinos eyaculados y epididimarios Evaluation of binding sperm-exogenous DNA in ejaculate and epididimary porcine spermatozoa

    Directory of Open Access Journals (Sweden)

    FA García-Vázquez

    2009-01-01

    , agriculture and biomedicine. Sperm mediated gene transfer (SMGT is an interesting tool for animal transgenesis consisting on the intrinsic ability of the spermatic cells to bind and internalize exogenous DNA and allow their transfer into oocytes after fertilization, to become part of the genome of the new embryo. The seminal plasma plays an important role acting as a natural barrier and protecting the spermatozoa from exogenous molecules that could compromise their integrity. So, the epididymal spermatozoa are a valuable model to explore the possible effect of seminal plasma components. The objective of this study was to evaluate the interaction among sperm and transgene using Epididymal (EP vs Ejaculated (EJ sperm without seminal plasma. Linealized plasmid (GFP (5.7 kb labelled with fluorescein was added (1x10(8 spermatozoa/ml + 5µg DNA/ml and incubated at 16 ºC. DNA binding and viability were measured simultaneously by flow cytometry during 120 minutes of incubation. The results showed that EP spermatozoa present a similar DNA-binding ability (12.63 ± 1.23% vs 10.94 ± 1.05%, P = 0.31 and viability throughout the incubation (14.64 ± 0.94% vs 13.42 ± 0.61%, P = 0.23 than EJ. We only detected a greater percentage of living DNA-bound spermatozoa in EP compared to EJ (2.10 ± 0.33% vs 1.05 ± 0.14%, P < 0.01. The DNA-binding was associated mainly to dead sperm or with low viability in both groups (EP: 10.53 ± 1.01% vs EJ: 9.89 ± 0.97%, P = 0.98. These results open new ways to explore and use epididymal spermatozoa in diverse applications (artificial insemination, in vitro fertilization and ICSI associated with SMGT method.

  18. Prolonged incubation of processed human spermatozoa will increase DNA fragmentation.

    Science.gov (United States)

    Nabi, A; Khalili, M A; Halvaei, I; Roodbari, F

    2014-05-01

    One of the causes of failure in ART is sperm DNA fragmentation which may be associated with long period of spermatozoa incubation at 37 °C. The objective was to evaluate the rate of sperm DNA fragmentation using the sperm chromatin dispersion (SCD) test after swim-up at different time intervals prior to use. In this prospective study, 21 normozoospermic specimens were analysed. The samples were incubated at 37 °C after preparation by direct swim-up. DNA fragmentation was assessed at different time intervals (0, 1, 2 and 3 h) using SCD test. Spermatozoa with no DNA fragmentation showed large- or medium-sized halos, and sperm cells with DNA fragmentation showed either a small halo or no halo. The rates of normal morphology and progressive motility after sperm processing were 72.33 ± 2.53% and 90 ± 1.02%, respectively. The rate of sperm DNA fragmentation was significantly higher after 2 h (8.81 ± 0.93%, P = 0.004) and 3 h (10.76 ± 0.89%, P fragmentation. Therefore, sperm samples intended for ART procedures should be used within 2 h of incubation at 37 °C. © 2013 Blackwell Verlag GmbH.

  19. Evaluation of protamines 1 and 2 transcript contents in spermatozoa from asthenozoospermic men.

    Directory of Open Access Journals (Sweden)

    Magdalena Depa-Martynow

    2008-04-01

    Full Text Available During mammalian spermatogenesis, the chromatin structure undergoes substantial condensation. The key role in this process is played by protamines 1 and 2 (PRM1, PRM2. We attempted to compare the levels of PRM1 and PRM2 transcripts in mature spermatozoa of normospermic and asthenozoospermic men. Human ejaculates from normozoospermic (n=70 and asthenozoospermic (n=100 donors were purified by centrifugation through discontinuous Percoll density gradient. RNA was isolated from spermatozoa according to the ChomczyĂąski and Sacchi method, treated with DNase I, and reverse-transcribed into cDNA. Using reverse transcription and real-time quantitative polymerase chain reaction analysis, we found a reduction in the levels of PRM1 and PRM2 transcripts in spermatozoa from asthenozoospermic men, as compared to controls (P<0.001. Our findings indicate that a reduction in contents of PRM1 and PRM2 transcripts in spermatozoa may be linked with asthenozoospermia.

  20. Physiology and Pharmacology of Ejaculation.

    Science.gov (United States)

    Clement, Pierre; Giuliano, François

    2016-10-01

    Ejaculation is the final stage of coitus in mammalian male and is mandatory for natural procreation. Two synchronized phases, emission and expulsion, form the ejaculatory response and involve specific organs and anatomical structures. The peripheral events leading to ejaculation are commanded by autonomic (sympathetic and parasympathetic) and somatic divisions of the nervous system. The autonomic and somatic motor efferents originate in spinal nuclei located in thoracolumbar and lumbosacral segments. Co-ordinated activation of autonomic and somatic spinal nuclei is orchestrated by a group of lumbar spinal interneurons defined as the spinal generator of ejaculation. The generator of ejaculation together with the autonomic and somatic spinal nuclei constitutes a spinal network that is under the strong influence of stimulating or inhibiting genital sensory and supraspinal inputs. A brain circuitry dedicated to ejaculation has been delineated that is part of a more global network controlling other aspects of the sexual response. This circuitry includes discrete neuronal populations distributed in all divisions of the brain. The corollary to the expanded CNS network is the variety of neurotransmitter systems participating in the ejaculatory process. Among them, serotonin neurotransmission plays a key role and its targeting led to the development of the first registered pharmacological treatment of premature ejaculation in human beings. Critical gaps remain in the understanding of neurophysiopharmacology of ejaculation and management of ejaculatory disorders in human beings needs improvement. Because the ejaculatory response in laboratory animals and in human beings shares many similarities, the use of animal models will certainly provide further advances in the field.

  1. Reduced levels of intracellular calcium releasing in spermatozoa from asthenozoospermic patients

    Directory of Open Access Journals (Sweden)

    García Juan F

    2009-02-01

    Full Text Available Abstract Background Asthenozoospermia is one of the most common findings present in infertile males characterized by reduced or absent sperm motility, but its aetiology remains unknown in most cases. In addition, calcium is one of the most important ions regulating sperm motility. In this study we have investigated the progesterone-evoked intracellular calcium signal in ejaculated spermatozoa from men with normospermia or asthenozoospermia. Methods Human ejaculates were obtained from healthy volunteers and asthenospermic men by masturbation after 4–5 days of abstinence. For determination of cytosolic free calcium concentration, spermatozoa were loaded with the fluorescent ratiometric calcium indicator Fura-2. Results Treatment of spermatozoa from normospermic men with 20 micromolar progesterone plus 1 micromolar thapsigargin in a calcium free medium induced a typical transient increase in cytosolic free calcium concentration due to calcium release from internal stores. Similar results were obtained when spermatozoa were stimulated with progesterone alone. Subsequent addition of calcium to the external medium evoked a sustained elevation in cytosolic free calcium concentration indicative of capacitative calcium entry. However, when progesterone plus thapsigargin were administered to spermatozoa from patients with asthenozoospermia, calcium signal and subsequent calcium entry was much smaller compared to normospermic patients. As expected, pretreatment of normospermic spermatozoa with both the anti-progesterone receptor c262 antibody and with progesterone receptor antagonist RU-38486 decreased the calcium release induced by progesterone. Treatment of spermatozoa with cytochalasin D or jasplakinolide decreased the calcium entry evoked by depletion of internal calcium stores in normospermic patients, whereas these treatments proved to be ineffective at modifying the calcium entry in patients with asthenozoospermia. Conclusion Our results suggest

  2. Assessment of reproductive tissues of gilts born from magnetic nanoparticles-treated boar spermatozoa

    Science.gov (United States)

    Semen ejaculates contain a heterogeneous population of spermatozoa that may interfere with male fertility. Indeed, poor quality semen generally translates into low fertility rates that are attributed to higher proportions of damaged or abnormal spermatozoa in the ejaculates. It is likely that their ...

  3. Cryopreservation of human spermatozoa. III. The effect of cryoprotectants on motility.

    Science.gov (United States)

    Critser, J K; Huse-Benda, A R; Aaker, D V; Arneson, B W; Ball, G D

    1988-08-01

    A series of experiments was conducted to examine potential toxic effects of cryoprotectants on motility of human spermatozoa. The data indicated that exposure of spermatozoa to cryoprotectant medium for as little as 15 minutes at room temperature caused a reduction in motility. This reduction in motility was caused by glycerol. Lowering glycerol concentrations from 7.5% to 5.0% improved sperm motility at 24 hours post-thaw. Sperm motility was not affected by either slow or abrupt cooling rates above -5 degrees C. Motility was greater in cryopreserved sperm at 24 hours post-thaw when glycerol was added at -5 degrees C rather than at room temperature. These data suggest that avoiding glycerol toxicity either by reducing the concentration used or by adding glycerol at a lower temperature, or both, may improve human sperm cryosurvival rates.

  4. The Effect of Fibronectin on the Fertilization Capacity of Human Spermatozoa

    Institute of Scientific and Technical Information of China (English)

    许复华; 崔晓; 杨抚华

    1996-01-01

    Fibronectin (Fn) is a major molecular glycoprotein that is known to play an important role in many systems involved in cell-to-cell interation and cell-to-matrix adhesion. We initially studied the effect of Fn on the fertilization capacity of human spermatozoa with a penetration test in vitro. Following co-incubation of Fn in a heterologous system (human spermatozoa and zona-free golden hamster oocytes ), we have noted a significant decrease in the rate of sperm penetration into oolemma at 2.8 μg Fn/ml,and a complete inhibition of fertilization at 15 μg Fn/ml. Besides, we noted a specific increase in the penetration rate during the co incubation of anti-Fn-serum in a heterologous system. These results suggest that extrinsic Fn binds with the Fn-receptor in the oolemma, thereby, competitively inhibiting fertilization.

  5. AMP-activated kinase in human spermatozoa: identification, intracellular localization, and key function in the regulation of sperm motility.

    Science.gov (United States)

    Calle-Guisado, Violeta; de Llera, Ana Hurtado; Martin-Hidalgo, David; Mijares, Jose; Gil, Maria C; Alvarez, Ignacio S; Bragado, Maria J; Garcia-Marin, Luis J

    2016-09-27

    AMP-activated kinase (AMPK), a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This work's aims are AMPK identification, intracellular localization, and their role in human spermatozoa function. Semen was obtained from healthy human donors. Sperm AMPK and phospho-Thr172-AMPK were analyzed by Western blotting and indirect immunofluorescence. High- and low-quality sperm populations were separated by a 40%-80% density gradient. Human spermatozoa motility was evaluated by an Integrated Semen Analysis System (ISAS) in the presence or absence of the AMPK inhibitor compound C (CC). AMPK is localized along the human spermatozoa, at the entire acrosome, midpiece and tail with variable intensity, whereas its active form, phospho-Thr172-AMPK, shows a prominent staining at the acrosome and sperm tail with a weaker staining in the midpiece and the postacrosomal region. Interestingly, spermatozoa bearing an excess residual cytoplasm show strong AMPK staining in this subcellular compartment. Both AMPK and phospho-Thr172-AMPK human spermatozoa contents exhibit important individual variations. Moreover, active AMPK is predominant in the high motility sperm population, where shows a stronger intensity compared with the low motility sperm population. Inhibition of AMPK activity in human spermatozoa by CC treatment leads to a significant reduction in any sperm motility parameter analyzed: percent of motile sperm, sperm velocities, progressivity, and other motility coefficients. This work identifies and points out AMPK as a new molecular mechanism involved in human spermatozoa motility. Further AMPK implications in the clinical efficiency of assisted reproduction and in other reproductive areas need to be studied.

  6. Spermatozoa-li ke cell invaders, nuclear vlimata, in human neoplasia

    OpenAIRE

    1993-01-01

    Spermatozoa-like cells (nuclear vlimata) have been identified in malignant cell cultures and embryonic cells, also common in the cytology and histology of all types of human neoplasia even after chemotherapy. A new mechanism of invasion of malignant cells has been described, according to which neoplastic cells behave and function as parasites using host-cells to divide, survive and eventually produce nuclear vlimata (bullets). Nuclear vlimata are the end ce...

  7. Effects of pentoxifylline treatment before freezing on motility, viability and acrosome status of poor quality human spermatozoa cryopreserved by the liquid nitrogen vapor method

    Directory of Open Access Journals (Sweden)

    S.C. Esteves

    2007-07-01

    Full Text Available The objective of the present study was to investigate the effects of the direct addition of pentoxifylline (PF to the ejaculates of men with poor sperm quality before freezing on post-thaw sperm motility, viability, acrosome integrity, and agonist-induced acrosome reaction. Semen specimens from 16 infertile men with impaired sperm count and motility (oligoasthenozoospermia were divided into two equal aliquots: one received no treatment (control while the other was incubated with 5 mM PF (treated. Both aliquots were cryopreserved by the liquid nitrogen vapor method. Motility was assessed according to WHO criteria. Acrosome integrity and spontaneous and calcium ionophore-induced acrosome reactions were assessed with fluorescein isothiocyanate-conjugated peanut agglutinin combined with a supra-vital dye (Hoechst-33258. Cryopreservation impaired sperm motility (percentage reduction: 87.4 (interquartile range, IQ: 70.3-92.9 vs 89.1 (IQ: 72.7-96.0%, viability (25.9 (IQ: 22.2-29.7 vs 25.6 (IQ: 19.7-40.3% and acrosome integrity (18.9 (IQ: 5.4-38.9 vs 26.8 (IQ: 0.0-45.2% to the same extent in both treated and control aliquots. However, PF treatment before freezing improved the acrosome reaction to ionophore challenge test scores in cryopreserved spermatozoa (9.7 (IQ: 6.6-19.7 vs 4.8 (IQ: 0.5-6.8%; P = 0.002. These data show that pre-freeze treatment of poor quality human sperm with pentoxifylline did not improve post-thaw motility or viability nor did it prevent acrosomal loss during the freeze-thaw process. However, PF, as used, improved the ability of thawed spermatozoa to undergo the acrosome reaction in response to calcium ionophore. The present data indicate that treatment of poor quality human sperm with PF may enhance post-thaw sperm fertilizing ability.

  8. In vitro incubation of human spermatozoa promotes reactive oxygen species generation and DNA fragmentation.

    Science.gov (United States)

    Cicaré, J; Caille, A; Zumoffen, C; Ghersevich, S; Bahamondes, L; Munuce, M J

    2015-10-01

    The aim of this study was to investigate the oxidative process associated with sperm capacitation and its impact on DNA fragmentation and sperm function. Redox activity and lipid peroxidation were analysed in human spermatozoa after 3, 6 and 22 h of incubation in Ham's F10 medium plus bovine albumin at 37° and 5% CO2 for capacitation. DNA status, tyrosine phosphorylation pattern and induced acrosome reaction were evaluated after capacitating conditions. At 22 h of incubation, there was a significant (P DNA fragmentation. These results indicate that when spermatozoa are incubated for several hours (22 h), a common practice in assisted reproductive techniques, an increase in oxidative sperm metabolism and in the proportion of fragmented DNA should be expected. However, there was no effect on any of the other functional parameters associated with sperm fertilising capacity. © 2014 Blackwell Verlag GmbH.

  9. Reactive oxygen species, lipid peroxidation and enzymatic defence systems in human spermatozoa.

    Science.gov (United States)

    Griveau, J F; Dumont, E; Renard, P; Callegari, J P; Le Lannou, D

    1995-01-01

    The reactive oxygen species, hydrogen peroxide (H2O2) and superoxide anion (O2o-), were generated with a xanthine-xanthine oxidase system and their effect on human sperm function was studied. The action of reactive oxygen species on selected human spermatozoa resulted in a decreased capacity for ionophore-induced acrosome reaction, a decrease in sperm motility, an increase in the concentration of lipid hydroperoxides and a loss of membrane polyunsaturated fatty acids. H2O2 was the key intermediate of the deleterious effects exerted by the xanthine and xanthine oxidase. Among these parameters, the acrosome reaction appeared most susceptible to the reactive oxygen species generated by the xanthine-xanthine oxidase system, and was decreased without sperm motility being affected. Treatment with H2O2 was shown to inactivate several enzymatic activities involved in the antioxidant defence of spermatozoa: glutathione peroxidase, superoxide dismutase and glucose-6-phosphate dehydrogenase. H2O2 and O2o- were shown to be involved in the lipid alterations triggered by the xanthine-xanthine oxidase system. Singlet oxygen is proposed to intervene in the lipoperoxidation process. The inefficacy of mannitol in protecting spermatozoa suggests that hydroxyl radicals were not produced in the extracellular medium.

  10. Feasibility of refreezing human spermatozoa through the technique of liquid nitrogen vapor

    Directory of Open Access Journals (Sweden)

    Sidney Verza Jr

    2004-12-01

    Full Text Available OBJECTIVE: To assess the feasibility of refreezing human semen using the technique of liquid nitrogen vapor with static phases. MATERIALS AND METHODS: Twenty samples from 16 subjects who required disposal of their cryopreserved semen were thawed, corresponding to 6 cancer patients and 10 participants in the assisted reproduction (AR program. Samples were refrozen using the technique of liquid nitrogen vapor with static phases, identical to the one used for the initial freezing, and thawed again after 72 hours. We assessed the concentration of motile spermatozoa, total and progressive percent motility and spermatic vitality, according to criteria of the World Health Organization (WHO, as well as spermatic morphology according to the strict Kruger criterion, after the first and after the second thawing. RESULTS: We observed a significant decrease in all the parameters evaluated between the first and the second thawing. Median values for the concentration of motile spermatozoa decreased from 2.0x10(6/mL to 0.1x10(6/mL (p < 0.01; total percent motility from 42% to 22.5% (p < 0.01; progressive percent motility from 34% to 9.5% (p < 0.01; vitality from 45% to 20% (p < 0.01; and morphology from 5% to 5% (p = 0.03. There was no significant difference in the spermatic parameters between the cancer and assisted reproduction groups, both after the first and after the second thawing. We observed that in 100% of cases there was retrieval of motile spermatozoa after the second thawing. CONCLUSIONS: Refreezing of human semen by the technique of liquid nitrogen vapor allows the retrieval of viable spermatozoa after thawing.

  11. Acrosome Reaction and Ca2+ Imaging in Single Human Spermatozoa: New Regulatory Roles of [Ca2+]i1

    Science.gov (United States)

    Sánchez-Cárdenas, Claudia; Servín-Vences, Martha Rocio; José, Omar; Treviño, Claudia Lydia; Hernández-Cruz, Arturo; Darszon, Alberto

    2014-01-01

    ABSTRACT The spermatozoa acrosome reaction (AR) is essential for mammalian fertilization. Few methods allow visualization of AR in real time together with Ca2+ imaging. Here, we show that FM4-64, a fluorescent dye used to follow exocytosis, reliably reports AR progression induced by ionomycin and progesterone in human spermatozoa. FM4-64 clearly delimits the spermatozoa contour and reports morphological cell changes before, during, and after AR. This strategy unveiled the formation of moving tubular appendages, emerging from acrosome-reacted spermatozoa, which was confirmed by scanning electron microscopy. Alternate wavelength illumination allowed concomitant imaging of FM4-64 and Fluo-4, a Ca2+ indicator. These AR and intracellular Ca2+ ([Ca2+]i) recordings revealed that the presence of [Ca2+]i oscillations, both spontaneous and progesterone induced, prevents AR in human spermatozoa. Notably, the progesterone-induced AR is preceded by a second [Ca2+]i peak and ∼40% of reacting spermatozoa also manifest a slow [Ca2+]i rise ∼2 min before AR. Our findings uncover new AR features related to [Ca2+]i. PMID:25100708

  12. Obesity and Bariatric Surgery Drive Epigenetic Variation of Spermatozoa in Humans

    DEFF Research Database (Denmark)

    Donkin, Ida; Versteyhe, Soetkin; Ingerslev, Lars R.

    2016-01-01

    Obesity is a heritable disorder, with children of obese fathers at higher risk of developing obesity. Environmental factors epigenetically influence somatic tissues, but the contribution of these factors to the establishment of epigenetic patterns in human gametes is unknown. Here, we hypothesized...... that weight loss remodels the epigenetic signature of spermatozoa in human obesity. Comprehensive profiling of the epigenome of sperm from lean and obese men showed similar histone positioning, but small non-coding RNA expression and DNA methylation patterns were markedly different. In a separate cohort...

  13. Physiological roles of semenogelin I and zinc in sperm motility and semen coagulation on ejaculation in humans.

    Science.gov (United States)

    Yoshida, Kaoru; Kawano, Natsuko; Yoshiike, Miki; Yoshida, Manabu; Iwamoto, Teruaki; Morisawa, Masaaki

    2008-03-01

    At ejaculation, human sperm are considered to be mechanically trapped and become immotile in the semen coagulum by binding to semenogelins (Sgs) from the seminal vesicle and zinc ions from the prostate. However, the physiological combined roles of the protein and heavy metal on sperm motility are unknown. Here, we have first demonstrated that Sg I alone, which does not form the semen coagulum without zinc, is an inhibitor of the motility of intact human sperm at physiological concentration. On the other hand, zinc ions alone had no effect on sperm motility, but confer recovery of sperm motility that has been inhibited by Sg I at a concentration equal to or less than 1 mg/ml. These observations suggest that the roles played by Sg I and zinc on sperm motility are not mechanical but physiological. Quartz crystal microbalance analysis suggests that the sperm extract first bind to Sg I and then zinc ions which subsequently increase the protein accumulation, suggesting that Sgs inhibit sperm motility by directly binding to the sperm surface. Further accumulation of Sg I mediated by zinc ions may entrap the quiescent sperm at semen ejaculation.

  14. Sperm fractions obtained following density gradient centrifugation in human ejaculates show differences in sperm DNA longevity

    Directory of Open Access Journals (Sweden)

    Jaime Gosálvez

    2014-06-01

    Conclusion: 1 Unnecessary incubation of spermatozoa prior to artificial insemination or in vitro fertilization, should be avoided, since sperm DNA longevity is significantly reduced after ex vivo sperm handling and 2 Although sperm selection by DCG significantly reduces the baseline levels of SDF of sperm in Fraction 3, sperm DNA longevity in this fraction was ultimately lower following 24 h incubation when compared to sperm recovered from non-centrifuged NSS.

  15. Mitochondrial permeability transition increases reactive oxygen species production and induces DNA fragmentation in human spermatozoa.

    Science.gov (United States)

    Treulen, Favián; Uribe, Pamela; Boguen, Rodrigo; Villegas, Juana V

    2015-04-01

    Does mitochondrial permeability transition (MPT) induced by calcium overload cause reactive oxygen species (ROS) production and DNA fragmentation in human spermatozoa? Studies conducted in vitro suggest that in human spermatozoa, MPT occurs in response to intracellular calcium increase and is associated with mitochondrial membrane potential (ΔΨm) dissipation, increased ROS production and DNA fragmentation. Oxidative stress is a major cause of defective sperm function in male infertility. By opening calcium-dependent pores in the inner mitochondrial membrane (IMM), MPT causes, among other things, increased ROS production and ΔΨm dissipation in somatic cells. MPT as a mechanism for generating oxidative stress and DNA fragmentation in human spermatozoa has not been studied. Human sperm were exposed to ionomycin for 1.5 h (n = 8) followed by analysis of sperm IMM permeability, ΔΨm, ROS production and DNA fragmentation. To evaluate the MPT in sperm cells, the calcein-AM and cobalt chloride method was used. The ΔΨm was evaluated by JC-1 staining, intracellular ROS production was evaluated with dihydroethidium and DNA fragmentation was evaluated by a modified TUNEL assay. Measurements were performed by fluorescence microscopy, confocal laser microscopy and flow cytometry. Decreased calcein fluorescence after treatment with ionomycin (P fragmentation. ROS production occurred prior to the decrease in ΔΨm. The study was carried out in vitro using motile sperm from healthy donors; tests on sperm from infertile patients were not carried out. We propose that the MPT, due to pores opening in sperm IMM, is an important mechanism of increased ROS and DNA fragmentation. Therefore, agents that modulate the opening of these pores might contribute to the prevention of damage by oxidative stress in human spermatozoa. This study was funded by grant DI12-0102 from the Universidad de La Frontera (J.V.V.) and a doctoral scholarship from CONICYT Chile (F.T.). The authors disclose

  16. Destabilization of acrosome and elastase influence mediate the release of secretory phospholipase A2 from human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Jacqueline Leβig; Uta Reibetanz; Jürgen Arnhold; Hans-Jürgen Glander

    2008-01-01

    Aim: To determine the cellular distribution of secretory phospholipase A2 (sPLA2) in dependence on the acrosomal state and under the action of elastase released under inflammatory processes from leukocytes. Methods: Acrosome reaction of spermatozoa was triggered by calcimycin. Human leukocyte elastase was used to simulate in flammatory conditions. To visualize the distribution of sPLA2 and to determine the acrosomal state, immunofluorescence tech-niques and lectin binding combined with confocal laser scanning fluorescence microscopy and flow cytometry were used. Results: Although sPLA2 was detected at the acrosome and tail regions in intact spermatozoa, it disappearedfrom the head region after triggering the acrosome reaction. This release of sPLA2 was associated with enhanced binding of annexin V-fluoroscein isothiocyanate (FITC) to spermatozoa surfaces, intercalation of ethidium-homodimer L and bnding of FITC-iabelled concanavalin A at the acrosomal region. Spermatozoa from healthy subjects treated with elastase were characterized by release of sPLA2, disturbance of acrosome structure, and loss of vitality. Conclusion:The ability of spermatozoa to release secretory phospholipase A2 is related to the acrosomal state. Premature destabi-lization of the acrosome and loss of sPLA2 can occur during silent inflammations in the male genital tract. The distribution pattern of sPLA2 in intact spermatozoa might be an additional parameter for evaluating sperm quality.

  17. Large Scale 7436-bp Deletions in Human Sperm Mitochondrial DNA with Spermatozoa Dysfunction and Male Infertility.

    Science.gov (United States)

    Ambulkar, Prafulla S; Waghmare, Jwalant E; Chaudhari, Ajay R; Wankhede, Vandana R; Tarnekar, Aaditya M; Shende, Moreshwar R; Pal, Asoke K

    2016-11-01

    Mitochondria and mitochondrial DNA are essential to sperm motility and fertility. It controls growth, development and differentiation through oxidation energy supply. Mitochondrial (mtDNA) deletions or mutation are frequently attributed to defects of sperm motility and finally these deletions lead to sperm dysfunction and causes infertility in male. To investigate the correlation between large scale 7436-bp deletions in sperm mtDNA and non-motility of sperm in asthenozoospermia and Oligoasthenoteratozoospermia (OAT) infertile men. The present prospective study was carried out in Human Genetic Division, Department of Anatomy, Mahatma Gandhi Institute of Medical Sciences, Sevagram from June 2014 to July 2016. We have studied 110 asthenozoospermia and OAT infertile men whose semen profile indicated abnormal motility and 50 normal fertile controls. Of 110 infertile men, 70 had asthenozoospermia and 40 had OAT. Fractionations of spermatozoa were done in each semen sample on the basis of their motility by percoll gradients discontinuous technique. Long-range PCR was used for detection of 7436-bp deletions in sperm mtDNA and was confirmed by primer shift technique. Overall eight subjects (8/110; 7.2%) of which six (6/70; 8.57%) asthenozoospermia and two (2/40; 5%) OAT had shown deletions of 7436-bp. In 40% percoll fraction had more non-motile spermatozoa than 80% percoll fraction. The non-motile spermatozoa in 40% percoll fractions showed more mtDNA deletions (7.2%) than the motile spermatozoa in 80% percoll fraction (2.7%). The sequencing of flanking regions of deleted mtDNA confirmed 7436-bp deletions. Interestingly, no deletions were found in control subjects. Though, the frequency of 7436-bp deletions in sperm mtDNA was low in infertile cases but meaningful indications were there when results were compared with controls. It is indicated that large scale deletions 7436-bp of mtDNA is associated with abnormal sperm motility. The 7436-bp deletions of mtDNA in spermatozoa

  18. Large Scale 7436-bp Deletions in Human Sperm Mitochondrial DNA with Spermatozoa Dysfunction and Male Infertility

    Science.gov (United States)

    Ambulkar, Prafulla S.; Waghmare, Jwalant E.; Chaudhari, Ajay R.; Wankhede, Vandana R.; Tarnekar, Aaditya M.; Shende, Moreshwar R.

    2016-01-01

    Introduction Mitochondria and mitochondrial DNA are essential to sperm motility and fertility. It controls growth, development and differentiation through oxidation energy supply. Mitochondrial (mtDNA) deletions or mutation are frequently attributed to defects of sperm motility and finally these deletions lead to sperm dysfunction and causes infertility in male. Aim To investigate the correlation between large scale 7436-bp deletions in sperm mtDNA and non-motility of sperm in asthenozoospermia and Oligoasthenoteratozoospermia (OAT) infertile men. Materials and Methods The present prospective study was carried out in Human Genetic Division, Department of Anatomy, Mahatma Gandhi Institute of Medical Sciences, Sevagram from June 2014 to July 2016. We have studied 110 asthenozoospermia and OAT infertile men whose semen profile indicated abnormal motility and 50 normal fertile controls. Of 110 infertile men, 70 had asthenozoospermia and 40 had OAT. Fractionations of spermatozoa were done in each semen sample on the basis of their motility by percoll gradients discontinuous technique. Long-range PCR was used for detection of 7436-bp deletions in sperm mtDNA and was confirmed by primer shift technique. Results Overall eight subjects (8/110; 7.2%) of which six (6/70; 8.57%) asthenozoospermia and two (2/40; 5%) OAT had shown deletions of 7436-bp. In 40% percoll fraction had more non-motile spermatozoa than 80% percoll fraction. The non-motile spermatozoa in 40% percoll fractions showed more mtDNA deletions (7.2%) than the motile spermatozoa in 80% percoll fraction (2.7%). The sequencing of flanking regions of deleted mtDNA confirmed 7436-bp deletions. Interestingly, no deletions were found in control subjects. Conclusion Though, the frequency of 7436-bp deletions in sperm mtDNA was low in infertile cases but meaningful indications were there when results were compared with controls. It is indicated that large scale deletions 7436-bp of mtDNA is associated with abnormal

  19. Mobile phone radiation induces reactive oxygen species production and DNA damage in human spermatozoa in vitro.

    Directory of Open Access Journals (Sweden)

    Geoffry N De Iuliis

    Full Text Available BACKGROUND: In recent times there has been some controversy over the impact of electromagnetic radiation on human health. The significance of mobile phone radiation on male reproduction is a key element of this debate since several studies have suggested a relationship between mobile phone use and semen quality. The potential mechanisms involved have not been established, however, human spermatozoa are known to be particularly vulnerable to oxidative stress by virtue of the abundant availability of substrates for free radical attack and the lack of cytoplasmic space to accommodate antioxidant enzymes. Moreover, the induction of oxidative stress in these cells not only perturbs their capacity for fertilization but also contributes to sperm DNA damage. The latter has, in turn, been linked with poor fertility, an increased incidence of miscarriage and morbidity in the offspring, including childhood cancer. In light of these associations, we have analyzed the influence of RF-EMR on the cell biology of human spermatozoa in vitro. PRINCIPAL FINDINGS: Purified human spermatozoa were exposed to radio-frequency electromagnetic radiation (RF-EMR tuned to 1.8 GHz and covering a range of specific absorption rates (SAR from 0.4 W/kg to 27.5 W/kg. In step with increasing SAR, motility and vitality were significantly reduced after RF-EMR exposure, while the mitochondrial generation of reactive oxygen species and DNA fragmentation were significantly elevated (P<0.001. Furthermore, we also observed highly significant relationships between SAR, the oxidative DNA damage bio-marker, 8-OH-dG, and DNA fragmentation after RF-EMR exposure. CONCLUSIONS: RF-EMR in both the power density and frequency range of mobile phones enhances mitochondrial reactive oxygen species generation by human spermatozoa, decreasing the motility and vitality of these cells while stimulating DNA base adduct formation and, ultimately DNA fragmentation. These findings have clear implications

  20. Image cytometer method for automated assessment of human spermatozoa concentration

    DEFF Research Database (Denmark)

    Egeberg, D L; Kjaerulff, S; Hansen, C

    2013-01-01

    to investigator bias. Here we show that image cytometry can be used to accurately measure the sperm concentration of human semen samples with great ease and reproducibility. The impact of several factors (pipetting, mixing, round cell content, sperm concentration), which can influence the read-out as well......In the basic clinical work-up of infertile couples, a semen analysis is mandatory and the sperm concentration is one of the most essential variables to be determined. Sperm concentration is usually assessed by manual counting using a haemocytometer and is hence labour intensive and may be subjected...... and easy measurement of human sperm concentration....

  1. Team swimming in ant spermatozoa.

    Science.gov (United States)

    Pearcy, Morgan; Delescaille, Noémie; Lybaert, Pascale; Aron, Serge

    2014-06-01

    In species where females mate promiscuously, competition between ejaculates from different males to fertilize the ova is an important selective force shaping many aspects of male reproductive traits, such as sperm number, sperm length and sperm-sperm interactions. In eusocial Hymenoptera (bees, wasps and ants), males die shortly after mating and their reproductive success is ultimately limited by the amount of sperm stored in the queen's spermatheca. Multiple mating by queens is expected to impose intense selective pressure on males to optimize the transfer of sperm to the storage organ. Here, we report a remarkable case of cooperation between spermatozoa in the desert ant Cataglyphis savignyi. Males ejaculate bundles of 50-100 spermatozoa. Sperm bundles swim on average 51% faster than solitary sperm cells. Team swimming is expected to increase the amount of sperm stored in the queen spermatheca and, ultimately, enhance male posthumous fitness.

  2. The opening of maitotoxin-sensitive calcium channels induces the acrosome reaction in human spermatozoa: differences from the zona pellucida

    Institute of Scientific and Technical Information of China (English)

    Julio C Chávez; Claudia L Trevi(n)o; Gerardo A de Blas; José L de la Vega-Beltrán; Takuya Nishigaki; Mayel Chirinos; María Elena González-González; Fernando Larrea; Alejandra Solís; Alberto Darszon

    2011-01-01

    The acrosome reaction(AR),an absolute requirement for spermatozoa and egg fusion,requires the influx of Ca2+into the spermatozoa through voltage-dependent Ca2+channels and store-operated channels.Maitotoxin(MTx),a Ca2+-mobilizing agent,has been shown to be a potent inducer of the mouse sperm AR,with a pharmacology similar to that of the zona pellucida(ZP),possibly suggesting a common pathway for both inducers.Using recombinant human ZP3(rhZP3),mouse ZP and two MTx channel blockers(U73122 and U73343),we investigated and compared the MTx-and ZP-induced ARs in human and mouse spermatozoa.Herein,we report that MTx induced AR and elevated intracellular Ca2+([Ca2+]1)in human spermatozoa,both of which were blocked by U73122 and U73343.These two compounds also inhibited the MTx-induced AR in mouse spermatozoa.In disagreement with our previous proposal,the AR triggered by rhZP3 or mouse ZP was not blocked by U73343,indicating that in human and mouse spermatozoa,the AR induction by the physiologicalligands or by MTx occurred through distinct pathways.U73122,but not U73343(inactive analogue),can block phospholipase C(PLC).Another PLC inhibitor,edelfosine,also blocked the rhZP3-and ZP-induced ARs.These findings confirmed the participation of a PLC-dependent signalling pathway in human and mouse zona protein-induced AR.Notably,edelfosine also inhibited the MTx-induced mouse sperm AR but not that of the human,suggesting that toxin-induced AR is PLC-dependent in mice and PLC-independent in humans.

  3. Protective effects of exogenous gangliosides on ROS-induced changes in human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Mirjana Gavella; Vaskresenija Lipovac

    2013-01-01

    This article summarizes the available evidence on the efficacy of gangliosides to reduce the degree of reactive oxygen species (ROS)-mediated damage.The antioxidative efficacy of exogenous gangliosides in protecting different cells encouraged us to examine their ability to protect human spermatozoa.Gangliosides are sialic acid-containing glycosphingolipids with strong amphiphilic character due to the bulky headgroup made of several sugar rings with sialic acid residues and the double-tailed hydrophobic lipid moiety.The amphiphilicity of gangliosides allows them to exist as micelles in aqueous media when they are present at a concentration above their critical micellar concentration.The protective effect of ganglioside micelles on spermatozoa is believed to stem from their ability to scavenge free radicals and prevent their damaging effects.In our study,we particularly focused our attention on the protective effect of ganglioside micelles on DNA in human spermatozoa exposed to cryopreservation.The results indicate that ganglioside micelles can modulate the hydrophobic properties of the sperm membrane to increase tolerance to DNA fragmentation,thus protecting the DNA from cryopreservation-induced damage.Further actions of ganglioside micelles,which were documented by biochemical and biophysical studies,included (i) the modulation of superoxide anion generation by increasing the diffusion barrier for membrane events responsible for signal translocation to the interior of the cell; (ii) the inhibition of iron-catalysed hydroxyl radical formation due to the iron chelation potential of gangliosides; and (iii) inhibition of hydrogen peroxide diffusion across the sperm membrane.

  4. Involvement of seminal leukocytes, reactive oxygen species, and sperm mitochondrial membrane potential in the DNA damage of the human spermatozoa.

    Science.gov (United States)

    Lobascio, A M; De Felici, M; Anibaldi, M; Greco, P; Minasi, M G; Greco, E

    2015-03-01

    Measurement of reactive oxygen species (ROS) producing leukocytes in semen has been a standard component of the semen analysis, but its true significance remains still unknown. In this study, we have correlated the number of seminal leukocytes to various semen parameters. We found a negative correlation between the leukocyte number and sperm concentration (rs  = -0.22; p = 0.01) and motility (rs  = -0.20; p = 0.02). In contrast, a positive correlation between the number of leukocytes and both seminal ROS (rs  = 0.70, p sperm mitochondrial membrane potential (MMP) (10% vs 35%, rs  = 0.25, p = 0.08; n = 50). Overall these results indicate that the presence of high number of leukocytes in the ejaculate negatively affects key semen parameters, as sperm concentration and motility, associated with infertility conditions. Moreover, they suggest that leukocytes are the major source of the seminal ROS and cause of sperm DNA fragmentation. However, the absence of a clear correlation between ROS and sperm DNA fragmentation, and spermatozoa with damaged DNA and MMP loss, suggest that ROS produced by leukocytes might be not the only cause of DNA damage in spermatozoa and that intrinsic mitochondrial-dependent apoptotic pathways might not have a major impact on sperm DNA fragmentation.

  5. Co-Incubation of Human Spermatozoa with Anti-VDAC Antibody Reduced Sperm Motility

    Directory of Open Access Journals (Sweden)

    Bianjiang Liu

    2014-01-01

    Full Text Available Background: Voltage-dependent anion channel (VDAC, a channel protein, exists in the outer mitochondrial membrane of somatic cells and is involved in multiple physiological and pathophysiological processes. Up until now, little has been known about VDAC in male germ cells. In the present study, the relationship between VDAC and human sperm motility was explored. Methods: Highly motile human spermatozoa were incubated in vitro with anti-VDAC antibody. Total sperm motility, straight line velocity (VSL, curvilinear velocity (VCL, and average path velocity (VAP were recorded. Intracellular free calcium concentration ([Ca2+]i, pH value (pHi, and ATP content were determined. Results: Co-incubation with anti-VDAC antibody reduced VSL, VCL, and VAP of spermatozoa. Co-incubation further reduced [Ca2+]i. Anti-VDAC antibody did not significantly alter total sperm motility, pHi and intracellular ATP content. Conclusion: The data suggest that co-incubation with anti-VDAC antibody reduces sperm motility through inhibition of Ca2+ transmembrane flow. In this way, VDAC participates in the modulation of human sperm motility through mediating Ca2+ transmembrane transport and exchange.

  6. Premature ejaculation

    Directory of Open Access Journals (Sweden)

    Chris G McMahon

    2007-01-01

    Full Text Available Premature ejaculation (PE is a common male sexual disorder. Recent normative data suggests that men with an intravaginal ejaculatory latency time (IELT of less than 1 minute have "definite" PE, while men with IELTs between 1 and 1.5 minutes have "probable" PE. Although there is insufficient empirical evidence to identify the etiology of PE, there is limited correlational evidence to suggest that men with PE have high levels of sexual anxiety and inherited altered sensitivity of central 5-HT (5-hydroxytryptamine, serotonin receptors. Pharmacological modulation of the ejaculatory threshold using off-label daily or on-demand selective serotonin re-uptake inhibitors is well tolerated and offers patients a high likelihood of achieving improved ejaculatory control within a few days of initiating treatment, consequential improvements in sexual desire and other sexual domains. Investigational drugs such as the ejaculo-selective serotonin transport inhibitor, dapoxetine represent a major development in sexual medicine. These drugs offer patients the convenience of on-demand dosing, significant improvements in IELT, ejaculatory control and sexual satisfaction with minimal adverse effects.

  7. Premature ejaculation.

    Science.gov (United States)

    McMahon, Chris G

    2007-04-01

    Premature ejaculation (PE) is a common male sexual disorder. Recent normative data suggests that men with an intravaginal ejaculatory latency time (IELT) of less than 1 minute have "definite" PE, while men with IELTs between 1 and 1.5 minutes have "probable" PE. Although there is insufficient empirical evidence to identify the etiology of PE, there is limited correlational evidence to suggest that men with PE have high levels of sexual anxiety and inherited altered sensitivity of central 5-HT (5-hydroxytryptamine, serotonin) receptors. Pharmacological modulation of the ejaculatory threshold using off-label daily or on-demand selective serotonin re-uptake inhibitors is well tolerated and offers patients a high likelihood of achieving improved ejaculatory control within a few days of initiating treatment, consequential improvements in sexual desire and other sexual domains. Investigational drugs such as the ejaculo-selective serotonin transport inhibitor, dapoxetine represent a major development in sexual medicine. These drugs offer patients the convenience of on-demand dosing, significant improvements in IELT, ejaculatory control and sexual satisfaction with minimal adverse effects.

  8. Malate dehydrogenase activity in human seminal plasma and spermatozoa homogenates

    Directory of Open Access Journals (Sweden)

    Hulya Leventerler

    2013-08-01

    Full Text Available Purpose: Malate Dehydrogenase is an important enzyme of the Krebs cycle, most cells require this enzyme for their metabolic activity. We evaluated the Malate Dehydrogenase (NAD/NADP activity in human seminal plasma and sperm homogenates in normozoospermic, fertile and infertile males. Also glucose and fructose concentrations were determined in the seminal plasma samples. Material and Methods: Malate Dehydrogenase (NAD/NADP activity in human seminal plasma and sperm homogenates of normozoospermic and infertile males was determined by spectrophotometric method. Semen analysis was considered according to the WHO Criteria. Results: Malat Dehydrogenase-NAD value in seminal plasma (the mean ± SD, mU/ml of asthenoteratospermic (40.0±25.7 and azospermic (38.0±43.6 groups were significantly lower than normozoospermic, (93.9±52.1 males. Malat Dehydrogenase-NAD value in sperm homogenates (the mean ± SD, mU/ 20x106 sperm of teratospermic group (136.8±61.8 was significantly higher compared to the normozoospermic (87.3±26.5 males. Glucose concentration (mg/dl in asthenoteratospermic (4.0±1.4 and azospermic (15.4±6.4 groups were significantly higher than fertile (2.0±2.1 males. Also fructose concentration (mg/dl in asthenoteratospermic (706.6±143.3 and azospermic (338.1±228.2 groups were significantly high compared to the normozoospermic (184.7±124.8 group. Conclusion: Sperm may be some part of the source of Malat Dehydrogenase activity in semen. Malat Dehydrogenase activity in seminal plasma has an important role on energy metabolism of sperm. Intermediate substrates of Krebs cycle might have been produced under the control of Malat Dehydrogenase and these substrates may be important for sperm motility and male infertility. [Cukurova Med J 2013; 38(4.000: 648-658

  9. Retrograde ejaculation, painful ejaculation and hematospermia

    OpenAIRE

    Parnham, Arie; Serefoglu, Ege Can

    2016-01-01

    Although there has been an increased interest on premature ejaculation in the recent years, our understanding regarding the disorders of retrograde ejaculation, painful ejaculation and hematospermia remain limited. All three of these conditions require a keen clinical acumen and willingness to engage in thinking outside of the standard established treatment paradigm. The development of novel investigational techniques and treatments has led to progress in the management of these conditions sy...

  10. PARK7 protein translocating into spermatozoa mitochondria in Chinese asthenozoospermia.

    Science.gov (United States)

    Sun, Yi; Zhang, Wen-Jia; Zhao, Xin; Yuan, Ren-Pei; Jiang, Hui; Pu, Xiao-Ping

    2014-09-01

    PARK7 (DJ1) is a multifunctional oxidative stress response protein that protects cells against reactive oxygen species (ROS) and mitochondrial damage. PARK7 defects are known to cause various physiological dysfunctions, including infertility. Asthenozoospermia (AS), i.e. low-motile spermatozoa in the ejaculate, is a common cause of human male infertility. In this study, we found that downregulation of PARK7 resulted in increased levels of lipid peroxide and ROS, decreased mitochondrial membrane potential, and reduced mitochondrial complex I enzyme activity in the spermatozoa from AS patients. Furthermore, it was observed that PARK7 was translocated into the mitochondria of damaged spermatozoa in AS. Finally, we examined the oxidative state of PARK7 and the results demonstrated the enhancement of oxidation, expressed by increased sulfonic acid residues, the highest form of oxidation, as the sperm motility decreased. Taken together, these results revealed that PARK7 deficiency may increase the oxidative stress damage to spermatozoa. Our present findings open new avenues of therapeutic intervention targeting PARK7 for the treatment of AS.

  11. Epigenetic Alterations in Density Selected Human Spermatozoa for Assisted Reproduction.

    Directory of Open Access Journals (Sweden)

    Bolan Yu

    Full Text Available Epidemiological evidence indicates that assisted reproductive technologies (ART may be associated with several epigenetic diseases such as Beckwith-Wiedemann syndrome (BWS or Silver-Russell syndrome (SRS. Selection of sperm by density-gradients in ART has improved DNA integrity and sperm quality; however, epigenetic alterations associated with this approach are largely unknown. In the present study, we investigated DNA methylation and histone retention profiles in raw sperm and selected sperm derived from the same individual and separated by using density-gradients. Results from a study group consisting of 93 males demonstrated that both global DNA methylation and histone retention levels decreased in density selected sperm. Compared to unselected raw sperm, histone transition rates decreased by an average of 27.2% in selected sperm, and the global methylation rate was 3.8% in unselected sperm and 3.3% in the selected sperm. DNA methylation and histone retention location profiling analyses suggested that these alterations displayed specific location patterns in the human genome. Changes in the pattern of hypomethylation largely occurred in transcriptional factor gene families such as HOX, FOX, and GATA. Histone retention increased in 67 genes, whereas it was significantly clustered in neural development-related gene families, particularly the olfactory sensor gene family. Although a causative relationship could not be established, the results of the present study suggest the possibility that sperm with good density also possess unique epigenetic profiles, particularly for genes involved in neural and olfactory development. As increasing evidence demonstrates that epigenetics plays a key role in embryonic development and offspring growth characteristics, the specific epigenetic alterations we observed in selected sperm may influence the transcriptional process and neural development in embryos.

  12. Human Spermatozoa Quantitative Proteomic Signature Classifies Normo- and Asthenozoospermia.

    Science.gov (United States)

    Saraswat, Mayank; Joenväärä, Sakari; Jain, Tushar; Tomar, Anil Kumar; Sinha, Ashima; Singh, Sarman; Yadav, Savita; Renkonen, Risto

    2017-01-01

    Scarcely understood defects lead to asthenozoospermia, which results in poor fertility outcomes. Incomplete knowledge of these defects hinders the development of new therapies and reliance on interventional therapies, such as in vitro fertilization, increases. Sperm cells, being transcriptionally and translationally silent, necessitate the proteomic approach to study the sperm function. We have performed a differential proteomics analysis of human sperm and seminal plasma and identified and quantified 667 proteins in sperm and 429 proteins in seminal plasma data set, which were used for further analysis. Statistical and mathematical analysis combined with pathway analysis and self-organizing maps clustering and correlation was performed on the data set.It was found that sperm proteomic signature combined with statistical analysis as opposed to the seminal plasma proteomic signature can differentiate the normozoospermic versus the asthenozoospermic sperm samples. This is despite the results that some of the seminal plasma proteins have big fold changes among classes but they fall short of statistical significance. S-Plot of the sperm proteomic data set generated some high confidence targets, which might be implicated in sperm motility pathways. These proteins also had the area under the curve value of 0.9 or 1 in ROC curve analysis.Various pathways were either enriched in these proteomic data sets by pathway analysis or they were searched by their constituent proteins. Some of these pathways were axoneme activation and focal adhesion assembly, glycolysis, gluconeogenesis, cellular response to stress and nucleosome assembly among others. The mass spectrometric data is available via ProteomeXchange with identifier PXD004098.

  13. Purification and characterization of 26S proteasomes from human and mouse spermatozoa.

    Science.gov (United States)

    Tipler, C P; Hutchon, S P; Hendil, K; Tanaka, K; Fishel, S; Mayer, R J

    1997-12-01

    We purified by fractionation on 10-40% glycerol gradients, 26S proteasomes from normal human spermatozoa. These proteasomes, which participate in the ATP-dependent degradation of ubiquitinated proteins, share a similar sedimentation coefficient to those purified from other human tissues. Fluorogenic peptide assays reveal they have chymotrypsin, trypsin and peptidyl-glutamyl-like peptide hydrolysing activities; the chymotrypsin activity is ablated by the specific 26S proteasome inhibitor MG132. Confirmation that these large proteases are 26S proteasomes is provided by detection of the 20S proteasome subunits HC2, XAPC7, RN3 and Z and regulatory ATPases MSS1, TBP1, SUG1 and SUG2 by Western analyses with monoclonal antisera. These antigens are found only in the gradient fractions enriched in proteolytic activities. We have also shown that, although mature spermatozoa from mice have considerably reduced amounts of a ubiquitin-conjugating enzyme (E2) and ubiquitin-protein conjugates in comparison with less mature germ cells, they retain relatively high values of 26S proteasome activity. This suggests that proteasomes may have further roles to play in normal sperm physiology.

  14. New insights on the origin and relevance of aneuploidy in human spermatozoa.

    Science.gov (United States)

    Templado, C; Uroz, L; Estop, A

    2013-10-01

    In humans, the most common chromosomal abnormality is aneuploidy. Because the majority of aneuploid conceptuses die during the early stages of embryonic development, an accurate estimate of the frequency of aneuploidy at conception can only be assessed by directly studying the gametes. The vast majority of aneuploidies arise de novo as a result of sporadic chromosome missegregation in paternal or maternal meiosis. In this review, we present the basic current knowledge about the incidence of aneuploidy in human spermatozoa in the general population and in patient populations where elevated levels of sperm aneuploidy are observed. These include infertile patients, patients with abnormal somatic karyotypes, and individuals exposed to certain environmental/lifestyle hazards. The clinical impact of increased levels of aneuploidy is discussed. We then focus on the non-disjunction mechanisms that cause aneuploidy during spermatogenesis and the factors that predispose to non-disjunction in male germ cells followed by an analysis of the sex differences in the incidence of gamete aneuploidy. Recent meiotic studies using multiplex-FISH on three fertile men have revealed that the frequency of conservative aneuploidy of metaphase II spermatocytes is similar to that observed in non-inseminated oocytes of young women. These findings suggest that the differences in the incidence of aneuploidy between spermatozoa and oocytes are not due to differences in chromosome segregation errors but rather to more effective checkpoint mechanisms in spermatogenesis than in oogenesis.

  15. Protective effect of butylated hydroxytoluene on sperm function in human spermatozoa cryopreserved by vitrification technique.

    Science.gov (United States)

    Merino, O; Aguagüiña, W E; Esponda, P; Risopatrón, J; Isachenko, E; Isachenko, V; Sánchez, R

    2015-03-01

    Butylhydroxytoluene (BHT), a synthetic analogue of vitamin E, shows antioxidant and antiviral properties and has been successfully used for mammalian sperm cryopreservation. In this study, BHT was included in a vitrification solution to determine its cryoprotective effect on human spermatozoa. Spermatozoa were selected by swim-up and vitrified in close sealed straw using either a combination of human tubal fluid (HTF), sucrose and BHT 1 mm (VMBHT), or only HTF and sucrose (VM). The optimal concentration of BHT was determined by the observation of preserved progressive sperm motility (PSM) after warming and detection of plasma membrane (PMI), membrane mitochondrial potential (ΔΨm) and DNA integrity. The presence of reactive oxygen species (ROS) was also detected. The PSM was significantly higher in the VMBHT group (80.86 ± 5.41%) compared with the VM group (68.9 ± 3.67%) (P < 0.05). Butylhydroxytoluene significantly preserved DNA integrity (4.0 ± 0.1% versus 6.1 ± 1.6%; P < 0.05) and reduced ROS production (5.5 ± 2.2 versus 8.6 ± 1.8%; P < 0.05). Plasma membrane and ΔΨm showed no statistical differences. One millimolar BHT effectively maintained cell function and due to its antioxidant and antiviral properties could be used in semen cryopreservation of patients with viral infections transmitted by seminal plasma.

  16. Cryoprotective effect of L-carnitine on motility, vitality and DNA oxidation of human spermatozoa.

    Science.gov (United States)

    Banihani, S; Agarwal, A; Sharma, R; Bayachou, M

    2014-08-01

    Successful cryopreservation for human spermatozoa markedly influences the reproductive outcomes of assisted reproductive technologies. But in spite of its usefulness, cryopreservation significantly decreases sperm quality. l-carnitine has been found to improve the quality of spermatozoa in selected cases with male infertility. Here, we examined the efficacy of l-carnitine in improving sperm motility and vitality and reducing sperm DNA oxidation during cryopreservation. Semen samples from infertile patients (n = 22) were collected and analysed. Cryopreservation medium supplemented with l-carnitine was mixed with the semen at a ratio of 1 : 1 (v/v). The final l-carnitine concentration in each cryovial was 0.5 mg ml(-1) per 5 × 10(6) cell ml(-1) . Controls were cryopreserved without addition of l-carnitine. After 24 h of cryopreservation, thawed sperm samples were analysed for motility, vitality and DNA oxidation. Sperm vitality was assessed by the eosin-nigrosin test, while sperm DNA oxidation was measured by flow cytometry. Addition of l-carnitine significantly improved sperm motility and vitality (P 0.05) in the levels of DNA oxidation between samples and controls. In conclusion, l-carnitine improves human sperm motility and vitality, but has no effect on sperm DNA oxidation after cryopreservation.

  17. [New progress in detection of antigens from human ejaculate in relationship with human sterility (extraction with N-cetylpyridinium chloride) (author's transl)].

    Science.gov (United States)

    Mettler, L; Steinhardt-Degola, N; Skrabei, H

    1980-01-01

    In an attempt to obtain certain sub-surface spermatozoa antigens, human spermatozoa were treated with N-cetylpyridinium chloride in basic buffer, The extract was further ultrasonated and separated with gel chromatography over a Sephadex--G--100 column. The obtained fractions were lyophilized and tested against human sperm agglutinating and sperm immobilizing sera, in order to detect their antigenicity. Positive and negative controls revealed intensive precipitations and no precipitations respectively. Fraction A4 reacted intensively with sperm immobilizing sera. The final objectives of these sperm antigens solubilization studies are twofold. On the one hand we are looking for a method to lower sperm antibodies titres in sterile females, and on the other hand we are seeking an immunological contraceptive method.

  18. In-vitro human spermatozoa nuclear decondensation assessed by flow cytometry.

    Science.gov (United States)

    Samocha-Bone, D; Lewin, L M; Weissenberg, R; Madgar, Y; Soffer, Y; Shochat, L; Golan, R

    1998-02-01

    The process of sperm chromatin decondensation occurs when a spermatozoon enters an ovum. Protamine disulphide bonds are reduced to SH and the polycationic protamines combine with the polyanionic egg protein, nucleoplasmin, thus being stripped from DNA which then combines with histones. Defective chromatin decondensation will thus prevent further development of the male pronucleus. In this study human sperm samples were incubated in vitro at 28 degrees C (using a medium in which the polyanion, heparin, substitutes for nucleoplasmin and beta-mercaptoethanol for egg glutathione) for 10, 20 and 30 min before stopping the reaction with formalin (to 3.6%). The DNA of the fixed cells was stained with Acridine Orange by a one-step method and subjected to flow cytometry and data analysis, in which a zone characteristic of condensed chromatin is outlined on red-green fluorescence contour plots. After 20 min of incubation 97% of the control spermatozoa that were in the mature window (WIN M) had decondensed and moved out of this region. Defects in sperm decondensation were seen in four semen samples of the 20 that were tested. In cases where spermatozoa fail to produce a fertilized egg the cause may lie with defective chromatin quality, including failure of the sperm chromatin to decondense. The method described here is a simple procedure for detecting sperm samples containing such defective cells.

  19. Cryopreservation—induced decrease in heat—shock protein 90 in human spermatozoa and its mechanism

    Institute of Scientific and Technical Information of China (English)

    Wen-LeiCAO; Yi-XinWANG; Zu-QiongXIANG; ZhengLI

    2003-01-01

    Aim:To study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism.Methods:In 18 healthy men,the seminal sperm motility and HSP90 levels were studied before and after cryopreservation using SDS-PAGE,Western blotting and computerized image analysis.Results:The sperm motility declined significantly after cryopreservation(P<0.01).The average grey level and the integrated grey level of sperm HSP90 before cooling were 34.1±3.2 and 243.0±21.6,respectively,while those after thawing were 23.2±2.5 and 105.7±28.5,respectively.Both parameters were decreased significantly(P<0.01).No HSP90 was found in the seminal plasma before and after cryopreservation.Conclusion:HSP90 in human spermatozoa was decreased substantially after cryopreservation.This may results from protein degradation,rather than leakage into the seminal plasma.

  20. Immobilising effect of Ruta graveolens L. on human spermatozoa: coumarin compounds are involved.

    Science.gov (United States)

    Naghibi Harat, Z; Lakpour, N; Sadeghipoor, H R; Kamalinejad, M; Eshraghian, M R; Naghibi, B; Akhondi, M M; Binaafar, S; Sadeghi, M R

    2015-12-01

    This study was designed to find out Ruta graveolens L. functional components, which have immobilisation effect on human spermatozoa for contraceptive use. A five-step fractionation method was used to derive different components from rue aqueous extract by using hexane, chloroform, ethanol, acetone and ultrapure water. Gas chromatography-mass spectrophotometery (GC-MS) of all fractions and the aqueous extract were performed to determine the chemical components. The immobilisation assay and membrane integrity test were also performed with four different coumarins, which were found in GC-MS in a concentration of 10 μm. Hexane, chloroform, acetone and ethanol fractions could significantly decrease motility of sperms within the first and the second hours. Hexane fraction had also significant immediate effect. The aqueous fraction had no effect on sperm motility. Meanwhile, GC-MS revealed that aqueous extract and effective fractions had similar coumarin compounds. We performed the immobilisation assay on four different coumarins, which were found in GC-MS in a concentration of 10 μm. Reduction of sperm motility was only significant for xanthotoxin. In the sperm viability and membrane integrity tests, hexane and ethanolic fractions could impair sperm vitality significantly, in contrast to coumarins. These results indicated that a part of immobilising effect of rue could be due to its coumarins. The possible mechanism could be blocking of spermatozoa potassium channels.

  1. Microfluorometric assessment of the DNA-DNP complex in human spermatozoa.

    Science.gov (United States)

    Johannisson, E; Jacolino, H

    1987-06-01

    During spermiogenesis, the DNA-nucleoprotein complex undergoes alterations that are reflected in a decreasing capacity for binding DNA-specific dyes, such as ethidium bromide (EB). Human spermatozoa with a low or high capacity for EB binding were depleted of RNA and most nuclear proteins by exposure to RNAse, EDTA and trypsin, with and without additional high salt buffer (HSB) treatment. When treated with RNAse, EDTA and trypsin only, the haploid DNA fluorescence value (calculated from the diploid value of the standard cell population) was found at EB concentrations of 6.5 to 12.5 micrograms/mL. At these EB concentrations, a significantly lower fluorescence was found in the material also treated with HSB, probably reflecting an unwinding of the highly negatively supercoiled DNA loops that are induced by HSB treatment. Maximal fluorescence was not found until a concentration of 50 micrograms EB/mL. This may be due to an overwinding of the DNA by the positive supercoiling caused by EB. The significant difference in EB uptake initially found between the two groups whose spermatozoa showed low and high capacities for EB binding disappeared after removal of the nucleoproteins, suggesting that this compartment of the nucleoprotein-DNA complex is responsible for the different uptakes of EB.

  2. On-chip cryopreservation: a novel method for ultra-rapid cryoprotectant-free cryopreservation of small amounts of human spermatozoa.

    Directory of Open Access Journals (Sweden)

    Yujie Zou

    Full Text Available Cryopreservation of human spermatozoa free from cryoprotectant can avoid toxicity caused by highly concentrated cryoprotectant and a series of specific carriers have been previously explored, except for PDMS chip. Our study is aimed at exploring a novel device for ultra-rapid cryopreservation of small numbers of spermatozoa without cryoprotectant based on polydimethylsiloxane (PDMS chips. Spermatozoa from 25 healthy men were involved in this study, comparing on-chip cryopreservation with different micro-channel height (group A: 10 µm height, group B: 50 µm height, group C: 100 µm height and conventional freezing (group D in liquid nitrogen for 72 h. The viability, motility, DNA integrity by comet assay and acrosome integrity by fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA staining of frozen-thawed spermatozoa of each group were compared. The motility and viability of post-thawed spermatozoa was significantly decreased than that of pre-freezing spermatozoa. There was no difference of viability and motility of frozen-thawed spermatozoa between group A and D, while viability and motility of group B and C decreased compared to group A. Comet assay showed that no matter for group A or D, there was no difference of CR, TL, TD and OTM between pre-frozen and post-thawed spermatozoa. There was no difference of CR, TL, TD and OTM of post-thawed spermatozoa between group A and group D neither, while spermatozoa DNA damage was more serious in group B and group C with increasing height of micro-channel compared with group A. The proportion of intact acrosome of post-thawed spermatozoa in group A was the highest when compared with group B and group C, though similar to that of group D. In conclusion, PDMS chip with 10 µm height micro-channel is ideal for ultra-rapid cryopreservation of small quantity of spermatozoa without cryoprotectant.

  3. Pharmacotherapy for premature ejaculation

    NARCIS (Netherlands)

    Waldinger, Marcel D

    PURPOSE OF REVIEW: As there are various drugs and different treatment strategies to delay ejaculation, a review of the current drug treatments for premature ejaculation is relevant for daily clinical practice. RECENT FINDINGS: There are four premature ejaculation subtypes: lifelong premature

  4. Pharmacotherapy for premature ejaculation

    NARCIS (Netherlands)

    Waldinger, Marcel D

    2014-01-01

    PURPOSE OF REVIEW: As there are various drugs and different treatment strategies to delay ejaculation, a review of the current drug treatments for premature ejaculation is relevant for daily clinical practice. RECENT FINDINGS: There are four premature ejaculation subtypes: lifelong premature ejacula

  5. Morphometric and kinematic sperm subpopulations in split ejaculates of normozoospermic men

    Directory of Open Access Journals (Sweden)

    Pilar Santolaria

    2016-01-01

    Full Text Available This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis (CASA-Mot, and of sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations (slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%], and three morphometric subpopulations (large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]. The distribution of kinematic sperm subpopulations was different among ejaculate fractions (P < 0.001, with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions (P < 0.001, with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.

  6. Morphometric and kinematic sperm subpopulations in split ejaculates of normozoospermic men

    Science.gov (United States)

    Santolaria, Pilar; Soler, Carles; Recreo, Pilar; Carretero, Teresa; Bono, Araceli; Berné, José M; Yániz, Jesús L

    2016-01-01

    This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3–5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis (CASA-Mot), and of sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations (slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%]), and three morphometric subpopulations (large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]). The distribution of kinematic sperm subpopulations was different among ejaculate fractions (P < 0.001), with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions (P < 0.001), with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications. PMID:27624985

  7. Interacting proteins on human spermatozoa: adaptive evolution of the binding of semenogelin I to EPPIN.

    Science.gov (United States)

    Silva, Erick J R; Hamil, Katherine G; O'Rand, Michael G

    2013-01-01

    Semenogelin I (SEMG1) is found in human semen coagulum and on the surface of spermatozoa bound to EPPIN. The physiological significance of the SEMG1/EPPIN interaction on the surface of spermatozoa is its capacity to modulate sperm progressive motility. The present study investigates the hypothesis that the interacting surface of SEMG1 and EPPIN co-evolved within the Hominoidea time scale, as a result of adaptive pressures applied by their roles in sperm protection and reproductive fitness. Our results indicate that some amino acid residues of SEMG1 and EPPIN possess a remarkable deficiency of variation among hominoid primates. We observe a distinct residue change unique to humans within the EPPIN sequence containing a SEMG1 interacting surface, namely His92. In addition, Bayes Empirical Bayes analysis for positive selection indicates that the SEMG1 Cys239 residue underwent positive selection in humans, probably as a consequence of its role in increasing the binding affinity of these interacting proteins. We confirm the critical role of Cys239 residue for SEMG1 binding to EPPIN and inhibition of sperm motility by showing that recombinant SEMG1 mutants in which Cys239 residue was changed to glycine, aspartic acid, histidine, serine or arginine have reduced capacity to interact to EPPIN and to inhibit human sperm motility in vitro. In conclusion, our results indicate that EPPIN and SEMG1 rapidly co-evolved in primates due to their critical role in the modulation of sperm motility in the semen coagulum, providing unique insights into the molecular co-evolution of sperm surface interacting proteins.

  8. Detection of spermatozoa following consensual sexual intercourse.

    Science.gov (United States)

    Astrup, Birgitte Schmidt; Thomsen, Jørgen Lange; Lauritsen, Jens; Ravn, Pernille

    2012-09-10

    In cases of sexual assault, the finding of semen can provide crucial evidence. The presence of spermatozoa serves as proof of a sexual act and may give the identity of the alleged perpetrator through DNA-profiling. In most western countries, there are guidelines for standardized examinations of sexual assault victims. For an objective evaluation of the findings, substantial knowledge of aspects regarding consensual sexual intercourse is crucial. The aim of this study was to examine detection frequencies and genital sampling sites of spermatozoa following consensual sexual intercourse. In a prospective setting, 60 women underwent forensic examination following consensual sexual intercourse. Specimens were obtained from the external genitalia, the posterior fornix and the cervical canal, and examined using the Papanicolau stain and standard light microscopy. We found that 88% of possible cases were positive for spermatozoa. The posterior fornix was significantly better than the other sites for detection of spermatozoa and the number of spermatozoa decreased significantly over time. In a large sub-group of women who reported that no intra-vaginal ejaculation had taken place during their latest intercourse, a significant number (14%) had spermatozoa in the vagina. Spermatozoa were best recovered from the posterior fornix, but spermatozoa were also present on swabs taken from the external genitalia. Detection of spermatozoa is thus possible in cases where a speculum examination is denied. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  9. Assessment of chromatin status (SCSA) in epididymal and ejaculated sperm in Iberian red deer, ram and domestic dog.

    Science.gov (United States)

    Garcia-Macias, Vanesa; Martinez-Pastor, Felipe; Alvarez, Mercedes; Garde, Jose Julian; Anel, Enrique; Anel, Luis; de Paz, Paulino

    2006-11-01

    Abnormal chromatin condensation is not detected using classical techniques for sperm analysis. SCSA has demonstrated its usefulness in sperm chromatin analysis in several species (human, bull, stallion and boar). In this work, we studied sperm samples from red deer, ram and dog to analyze the differentiation of chromatin structure applying SCSA in epididymal and ejaculated spermatozoa. Epididymal samples were obtained from the caput, corpus and cauda by means of cuts, and ejaculated ones were obtained by electroejaculation (deer), artificial vagina (ram) and digital manipulation (dog). SCSA results suggested different critical points in sperm maturation (spermatozoa with loose chromatin to more condensed chromatin) among species: from corpus to cauda in ram and from caput to corpus in deer and dog. Moreover, we also detected differences in ruminants and dog, reflected in the appearance of SCSA plots. Indeed, ram and deer samples rendered two peaks within the sperm main population (sperm with condensed chromatin), whereas only one was detected in dog. Although some differences were observed between cauda and ejaculated samples, SCSA parameters indicated good chromatin condensation, making these samples suitable for germplasm banking. Some species-dependent modifications in the analysis of the results may be necessary to take full advantage of its analytical power.

  10. Semen characteristics in pubertal boys. I. Semen quality after first ejaculation.

    Science.gov (United States)

    Janczewski, Z; Bablok, L

    1985-01-01

    Semen specimens from 134 pubertal boys were examined, and some 274 assays were made. An analysis of the biological quality of semen in relation to the period of time after first ejaculation brings high values of statistical dependence of the volume of semen, its liquefaction, spermatozoal concentration, percentage of morphologically normal forms of spermatozoa, and normal spermatozoal motility on the period of time after first ejaculation. Normal figures for semen volume, semen liquefaction, spermatozoal concentration, and morphology are observed 12-14 months after first ejaculation. The percentage of normally motile spermatozoa becomes standard 21-23 months after first ejaculation. There were changes in semen characteristics from azoospermia through cryptozoospermia, oligozoospermia, and asthenozoospermia to normospermia. Azoospermia dominates until the fifth month after the first ejaculation, oligozoospermia from the sixth to the eleventh month, asthenozoospermia from the twelfth to the twentieth month, and normospermia from the twenty-first month.

  11. Human Herpesvirus-6A/B Binds to Spermatozoa Acrosome and Is the Most Prevalent Herpesvirus in Semen from Sperm Donors

    DEFF Research Database (Denmark)

    Kaspersen, Maja Døvling; Larsen, Peter; Kofod-Olsen, Emil;

    2012-01-01

    ejaculate that was positive for one or more human herpesvirus. Of these 27.3% (n = 15) had a double herpesvirus infection. If corrected for the presence of multiple ejaculates from some donors, the adjusted frequency of herpesviruses in semen was 27.2% with HSV-1 in 0.4%; HSV-2 in 0.1%; EBV in 6.3%; HCMV...

  12. Retrograde ejaculation, painful ejaculation and hematospermia.

    Science.gov (United States)

    Parnham, Arie; Serefoglu, Ege Can

    2016-08-01

    Although there has been an increased interest on premature ejaculation in the recent years, our understanding regarding the disorders of retrograde ejaculation, painful ejaculation and hematospermia remain limited. All three of these conditions require a keen clinical acumen and willingness to engage in thinking outside of the standard established treatment paradigm. The development of novel investigational techniques and treatments has led to progress in the management of these conditions symptoms; however, the literature almost uniformly is limited to small series and rare randomised trials. Further investigation and randomised controlled trials are needed for progress in these often challenging cases.

  13. Animal models of premature and retarded ejaculation.

    Science.gov (United States)

    Waldinger, Marcel D; Olivier, Berend

    2005-06-01

    Most of our current understanding of the neurobiology of sexual behavior and ejaculatory function has been derived from animal studies using rats with normal sexual behaviour. However, none of these proposed models adequately represents human ejaculatory disorders. Based on the "ejaculation distribution theory", which postulates that the intravaginal ejaculation latency time in men is represented by a biological continuum, we have developed an animal model for the research of premature and delayed ejaculation. In this model, a large number of male Wistar rats are investigated during 4-6 weekly sexual behavioural tests. Based on the number of ejaculations during 30 min tests, rapid and sluggish ejaculating rats are distinguished, each representing approximately 10% at both ends of a Gaussian distribution. Together with other parameters, such as ejaculation latency time, these rats at either side of the spectrum resemble men with premature and delayed ejaculation, respectively. Comparable to the human situation, in a normal population of rats, endophenotypes exist with regard to basal sexual (ejaculatory) performance.

  14. Autophagy and apoptosis have a role in the survival or death of stallion spermatozoa during conservation in refrigeration.

    Science.gov (United States)

    Gallardo Bolaños, Juan M; Miró Morán, Álvaro; Balao da Silva, Carolina M; Morillo Rodríguez, Antolín; Plaza Dávila, María; Aparicio, Inés M; Tapia, José A; Ortega Ferrusola, Cristina; Peña, Fernando J

    2012-01-01

    Apoptosis has been recognized as a cause of sperm death during cryopreservation and a cause of infertility in humans, however there is no data on its role in sperm death during conservation in refrigeration; autophagy has not been described to date in mature sperm. We investigated the role of apoptosis and autophagy during cooled storage of stallion spermatozoa. Samples from seven stallions were split; half of the ejaculate was processed by single layer centrifugation, while the other half was extended unprocessed, and stored at 5°C for five days. During the time of storage, sperm motility (CASA, daily) and membrane integrity (flow cytometry, daily) were evaluated. Apoptosis was evaluated on days 1, 3 and 5 (active caspase 3, increase in membrane permeability, phosphatidylserine translocation and mitochondrial membrane potential) using flow cytometry. Furthermore, LC3B processing was investigated by western blotting at the beginning and at the end of the period of storage. The decrease in sperm quality over the period of storage was to a large extent due to apoptosis; single layer centrifugation selected non-apoptotic spermatozoa, but there were no differences in sperm motility between selected and unselected sperm. A high percentage of spermatozoa showed active caspase 3 upon ejaculation, and during the period of storage there was an increase of apoptotic spermatozoa but no changes in the percentage of live sperm, revealed by the SYBR-14/PI assay, were observed. LC3B was differentially processed in sperm after single layer centrifugation compared with native sperm. In processed sperm more LC3B-II was present than in non-processed samples; furthermore, in non-processed sperm there was an increase in LC3B-II after five days of cooled storage. These results indicate that apoptosis plays a major role in the sperm death during storage in refrigeration and that autophagy plays a role in the survival of spermatozoa representing a new pro-survival mechanism in

  15. Evolutionary conservation of mammalian sperm proteins associates with overall, not tyrosine, phosphorylation in human spermatozoa.

    Science.gov (United States)

    Schumacher, Julia; Ramljak, Sanja; Asif, Abdul R; Schaffrath, Michael; Zischler, Hans; Herlyn, Holger

    2013-12-06

    We investigated possible associations between sequence evolution of mammalian sperm proteins and their phosphorylation status in humans. As a reference, spermatozoa from three normozoospermic men were analyzed combining two-dimensional gel electrophoresis, immunoblotting, and mass spectrometry. We identified 99 sperm proteins (thereof 42 newly described) and determined the phosphorylation status for most of them. Sequence evolution was studied across six mammalian species using nonsynonymous/synonymous rate ratios (dN/dS) and amino acid distances. Site-specific purifying selection was assessed employing average ratios of evolutionary rates at phosphorylated versus nonphosphorylated amino acids (α). According to our data, mammalian sperm proteins do not show statistically significant sequence conservation difference, no matter if the human ortholog is a phosphoprotein with or without tyrosine (Y) phosphorylation. In contrast, overall phosphorylation of human sperm proteins, i.e., phosphorylation at serine (S), threonine (T), and/or Y residues, associates with above-average conservation of sequences. Complementary investigations suggest that numerous protein-protein interactants constrain sequence evolution of sperm phosphoproteins. Although our findings reject a special relevance of Y phosphorylation for sperm functioning, they still indicate that overall phosphorylation substantially contributes to proper functioning of sperm proteins. Hence, phosphorylated sperm proteins might be considered as prime candidates for diagnosis and treatment of reduced male fertility.

  16. Detection of spermatozoa following consensual sexual intercourse

    DEFF Research Database (Denmark)

    Astrup, Birgitte Schmidt; Thomsen, Jørgen Lange; Lauritsen, Jens

    2012-01-01

    examinations of sexual assault victims. For an objective evaluation of the findings, substantial knowledge of aspects regarding consensual sexual intercourse is crucial. The aim of this study was to examine detection frequencies and genital sampling sites of spermatozoa following consensual sexual intercourse....... METHODS: In a prospective setting, 60 women underwent forensic examination following consensual sexual intercourse. Specimens were obtained from the external genitalia, the posterior fornix and the cervical canal, and examined using the Papanicolau stain and standard light microscopy. RESULTS: We found...... that 88% of possible cases were positive for spermatozoa. The posterior fornix was significantly better than the other sites for detection of spermatozoa and the number of spermatozoa decreased significantly over time. In a large sub-group of women who reported that no intra-vaginal ejaculation had taken...

  17. Ejaculate traits in the Namibian cheetah (Acinonyx jubatus): influence of age, season and captivity.

    Science.gov (United States)

    Crosier, Adrienne E; Marker, Laurie; Howard, JoGayle; Pukazhenthi, Budhan S; Henghali, Josephine N; Wildt, David E

    2007-01-01

    The objective was to examine the influence of animal age, season and captivity status on seminal quality in wild-born cheetahs (Acinonyx jubatus) in Namibia, Africa. Animals were divided into three age categories: juvenile (14-24 months; n = 16 males, 23 ejaculates); adult (25-120 months; n = 76 males, 172 ejaculates); and aged (>120 months; n = 5 males, 5 ejaculates). Seasons were categorised into hot-wet (January-April), cold-dry (May-August) and hot-dry (September-December). A comparison between freshly wild-caught (n = 29 males, 41 ejaculates) and captive-held cheetahs (n = 68 males, 159 ejaculates) was also conducted. Raw ejaculates contained 69.0 +/- 1.1% motile spermatozoa (mean +/- s.e.m.) with 73.6 +/- 1.5% of these cells containing an intact acrosome. Overall, 18.4 +/- 0.9% of spermatozoa were morphologically normal, with midpiece anomalies being the most prevalent (approximately 39%) defect. Juvenile cheetahs produced ejaculates with poorer sperm motility, forward progressive status, lower seminal volume and fewer total motile spermatozoa than adult and aged animals. Spermatogenesis continued unabated throughout the year and was minimally influenced by season. Proportions of sperm malformations were also not affected by season. Ejaculates from captive cheetahs had increased volume and intact acrosomes, but lower sperm density than wild-caught counterparts. In summary, Namibian cheetahs produce an extraordinarily high proportion of pleiomorphic spermatozoa regardless of age, season or living (captive versus free-ranging) status. Young males less than 2 years of age produce poorer ejaculate quality than adult and aged males. Because (1) all study animals were wild born and (2) there was little difference between freshly caught males and those maintained in captivity for protracted periods, our results affirm that teratospermia in the cheetah is mostly genetically derived. It also appears that an ex situ environment for the Namibian cheetah can ensure sperm

  18. Protective features of resveratrol on human spermatozoa cryopreservation may be mediated through 5' AMP-activated protein kinase activation.

    Science.gov (United States)

    Shabani Nashtaei, M; Amidi, F; Sedighi Gilani, M A; Aleyasin, A; Bakhshalizadeh, Sh; Naji, M; Nekoonam, S

    2017-03-01

    Biochemical and physical modifications during the freeze-thaw process adversely influence the restoration of energy-dependent sperm functions required for fertilization. Resveratrol, a phytoalexin, has been introduced to activate 5' AMP-activated protein kinase which is a cell energy sensor and a cell metabolism regulator. The cryoprotection of resveratrol on sperm cryoinjury via activation of AMP-activated protein kinase also remains to be elucidated. Our aim, thus, was to investigate: (i) the presence and intracellular localization of AMP-activated protein kinase protein; (ii) whether resveratrol may exert a protective effect on certain functional properties of fresh and post-thaw human spermatozoa through modulation of AMP-activated protein kinase. Spermatozoa from normozoospermic men were incubated with or without different concentrations of Compound C as an AMP-activated protein kinase inhibitor or resveratrol as an AMP-activated protein kinase activator for different lengths of time and were then cryopreserved. AMP-activated protein kinase is expressed essentially in the entire flagellum and the post-equatorial region. Viability of fresh spermatozoa was not significantly affected by the presence of Compound C or resveratrol. However, although Compound C caused a potent inhibition of spermatozoa motility parameters, resveratrol did not induce negative effect, except a significant reduction in motility at 25 μm for 1 h. Furthermore, resveratrol significantly increased AMP-activated protein kinase phosphorylation and mitochondrial membrane potential and decreased reactive oxygen species and apoptosis-like changes in frozen-thawed spermatozoa. Nevertheless, it was not able to compensate decreased sperm viability and motility parameters following cryopreservation. In contrast, Compound C showed opposite effects to resveratrol on AMP-activated protein kinase phosphorylation, reactive oxygen species, apoptosis-like changes, mitochondrial membrane potential, and

  19. Determination of selenium in human spermatozoa and prostasomes using base digestion and electrothermal atomic absorption spectrophotometry.

    Science.gov (United States)

    Suistomaa, U; Saaranen, M; Vanha-Perttula, T

    1987-10-15

    A method for the determination of selenium in human spermatozoa and prostasomes is described. The samples were digested with 25% (w/v) tetramethylammonium hydroxide (TMAH) in methanol and analyzed by atomic absorption spectrometry with electrothermal atomization and Zeeman background correction (ET-AAS). Nickel was used as a matrix modifier. Calibration was performed using the matrix-based calibration curve. The TMAH-digestion method agreed well with a conventional digestion procedure using concentrated nitric acid. The TMAH-digestion does not require heating or strong acids and it was suitable for small biological samples. The average recovery of added selenium in spermatozoan digests was 95.1 +/- 5.2% (n = 5). The coefficient of variation was 9.1% (n = 21). The accuracy of the method tested with the NBS standard 1577 (bovine liver, certified at 1.1 +/- 0.1 micrograms Se/g) resulted in a value of 0.98 +/- 0.10 micrograms Se/g (n = 16). The method was further tested in an interlaboratory comparison study.

  20. Effects of exposure of epididymal boar spermatozoa to seminal plasma on the binding of zona pellucida proteins during in vitro capacitation

    NARCIS (Netherlands)

    Harkema, W.; Colenbrander, B.; Engel, B.; Woelders, H.

    2004-01-01

    The purpose of the investigation was to determine whether seminal plasma plays a role in the increase during in vitro capacitation of the number of boar spermatozoa with enhanced binding of zona pellucida proteins. Ejaculated spermatozoa and spermatozoa collected from the caudae epididymides of boar

  1. Changes in the expression of transient receptor potential canonical type 1 channel mRNA in asthenospermia ejaculated spermatozoa and its significance%弱精子症患者精子中TRPC1基因表达的变化及意义

    Institute of Scientific and Technical Information of China (English)

    李世林; 王怀鹏; 蒲小勇

    2012-01-01

    Objective To investigate the relationship between the expression of transient receptor potential canonical type 1 channel (TRPC1) mRNA in asthenospermia ejaculated spermatozoa and asthenospermia. Methods Semen samples were obtained from the healthy volunteers and patients with asthenospermia. Ejaculated spermatozoa were separated by discontinuous Percoll Gradient centrifugation. The movement parameters of spermatozoa were measured by computer-aided sperm analysis (CASA). Expressions of TRPClmRNA in the sperm of healthy men and asthenospermia patients were detected using the reverse transcription polymerase chain reaction (RT-PCR) and realtime quantitative RT-PCR. Results The relative expression of TRPC1 mRNA in the sperm of healthy men and asthenospermia patients were (0.91±0.15) and (0.32±0.08) respectively, which showed a significant difference (P<0.01). The movement parameters of spermatozoa in asthenospermia decreased obviously compared with that of the healthy group. The Pearson correlation analysis showed the expression of TRPC1 mRNA in spermatozoa was strongly and positively correlated with the linear velocity(r=0.895, PO.01), whipping frequency (r=0.742, P<0.01), forwardness (r=0.782, P<0.01), linearity (r=0.721, P<0.05), and the amplitude of the side pendulum of spermatozoa(r=0.809, P<0.01), whereas it was weakly and positively relative to the curve velocity(r=0.254, P<0.05) and average path velocity (r=0.364, P<0.05). Conclusion The abnormal expression of TRPC1 mRNA might be one of the causes of asthenospermia. It has provided us with a new approach for asthenospermia study.%目的 探讨弱精子症与瞬时感受器电位C1型(transient receptor potential canonical type 1,TRPC1)基因表达的关系.方法 收集40例A、B级精子活力低于20%的弱精子症患者(实验组)和40例精子活力正常健康成年男性(对照组)的精液,采用Percoll非连续梯度离心法分离精液精子,计算机辅助精液分析(CASA

  2. Expression of TEKT4 protein decreases in the ejaculated spermatozoa of idiopathic asthenozoospermic men%TEKT4蛋白在特发性弱精子症患者精子中的表达

    Institute of Scientific and Technical Information of China (English)

    武文斌; 李玉山; 吉晓菲; 王全先; 高学敏; 杨险峰; 潘周辉; 冯晓霞

    2012-01-01

    目的:探讨TEKT4蛋白在特发性弱精子症发生机制中的作用. 方法:采用非连续密度梯度离心分离和纯化特发性弱精子症患者和正常男性精子,采用RT-PCR和Western印迹方法,从mRNA和蛋白水平检测TEKT4的表达及其差异. 结果:RT-PCR结果表明,TEKT4 mRNA在特发性弱精子症患者精子中的表达水平与正常男性相比显著降低(0.59±0.13 vs 0.75±0.15,t=4.325,P<0.05);Western印迹结果与RT-PCR结果一致,TEKT4蛋白在特发性弱精子症患者精于中的表达水平与正常男性相比亦显著降低(0.48±0.14 vs 0.69±0.13,t=5.939,P<0.05). 结论:特发性弱精子症患者精子中TEKT4表达水平显著降低,可能是导致弱精子症发生的重要环节之一.%Objective: To investigate the role of the TEKT4 protein in the pathogenesis of idiopathic asthenozoospermia. Methods; We separated and purified the ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men by Percoll discontinuous density gradients, and detected the distribution and the expressions of TEKT4 mRNA and TEKT4 protein by RT-PCR and Western blot. Results-. RT-PCR revealed that the expression of TEKT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (0.59 ±0.13 isO. 75 ±0.15, t=4.325, P<0.05), and Western blot confirmed the results of RT-PCR (0.48 ±0.14 iv 0.69 ±0.13, (=5.939, P<0.05). Conclusion: The expression of TEKT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.

  3. Superoxide anion production by human spermatozoa as a part of the ionophore-induced acrosome reaction process.

    Science.gov (United States)

    Griveau, J F; Renard, P; Le Lannou, D

    1995-04-01

    The involvement of superoxide anion (O2o-) in human sperm capacitation and/or acrosome reaction was investigated. Addition of superoxide dismutase (SOD) to the medium at the beginning of the capacitation process or 15 min before induction of the acrosome reaction, decreased the level of ionophore-induced acrosome reaction. Hyperactivation was unaffected by the presence of SOD during the capacitation process. Addition of calcium ionophore to the sperm suspension increased production of O2o- by the spermatozoa by four to five-fold and induced the acrosome reaction. In the presence of SOD, superoxide anion could not be detected in the medium and the rate of induced-acrosome reaction was decreased greatly. The presence of an inhibitor of protein kinase C inhibited the production of O2o- in the medium and reduced the induced-acrosome reaction. The production of O2o- and the acrosome reaction were also increased by exposure of spermatozoa to 12-myristate 13-acetate phorbol ester, a specific activator of protein kinase C. While the level of spontaneous acrosome reaction was not increased by the direct addition of O2o- to the medium, its presence induced the release of unesterified fatty acids from membrane phospholipids. These findings suggest that the production of O2o- by spermatozoa could be involved in the ionophore-induced acrosome reaction, possibly through the de-esterification of membrane phospholipids. However, this production of superoxide anion is not sufficient on its own to induce the acrosome reaction.

  4. In vitro effect of zinc on oxidative changes in human semen.

    Science.gov (United States)

    Gavella, M; Lipovac, V

    1998-11-01

    The in vitro effect of zinc on superoxide anion (O2-) generation and on experimentally induced lipid peroxidation (LPO) in spermatozoa of infertile men was investigated. Washed spermatozoa pre-incubated for 30 min at 37 degrees C in the presence of 1 or 3 mmol l-1 zinc, released less superoxide anions (P spermatozoa. Similar results were obtained using activated polymorphonuclear leukocytes (1 x 10(6) cells ml-1) in the presence of 1 or 3 mmol l-1 Zn (P spermatozoa and leukocytes indicates that zinc may act in vivo as a scavenger of excessive O2- production by defective spermatozoa and/or leukocytes in semen after ejaculation. A significant stimulatory effect of Zn (3 mmol l-1) on iron-induced lipid peroxidation, measured by the formation of thiobarbituric acid reactive substances (TBARS), was detected in the spermatozoa of 16 normo- and 17 asthenozoospermic males (P spermatozoa), no effect of zinc on the LPO rate was found. The observed inhibitory effect of zinc on superoxide anion regardless of the initial O2- level and stimulatory effect of zinc depending on the initial LPO rate in human spermatozoa suggests that this metal ion participates in the oxidative changes occurring after ejaculation and thus may modulate the properties of germ cells.

  5. Effects of intracellular zinc depletion on the abundance of VDAC mRNA in cultured human spermatozoa%Zn缺乏对人精子VDAC mRNA的影响

    Institute of Scientific and Technical Information of China (English)

    赵丹; 吴炜; 章轶; 王增军; 王心如

    2012-01-01

    目的:探讨Zn离子缺乏对成熟精子活动能力和电压依赖性离子通道(voltage-dependent anion channel,VDAC)的mRNA丰度的影响.方法:从3个正常生育者捐献所得的精液经非连续Percoll梯度离心获得精子样本,并将其分3组连续培养24h:对照组;加入Zn离子螯合剂的TPEN组(2 μmol/L);同时加入Zn离子螯合剂N,N,N',N'-tetrakis (2-pyridylmethy1)Ethylenediamine (TPEN)和Zn离子(5μmol/L)组.行CASA和荧光定量PCR(qPCR)分别分析不同时间的精子活动能力和24h后各VDAC亚型mRNA丰度.结果:CASA显示:培养3h,TPEN组与对照组比较,精子活力明显下降(P<0.05).qPCR结果显示:培养24h后,TPEN组VDAC3 mRNA丰度明显低于对照组(P<0.05).而同时加入TPEN和Zn离子组与对照组比较,差异无统计学意义.结论:Zn离子缺乏使VDAC3 mRNA丰度降低,可能影响到精子的正常功能.%Objective:To investigate whether intracellular zinc depletion can actually change spermatozoa motiJity and the abundance of voltage-dependent anion channel (VDAC) mRNA in cultured human spermatozoa. Methods; Spermatozoa were separated from ejaculates of three donors using a discontinuous Percoll gradient centrifugation. Each spermatozoa specimen were divided into 3 groups,one was exposed to a cell membrane-permeable zinc chelator N,N,N',N'-tetrakis (2-pyridylmethyl) Ethylenediamine (TPEN) (2 μmol/L) for 24 hours,one was to TPEN plus zinc sulfate (5 μmol/L) for 24 hours,and one was as control. CASA and quantitative real-time PCR (qPCR) were performed to detect sperm motility characteristics,mRNA abundance and difference of the three VDAC subtypes between three different treatment groups respectively. Results;The results of CASA demonstrated that exposure of spermatozoa to TPEN for three hours significantly decreased sperm motility compared with the control group (P = 0.049). qPCR demonstrated that VDAC3 mRNA level of TPEN group were significantly lower than that of control group,while no

  6. Abnormal expression of centrosome protein (centrin) in spermatozoa of male human infertility

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    To study the relations between male infertility and centrosome protein (centrin) and the functions of centrin in spermatogenesis, the matured spermatozoa of 10 normal male people and 18 male infertility patients were stained by immunofluorescence labeling antibody against centrin. The results showed that two fluorescence signal dots appeared in the normal male spermatozoa and were located at the base of flagellum. They are proximal centriole and distal centriole. However, in some spermatozoa of the male infertility, centrin protein was located abnormally at the base of flagellum and its staining signals were spread, the normal proximal and distal centrioles were confused and could not be recognized separately. These results suggest that abnormality of centrosome protein may be related to male infertility. This discovery may be used as a marker of abnormal sperm and male infertility.

  7. Microphotokinesigraphic analysis of buffalo spermatozoa II. Photokinesigraphic characteristics of buffalo vibrating spermatozoa

    Directory of Open Access Journals (Sweden)

    M. Boichev

    2010-02-01

    Full Text Available The study concerns images of vibrating spermatozoa in the ejaculation of buffalo bulls from Bulgarian murrah breed through the method of Tchakarov and Natchev [1962], obtained at 5 sec exposures, optimal for this particular species. The negative registration material has been developed with high-equalising developer by Windisch, which reduces the glow due to overexposure around highaktinic parts of spermatozoa. On the basis of the positive graphic material – kinesigrams, we have established and analysed the main movement patterns of vibrating spermatozoa: with vibrating tail; with vibrating head; with a vibrating tail and head; with whole bilateral vibration; vibrating in circle; chaotically vibrating. We have described the differences in photokinesigraphic images of spermatozoa within given subclass.

  8. Does the microbial flora in the ejaculate affect the freezeability of stallion sperm?

    Science.gov (United States)

    Ortega-Ferrusola, C; González-Fernández, L; Muriel, A; Macías-García, B; Rodríguez-Martínez, H; Tapia, J A; Alonso, J M; Peña, F J

    2009-06-01

    In an attempt to evaluate the possible relationship between the microbial flora in the stallion ejaculate and its ability to freeze,three ejaculates from five stallions were frozen using a standard protocol. Before freezing, an aliquot was removed for bacteriological analysis. Bacterial growth was observed in all the ejaculates studied. The isolated microorganisms were:Staphylococcus spp. and Micrococcus spp. (in all the stallions), beta-haemolytic Streptococcus (in stallions 3 and 4), Corynebacterium spp. (in stallions 1, 3-5), Rhodococcus spp. (in stallion number 2), Pseudomonas spp. (in stallion number 1) and Klebsiella spp. (in stallions 1, 3 and 5). The presence and richness of Klebsiella and beta-haemolytic Streptococcus in the ejaculate were related to two sperm variables post-thaw,namely the proportion of dead spermatozoa (ethidium+ cells; r = 0.55, p < 0.05) and the amplitude of lateral displacement of the sperm head (ALH, microm; r = -0.56, p < 0.05), respectively.The degree of growth of Corynebacterium spp. in the ejaculate was positively correlated with the percentage of spermatozoa showing high caspase activity post-thaw(r = 0.62, p < 0.05). The presence and number of colonies of beta-haemolytic Streptococcus were negatively correlated (r = -0.55, p < 0.05) with low sperm caspase activity. It is concluded that the microbial flora of the equine ejaculate maybe responsible for some of the sublethal damage experimented by the spermatozoa during cryopreservation.

  9. Premature ejaculation: A review

    Directory of Open Access Journals (Sweden)

    Sukumar Reddy Gajjala

    2014-01-01

    Full Text Available Premature ejaculation (PE is a common male sexual disorder. It is defined by the Diagnostic and statistical manual of mental disorders as "ejaculation occurring, without control, on or shortly after penetration and before the person wishes it, causing marked distress or interpersonal difficulty. [1] Although the timing of intravaginal ejaculatory latency time (IELT (i.e., time from penetration to ejaculation is not included in this definition, an IELT of <2 min, or ejaculation occurring before penetration, has been considered consistent with PE. [2] Management involves both the patient and his partner. Therapeutic options should suit both partners and be appropriate to their habit in planning and frequency of intercourse. Follow-up at appropriate intervals to judge efficacy, titrate dosage of pharmacological treatments and ascertain side effects is mandatory.

  10. Cooperative functions of manganese and thiol redox system against oxidative stress in human spermatozoa

    Directory of Open Access Journals (Sweden)

    Amrit Kaur Bansal

    2009-01-01

    Full Text Available Aims: In this study, the effects of 0.1 mM Mn 2+ on thiol components (total thiols [TSH], glutathione reduced [GSH], glutathione oxidized [GSSG] and redox ratio [GSH/ GSSG] have been determined in human spermatozoa. Settings and Design: The subjects of the study were healthy males having more than 75% motility and 80 x 10 6 sperms/mL. Materials and Methods: Fresh semen was suspended in phosphate-buffered saline (PBS (pH 7.2 and this suspension was divided into eight equal fractions. All fractions, control (containing PBS and experimental (treated/untreated with [ferrous ascorbate, FeAA - 200 FeSO 4 μM, 1000 μM ascorbic acid, nicotine (0.5 mM and FeAA + nicotine], supplemented/unsupplemented with Mn 2+ [0.1 mM], were incubated for 2 h at 378C. These fractions were assessed for determining the thiol components. Statistical Analysis: The data were statistically analyzed by Students " t" test. Results and Conclusions: Ferrous ascorbate, nicotine and ferrous ascorbate + nicotine induced oxidative stress and decreased GSH and redox ratio (GSH/GSSG ratio but increased the TSH and GSSG levels. Mn 2+ supplementation improved TSH, GSH and redox ratio (GSH/GSSG but decreased the GSSG level under normal and oxidative stress conditions. Thiol groups serve as defense mechanisms of sperm cells to fight against oxidative stress induced by stress inducers such as ferrous ascorbate, nicotine and their combination (ferrous ascorbate + nicotine. In addition, Mn 2+ supplementation maintains the thiol level by reducing oxidative stress.

  11. Structure of chromatin in spermatozoa.

    Science.gov (United States)

    Björndahl, Lars; Kvist, Ulrik

    2014-01-01

    The specialized structure of the sperm chromatin has a dual function - first to protect the DNA from damage during storage and transport to the oocyte, and then to enable a rapid and complete unpacking of the undamaged paternal genome in the ooplasm. It is evident that zinc has a pivotal role in maintaining the structural stability and in enabling a rapid decondensation at the appropriate time. It is important for the sperm chromatin structure that the spermatozoa are ejaculated together with the zinc-rich prostatic secretion. Early exposure to zinc-binding seminal vesicular fluid can deplete the sperm chromatin of zinc and most likely induce surplus formation of disulfide bridges, likely to cause incomplete and delayed decondensation of the sperm chromatin in the oocyte. A premature decrease in sperm chromatin structure stability is likely to increase the risk for damage to the DNA due to increased access to the genome for DNA damaging compounds. The status of the sperm chromatin structure can vary in vitro depending on the exposure to zinc-depleting conditions when spermatozoa are stored in semen after ejaculation. When sperm DNA damage tests are evaluated and validated, it is therefore essential to also take into account the dynamics of zinc-dependent and zinc-independent sperm chromatin stability.

  12. Complete staining of human spermatozoa and immature germ cells combined with phase contrast microscopy

    DEFF Research Database (Denmark)

    Michael, A Y; Drejer, J O; Bagger, P V

    1987-01-01

    A method combining Janus green B and Thymol blue stains the anterior part of the head, the nuclear membrane, middle piece, and tail of spermatozoa light green and the nucleus deep purple. The method provides excellent stained preparations for the evaluation of sperm morphology by phase contrast...

  13. Cryopreservation of human spermatozoa. I. Effects of holding procedure and seeding on motility, fertilizability, and acrosome reaction.

    Science.gov (United States)

    Critser, J K; Huse-Benda, A R; Aaker, D V; Arneson, B W; Ball, G D

    1987-04-01

    Three experiments were conducted to evaluate effects of holding semen at +5.0 degrees C for 30 minutes or -5.0 degrees C for 10 minutes and ice crystal induction (seeding) on frozen-thawed human spermatozoa. In experiment 1, spermatozoa were frozen, and postthaw motility was evaluated immediately (0 hour) and 24 hours later. At both 0 and 24 hours, nonfrozen control samples had higher motility than all other treatment groups. At 0 hour postthaw, motility was higher in samples held at -5.0 degrees C for 10 minutes with no significant effect of seeding. At 24 hours, samples held at -5.0 degrees C for 10 minutes and seeded, but not samples held at -5.0 degrees C and not seeded, had higher motility than samples held at +5.0 degrees C. In experiment 2, semen samples were frozen, and fertilizability was evaluated in a zona-free hamster egg penetration assay. Seeded samples had a higher frequency of sperm penetration than either nonfrozen or nonseeded samples. In experiment 3, nonfrozen controls and frozen treatment groups were evaluated for the frequency of survival and acrosomal integrity. Seeded samples had higher frequencies of survival and loss of acrosomal integrity than nonseeded samples. All frozen-thawed samples had a lower frequency of survival and a higher frequency of loss of acrosomal integrity than nonfrozen controls. Although altered patterns of fertilizability and acrosomal integrity are induced, collectively these data suggest that incorporating a holding temperature of -5.0 degrees C for 10 minutes and seeding may result in a superior protocol for freezing human spermatozoa.

  14. Propolis protects human spermatozoa from DNA damage caused by benzo[a]pyrene and exogenous reactive oxygen species.

    Science.gov (United States)

    Russo, A; Troncoso, N; Sanchez, F; Garbarino, J A; Vanella, A

    2006-02-23

    Many environmental, physiological and genetic factors have been implicated in defective sperm function, the most common cause of infertility. In addition, sperm preparation techniques such as centrifugation, used prior to in vitro fertilization, are associated with the generation of reactive oxygen species (ROS) and an increase in the level of DNA damage. Factors that can offer spermatozoa protection are, therefore, of great importance. This study was designed to examine in vitro the effect of a Chilean propolis ethanolic extract on human spermatozoa treated with benzo[a]pyrene and exogenous reactive oxygen species. Our experimental evidence demonstrated that the natural drug under investigation is able to protect genomic DNA by damage induced by benzo[a]pyrene, hydrogen peroxide (H2O2) and hydrogen peroxide in combination with adenosine 5'-diphosphate (ADP) and ferrous sulfate (FeSO4), determining a significant reduction of the intracellular oxidants. An increase in membrane damage, measured by monitoring the formation of thiobarbituric acid-reactive substances (TBARS) and lactic dehydrogenase (LDH) release, was observed only in sperm treated with H2O2, ADP and FeSO4. The propolis extract was shown to possess the capacity to protect sperm membrane from the deleterious action of oxidative attack, reducing TBARS formation and LDH release. In summary, our results evidence that the protective effect exhibited by this natural compound in human spermatozoa is correlated, at least in part, to the antioxidant capacity of its active components, and suggest that propolis may have a role in protection against male infertility.

  15. Early apoptotic changes in human spermatozoa and their relationships with conventional semen parameters and sperm DNA fragmentation

    Institute of Scientific and Technical Information of China (English)

    Hao-Bo Zhang; Shao-Ming Lu; Chun-Yan Ma; Li Wang; Xiao Li; Zi-Jiang Chen

    2008-01-01

    Aim: To investigate whether early apoptotic changes in spermatozoa can be significant markers for sperm quality.Methods: Two early apoptotic changes in the semen of 56 men were assessed using Annexin V (AN)/propidium iodide (PI) staining for phosphatidylserine externalization and JC-1 staining for mitochondrial membrane potential (MMP). The results were compared with conventional semen parameters and DNA fragmentation identified using the TUNEL assay. Results: The different labeling patterns in the bivariate Annexin V/PI analysis identified four distinc-tive spermatozoa populations. The percentage of AN-/PI- spermatozoa positively correlated with conventional semen parameters and MMP, but negatively correlated with TUNEL (+) spermatozoa. As for the AN-/PI+ fraction, we found an opposite result in comparison to AN-/PI- spermatozoa. The level of early apoptotic AN+/PI- spermatozoa negatively correlated with MMP and sperm motility. The level of late apoptotic AN+/PI+ spermatozoa negatively correlated with conventional semen parameters and MMP, and positively correlated with TUNEL (+) spermatozoa. MMP positively correlated with conventional semen parameters, but negatively correlated with TUNEL (+) spermatozoa. Conclusion:Although early apoptotic AN+/PI- spermatozoa only negatively correlates with sperm motility, the differences in proportion of each subpopulation of spermatozoa (especially, the percentage of AN-/PI-spermatozoa), and decreased MMP might be significant markers for diagnosing male infertility. They possibly bring additional information to predict the outcome of in vitro fertilization.

  16. Changes of IZUMO1 in bull spermatozoa during the maturation, acrosome reaction, and cryopreservation.

    Science.gov (United States)

    Fukuda, Masaki; Sakase, Mitsuhiro; Fukushima, Moriyuki; Harayama, Hiroshi

    2016-12-01

    To obtain basic information of bull IZUMO1 (a sperm protein essential for sperm-egg fusion) and disclose possible causes for the impaired fertilizing ability in bull cryopreserved spermatozoa, we investigated this protein in bull spermatozoa collected from various regions of epididymides, freshly ejaculated spermatozoa, acrosome-reacted spermatozoa, and cryopreserved spermatozoa by Western blotting and the triple staining with the anti-IZUMO1 antibody, fluorescein isothiocyanate-peanut agglutinin, and 4',6-diamidino-2-phenylindole. In the cauda epididymal spermatozoa and freshly ejaculated spermatozoa, bull IZUMO1 was detected mainly as a 45-kDa major form. This major form was derived probably from a 52-kDa precursor form in the epididymis. Bull IZUMO1 was immunolocalized along the border between the principal and equatorial segments of the acrosomal region (pattern P1 of IZUMO1) in the most of epididymal and freshly ejaculated spermatozoa with normal acrosomes. In the samples after the treatments to induce the acrosome reaction, the percentages of spermatozoa without acrosomes and with IZUMO1 in whole equatorial segment (pattern P2 of IZUMO1) significantly increased. These results indicate that bull IZUMO1 undergoes maturation-related changes during sperm transit through the epididymis and that it is translocated to the equatorial segment of acrosomal region during the acrosome reaction. On the other hand, severe damages were observed in the acrosomes of 60% of the cryopreserved spermatozoa. Localization of IZUMO1 in these spermatozoa was pattern P2 (IZUMO1 in whole equatorial segment), P3 (IZUMO1 in whole acrosomal region), or P4 (IZUMO was lost). Moreover, after the incubation to compare the stability of acrosomes and IZUMO1 localization between cryopreserved spermatozoa and freshly ejaculated spermatozoa, much more spermatozoa lost acrosomes and IZUMO1 in the cryopreserved samples compared with freshly ejaculated samples. These findings indicate that impaired

  17. Epidemiology of delayed ejaculation.

    Science.gov (United States)

    Di Sante, Stefania; Mollaioli, Daniele; Gravina, Giovanni Luca; Ciocca, Giacomo; Limoncin, Erika; Carosa, Eleonora; Lenzi, Andrea; Jannini, Emmanuele A

    2016-08-01

    A large body of literature on diminished ejaculatory disorders has been generated without the use of a clear diagnostic definition. Many studies have not distinguished between the orgasm and ejaculation disorders leading to doubtful results. Delayed ejaculation (DE) is one of the diminished ejaculatory disorders, which range from varying delays in ejaculatory latency to a complete inability to ejaculate. The present review is aimed at providing a comprehensive overview of the current knowledge on the definition and epidemiology of diminished ejaculatory disorders. We focus on the acquired diseases, such as benign prostatic hyperplasia (BPH) and specific drug regimens that may cause an iatrogenic form of ejaculatory disorder. In addition, the impact of aging is discussed since the prevalence of DE appears to be moderately but positively related to age. Finally, we also focus on the importance of the hormonal milieu on male ejaculation. To date, evidence on the endocrine control of ejaculation is derived from small clinical trials, but the evidence suggests that hormones modulate the ejaculatory process by altering its overall latency.

  18. Modeling of progesterone-induced intracellular calcium signaling in human spermatozoa.

    Science.gov (United States)

    Li, Long-Fei; Xiang, Cheng; Zhu, Ya-Bing; Qin, Kai-Rong

    2014-06-21

    Calcium ion is a secondary messenger of mammalian spermatozoa. The dynamic change of its concentration plays a vital role in the process of sperm motility, capacitation, acrosome and fertilization. Progesterone released by the cumulus cells, as a potent stimulator of fertilization, can activate the calcium channels on the plasma membrane, which in turn triggers the dynamic change of intracellular calcium concentration. In this paper, a mathematical model of calcium dynamic response in mammalian spermatozoa induced by progesterone is proposed and numerical simulation of the dynamic model is conducted. The results show that the dynamic response of calcium concentration predicted by the model is in accordance with experimental evidence. The proposed dynamic model can be used to explain the phenomena observed in the experiments and predict new phenomena to be revealed by experimental investigations, which will provide the basis to quantitatively investigate the fluid mechanics and biochemistry for the sperm motility induced by progesterone.

  19. Shortening of alkaline DNA unwinding time does not interfere with detecting DNA damage to mouse and human spermatozoa in the comet assay

    Institute of Scientific and Technical Information of China (English)

    Hirokazu Kusakabe; Hiroyuki Tateno

    2011-01-01

    The comet assay was performed on mouse and human spermatozoa to examine the effect of alkaline DNA unwinding time.The spermatozoa were treated in vitrowith the DNA-damaging agents,methyl methanesulfonate(MMS)or hydrogen peroxide(H2O2),and then embedded in agarose gel on glass slides.The slides were immersed in alkaline solution(>pH 1.3)for 1,5,10 and 20 min,and then subjected to the electrophoresis under neutral conditions.In mouse spermatozoa,comet tails seen in solvent controls became brighter and longer as the alkaline DNA unwinding time increased.However,in the MMS-treated mouse spermatozoa,a smaller difference in the damage from that in the solvent control was seen with time within a dose.DNA damage induced by H2O2 could also be detected accurately after alkali treatment for 1-20 min.In human spermatozoa,DNA damage induced by MMS and H2O2 could be detected in a dose-dependent manner after alkali treatment for 1 min.The ability of the comet assay to detect DNA damage was not adversely affected by the short period(1 min)of the alkaline DNA unwinding time.

  20. Toxic effect of lead on human spermatozoa: A study among pigment factory workers

    Directory of Open Access Journals (Sweden)

    Naha N

    2005-01-01

    Full Text Available Occupational lead exposure caused male reproductive impairment, but information on spermatozoa activity, motility, and maturation is limited. In the above perspective, spermatozoa morphology, motility, activity, and nutritional status in lead exposed workers (7-15 years exposure were assessed. Low sperm velocity, gross, and forward progressive motility with high stationary motile spermatozoa revealed lowering of sperm cell activity after exposure (P < 0.001, which was supported by higher seminal fructose and reduced sperm ATPase activity. Lowering of seminal plasma total protein with concomitant increase in free amino acid was prevalent as exposure increased (P < 0.001, suggesting disturbance in cellular nutritional status. Prolonged liquefaction time, reduced semen volume, viscosity, seminal plasma protein, fructose, and cholesterol level among workers indicated accessory sex gland dysfunction after occupational lead exposure (P < 0.001. Deterioration of sperm density and morphology was associated with high blood and semen lead of workers (P < 0.001 leading to infertility without altering FSH, LH, and testosterone level.

  1. Cauda Epididymis Spermatozoa: Cryopreservation and Utilization for Artificial Insemination and In Vitro Fertilization

    Directory of Open Access Journals (Sweden)

    Fitra Aji Pamungkas

    2012-12-01

    Full Text Available Genetic material either from animals of economical interest or from wildlife conservation can be lost anytime by unexpected death of the animal, low libido, or disorder at reproduction. In this case, an effort can be made occur to avoid the total lost of that genetic material by using an epididymis spermatozoa. Cauda epididymis spermatozoa generally motile, mature and can be used to fertilize oocytes as well as ejaculated spermatozoa. Some research indicated that cryopreservation of cauda epididymis spermatozoa for the purpose of artificial insemination and in vitro fertilization showed the ability to fertilize oocytes and produce offspring.

  2. Cryopreservation of human spermatozoa. II. Postthaw chronology of motility and of zona-free hamster ova penetration.

    Science.gov (United States)

    Critser, J K; Arneson, B W; Aaker, D V; Huse-Benda, A R; Ball, G D

    1987-06-01

    Postthaw dynamics of motility maintenance and ability to penetrate zona-free hamster ova were examined with human sperm. Ten semen samples were each divided into two equal volumes; one was cryopreserved while the other half remained untreated. Frozen samples were thawed, and initial evaluations for motility and hamster egg penetration were made on both untreated and frozen-thawed samples. The time difference between the initial evaluations for the two treatment groups was approximately 30 minutes as a result of the time required to freeze and thaw aliquots. Subsequent evaluations were made 6, 12, 24, and 48 hours later. Over all times both the motility and fertilizability of cryopreserved spermatozoa were significantly reduced (P less than 0.05) when compared with those of untreated sperm. The pattern of motility loss over time was similar between untreated and frozen-thawed sperm (P greater than 0.10). Conversely, differences between untreated and frozen-thawed sperm in fertilizability patterns were dramatic (P less than 0.05). This was evidenced by penetration rates for cryopreserved sperm highest at 0 hour and decreasing over time, whereas penetration by untreated spermatozoa was lowest at 0 hour, increasing to a maximum at 24 hours. These observations may be important in the development of laboratory protocols for freezing and clinical protocols for using frozen-thawed sperm.

  3. Differential analysis of two-dimensional gel electrophoresis profiles of spermatozoa protein in human normal motility sperm and idiopathic asthenospermia

    Institute of Scientific and Technical Information of China (English)

    SHEN Shu-lin; HE Da-lin; LUO Yong; NING Liang

    2006-01-01

    Objective: To evaluate the application of two-dimensional electrophoresis in the research of differentially expressed proteins in the human asthenospermia. Methods: Two-dimensional gel electrophoresis was performed on 4 normal sperm samples from healthy men and 4 sperm samples from 4 asthenospermia patients. After silver staining, the differential expression proteins were analyzed by PDQuest 2D analysis software. Results: Six differential protein spots were identified. Four spots showed increased expression in the control gels compared with the patient gels. Conclusion: The protein profiles of differential expression between the normal spermatozoa and idiopathic asthenospermia were established and some differential proteins were found. The data of this study would establish the better fundament for further isolation and identification of differentially expressed proteins in human asthenospermia sperm.

  4. Expression of the vitamin D metabolizing enzyme CYP24A1 at the annulus of human spermatozoa may serve as a novel marker of semen quality

    DEFF Research Database (Denmark)

    Jensen, Martin Blomberg; Jørgensen, A; Nielsen, J E;

    2012-01-01

    Vitamin D (VD) is important for male reproduction in mammals and the VD receptor (VDR) and VD-metabolizing enzymes are expressed in human spermatozoa. The VD-inactivating enzyme CYP24A1 titrates the cellular responsiveness to VD, is transcriptionally regulated by VD, and has a distinct expression...

  5. DESCRIPTION OF ULTRASTRUCTURAL DAMAGES IN FROZEN-THAWED CANINE SPERMATOZOA DESCRIÇÃO DE DANOS ULTRAESTRUTURAIS EM ESPERMATOZOIDES CANINOS CONGELADOS-DESCONGELADOS

    OpenAIRE

    2009-01-01

    In spite of the advances in transmission electronic microscopy (TEM), there are few studies presenting a systematic description of the canine spermatozoa and they are only focused on sperm cell heads. The goal of the present study was to describe ultrastructural appearance of canine fresh and frozen-thawed spermatozoa, focusing on damages induced by freezing-thawing in different sperm regions. Ten ejaculates from five proven stud dogs (two ejaculates/dog) were collected, evaluated, extended i...

  6. Effect of zinc on decondensation of human spermatozoa nuclei by heparin.

    Science.gov (United States)

    Reyes, R; Magdaleno, V M; Hernández, O; Rosado, A; Delgado, N M

    1983-05-01

    Swelling sperm nuclei were assayed, exposing them to the combine action in increasing amounts of seminal plasma 0.1-1.0 ml (124 +/- 21 micrograms of Zn++/ml). An inhibition of almost 30% in the swollen spermatozoa nuclei was observed with 0.1 ml of seminal plasma (12 micrograms of Zn++/ml) reaching to 71% with 74-124 micrograms of Zn++/ml. Inactivated seminal plasma (boiling) induced the same percentage of inhibition (73%) than normal seminal plasma. Dialyzed seminal plasma (31 micrograms of Zn++/ml) produced an inhibition of 88% of swollen spermatozoa nuclei, the same percentage produced with 30-35 micrograms of zinc ions (ZnCl2). Previous release of sperm zinc by preincubation with EDTA 6 mM changed the decondensation kinetics, making sperm nuclei more susceptible to the action of the glycosaminoglycan. No effect was observed in the presence of calcium ions. Therefore, zinc, among its several physiological roles, may act as a nuclear chromatin stabilizer.

  7. The quality of sperm preparation medium affects the motility, viability, and DNA integrity of human spermatozoa

    Science.gov (United States)

    Anbari, Fatemeh; Halvaei, Iman; Nabi, Ali; Ghazali, Shahin; Khalili, Mohammad Ali; Johansson, Lars

    2016-01-01

    AIM: The goal was to compare the effects of three different sperm preparation media on sperm motility, viability, and DNA integrity of semen samples from normozoospermic men. METHODS: A total of 15 normozoospermic males were included in the study. The semen analysis (SA) was performed in accordance with the WHO guidelines (2010). After SA, each sample was divided into three aliquots, and swim-up was performed with three different sperm preparation media (Sperm Preparation Media, Origio, Denmark; Ham's F10, Biochrome, Berlin, Germany; and VitaSperm™, Innovative Biotech, Iran). Sperm motility, viability, and DNA fragmentation were evaluated at 0, 1, 2, and 24 h after swim-up. RESULTS: There were no significant differences, at any time intervals, in the total sperm motility between the different sperm preparation media. However, the rate of progressive motility was significantly higher in spermatozoa prepared using the media from Origio in comparison with VitaSperm™ (P = 0.03), whereas no significant difference was found against Ham's F10 medium. No significant differences in sperm viability were seen between the media products. However, 1 h after swim-up, the extent of sperm DNA fragmentation was lower in the medium from Origio versus VitaSperm™ (P = 0.02). CONCLUSIONS: The data showed that the quality of medium for preparation of semen samples from normozoospermic men significantly affects the performance of spermatozoa in assisted conception programs. PMID:28216914

  8. The quality of sperm preparation medium affects the motility, viability, and DNA integrity of human spermatozoa

    Directory of Open Access Journals (Sweden)

    Fatemeh Anbari

    2016-01-01

    Full Text Available Aim: The goal was to compare the effects of three different sperm preparation media on sperm motility, viability, and DNA integrity of semen samples from normozoospermic men. Methods: A total of 15 normozoospermic males were included in the study. The semen analysis (SA was performed in accordance with the WHO guidelines (2010. After SA, each sample was divided into three aliquots, and swim-up was performed with three different sperm preparation media (Sperm Preparation Media, Origio, Denmark; Ham′s F10, Biochrome, Berlin, Germany; and VitaSperm TM , Innovative Biotech, Iran. Sperm motility, viability, and DNA fragmentation were evaluated at 0, 1, 2, and 24 h after swim-up. Results: There were no significant differences, at any time intervals, in the total sperm motility between the different sperm preparation media. However, the rate of progressive motility was significantly higher in spermatozoa prepared using the media from Origio in comparison with VitaSperm TM (P = 0.03, whereas no significant difference was found against Ham′s F10 medium. No significant differences in sperm viability were seen between the media products. However, 1 h after swim-up, the extent of sperm DNA fragmentation was lower in the medium from Origio versus VitaSperm TM (P = 0.02. Conclusions: The data showed that the quality of medium for preparation of semen samples from normozoospermic men significantly affects the performance of spermatozoa in assisted conception programs.

  9. The quality of sperm preparation medium affects the motility, viability, and DNA integrity of human spermatozoa.

    Science.gov (United States)

    Anbari, Fatemeh; Halvaei, Iman; Nabi, Ali; Ghazali, Shahin; Khalili, Mohammad Ali; Johansson, Lars

    2016-01-01

    The goal was to compare the effects of three different sperm preparation media on sperm motility, viability, and DNA integrity of semen samples from normozoospermic men. A total of 15 normozoospermic males were included in the study. The semen analysis (SA) was performed in accordance with the WHO guidelines (2010). After SA, each sample was divided into three aliquots, and swim-up was performed with three different sperm preparation media (Sperm Preparation Media, Origio, Denmark; Ham's F10, Biochrome, Berlin, Germany; and VitaSperm™, Innovative Biotech, Iran). Sperm motility, viability, and DNA fragmentation were evaluated at 0, 1, 2, and 24 h after swim-up. There were no significant differences, at any time intervals, in the total sperm motility between the different sperm preparation media. However, the rate of progressive motility was significantly higher in spermatozoa prepared using the media from Origio in comparison with VitaSperm™ (P = 0.03), whereas no significant difference was found against Ham's F10 medium. No significant differences in sperm viability were seen between the media products. However, 1 h after swim-up, the extent of sperm DNA fragmentation was lower in the medium from Origio versus VitaSperm™ (P = 0.02). The data showed that the quality of medium for preparation of semen samples from normozoospermic men significantly affects the performance of spermatozoa in assisted conception programs.

  10. Serotonin & Ejaculation : a psychopharmacological and neuroanatomical approach

    NARCIS (Netherlands)

    Jong, T.R. de

    2005-01-01

    Ejaculatory dysfunctions, such as premature or retarded ejaculation, are common human disorders that can be very stressful for patients and their partners. In recent years it became clear that ejaculatory disorders have a neurobiological rather than a psychological cause, indicating that psychopharm

  11. Autophagy-related proteins are functionally active in human spermatozoa and may be involved in the regulation of cell survival and motility

    Science.gov (United States)

    Aparicio, I. M.; Espino, J.; Bejarano, I.; Gallardo-Soler, A.; Campo, M. L.; Salido, G. M.; Pariente, J. A.; Peña, F. J.; Tapia, J. A.

    2016-01-01

    Macroautophagy (hereafter autophagy) is an evolutionarily highly conserved cellular process that participates in the maintenance of intracellular homeostasis through the degradation of most long-lived proteins and entire organelles. Autophagy participates in some reproductive events; however, there are not reports regarding the role of autophagy in the regulation of sperm physiology. Hence, the aim of this study was to investigate whether autophagy-related proteins are present and functionally active in human spermatozoa. Proteins related to autophagy/mitophagy process (LC3, Atg5, Atg16, Beclin 1, p62, m-TOR, AMPKα 1/2, and PINK1) were present in human spermatozoa. LC3 colocalized with p62 in the middle piece of the spermatozoa. Autophagy activation induced a significant increase in motility and a decrease in PINK1, TOM20 expression and caspase 3/7 activation. In contrast, autophagy inhibition resulted in decreased motility, viability, ATP and intracellular calcium concentration whereas PINK1, TOM20 expression, AMPK phosphorylation and caspase 3/7 activation were significantly increased. In conclusion our results show that autophagy related proteins and upstream regulators are present and functional in human spermatozoa. Modification of mitochondrial proteins expression after autophagy activation/inhibition may be indicating that a specialized form of autophagy named mitophagy may be regulating sperm function such as motility and viability and may be cooperating with apoptosis. PMID:27633131

  12. THE EFFECT OF DEOXYNIVALENOL ON RABBIT SPERMATOZOA MOTILITY IN VITRO

    Directory of Open Access Journals (Sweden)

    Marína Medveďová

    2012-08-01

    Full Text Available In this in vitro study the effects of mycotoxin deoxynivalenol (DON on the motility parameters of rabbit spermatozoa were investigated. The spermatozoa motility was evaluated using CASA assay. Different concentrations of DON in the ejaculate was divided into four experimental group: 0 ng/mL, 10 ng/mL, 100 ng/mL and 1000 ng/mL. Significant differences have not been detected between control group and experimental groups after the detailed analysis of certain motility parameters – total motile spermatozoa (%, progressively motile spermatozoa (%, average path distance (DAP, µm, average path velocity (VAP, µm/s and amplitude of lateral head displacement (ALH, µm. Observed data suggested that further experiments are needed to be done because of a lack of the evidence toxinogenic effects of mycotoxin DON during its constant detection in feed and food.

  13. Impact of estrogenic compounds on DNA integrity in human spermatozoa: Evidence for cross-linking and redox cycling activities

    Energy Technology Data Exchange (ETDEWEB)

    Bennetts, L.E.; De Iuliis, G.N.; Nixon, B.; Kime, M.; Zelski, K. [ARC Centre of Excellence in Biotechnology and Development and Discipline of Biological Sciences, University of Newcastle, NSW (Australia); McVicar, C.M.; Lewis, S.E. [Obstetrics and Gynaecology, Queen' s University, Belfast (United Kingdom); Aitken, R.J. [ARC Centre of Excellence in Biotechnology and Development and Discipline of Biological Sciences, University of Newcastle, NSW (Australia)], E-mail: jaitken@mail.newcastle.edu.au

    2008-05-10

    A great deal of circumstantial evidence has linked DNA damage in human spermatozoa with adverse reproductive outcomes including reduced fertility and high rates of miscarriage. Although oxidative stress is thought to make a significant contribution to DNA damage in the male germ line, the factors responsible for creating this stress have not been elucidated. One group of compounds that are thought to be active in this context are the estrogens, either generated as a result of the endogenous metabolism of androgens within the male reproductive tract or gaining access to the latter as a consequence of environmental exposure. In this study, a wide variety of estrogenic compounds were assessed for their direct effects on human spermatozoa in vitro. DNA integrity was assessed using the Comet and TUNEL assays, lesion frequencies were quantified by QPCR using targets within the mitochondrial and nuclear ({beta}-globin) genomes, DNA adducts were characterized by mass spectrometry and redox activity was monitored using dihydroethidium (DHE) as the probe. Of the estrogenic and estrogen analogue compounds evaluated, catechol estrogens, quercetin, diethylstilbestrol and pyrocatechol stimulated intense redox activity while genistein was only active at the highest doses tested. Other estrogens and estrogen analogues, such as 17{beta}-estradiol, nonylphenol, bisphenol A and 2,3-dihydroxynaphthalene were inactive. Estrogen-induced redox activity was associated with a dramatic loss of motility and, in the case of 2-hydroxyestradiol, the induction of significant DNA fragmentation. Mass spectrometry also indicated that catechol estrogens were capable of forming dimers that can cross-link the densely packed DNA strands in sperm chromatin, impairing nuclear decondensation. These results highlight the potential importance of estrogenic compounds in creating oxidative stress and DNA damage in the male germ line and suggest that further exploration of these compounds in the aetiology of

  14. High rates of de novo 15q11q13 inversions in human spermatozoa

    Directory of Open Access Journals (Sweden)

    Molina Òscar

    2012-02-01

    Full Text Available Abstract Low-Copy Repeats predispose the 15q11-q13 region to non-allelic homologous recombination. We have already demonstrated that a significant percentage of Prader-Willi syndrome (PWS fathers have an increased susceptibility to generate 15q11q13 deletions in spermatozoa, suggesting the participation of intrachromatid exchanges. This work has been focused on assessing the incidence of de novo 15q11q13 inversions in spermatozoa of control donors and PWS fathers in order to determine the basal rates of inversions and to confirm the intrachromatid mechanism as the main cause of 15q11q13 anomalies. Semen samples from 10 control donors and 16 PWS fathers were processed and analyzed by triple-color FISH. Three differentially labeled BAC-clones were used: one proximal and two distal of the 15q11-q13 region. Signal associations allowed the discrimination between normal and inverted haplotypes, which were confirmed by laser-scanning confocal microscopy. Two types of inversions were detected which correspond to the segments involved in Class I and II PWS deletions. No significant differences were observed in the mean frequencies of inversions between controls and PWS fathers (3.59% ± 0.46 and 9.51% ± 0.87 vs 3.06% ± 0.33 and 10.07% ± 0.74. Individual comparisons showed significant increases of inversions in four PWS fathers (P Results suggest that the incidence of heterozygous inversion carriers in the general population could reach significant values. This situation could have important implications, as they have been described as predisposing haplotypes for genomic disorders. As a whole, results confirm the high instability of the 15q11-q13 region, which is prone to different types of de novo reorganizations by intrachromatid NAHR.

  15. Nerve growth factor promotes human sperm motility in vitro by increasing the movement distance and the number of A grade spermatozoa.

    Science.gov (United States)

    Lin, Kai; Ding, Xue-Feng; Shi, Cui-Ge; Zeng, Dan; QuZong, SuoLang; Liu, Shu-Hong; Wu, Yan; LuoBu, GeSang; Fan, Ming; Zhao, Y-Q

    2015-11-01

    Nerve growth factor (NGF) was first found in the central nervous system and is now well known for its multiple pivotal roles in the nervous system and immune system. However, more and more evidences showed that NGF and its receptors TrkA and p75 were also found in the head and tail of spermatozoa, which indicate the possible effect of NGF on the sperm motility. Nevertheless, the exact role of NGF in the human sperm motility remains unclear until now. In this study, we investigated the effect of NGF on human sperm motility, and the results showed that NGF could promote human sperm motility in vitro by increasing the movement distance and the number of A grade spermatozoa. Further analysis demonstrated that NGF promoted the sperm motility in a dose-dependent manner in vitro. These results may facilitate the further studies on human fertility and assisted reproduction techniques.

  16. Characteristics of donkey spermatozoa along the length of the epididymis.

    Science.gov (United States)

    Contri, A; Gloria, A; Robbe, D; De Amicis, I; Carluccio, A

    2012-01-01

    In mammals, the epididymis has numerous interrelated functions including absorptive and secretory activity that affect luminal environment and cell membrane, and the maturation and storage of sperm. Spermatozoa acquire their motility and fertilizing ability during their passage through the epididymis and the motility of epididymal spermatozoa should be a balance between the maturation of flagellum and the inhibition of the flagellar machinery. In this study maturational change in sperm characteristics were evaluated in the epididymis of donkey. Spermatozoa collected from four portions of the epididymis (head, cranial corpus, caudal corpus, tail) were compared before and after ejaculation for viability, mitochondrial activity, kinetic parameters, and morphology. A significant increase in the mitochondrial activity along the epididymis was reported, suggesting a possible involvement in the motion mechanism. This should be corroborated by the significant correlation between mitochondrial activity and the total and progressive motility and the increase in velocities of spermatozoa recorded by computer-assisted sperm analysis. The percentage of most of the abnormal spermatozoa were similar in all tracts, with a great variability between jackasses. Only the bent midpiece percentage decreased significantly along epididymis. A significant increase in the percentage of distal cytoplasmic droplets (DCD), and a simultaneous decrease in the proximal cytoplasmic droplets (PCD), was found. The DCD fell down after ejaculation suggesting the late loss of the cytoplasmic residual (DCD) in the donkey, as hypothesized in the stallion. Because the prevalence of PCD were similar in both tail epididymal and ejaculated spermatozoa, a defect of the maturative process in the PCD sperm should be speculated.

  17. Effect of cholesterol supplementation on cryosurvival of goat spermatozoa

    Directory of Open Access Journals (Sweden)

    Sunita Behera

    2015-12-01

    Full Text Available Aim: Sperm membrane cholesterol influences cryodamage during cryopreservation. The present study was carried out to evaluate the effect of varying cholesterol levels in Tris based extenders on the freezability of sexually healthy Malabari buck semen. Materials and Methods: A total of 48 ejaculates from two adults healthy sexually healthy Malabari bucks were utilized for the study. The collected and pooled ejaculates were divided into four groups with Group I serving as Control - I, Group II and III were treated with 1 mg and 2 mg of cholesterol-loaded-cyclodextrin (CLC/120 × 106 spermatozoa, respectively, and Group IV treated with 1 mg methyl-β-cyclodextrin (MβCD served as Control - II. Manual freezing was carried out to cryopreserve the treated and control spermatozoa. Results: Treatment of semen samples with CLC resulted in improved maintenance of sperm motility at pre-freeze and post-thaw stages of cryopreservation without affecting hypo-osmotic swelling response. Treatment of semen with 1 mg of CLC/120 × 106 spermatozoa was observed to be better than treatment with 2 mg of CLC/120 × 106 spermatozoa. In general, MβCD treatment was found to result in significantly lower sperm characteristics than those of Control - I and CLC treatment at pre-feeze and post-thaw stages and when incubated up to 4 h. Conclusion: Cholesterol treatment of sexually healthy Malabari buck semen was found to hold promise for improving cryopreser-vability of spermatozoa.

  18. Sources of spermatozoa loss during collection and artificial insemination of horses.

    Science.gov (United States)

    Côté, Michelle A; Blum, Katherine M; Burd, Matthew A

    2011-07-01

    During artificial insemination of horses, it is important to accurately estimate the number of spermatozoa in each insemination dose. However, little research exists regarding sources of spermatozoa loss during collection and artificial insemination. Therefore, spermatozoal losses were quantified in the dismount loss (187.6×10(6)±62.5×10(6)spermatozoa), gel fraction (179.8×10(6)±61.7×10(6)spermatozoa), and the collection receptacle (136.1×10(6)±26.9×10(6)spermatozoa). Spermatozoal losses were examined in the centrifuge tube (25.8×10(6)±2.1×10(6)spermatozoa), AI pipette during the air removal (90.9×10(6)±8.5×10(6)spermatozoa), and spermatozoa remaining in the AI pipette after insemination (342.9×10(6)±21.4×10(6)spermatozoa). The average cumulative loss was 14.2±2.9% of the total spermatozoa ejaculated with approximately half of the loss due to the process of semen collection and half due to the process of artificial insemination. Spermatozoa retained in the AI pipette, after insemination with extended semen, represented the greatest source of loss.

  19. Chromosomal aneuploidies and DNA fragmentation of human spermatozoa from patients exposed to perfluorinated compounds.

    Science.gov (United States)

    Governini, L; Guerranti, C; De Leo, V; Boschi, L; Luddi, A; Gori, M; Orvieto, R; Piomboni, P

    2015-11-01

    This study investigated chromosomal aneuploidies and DNA damage in spermatozoa from male patients contaminated by perfluorinated compounds (PFCs) in whole blood and seminal plasma. Sperm aneuploidy and diploidy rate for chromosomes 18, X and Y were evaluated by FISH; sperm DNA fragmentation was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling technique coupled to flow cytometry. Our results indicated that PFC contamination was present in 58% of subjects included in the study. A significant increase in alterations of sperm parameters was observed in PFC-positive subjects compared to PFC-negative subjects. As regards the sperm aneuploidy, both disomy and diploidy rates resulted significantly increased in subjects positive for PFC contamination compared to PFC-negative samples. In addition, sperm DNA fragmentation index resulted significantly increased in PFC-contaminated subjects compared to PFC-non-contaminated subjects, with a significant increased level of dimmer DNA fragmentation index. Our results clearly indicate that PFC contamination may detrimentally affect spermatogenesis, disturbing both meiotic segregation and DNA integrity. We could therefore suggest cautions to reduce or eliminate any contact with these compounds because the long-term effects of PFC accumulation in the body are not predictable. © 2014 Blackwell Verlag GmbH.

  20. Efficacy of Hyaluronic Acid in The Selection of Human Spermatozoa with Intact DNA by The Swim-up Method

    Directory of Open Access Journals (Sweden)

    Aslihan Saylan

    2016-04-01

    Full Text Available Objective: In 2014, enrolled 20 patients who applied to the Unit of Assisted Reproduction Techniques, Konya Necmettin Erbakan University. Based on the presence of hyaluronic acid (HA in the oocyte-cumulus cell complex, sperm attached to HA in vivo were modeled in vitro. Available healthy sperm obtained in the swim-up procedure using HA were investigated. Materials and Methods: This observational cohort study, a routine analysis was conducted on the ejaculation samples obtained from 20 patients. We divided each sample into two groups and the swim-up method was applied. Human serum albumin (HSA, 0.5% was added to samples from the first group. HA (10% was added to samples from the second group. We determined the floating linear and non-linear sperm concentrations of both groups annexin V was used to determine the rate of apoptosis of these sperm. Results: Following swim-up, linear and non-linear sperm concentrations were higher in the group that contained HA compared to the group with HSA. However, there was a significantly higher apoptosis rate in the HSA group compared to the HA group. Conclusion: The addition of HA to the medium in the swim-up procedure positively affected sperm parameters. Thus, healthier sperm cells were obtained without DNA damage and with high motility.

  1. Studies on the viability and membrane integrity of human spermatozoa treated with essential oil of Trachyspermum ammi (L.) Sprague ex Turrill fruit.

    Science.gov (United States)

    Paul, S; Kang, S C

    2012-05-01

    The present study aimed at investigating the effects of essential oil of Trachyspermum ammi fruits, an oil-bearing plant of Apiaceae family, on human sperm viability and membrane integrity. Chemical compositions of the oil were analysed by GC-MS. Thirty compounds representing 91.39% of the total oil were identified. The viability and membrane integrity of human spermatozoa were assessed using minimum effective dose (MED) concentration (125 μg ml(-1)) of the oil. Sperm treated with essential oil showed a significant decrease (P < 0.05) in viability assessed by eosin-nigrosin and fluorescence dual staining. Moreover, the treated sperm also showed a significant loss (P < 0.05) of functional mitochondria and antioxidant enzyme, catalase (EC 1.11.1.6, CAT), when compared to control. The cholesterol:phospholipid ratio was also increased (P < 0.05) in treated sperm when compared to control, which is an indicator of loss of binding ability of human spermatozoa to the zona pellucida. The scanning electron microscopic studies demonstrated the loss of membrane integrity in essential oil-treated human spermatozoa, which showed vacuolation, swelling of acrosomal cap, detachment of head portion and tail coiling. Present observations indicate the spermicidal property of essential oil of T. ammi fruits, which could be helpful to develop medicinal preparations as a male contraceptive.

  2. Phospholipid hydroperoxide glutathione peroxidase in bull spermatozoa provides a unique marker in the quest for semen quality analysis.

    Science.gov (United States)

    Stradaioli, G; Sylla, L; Monaci, M; Maiorino, M

    2009-07-01

    Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a selenoperoxidase accounting for most of the selenium content in mammalian testis, which has been found to be linked to fertility in humans. In this study, we addressed the issue whether PHGPx content in spermatozoa could be a predictive index of fertilization capacity for sire selection in bulls. Measurement of PHGPx in spermatozoa of 92 yearling bulls of three different Italian breeds (Chianina, Romagnola, and Marchigiana) revealed the presence of two quite well separated populations. A PHGPx activity of 130 mU/mg separated the high-PHGPx group (H-PHGPx, n=73) from the low-PHGPx group (L-PHGPx, n=19). Forward motility was markedly higher in the H-PHGPx group, which also contained a lower percentage of detached heads, abnormal midpiece, and proximal droplets. On the other hand, differently from the human studies, no correlation was observed between PHGPx activity and number of spermatozoa in the ejaculate. Apart from sperm count, which typically differed among breeds, and number of detached heads in the L-PHGPx group, which correlated with higher sperm count, no other significant difference in seminal parameters among breeds was apparent. The assay for sperm PHGPx activity therefore emerges as a unique tool to evaluate semen quality for sire selection.

  3. Secretory expression of a novel human spermatozoa antigen in E. coli and its application to a protein chip.

    Science.gov (United States)

    Wang, Ming-Zhu; Qiu, Zhuo-Lin; Cai, Xiang-Sheng; Li, Jing-Jing; She, Miao-Qin; Xu, Yuan-Feng; Wu, Ying-Song

    2017-06-09

    To produce a recombinant spermatozoa antigen peptide using the E. coli: PhoA system on a protein chip for screening anti-sperm antibodies (ASA). The purity of the recombinant spermatozoa antigen exceeded 95% after two-step purification, as assessed using SDS-PAGE and HPLC. The diagnostic performance of a protein chip coated with the recombinant antigen peptide was evaluated by examining ASA in 51 infertile patients in comparison with a commercial ELISA kit. The area under the receiver operating characteristic curve (AUC) was 0.944, which indicated that the protein chip coated with recombinant spermatozoa antigen peptide was consistent with ELISA for ASA detection. A recombinant spermatozoa antigen was expressed in the E. coli PhoA secretory expression system and its potential application for clinical ASA detection was validated.

  4. Boar sperm quality in relation to presence of sp32-like protein in spermatozoa - preliminary studies

    Directory of Open Access Journals (Sweden)

    Orzołek Aleksandra

    2015-06-01

    Full Text Available The aim of the study was to analyse sperm proteomes of ejaculates from Polish Large White (PLW and Polish Landrace (PL boars and to identify differences which putatively influence semen quality. Spermatozoa protein profiles were analysed by electrophoretic methods followed by selected techniques to evaluate semen quality on the following factors: sperm motility, lipid peroxidation levels (MDA production, ATP content, activities of superoxide dismutase (SOD and catalase (CAT, total antioxidant status (TAS, and total oxidant status (TOS of seminal plasma. A protein with an estimated molecular weight of 30 kDa was found in spermatozoa of selected ejaculates. Mass spectrometry demonstrated that this polypeptide is most similar to proacrosin binding protein (sp32. The presence of the protein was more frequently observed in sperm extracts obtained in spring-summer period. Ejaculates containing sp32-like protein demonstrated significantly higher spermatozoa motility, lower inhibition of MDA production by seminal plasma, and higher SOD activity in seminal plasma. Boar semen which included sp32-like protein also demonstrated lower ATP levels in spermatozoa as well as higher TAS and lower TOS of seminal plasma, though the differences were not statistically significant. Ejaculates from PLW boars, with sp32-like protein present in sperm, were characterised by significantly higher sperm motility, lower ATP content in spermatozoa, and higher TAS of seminal plasma. The diminished parameters of semen quality were observed in ejaculates from PL boars that also contained the discussed protein, but the differences were not statistically significant. These findings suggest that the presence of sp32-like protein in boar spermatozoa could influence semen quality

  5. DNA damage in human spermatozoa is highly correlated with the efficiency of chromatin remodeling and the formation of 8-hydroxy-2'-deoxyguanosine, a marker of oxidative stress.

    Science.gov (United States)

    De Iuliis, Geoffry N; Thomson, Laura K; Mitchell, Lisa A; Finnie, Jane M; Koppers, Adam J; Hedges, Andrew; Nixon, Brett; Aitken, R John

    2009-09-01

    DNA damage in human spermatozoa has been associated with a range of adverse clinical outcomes, including infertility, abortion, and disease in the offspring. We have advanced a two-step hypothesis to explain this damage involving impaired chromatin remodeling during spermiogenesis followed by a free radical attack to induce DNA strand breakage. The objective of the present study was to test this hypothesis by determining whether impaired chromatin protamination is correlated with oxidative base damage and DNA fragmentation in human spermatozoa. DNA fragmentation, chromatin protamination, mitochondrial membrane potential, and formation of the oxidative base adduct, 8-hydroxy-2'-deoxyguanosine (8OHdG), were monitored by flow cytometry/fluorescence microscopy. Impairment of DNA protamination during late spermatogenesis was highly correlated (P human spermatozoa. The disruption of chromatin remodeling also was associated with a significant elevation in the levels of 8OHdG (P < 0.001), and the latter was itself highly correlated with DNA fragmentation (P < 0.001). The significance of oxidative stress in 8OHdG formation was demonstrated experimentally using H2O2/Fe2+ and by the correlation observed between this base adduct and superoxide generation (P < 0.001). That 8OHdG formation was inversely associated with mitochondrial membrane potential (P < 0.001) suggested a possible role for these organelles in the creation of oxidative stress. These results clearly highlight the importance of oxidative stress in the induction of sperm DNA damage and carry significant implications for the clinical management of this condition.

  6. Diagnostic and treatment of retrograde ejaculation as a manifestation of urogenital form of autonomic diabetic polyneuropathy

    Directory of Open Access Journals (Sweden)

    Dmitry Gennadyevich Kurbatov

    2015-07-01

    Full Text Available Background. Retrograde ejaculation in patients with type 1 diabetes mellitus is a complication of autonomic neuropathy that causes excretory infertility. It can be partial (reduction of ejaculate or total (absence of ejaculate and occurs in 10%–20% of men with type 1 diabetes mellitus.Aim. The aim of the study was to assess the effectiveness of a new endoscopic method for retrograde ejaculation correction and antegrade ejaculation restoration.Materials and methods. We included 30 patients with type 1 diabetes mellitus who had spermatozoa present in their post-orgasmic urine and ultrasonographic evidence of impaired or absent bladder neck closure. The mean age of participants was 32 (30–35 years, mean duration of diabetes was 17 (12–22 years and mean preoperative glycated haemoglobin level was 7.4% (6.9%–8.0%. All participants had total retrograde ejaculation. We used conventional irrigated urethrocystoscopy under local anaesthesia. During urethroscopy, bladder neck gaping was observed in all cases. Biocompatible material was injected at three points under the mucous layer of the posterior urethra, reaching the closing of the opposite edges of the urethra. A spermiogram was examined 1 week after the operation.Results. Restoration of antegrade ejaculation was achieved for 22 patients (73%, and the effects persisted for a mean of 7 (2–12 months. The spouses of four men became pregnant after surgery. In one case, the pregnancy resulted in a spontaneous abortion at gestational week 8, but the other three cases continued normally.Conclusion.This new method provides a highly effective means of restoring the physiological passage of the ejaculate. The operation is a low-invasive endoscopic procedure that does not disrupt urination, and it is possible to receive ejaculate of sufficient.

  7. Binding of free and protein-associated zinc to rat spermatozoa.

    Science.gov (United States)

    Sansone, G; Martino, M; Abrescia, P

    1991-01-01

    1. The zinc content of rat spermatozoa increases, upon ejaculation, from 0.035 to 1.055 micrograms/10(6) cells. 2. The rat seminal plasma holds zinc both as free ion and as protein-bound forms. 3. Zinc-free ions bind in vitro to rat epididymal spermatozoa. 4. Zinc-protein complexes can be isolated, by a chromatographic procedure, from the dorsolateral lobe of rat prostate. 5. The isolated zinc-protein complexes bind in vitro to rat epididymal spermatozoa.

  8. The effect of Ureaplasma urealyticum on the level of P34H expression, the activity of hyaluronidase, and DNA fragmentation in human spermatozoa.

    Science.gov (United States)

    Ma, Xiao-Ping; Gao, Xiao-Qin

    2017-01-01

    To study the effect of Ureaplasma urealyticum on the level of P34H expression, the activity of hyaluronidase, and the DNA fragmentation in human spermatozoa. Western blot was used to detect the level of P34H expression on spermatozoa.The localization of this protein on human spermatozoa was determined by indirect immunofluorescent and observed by laser scanning confocal microscope. The activity of hyaluronidase was examined by improved fixed-substrate film method. The DNA fragmentation was assayed with the use of TUNEL. There were significant differences in the level of P34H protein expression, the percentage of the P34H-positive rate, the activity of HYD, and the percentage of DNA fragmentation between each infertile group and the control (Pfragmentation. Ureaplasma urealyticum infection may affect the level of P34H protein expression, the percentage of the P34H-positive rate, the activity of HYD, and DNA fragmentation that influence fertility. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. Sperm morphometry: a tool for detecting biophysical changes associated with viability in cryopreserved bovine spermatozoa.

    Science.gov (United States)

    García-Herreros, M; Leal, C L V

    2014-09-01

    The aim of this study was to determine whether computerised sperm head morphometric analysis can be used as a diagnostic tool for detecting biophysical changes associated with sperm viability in frozen-thawed bovine spermatozoa. Ejaculates from five bulls (4 ejaculates/bull) were pooled and processed for computerised morphometric analysis, and SYBR-14 green/ethidium homodimer-1 fluorescence-based live/dead viability assay was used simultaneously to confirm the viability index of frozen-thawed spermatozoa. Sperm samples were assigned to three experimental groups. The first group was enriched in live spermatozoa (after a double Percoll selection), the second group was enriched in dead spermatozoa (after a refreeze-thaw procedure), and the last group was a 50 : 50 pool of live/dead spermatozoa (from first and second group samples). There were significant differences (P sperm morphometric dimensional parameters among the three groups analysed, being the lowest overall sperm head dimension found in the second (dead spermatozoa) group. In conclusion, sperm head morphometry can be used as a potential diagnostic tool for detecting biophysical changes associated with sperm viability in frozen-thawed bovine spermatozoa.

  10. Increased frequency of aneuploidy in long-lived spermatozoa.

    Science.gov (United States)

    You, Young-Ah; Park, Yoo-Jin; Kwon, Woo-Sung; Yoon, Sung-Jae; Ryu, Buom-Yong; Kim, Young Ju; Pang, Myung-Geol

    2014-01-01

    Aneuploidy commonly causes spontaneous abortions, stillbirths, and aneuploid births in humans. Notably, the majority of sex chromosome aneuploidies in live births have a paternal origin. An increased frequency of aneuploidy is also associated with male infertility. However, the dynamics and behavior of aneuploid spermatozoa during fertilization in humans have not been studied in detail. Therefore, we compared the frequency of aneuploidy and euploidy in live spermatozoa from normozoospermic men over a 3-day period. To assess the dynamics and behavior of aneuploid spermatozoa, we simultaneously evaluated sperm viability using the hypo-osmotic swelling test and sperm aneuploidy using fluorescence in situ hybridization. Whereas the frequency of viable euploid spermatozoa significantly decreased over 3 days, the frequency of viable spermatozoa with aneuploidy interestingly showed a time-dependent increase. In addition, spermatozoa with abnormal sex chromosomes survived longer. To compared with spermatozoa with other swelling patterns, those with tail-tip swelling patterns had a lower frequency of aneuploidy at all time points. This study revealed the novel finding that the frequency of aneuploid spermatozoa with fertilization capability significantly increased compared to that of euploid spermatozoa over 3 days, suggesting that aneuploid spermatozoa can survive longer than euploid spermatozoa and have a greater chance of fertilizing oocytes.

  11. Increased frequency of aneuploidy in long-lived spermatozoa.

    Directory of Open Access Journals (Sweden)

    Young-Ah You

    Full Text Available Aneuploidy commonly causes spontaneous abortions, stillbirths, and aneuploid births in humans. Notably, the majority of sex chromosome aneuploidies in live births have a paternal origin. An increased frequency of aneuploidy is also associated with male infertility. However, the dynamics and behavior of aneuploid spermatozoa during fertilization in humans have not been studied in detail. Therefore, we compared the frequency of aneuploidy and euploidy in live spermatozoa from normozoospermic men over a 3-day period. To assess the dynamics and behavior of aneuploid spermatozoa, we simultaneously evaluated sperm viability using the hypo-osmotic swelling test and sperm aneuploidy using fluorescence in situ hybridization. Whereas the frequency of viable euploid spermatozoa significantly decreased over 3 days, the frequency of viable spermatozoa with aneuploidy interestingly showed a time-dependent increase. In addition, spermatozoa with abnormal sex chromosomes survived longer. To compared with spermatozoa with other swelling patterns, those with tail-tip swelling patterns had a lower frequency of aneuploidy at all time points. This study revealed the novel finding that the frequency of aneuploid spermatozoa with fertilization capability significantly increased compared to that of euploid spermatozoa over 3 days, suggesting that aneuploid spermatozoa can survive longer than euploid spermatozoa and have a greater chance of fertilizing oocytes.

  12. Impact of cigarette smoking on histone (H2B) to protamine ratio in human spermatozoa and its relation to sperm parameters.

    Science.gov (United States)

    Hamad, M F; Shelko, N; Kartarius, S; Montenarh, M; Hammadeh, M E

    2014-09-01

    Smoking is strongly associated with abnormalities in histone-to-protamine transition and with alteration of protamine expression in human spermatozoa. A proper protamine to histone ratio is, however, essential for sperm chromatin maturity and DNA integrity. Alterations in these sperm nuclear proteins were observed in infertile men. The present prospective study is aimed at evaluating the possible relationship among smoking, semen quality and the histone-to-protamine transition ratio in mature spermatozoa. Histone H2B and protamine 1 (P1) and 2 (P2) were quantified using acid-urea polyacrylamide gel electrophoresis in the spermatozoa of 35 smokers and 19 non-smokers. Levels of lipid peroxidation marker malondialdehyde (MDA) were measured in seminal plasma by thiobarbituric acid assay. Cotinine concentrations were determined in seminal plasma using an enzyme-linked immunosorbent assay. Histone H2B levels in smokers (292.27 ± 58.24 ng/10(6)) were significantly higher (p = 0.001) than that of non-smokers (109.1 ± 43.70 ng/10(6)), besides, a significant difference (p > 0.0001) was found for the P1 and P2 ratio between smokers (1.71 ± 0.071) and non-smokers (1.05 ± 0.033). The H2B/(H2B+P1 + P2) ratio (0.29 ± 0.71) of smokers were significantly higher (p = sperm count, motility (p = 0.018), vitality (p = 0.009) and membrane integrity (p = 0.0001) than non-smokers. These results reveal that patients who smoke possess a higher proportion of spermatozoa with an alteration of the histone to protamine ratio than patients who do not smoke, and suggest that cigarette smoking may inversely affect male fertility.

  13. Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection.

    Science.gov (United States)

    Yeoman, Richard R; Mitalipov, Shoukhrat; Gerami-Naini, Behzad; Nusser, Kevin D; Wolf, Don P

    2005-06-01

    The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen-thawed rhesus monkey spermatozoa and fertility via intracytoplasmic spermatozoa injection (ICSI) of sibling oocytes. Washed spermatozoa were frozen in straws or in pellets using different cryoprotective media and processed post-thaw with or without a density gradient centrifugation step. Among the four study series, motility post-thaw was improved with density gradient centrifugation (17-24% versus 75%, P<0.01) achieving levels similar to fresh spermatozoa. Spermatozoa injected oocytes (total n=377) were co-cultured on BRL cells and observed for fertilization and development. With spermatozoa frozen in straws in liquid nitrogen vapors, the fertilization rate after ICSI was lower than with fresh spermatozoa (40-44% versus 77-86%, P<0.05), even with the Percoll-enriched fraction that exhibited robust motility. In contrast, somewhat slower freezing of spermatozoa in pellets on dry ice supported fertilization rates (73%) that were similar to the fresh counterpart. Developmental rates of fertilized eggs were similar in all experiments. A total of 106 embryo transfers has resulted in the first primate born after ICSI with F/T ejaculated spermatozoa plus 22 other infants to date. Additionally, a 3-4 h incubation after thawing improved the fertilization rate with spermatozoa from a male with poor post-thaw recovery of sperm motility. In conclusion, an acceptable fertilization rate after ICSI with motile, frozen-thawed primate spermatozoa was observed comparable to that obtained with fresh spermatozoa allowing small quantities of competent spermatozoa to be used with ICSI to facilitate propagation of desirable primate genotypes.

  14. Hypo-osmotic test in cat spermatozoa.

    Science.gov (United States)

    Comercio, E A; Monachesi, N E; Loza, M E; Gambarotta, M; Wanke, M M

    2013-10-01

    The hypo-osmotic (HOS) test has been used in other species as an indicator of the fertilising capacity of spermatozoa. The aims of this study were to assess the response of domestic cat spermatozoa to the hypo-osmotic test, to determine the type of solution, concentration and time of incubation needed to obtain a maximum percentage of swelling, to correlate the selected combination with the percentages of progressive motility and to evaluate whether dilution of the ejaculate alters the results. Incubation for 30 and 45 min in solutions of fructose and of citrate of 50 and 100 mOsmol kg⁻¹ was evaluated. The highest percentage of swelling was obtained using the 50 mOsmol kg⁻¹ solution, and no significant differences were observed between the times of exposure to the solutions. A positive correlation was observed between the percentage of individual progressive motility and the percentage of sperm swelling in a 50 mOsmol kg⁻¹ fructose solution, with no significant differences being observed between raw and diluted semen samples. The results of this study suggest that the HOS test could be useful for evaluating membrane function in domestic cat spermatozoa, both in raw semen and in samples diluted in the EZ Mixin® commercial extender, and thus could be incorporated into routine semen evaluation protocols.

  15. Cold-preserved human spermatozoa in electrolyte-free solution retain their penetration capacity

    Institute of Scientific and Technical Information of China (English)

    全松; 周海宽; 山野修司; 中坂尚代; 青野敏博

    2003-01-01

    Objective: To evaluate penetration capacity of human sperm preserved in electrolyte-free (EF) solution at 4 ℃.Methods: The motility, acrosomal status penetration rate and fertility index of human sperm were assessed before and after cold-preservation in EF solution, respectively.Results: The motility of human sperm cold-preserved in EF solution for 1 week was significantly higher than that of human sperm cold-preserved in modified human tubal fluid (mHTF) (43.4%±7.9% vs 9.5%±2.5%, P0.05), the percentage of capacitated and acrosome-reacted sperm in the EF solution significantly increased after reinitiation (capacitated sperm: 16.0%±2.3% vs 7.6±1.8%, acrosome-reacted sperm: 9.4%±2.1% vs 3.0%±1.7%, P0.05).Conclusion: Cold-preservation did not induce capacitation and acrosome reaction of human sperm in the EF solution, but human sperm cold-preserved in the EF solution for 1 week possesses as much penetration capacity as fresh sperm.

  16. Ejaculation training, seminal alkaline phosphatase and semen preservation through cooling in a milk-based extender in domestic cats.

    Science.gov (United States)

    Valiente, Carla; de la Sota, Pablo E; Arauz, Sandra; Gobello, Cristina

    2014-04-01

    The purpose of this report is to describe (1) the training of domestic cats in ejaculation into an artificial vagina (AV), (2) alkaline phosphatase (AP) concentrations in whole ejaculates, and (3) the in vitro effect of a skimmed-milk plus egg yolk (SM-Y) extender on feline spermatozoa incubated at 4ºC. Five post-pubertal cats were trained to ejaculate into an AV three times a week for 20 mins in the presence of a teaser queen. Fifty AV-obtained ejaculates were macro- and microscopically assessed, and the AP therein measured by optimized colorimetry. Eighty AV-obtained ejaculates were pooled, diluted in SM-Y extender [80% (v/v) skimmed milk, 20% (v/v) egg yolk, and antibiotics], stored at 4°C and evaluated daily for 6 days. All the animals could be trained to ejaculate, although the interval up to the first AV ejaculation varied from 1.5 to 5.5 months (mean 3.9 months). The final performance at collection ranged from excellent to poor and was inversely related to the training period required in all cases. The mean AP concentration in whole ejaculates was 20,645.6 ± 4405U/l, which was not correlated with the concentration of spermatozoa. Most seminal parameters [(%); total (77 ± 2.3) and progressive (62.7 ± 3.4) motility, live sperm (91.8 ± 1.2), intact plasmalemma (83.5 ± 2.6), normal acrosomes (83.5 ± 2.6), pH (6.6 ± 0.0) and osmolarity (mOsm/l; 321 ± 5.2)], though decreasing during storage in the cold, remained within values compatible with in vivo fertilization for 2 days.

  17. Comparative analysis of the nuclear basic proteins in rat, human, guinea pig, mouse and rabbit spermatozoa.

    Science.gov (United States)

    Calvin, H I

    1976-06-15

    Cysteine-rich protamines (Arg = 47-61%, Cys = 8-16%) were isolated from the sperm of an individual guinea pig, human and rabbit and from pooled samples of mouse and rat sperm. Appreciable concentrations of histones were not found in the sperm nuclei of these species. In addition to the protamines, a substance of relatively low molecular weight, which reacted with the Lowry reagent, appeared in crude acid-soluble extracts of sperm nucleoprotein. This unidentified contaminent was resolved from the protamines by chromatography on Bio-Rex 70. Heterogeneity of human and mouse protamines was revealed by electrophoresis at pH 2.7, in the presence of 2.5 M urea, and confirmed by amino acid analysis, which also suggested the presence of 2 or more species of protamine in the rabbit. By contrast, the guinea pig and rat preparations displayed nearly stoichiometric ratios of amino acid residues, approaching homogeneity by this criterion. The functional consequences of crosslinks between cysteine residues of these proteins and the possible species-specific significance of their differing percentages of histidine are discussed. Potentially analogous functions are suggested for phosphorylated serine and threonine, and for ionized cysteine and tyrosine, within the protamines of developing spermatids. Their amino acid compositions indicate that the protamines of eutherian mammals are coded by a C.G-rich genome which has been unusually susceptible to genetic drift. An especially high rate of G leads to A transitions seems to have occurred in the human protamine genes.

  18. Identification of multiple HPV types on spermatozoa from human sperm donors

    DEFF Research Database (Denmark)

    Kaspersen, Maja D; Larsen, Peter B; Ingerslev, Hans Jakob

    2011-01-01

    Human papillomaviruses (HPV) may cause sexually transmitted disease. High-risk types of HPV are involved in the development of cervical cell dysplasia, whereas low-risk types may cause genital condyloma. Despite the association between HPV and cancer, donor sperm need not be tested for HPV...... contain HPV, most of them of high-risk types binding to the equatorial segment of the sperm cell. Most HPV-positive sperm showed decreased staining with DAPI, indicative of reduced content of DNA. Our data demonstrate that oncogenic HPV types are frequent in men....... according to European regulations. Consequently, the potential health risk of HPV transmission by donor bank sperm has not been elucidated, nor is it known how HPV is associated with sperm. The presence of 35 types of HPV was examined on DNA from semen samples of 188 Danish sperm donors using a sensitive...

  19. [Herpetic infection of spermatozoa in failure of reproductive technologies and spontaneous abortion].

    Science.gov (United States)

    Bocharova, E N; Bragina, E E; Gusak, Iu K; Zotov, V V; Tereshchenko, A B; Shileĭko, L V; Kurilo, L F; Klimova, R R; Kushch, A A

    2007-01-01

    Herpes simplex virus (HSV) was studied in ejaculate of 36 men whose wives had the history of spontaneous abortions, or failure of artificial insemination (AI), or extracorporeal fertilization (ECF). The control group consisted of 222 patients examined prophylactically after urogenital inflammation or for primary infertility. HSV was detected with immunofluorescence (IF) reaction with monoclonal antibodies to HSV antigens, rapid culture method (RCM) and transmission electronic microscopy. HSV antigens were detected in fraction of mobile spermatozoa in 13 (65%) of 20 samples from patients of the study group. HSV detectability in the study group was significantly higher than in the controls (p = 0.003). Intragametic capsides of HSV were detected at ultrastructural investigation in 13 of 19 (68%) ejaculate samples from patients of the study group in whom IF and/ or RCM results were positive. The study of 6 ejaculate samples of this group in which HSV was not detected by IF or RCM and 24 control samples HSV capsides were not detected. The following conclusions can be made: mobile spermatozoa of normal morphology can be infected with HSV; as shown morphologically, samples of ejaculate from men whose wives had spontaneous abortions contained intragametic HSV capsides; intragametic HSV infection of spermatozoa had a negative effect on the course of pregnancy and/or embryon vitality.

  20. Identification of multiple HPV types on spermatozoa from human sperm donors.

    Science.gov (United States)

    Kaspersen, Maja D; Larsen, Peter B; Ingerslev, Hans Jakob; Fedder, Jens; Petersen, Gert Bruun; Bonde, Jesper; Höllsberg, Per

    2011-03-29

    Human papillomaviruses (HPV) may cause sexually transmitted disease. High-risk types of HPV are involved in the development of cervical cell dysplasia, whereas low-risk types may cause genital condyloma. Despite the association between HPV and cancer, donor sperm need not be tested for HPV according to European regulations. Consequently, the potential health risk of HPV transmission by donor bank sperm has not been elucidated, nor is it known how HPV is associated with sperm. The presence of 35 types of HPV was examined on DNA from semen samples of 188 Danish sperm donors using a sensitive HPV array. To examine whether HPV was associated with the sperm, in situ hybridization were performed with HPV-6, HPV-16 and -18, and HPV-31-specific probes. The prevalence of HPV-positive sperm donors was 16.0% and in 66.7% of these individuals high-risk types of HPV were detected. In 5.3% of sperm donors, two or more HPV types were detected. Among all identified HPV types, 61.9% were high-risk types. In situ hybridization experiments identified HPV genomes particularly protruding from the equatorial segment and the tail of the sperm. Semen samples from more than one in seven healthy Danish donors contain HPV, most of them of high-risk types binding to the equatorial segment of the sperm cell. Most HPV-positive sperm showed decreased staining with DAPI, indicative of reduced content of DNA. Our data demonstrate that oncogenic HPV types are frequent in men.

  1. The pathophysiology of acquired premature ejaculation

    OpenAIRE

    McMahon, Chris G; Jannini, Emmanuele A.; Serefoglu, Ege C.; Hellstrom, Wayne J.G.

    2016-01-01

    The second Ad Hoc International Society for Sexual Medicine (ISSM) Committee for the Definition of Premature Ejaculation defined acquired premature ejaculation (PE) as a male sexual dysfunction characterized by a the development of a clinically significant and bothersome reduction in ejaculation latency time in men with previous normal ejaculatory experiences, often to about 3 minutes or less, the inability to delay ejaculation on all or nearly all vaginal penetrations, and the presence of ne...

  2. 人精子在附睾中的成熟%Epididymal Maturation of Human Spermatozoa

    Institute of Scientific and Technical Information of China (English)

    库卓华

    2001-01-01

    精子在人附睾内转运过程中获得使卵子受精的主要功能。这些功能包括:相对活率、一致的形态、结合透明带的能力、发生顶体反应的能力、与卵黄和致密染色质融合的能力、体内和体外使卵子受精的能力、维持正常胚胎发育的能力等。已经鉴定了许多附睾分泌的糖蛋白,它们与特定的精子膜区域结合。这些蛋白可能与促进精-卵作用有关。作为辅助生殖技术的结果,取自附睾近端的“未成熟”精子或睾丸生殖细胞启动胚胎发育的能力依赖于精子在附睾内的转运、在女性生殖道中存活、输送直至被卵子包围。%All of the major functions of sperm that are required for them tofertilise an egg (relevant motility, uniform morphology, ability to bind to the zona pellucida, capacity to undergo the acrosome reaction, capacity to fuse with the vitellus, condensed chromatin, the ability to fertilise eggs in vivo and in vitro and the ability to support normal embryo development) are acquired within the human epididymal canal. Many secreted epididymal glycoproteins have been identified that bind to particular sperm membrane domains and appear to be involved in facilitating sperm-ovum interaction. The ability of “immature” sperm from the proximal epididymis or germ cells from the testis to initiate embryo development as a result of artificial reproductive technologies (ART) depends on the bypassing of the very steps that require epididymal input; that of permitting survival in the female tract and transport to and through the egg investments. Ntl J Androl,2001,7(1):1~9

  3. Heat Sensing Receptor TRPV1 Is a Mediator of Thermotaxis in Human Spermatozoa

    Science.gov (United States)

    Menegazzo, Massimo; Magagna, Sabina; Di Nisio, Andrea; Šabović, Iva; Rocca, Maria Santa; Scattolini, Valentina; Filippi, Andrea; Foresta, Carlo

    2016-01-01

    The molecular bases of sperm thermotaxis, the temperature-oriented cell motility, are currently under investigation. Thermal perception relies on a subclass of the transient receptor potential [TRP] channels, whose member TRPV1 is acknowledged as the heat sensing receptor. Here we investigated the involvement of TRPV1 in human sperm thermotaxis. We obtained semen samples from 16 normozoospermic subjects attending an infertility survey programme, testis biopsies from 6 patients with testicular germ cell cancer and testis fine needle aspirates from 6 patients with obstructive azoospermia undergoing assisted reproductive technologies. Expression of TRPV1 mRNA was assessed by RT-PCR. Protein expression of TRPV1 was determined by western blot, flow cytometry and immunofluorescence. Sperm motility was assessed by Sperm Class Analyser. Acrosome reaction, apoptosis and intracellular-Ca2+ content were assessed by flow cytometry. We found that TRPV1 mRNA and protein were highly expressed in the testis, in both Sertoli cells and germ-line cells. Moreover, compared to no-gradient controls at 31°C or 37°C (Ctrl 31°C and Ctrl 37°C respectively), sperm migration towards a temperature gradient of 31–37°C (T gradient) in non-capacitated conditions selected a higher number of cells (14,9 ± 4,2×106 cells T gradient vs 5,1± 0,3×106 cells Ctrl 31°C and 5,71±0,74×106 cells Ctrl 37°C; P = 0,039). Capacitation amplified the migrating capability towards the T gradient. Sperms migrated towards the T gradient showed enriched levels of both TRPV1 protein and mRNA. In addition, sperm cells were able to migrate toward a gradient of capsaicin, a specific agonist of TRPV1, whilst capsazepine, a specific agonist of TRPV1, blocked this effect. Finally, capsazepine severely blunted migration towards T gradient without abolishing. These results suggest that TRPV1 may represent a facilitating mediator of sperm thermotaxis. PMID:27992447

  4. Acrosome membrane integrity and cryocapacitation are related to cholesterol content of bull spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Srivastava N; Srivastava SK; Ghosh SK; Amit Kumar; Perumal P; Jerome A

    2013-01-01

    Objective: To evaluate the cryoinjury prediction of spermatozoa in relation to its cholesterol content at fresh and frozen-thaw stages. Methods: Ejaculates (n=12) were processed for cryopreservation, acrosome integrity (fluorescent and giemsa stains), cryoinjury (distribution of non capacitated, capacitated and acrosome reacted, pattern F, B and AR, respectively of Chlortetracycline, CTC assay), in vitro fertiltiy (IVF) and cholesterol content of spermatozoa at fresh, pre-freeze and frozen-thaw stages were evaluated. Values were fitted in prediction equation to predict acrosome integrity (AI) and cryoinjury. Results: Study indicated that cholesterol content of fresh spermatozoa can be used to predict cholesterol content of spermatozoa at pre-freeze and frozen-thaw stages of cryopreservation protocol with medium to high level of accuracy (P<0.05 and P<0.01, respectively). Cholesterol content of fresh spermatozoa can be used to predict AI, pattern B and AR and Penetration Index (PI) of IVF with medium level of accuracy (P<0.05) at frozen-thaw but not at pre-freeze stage. Similarly cholesterol content of frozen-thaw spermatozoa can be used to predict AI and pattern AR of frozen-thaw spermatozoa with medium level of accuracy (P<0.05). Conclusion: This study revealed strong evidence that cholesterol content of fresh as well as frozen-thaw bull spermatozoa can be a good predictor of level of cryoinjury following preservation at ultra low temperature.

  5. Longevity of spermatozoa in the post-ejaculatory urine of fertile men.

    Science.gov (United States)

    Engelbertz, Fabian; Korda, Joanna B; Engelmann, Udo; Rothschild, Markus; Banaschak, Sibylle

    2010-01-30

    Many scientists of varying clinical backgrounds have described the phenomenon of spermaturia in animals, adolescents as well as fertile and infertile men. Nevertheless, research for an expert opinion on a law case in the field of forensic medicine revealed a lack of valid information about the longevity of spermatozoa in post-ejaculatory urine (PEU) of fertile men. Our goal was to measure the appearance of vivid sperm in PEU while considering the factor of time in order to predict a realistic interval, in which positive sperm findings might occur. Therefore ten healthy, young men donated their sperm for fertility analysis and a urine sample prior to and after ejaculation. The time intervals between ejaculation and the first micturition were preset ranging between 30 min and maximal 11h. Each ejaculate underwent a semen analysis. The pre- and post-ejaculatory urine samples were screened for the presence of viable and motile spermatozoa. Semen parameters were determined and related to the sperm findings in the precipitate of the urine samples. The amount, the viability and motility status of the detected spermatozoa were recorded after each preset time interval. The results showed that none of the 10 participants had sperm in their urine samples prior to ejaculation. The average sperm concentration was 50.1+/-25.8 million/ml. After a time span of 30 min 59.5% of the first fractions of PEU samples were sperm positive, after 2 and 4h still 70%, and after 5h sperm were no longer detected. The last motile spermatozoa could be found after 4.5h. It seems that remaining sperm in the urethra are washed out with the first micturition in the majority of fertile men, however, the conclusion as to whether sperm findings >5h after ejaculation are improbable needs to be confirmed by further investigations.

  6. Enhanced fertility prediction of cryopreserved boar spermatozoa using novel sperm function assessment.

    Science.gov (United States)

    Daigneault, B W; McNamara, K A; Purdy, P H; Krisher, R L; Knox, R V; Rodriguez-Zas, S L; Miller, D J

    2015-05-01

    Due to reduced fertility, cryopreserved semen is seldom used for commercial porcine artificial insemination (AI). Predicting the fertility of individual frozen ejaculates for selection of higher quality semen prior to AI would increase overall success. Our objective was to test novel and traditional laboratory analyses to identify characteristics of cryopreserved spermatozoa that are related to boar fertility. Traditional post-thaw analyses of motility, viability, and acrosome integrity were performed on each ejaculate. In vitro fertilization, cleavage, and blastocyst development were also determined. Finally, spermatozoa-oviduct binding and competitive zona-binding assays were applied to assess sperm adhesion to these two matrices. Fertility of the same ejaculates subjected to laboratory assays was determined for each boar by multi-sire AI and defined as (i) the mean percentage of the litter sired and (ii) the mean number of piglets sired in each litter. Means of each laboratory evaluation were calculated for each boar and those values were applied to multiple linear regression analyses to determine which sperm traits could collectively estimate fertility in the simplest model. The regression model to predict the percent of litter sired by each boar was highly effective (p boar was also effective (p boar spermatozoa can be predicted effectively by including traditional and novel laboratory assays that consider functions of spermatozoa.

  7. Fertilization potential and qualitative characteristics of human spermatozoa after short-term cryostorage at 5 degrees C in two different TEST-yolk buffer preparations.

    Science.gov (United States)

    Zavos, P M; Correa, J R; Foster, C L; Massey, J B; Zarmakoupis-Zavos, P N

    1998-02-01

    preparations and further prepared via filtration, was assessed by the sperm penetration assay (SPA) using zona-free hamster oocytes. The average penetration rate (PR) and penetration index (PI) were significantly better for the NT-TYB than for the T-TYB. The PR was 54% vs. 25%, and the PI 0.78 and 0.27 for spermatozoa incubated in the NT-TYB vs. T-TYB. The range of penetration was also much lower for the T-TYB (6 to 100%) preparation when compared to the NT-TYB (22 to 100%). The highest penetrator showed 100% for both preparations. However, the lowest penetrator showed 6% for the T-TYB and 22% for the NT-TYB. The data obtained in this study suggest that both TYB preparations can be employed in short-term cryostorage (5 degrees C) of human spermatozoa and can adequately maintain the qualitative characteristics of those spermatozoa. The data also showed that the NT-TYB preparation yielded sperm samples of higher fertilization potential, thus possibly establishing the superior usefulness of the NT-TYB in an ART program.

  8. Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem.

    Science.gov (United States)

    Kaabi, M; Paz, P; Alvarez, M; Anel, E; Boixo, J C; Rouissi, H; Herraez, P; Anel, L

    2003-10-15

    Post-mortem spermatozoa recovery is an important technique for obtaining germplasm reserves from genetically valuable animals or endangered species. However, there are many factors that influence the outcome of this technique. We have studied the effect of the interval between animal's death and sperm recovery (0, 24 or 48 h) on the quality and freezability of ram spermatozoa from cauda epididymidis. Storage temperature of epididymis (room temperature or 5 degrees C) was also analysed. Spermatozoa were diluted with Tes-Tris-Fructose solution supplemented with egg yolk (10%) and glycerol (4%), and frozen using a programmable biofreezer (-20 degrees C/min). Pre-freeze and post-thaw sperm samples showed viable spermatozoa up to 48 h after the animal's death, although their quality declined significantly as post-mortem storage time increased. Epididymis sperm stored at 5 degrees C showed better motility and a lower percentage of abnormal forms than epididymis stored at room temperature after 24 and 48 h. The fertilizing ability of cauda epididymis ram spermatozoa obtained at 0 and 24h after the animal's death is similar to that of ejaculated spermatozoa. Therefore, a good protocol for post-mortem semen collection in rams when epididymal spermatozoa cannot be collected immediately, is to preserve the epididymis at 5 degrees C and process the samples in the first 24h after the animal's death.

  9. Comparative analysis of boar seminal plasma proteome from different freezability ejaculates and identification of Fibronectin 1 as sperm freezability marker.

    Science.gov (United States)

    Vilagran, I; Yeste, M; Sancho, S; Castillo, J; Oliva, R; Bonet, S

    2015-03-01

    Variation in boar sperm freezability (i.e. capacity to withstand cryopreservation) between ejaculates is a limitation largely reported in the literature. Prediction of sperm freezability and classification of boar ejaculates into good (GFEs) and poor freezability ejaculates (PFEs) before cryopreservation takes place may increase the use of frozen-thawed spermatozoa. While markers of boar sperm freezability have been found from sperm cell extracts, little attention has been paid to seminal plasma. On this basis, the present study compared the fresh seminal plasma proteome of 9 GFEs and 9 PFEs through two-dimensional difference gel electrophoresis (2D-DIGE) and liquid chromatography mass spectrometry (LC-MS/MS). The ejaculates were previously classified as GFE or PFE upon their sperm viability and progressive motility assessments at 30 and 240 min post thawing. From a total of 51 spots, four were found to significantly (p sperm quality parameters. Results confirmed that FN1 is a reliable marker of boar sperm freezability, because GFEs presented significantly (p boar sperm freezability marker. We can thus conclude that levels of FN1 in fresh seminal plasma from boar semen may be used as a sperm freezability marker, thereby facilitating the use of frozen-thawed boar spermatozoa.

  10. Human sperm chromatin stabilization: a proposed model including zinc bridges.

    Science.gov (United States)

    Björndahl, Lars; Kvist, Ulrik

    2010-01-01

    The primary focus of this review is to challenge the current concepts on sperm chromatin stability. The observations (i) that zinc depletion at ejaculation allows a rapid and total sperm chromatin decondensation without the addition of exogenous disulfide cleaving agents and (ii) that the human sperm chromatin contains one zinc for every protamine for every turn of the DNA helix suggest an alternative model for sperm chromatin structure may be plausible. An alternative model is therefore proposed, that the human spermatozoon could at ejaculation have a rapidly reversible zinc dependent chromatin stability: Zn(2+) stabilizes the structure and prevents the formation of excess disulfide bridges by a single mechanism, the formation of zinc bridges with protamine thiols of cysteine and potentially imidazole groups of histidine. Extraction of zinc enables two biologically totally different outcomes: immediate decondensation if chromatin fibers are concomitantly induced to repel (e.g. by phosphorylation in the ooplasm); otherwise freed thiols become committed into disulfide bridges creating a superstabilized chromatin. Spermatozoa in the zinc rich prostatic fluid (normally the first expelled ejaculate fraction) represent the physiological situation. Extraction of chromatin zinc can be accomplished by the seminal vesicular fluid. Collection of the ejaculate in one single container causes abnormal contact between spermatozoa and seminal vesicular fluid affecting the sperm chromatin stability. There are men in infertile couples with low content of sperm chromatin zinc due to loss of zinc during ejaculation and liquefaction. Tests for sperm DNA integrity may give false negative results due to decreased access for the assay to the DNA in superstabilized chromatin.

  11. Differential Genes Expression between Fertile and Infertile Spermatozoa Revealed by Transcriptome Analysis.

    Directory of Open Access Journals (Sweden)

    Sandeep Kumar Bansal

    Full Text Available It was believed earlier that spermatozoa have no traces of RNA because of loss of most of the cytoplasm. Recent studies have revealed the presence of about 3000 different kinds of mRNAs in ejaculated spermatozoa. However, the correlation of transcriptome profile with infertility remains obscure.Total RNA from sperm (after exclusion of somatic cells of 60 men consisting of individuals with known fertility (n=20, idiopathic infertility (normozoospermic patients, n=20, and asthenozoospermia (n=20 was isolated. After RNA quality check on Bioanalyzer, AffymetrixGeneChip Human Gene 1.0 ST Array was used for expression profiling, which consisted of >30,000 coding transcripts and >11,000 long intergenic non-coding transcripts.Comparison between all three groups revealed that two thousand and eighty one transcripts were differentially expressed. Analysis of these transcripts showed that some transcripts [ribosomal proteins (RPS25, RPS11, RPS13, RPL30, RPL34, RPL27, RPS5, HINT1, HSP90AB1, SRSF9, EIF4G2, ILF2] were up-regulated in the normozoospermic group, but down-regulated in the asthenozoospermic group in comparison to the control group. Some transcripts were specific to the normozoospermic group (up-regulated: CAPNS1, FAM153C, ARF1, CFL1, RPL19, USP22; down-regulated: ZNF90, SMNDC1, c14orf126, HNRNPK, while some were specific to the asthenozoospermic group (up-regulated: RPL24, HNRNPM, RPL4, PRPF8, HTN3, RPL11, RPL28, RPS16, SLC25A3, C2orf24, RHOA, GDI2, NONO, PARK7; down-regulated: HNRNPC, SMARCAD1, RPS24, RPS24, RPS27A, KIFAP3. A number of differentially expressed transcripts in spermatozoa were related to reproduction (n = 58 and development (n= 210. Some of these transcripts were related to heat shock proteins (DNAJB4, DNAJB14, testis specific genes (TCP11, TESK1, TSPYL1, ADAD1, and Y-chromosome genes (DAZ1, TSPYL1.A complex RNA population in spermatozoa consisted of coding and non-coding RNAs. A number of transcripts that participate in a host of

  12. The effects of ZAG on the acrosome reaction (AR) of human spermatozoa%锌-α2糖蛋白在人精子顶体反应过程中的作用

    Institute of Scientific and Technical Information of China (English)

    郭毅; 刘悦; 王羽; 滕晓明

    2013-01-01

    Objective To explore the effects of ZAG(Zn-alpha2-glycoprotein) on the process of AR. Methods Indirect immunofluorescence and spermatozoa-ZP binding tests were used to analyze the effects of ZAG on human sperm acrosome reaction. Results Immunofluorescence staining revealed intensive immunoreactivity of ZAG in the acrosomal region was largely reduced in acrosome reacted spermatozoa, and human spermatozoa-ZP binding test indicated the spermatozoa’ s ability binding to ZP significantly decreased in spermatozoa treated with anti-ZAG antibody. Conclusion ZAG has significant effects on the acrosome reaction and the spermatozoa-ZP binding.%目的:初步探讨锌-α2糖蛋白(ZAG)在人精子顶体反应中的作用。方法采用ZAG抗体对人精子进行免疫荧光染色,激光共聚焦显微镜观察ZAG在顶体反应前后定位的变化,并通过精子透明带结合实验进一步证明ZAG在顶体反应中的作用。结果顶体反应后精子顶体头部的ZAG荧光明显下降甚至消失。精子经ZAG抗体处理后,其透明带结合能力显著下降。结论 ZAG在人精子顶体反应中发挥一定的作用,并影响人精子的透明带结合能力。本研究为探索ZAG在受精过程中的功能及分子作用机制奠定基础。

  13. Dynamic quantification of intracellular calcium and protein tyrosine phosphorylation in cryopreserved boar spermatozoa during short-time incubation with oviductal fluid.

    Science.gov (United States)

    Kumaresan, A; González, R; Johannisson, A; Berqvist, A-S

    2014-11-01

    Freshly ejaculated boar spermatozoa require several hours of exposure to capacitating conditions to undergo capacitation. We hypothesized that cryopreserved boar spermatozoa might elicit a capacitation response after a relatively shorter time of exposure to capacitating conditions. Washed, frozen-thawed boar spermatozoa were incubated separately with pre-ovulatory isthmic oviductal fluid (EODF), post-ovulatory ODF (MODF), capacitation medium (CM), and noncapacitating medium (NCM) for 60 minutes. Aliquots of spermatozoa were taken at 0, 5, 15, 30, and 60 minutes during incubation and sperm kinematics, intracellular calcium [Ca2(+)]i content, and protein tyrosine phosphorylation (PTP) were studied. The proportion of motile spermatozoa increased significantly after 5 minutes of incubation with EODF. A similar increase was not observed in the other groups. During the initial 5 minutes of incubation, the proportion of spermatozoa with high [Ca(2+)]i decreased significantly in all four groups. The proportion of tyrosine phosphorylated spermatozoa increased from 6.49 ± 1.93% to 15.42 ± 3.58% and 18.41 ± 1.57% in EODF and MODF groups, respectively, at 5 minutes of incubation. Neither CM nor NCM elicited any immediate effect on PTP in spermatozoa. There was a positive and significant correlation between [Ca(2+)]i and sperm motility (P = 0.009). It may be concluded that frozen-thawed boar spermatozoa undergo capacitation-associated changes after a relatively short exposure to EODF, and there are some subpopulations of spermatozoa that undergo PTP despite possessing low [Ca(2+)]i.

  14. [Premature ejaculation: pills or sexology?].

    Science.gov (United States)

    Wisard, M; Audette, N

    2008-03-26

    Premature ejaculation (PE) is a frequent male sexual complaint that affects 20 to 30% of men. The exact aetiology is unknown: psychological/behavioristic and biogenic etiologies have been proposed. The introduction of selective serotonin reuptake inhibitors (SSRI) was revolutionary in the medical treatment of PE. However precautions should be taken because of potential adverse side effects. There is no clear consensus as to whether SSRI may represent an eventual cure of PE or will be required for life. The sexocorporal approach is an other treatment of PE, but convincing scientific treatment data are also lacking.

  15. Current therapies for premature ejaculation.

    Science.gov (United States)

    Gur, Serap; Kadowitz, Philip J; Sikka, Suresh C

    2016-07-01

    Premature ejaculation (PE) subjectively affects 20-30% of men globally. Until recently, understanding of PE was hampered by the absence of a widely accepted definition, paucity of evidence-based clinical studies, and the absence of an appropriate animal model. Here, we elaborate on the current definition of PE, its pathogenesis, currently available therapies, and future treatment prospects. Most treatments for PE are 'off-label' and include selective serotonin reuptake inhibitors (SSRIs), topical anesthetics, tramadol, and phosphodiesterase type 5 (PDE5) inhibitors. Such knowledge of the benefit and limitations of each treatment will help to direct future drug design and formulations.

  16. Rehabilitation for severe delayed ejaculation (intravaginal ejaculation disorder) with use of a masturbation aid

    Institute of Scientific and Technical Information of China (English)

    Yoshitomo Kobori; Hiroaki Aoki; Koujiro Nishio; Ryo Sato; Yoshio Ashizawa; Hiroshi Yagi; Shigehiro So; Gaku Arai; Hiroshi Okada

    2012-01-01

    Objective:To overcome intravaginal ejaculation disorder with the help of a masturbation aid (TENGA®). Methods:A total of 10 patients with intravaginal ejaculation disorder underwent rehabilitation using TENGA. Patients' satisfaction, achievement of intravaginal ejaculation and pregnancy were evaluated through a questionnaire. Results:Seven of the patients (70%) achieved ejaculation in the masturbation aid (TENGA) of which two patients (20%) succeeded in ejaculation within a partner's vagina after rehabilitation. One case achieved spontaneous pregnancy. Conclusions:The use of a masturbation aid (TENGA) has shown to be a useful tool in correcting the methods of masturbation and achieve normal intravaginal ejaculation, and could be an effective option for the treatment of intravaginal ejaculation disorder.

  17. Evaluation of epididymal function through specific protein on spermatozoa.

    Science.gov (United States)

    Del Río, A G; De Sánchez, L Z; Sirena, A

    1984-01-01

    Investigations were focused on the characterization of specific epididymal proteins on the human spermatozoa as a representative parameter for epididymal function. An easy and attainable method, suitable for investigators and clinical use, is proposed in this article.

  18. Determination of fatty acid profile in ram spermatozoa and seminal plasma.

    Science.gov (United States)

    Díaz, R; Torres, M A; Bravo, S; Sanchez, R; Sepúlveda, N

    2016-08-01

    Fatty acids are important in male reproductive function because they are associated with membrane fluidity, acrosome reaction, sperm motility and viability, but limited information exists about the fatty acid profile of ram semen. Our aim was to determine the fatty acid composition in ram spermatozoa and seminal plasma. Sixty ejaculates were obtained from three ram (20 ejaculates/ram) using artificial vagina. Ram spermatozoa (RS) and seminal plasma (SP) were separated using centrifugation, and the fatty acids were analysed by gas chromatography. Total lipids obtained in ram spermatozoa were 1.8% and 1.6% in seminal plasma. Saturated fatty acid (SFA) was proportionally major in SP (66.6%) that RS (49.9%). The highest proportions of SFA corresponded to C4:0 (RS = 16.3% and SP = 28.8%) and C16:0 (RS = 16.3% and PS = 20%). The most important unsaturated fatty acid (UFA) was docosahexaenoic acid (DHA), 44.9% in RS and 31.5% in SP. The profile of fatty acid and their proportions showed differences between spermatozoa and seminal plasma.

  19. Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa

    Directory of Open Access Journals (Sweden)

    N. Prapaiwan

    2016-05-01

    Full Text Available Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05 than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05. In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.

  20. Aquaporins 7 and 11 in boar spermatozoa: detection, localisation and relationship with sperm quality.

    Science.gov (United States)

    Prieto-Martínez, Noelia; Vilagran, Ingrid; Morató, Roser; Rodríguez-Gil, Joan E; Yeste, Marc; Bonet, Sergi

    2016-04-01

    Aquaporins (AQPs) are integral membrane water channels that allow transport of water and small solutes across cell membranes. Although water permeability is known to play a critical role in mammalian cells, including spermatozoa, little is known about their localisation in boar spermatozoa. Two aquaporins, AQP7 and AQP11, in boar spermatozoa were identified by western blotting and localised through immunocytochemistry analyses. Western blot results showed that boar spermatozoa expressed AQP7 (25kDa) and AQP11 (50kDa). Immunocytochemistry analyses demonstrated that AQP7 was localised in the connecting piece of boar spermatozoa, while AQP11 was found in the head and mid-piece and diffuse labelling was also seen along the tail. Despite differences in AQP7 and AQP11 content between boar ejaculates, these differences were not found to be correlated with sperm quality in the case of AQP7. Conversely, AQP11 content showed a significant correlation (Psperm membrane integrity and fluidity and sperm motility. In conclusion, boar spermatozoa express AQP7 and AQP11, and the amounts of AQP11 but not those of AQP7 are correlated with sperm motility and membrane integrity.

  1. Easy sperm processing technique allowing exclusive accumulation and later usage of DNA-strandbreak-free spermatozoa.

    Science.gov (United States)

    Ebner, T; Shebl, O; Moser, M; Mayer, R B; Arzt, W; Tews, G

    2011-01-01

    Sperm DNA fragmentation is increased in poor-quality semen samples and correlates with failed fertilization, impaired preimplantation development and reduced pregnancy outcome. Common sperm preparation techniques may reduce the percentage of strandbreak-positive spermatozoa, but, to date, there is no reliable approach to exclusively accumulate strandbreak-free spermatozoa. To analyse the efficiency of special sperm selection chambers (Zech-selectors made of glass or polyethylene) in terms of strandbreak reduction, 39 subfertile men were recruited and three probes (native, density gradient and Zech-selector) were used to check for strand breaks using the sperm chromatin dispersion test. The mean percentage of affected spermatozoa in the ejaculate was 15.8 ± 7.8% (range 5.0–42.1%). Density gradient did not significantly improve the quality of spermatozoa selected(14.2 ± 7.0%). However, glass chambers completely removed 90% spermatozoa showing strand breaks and polyethylene chambers removed 76%. Both types of Zech-selectors were equivalent in their efficiency, significantly reduced DNA damage (P sperm preparation technique concentrating spermatozoa unaffected in terms of DNA damage. The special chambers most probably select for sperm motility and/or maturity.

  2. Medical therapy for premature ejaculation

    Science.gov (United States)

    Mohee, Amar; Eardley, Ian

    2011-01-01

    Premature ejaculation (PE) is a common male sexual dysfunction. Advances in PE research have been hampered owing to a nonstandardized definition of PE, until the definition by the International Society of Sexual Medicine (ISSM) in 2009. Once the diagnosis of PE is established through a thorough history, a variety of medical therapies is available, including tricyclic antidepressants, selective serotonin reuptake inhibitors (SSRIs), centrally acting opiates, phosphodiesterase 5 inhibitors and topical desensitizing creams. Most of these treatments increase the intravaginal ejaculation latency time (IELT) and patient satisfaction scores, with the most convincing evidence for SSRIs and topical creams. Daily SSRIs such as paroxetine, although efficacious, do have a substantial and prolonged side effect profile. Dapoxetine, which is a on-demand SSRI, is the only licensed drug for the treatment of PE, increasing IELT by a factor of 2.5 to 3 with limited and tolerable side effects. In the near future, the topical aerosol PSD502 is due to be licensed for the treatment of PE, increasing IELT by up to a factor of 6 but having minimal local and negligible systemic side effects. PMID:22046199

  3. Ejaculate traits and sperm cryopreservation in the endangered Baird's tapir (Tapirus bairdii).

    Science.gov (United States)

    Pukazhenthi, Budhan S; Togna, Gina Della; Padilla, Luis; Smith, Diorene; Sanchez, Carlos; Pelican, Katey; Sanjur, Oris I

    2011-01-01

    There is little information on the reproductive biology of the male Baird's tapir (Tapirus bairdii). In this study, we characterized the ejaculate traits and evaluated the efficacy of 2 cryodiluents on sperm cryosurvival. Ejaculates were assessed for volume, pH, sperm motility, forward progression, osmolality, sperm concentration, sperm morphology, and acrosomal integrity. For cryopreservation, ejaculates with >50% total sperm motility were washed, and sperm pellets were resuspended in either Botu-Crio (CryoVital, Grandau, Germany) or INRA 96 containing 2% egg yolk and 2.5% each of methyl- and dimethylformamide (INRA 96), and they were cryopreserved over liquid nitrogen vapor. Thawed samples were incubated in vitro (25 °C) and evaluated for percent total sperm motility, forward progression, and acrosomal integrity at hourly intervals for 4 hours. Spermic ejaculates were obtained from all males, and the mean seminal volume, sperm concentration per milliliter, percent sperm motility, progressive status, and percent morphologically normal cells were 20.4 ± 4.3 mL, 101.2 ± 24.0 × 10(6)/mL, 46.1% ± 5.0%, 2.9 ± 0.1, and 6.9% ± 1.4%, respectively. There was a positive significant correlation between percent normal sperm and animal age (r = 0.66; P < .004). Cryopreservation in either Botu-Crio or INRA 96 resulted in a decline (P < .05) in percent sperm motility and acrosomal integrity. Sperm forward progression remained unaffected immediately after thawing in INRA 96 but continued to decline over time. These results characterize, for the first time, the ejaculate traits of the tapir; demonstrate that tapir spermatozoa can be cryopreserved in diluents containing amides alone or in combination with glycerol; and provide fundamental information critical for development of assisted reproductive technologies for the Baird's tapir.

  4. Short communication. Stallion sperm quality after combined ejaculate fractionation and colloidal centrifugation

    Directory of Open Access Journals (Sweden)

    Francisco Crespo

    2015-12-01

    Full Text Available This study investigated the possible additive benefit of ejaculate fractionation and colloidal centrifugation on stallion sperm quality. Using an open-end artificial vagina, the sperm-rich fraction (FRAC-1 was separated from the rest of the ejaculate (FRAC-2 and a third sperm sample representing the combined ejaculate was reconstituted post-ejaculation (RAW. Each semen sample was processed for colloidal centrifugation. The percentage of abnormal spermatozoa was 17.8 ± 7.0% in RAW and 14.6 ± 9.5% in FRAC-1 but decreased to 11.4 ± 4.7% and 9.6 ± 6.9% respectively, after colloidal centrifugation. A sperm DNA fragmentation index of 10.9 ± 5.1% was observed in RAW and 7.5 ± 2.4% in FRAC-1 semen collected with the AV but this decreased to 7.8 ± 2.8% and 5.2 ± 2.3% after colloidal centrifugation. The rate of increase in sperm DNA fragmentation during the first 6 h of incubation at 37 ºC was 1.8 ± 0.9% per hour in RAW semen and 2.0 ± 2.0% per hour in FRAC-1 but this significantly decreased to 1.3 ± 1.4% and 0.9 ± 0.8% respectively after colloidal centrifugation. While stallion seminal characteristics can be improved using colloidal centrifugation, further enhancement is possible if the ejaculate is initially fractionated.

  5. The associations among semen quality, oxidative DNA damage in human spermatozoa and concentrations of cadmium, lead and selenium in seminal plasma.

    Science.gov (United States)

    Xu, De-Xiang; Shen, Han-Ming; Zhu, Qi-Xing; Chua, Laiha; Wang, Qu-Nan; Chia, Sin-Eng; Ong, Choon-Nam

    2003-01-10

    To explore the associations among semen quality, oxidative DNA damage in human spermatozoa and concentrations of cadmium, lead and selenium in seminal plasma, 56 non-smoking subjects were asked to collect semen by masturbation into a sterile wide-mouth metal-free plastic container after 3 days of abstinence. The conventional semen parameters were analysed. The concentrations of Cd, Pb and Se in seminal plasma were detected using atomic absorption spectrophotometer. 8-OHdG levels in sperm DNA were measured using HPLC-EC. The results showed that the geometric mean concentrations of Cd, Pb and Se were 0.78, 7.8 and 51.4 microg/l, respectively. The geometric mean of 8-OHdG/10(6) dG was 51.4 (95% CI: 21.5-123.0). A significant inverse correlation exists between Cd and sperm density (r=-0.28, Pspermatozoa. Se could protect against oxidative DNA damage in human sperm cells. Pb did not appear to have any association with the semen quality when concentration of Pb in seminal plasma was below 10 microg/l.

  6. Cryotolerance of stallion spermatozoa is related to ROS production and mitochondrial membrane potential rather than to the integrity of sperm nucleus.

    Science.gov (United States)

    Yeste, M; Estrada, E; Rocha, L G; Marín, H; Rodríguez-Gil, J E; Miró, J

    2015-03-01

    Although cryopreservation of stallion spermatozoa allows long-term preservation of spermatozoa from particular stallions and facilitates international trade, it is understood to inflict damages on sperm cells that may finally reduce their fertilizing ability. In addition, individual differences are known to exist in the sperm ability to withstand freeze-thawing protocols. To date, these differences have mainly been reported on the basis of sperm motility and membrane integrity. For this reason, the present work sought to determine differences between good (good freezability ejaculates: GFE) and poor (poor freezability ejaculates: PFE) freezability stallion ejaculates in other sperm parameters, including peroxide and superoxide levels, potential of mitochondrial membrane and nuclear integrity. With this purpose, a total of 24 stallion ejaculates were cryopreserved and classified into two groups (GFE vs. PFE), depending on their sperm membrane integrity and motility after freeze-thawing. From the total of 24 ejaculates, 13 were classified as GFE and the other 11 were classified as PFE. Apart from differences in sperm membrane permeability and lipid disorder after freeze-thawing, GFE presented significantly (p sperm head proteins, no significant differences between GFE and PFE were seen. We can thus conclude that good and poor freezability stallion ejaculates differ in their reactive oxygen species levels after cryopreservation, but not in the damage extent on sperm nucleus.

  7. Studies on liquefaction and storage of ejaculated dromedary camel (Camelus dromedarius) semen.

    Science.gov (United States)

    Wani, N A; Billah, M; Skidmore, J A

    2008-12-01

    The purpose of this study was to evaluate seminal liquefaction and quality of ejaculated camel semen during storage in different extenders at room (23 degrees C) and refrigeration (4 degrees C) temperature. Semen was collected using an artificial vagina and diluted immediately (1:1), using a split-sample technique, in five extenders [(1) Tris-tes egg yolk, (2) Tris-lactose egg yolk, (3) citrate egg yolk, (4) sucrose egg yolk and (5) Tris-fructose egg yolk], while one fraction was kept without an extender to act as control. The semen was transported to the lab at 37 degrees C, in a portable incubator within half an hour, and thereafter liquefaction of semen was monitored every 15 min. After complete liquefaction of the semen it was evaluated for sperm concentration and morphology and then was extended to a final ratio of 1:3. Aliquots of each semen sample were then stored at refrigeration and room temperature. The average volume of an ejaculate was 4.3+/-0.4 mL and it had a very viscous consistency. The average concentration of spermatozoa was 230.4+/-10.7 x 10(6)mL(-1) and the proportion of spermatozoa with protoplasmic droplets averaged 1.02+/-0.2, while 2.7+/-0.6 and 9.7+/-2.9% had mid-piece and tail abnormalities, respectively. All extended semen samples liquefied within 1.5h at 37 degrees C, however, there was slow liquefaction in the sample without an added extender (control). Best liquefaction was observed in Tris-lactose extender followed by Tris-fructose and citrate egg yolk diluents whereas in the other two extenders there was head-to-head agglutination of the spermatozoa. There was no difference in the initial motility of the spermatozoa in extenders 1-5 after its liquefaction, however, after 24 and 48 h of storage a higher proportion of spermatozoa were motile in extenders 1, 2 and 4 (Pdromedary semen, when added to an extender (1:1) immediately after collection, liquefies within 60-90 min at 37 degrees C. It maintains a high proportion of motile and

  8. Orchid sexual deceit provokes ejaculation.

    Science.gov (United States)

    Gaskett, A C; Winnick, C G; Herberstein, M E

    2008-06-01

    Sexually deceptive orchids lure pollinators by mimicking female insects. Male insects fooled into gripping or copulating with orchids unwittingly transfer the pollinia. The effect of deception on pollinators has been considered negligible, but we show that pollinators may suffer considerable costs. Insects pollinating Australian tongue orchids (Cryptostylis species) frequently ejaculate and waste copious sperm. The costs of sperm wastage could select for pollinator avoidance of orchids, thereby driving and maintaining sexual deception via antagonistic coevolution or an arms race between pollinator learning and escalating orchid mimicry. However, we also show that orchid species provoking such extreme pollinator behavior have the highest pollination success. How can deception persist, given the costs to pollinators? Sexually-deceptive-orchid pollinators are almost exclusively solitary and haplodiploid species. Therefore, female insects deprived of matings by orchid deception could still produce male offspring, which may even enhance orchid pollination.

  9. Localization of porcine seminal plasma (PSP) proteins in the boar reproductive tract and spermatozoa.

    Science.gov (United States)

    Manásková, P; Jonáková, V

    2008-06-01

    Spermadhesins are proteins containing a characteristic CUB domain, originally isolated from seminal plasma and ejaculated spermatozoa in domestic animals. Boar spermadhesins are multifunctional proteins exhibiting ligand-binding abilities with various endogenous ligands present in the male and female reproductive tracts and may play a role in the reproduction process. Porcine spermadhesins (AQN, AWN, PSP protein families) are secreted mainly by the seminal vesicles, but their mRNAs have been found also in the cauda epididymis and prostate. Unlike AQN and AWN spermadhesins, localization of porcine seminal plasma (PSP) proteins in the boar reproductive tract has not been completely resolved. This work has focused on PSP protein expression and localization in the boar reproductive organs and on spermatozoa. Using specific rabbit polyclonal antibodies (anti-PSP I and anti-PSP II), PSP I and PSP II proteins were immunodetected in tissue extracts and in secretory tissues of cauda epididymis, prostate, seminal vesicles and Cowper's glands on the blots and by an indirect immunofluorescence technique, respectively. Moreover, the ability of PSP proteins to bind to epididymal spermatozoa indicated their presence on cauda epididymal and ejaculated spermatozoa. Porcine seminal plasma proteins bind to the sperm surface at ejaculation and may modulate several aspects of sperm activity during reproduction. PSP proteins are produced not only by seminal vesicles and prostate, but also by epididymis. However, their prospective role in sperm epididymal maturation is not clear. Further characterization of seminal plasma protein forms expressed in the individual reproductive organs will help to understand their subsequent role in the reproduction process.

  10. DNA fragmentation in spermatozoa

    DEFF Research Database (Denmark)

    Rex, A S; Aagaard, J.; Fedder, J

    2017-01-01

    Sperm DNA Fragmentation has been extensively studied for more than a decade. In the 1940s the uniqueness of the spermatozoa protein complex which stabilizes the DNA was discovered. In the fifties and sixties, the association between unstable chromatin structure and subfertility was investigated....... In the seventies, the impact of induced DNA damage was investigated. In the 1980s the concept of sperm DNA fragmentation as related to infertility was introduced as well as the first DNA fragmentation test: the Sperm Chromatin Structure Assay (SCSA). The terminal deoxynucleotidyl transferase nick end labelling...... (TUNEL) test followed by others was introduced in the nineties. The association between DNA fragmentation in spermatozoa and pregnancy loss has been extensively investigated spurring the need for a therapeutic tool for these patients. This gave rise to an increased interest in the aetiology of DNA damage...

  11. Contemporary Management of Disorders of Male Orgasm and Ejaculation.

    Science.gov (United States)

    Althof, Stanley E; McMahon, Chris G

    2016-07-01

    Ejaculatory disorders lie along a conceptual continuum with premature ejaculation anchoring one end, normal ejaculation in the center, and difficulties with delayed or anejaculation at the opposite end. Retrograde ejaculation, painful ejaculation, and postorgasmic illness syndrome can occur at any point on the continuum. This manuscript defines the ejaculatory dysfunctions, reviews the anatomy and physiology of orgasm and ejaculation, and summarizes the pharmacological, psychological, and combined treatment approaches to ejaculatory dysfunctions.

  12. Ejaculatory training lengthens the ejaculation latency and facilitates the functioning of the spinal generator for ejaculation of rats with rapid ejaculation.

    Science.gov (United States)

    Rodríguez-Peña, M de L; Rodríguez-Manzo, G; Carro-Juárez, M

    2017-01-01

    A spinal pattern generator controls the ejaculatory response. Central pattern generators (CPGs) may be entrained to improve the motor patterns under their control. In the present study we tested the hypothesis that training of the spinal generator for ejaculation (SGE) by daily copulation until ejaculation, could promote substantive changes in its functioning permitting a better SGE control of the genital motor pattern of ejaculation (GMPE) and, as a consequence, a normalization of the ejaculation latency of rats with rapid ejaculation. To that aim, we evaluated in sexually experienced male rats with rapid ejaculation (1) the effects of daily copulation to ejaculation, following different entrainment schedules, on their ejaculation latencies, (2) the impact of these different ejaculatory entrainment schedules upon the parameters of the GMPE and (3) the possible emergence of persistent changes in the functioning of the SGE associated to the daily ejaculation entrainment schedules. The data obtained show that intense ejaculatory training of rats with rapid ejaculation lengthens the ejaculation latency during copulation and augments the ejaculatory capacity of the SGE in this population when spinalized. Thus, present data reveal that like other CPGs, the SGE can be trained and put forward that training of the SGE by daily copulation to ejaculation might be a promising alternative that should be taken into consideration for the treatment of premature ejaculation.

  13. Urethral anatomy and semen flow during ejaculation

    Science.gov (United States)

    Kelly, Diane

    2016-11-01

    Ejaculation is critical for reproductive success in many animals, but little is known about its hydrodynamics. In mammals, ejaculation pushes semen along the length of the penis through the urethra. Although the urethra also carries urine during micturition, the flow dynamics of micturition and ejaculation differ: semen is more viscous than urine, and the pressure that drives its flow is derived primarily from the rhythmic contractions of muscles at the base of the penis, which produce pulsatile rather than steady flow. In contrast, Johnston et al. (2014) describe a steady flow of semen through the crocodilian urethral groove during ejaculation. Anatomical differences of tissues associated with mammalian and crocodilian urethral structures may underlie these differences in flow behavior.

  14. Effect of glutamine supplementation and replacement of tris-egg yolk based extender with defatted cow milk on spermatozoa quality after equilibration and thawing

    Directory of Open Access Journals (Sweden)

    Vasundhara Dawra

    2015-08-01

    Full Text Available Aim: The present study was designed to evaluate the effect of supplementation of glutamine and replacement of Tris-egg yolk (TE based buffer with defatted cow milk on the spermatozoa quality after equilibration and thawing. Materials and Methods: Semen was collected from five Bhadawari bulls biweekly, and a total of 30 ejaculates were taken. The semen ejaculates were pooled and divided into three equal parts. The pooled semen was diluted by TE based extender (control, TE + glutamine (8 mM (T1 and 50% TE + 50% deffated cow milk + glutamine (8 mM (T2. At two stages viz. after equilibration and after 12 h of cryopreservation (thawed samples, progressive motility, percent live, and percent acrosomal damage of the spermatozoa was assessed. Results: Supplementation of glutamine improved (p<0.05 the spermatozoa quality with respect to the progressive motility, percent live and acrosomal damage both post-equilibration and post-thaw. T2 improved (p<0.05 the spermatozoa quality as compared to control, however; it was less (p<0.05 effective as compared T1 both post-equilibration and post-thaw. Conclusion: From the results of present study it can be concluded that glutamine supplementation was effective in maintaining post-equilibration and post-thaw spermatozoa quality whereas defatted cow milk was not as effective as TE based buffer in the extender in improving the spermatozoa quality.

  15. Different patterns of metabolic cryo-damage in domestic cat (Felis catus) and cheetah (Acinonyx jubatus) spermatozoa.

    Science.gov (United States)

    Terrell, Kimberly A; Wildt, David E; Anthony, Nicola M; Bavister, Barry D; Leibo, S P; Penfold, Linda M; Marker, Laurie L; Crosier, Adrienne E

    2012-04-01

    Felid spermatozoa are sensitive to cryopreservation-induced damage, but functional losses can be mitigated by post-thaw swim-up or density gradient processing methods that selectively recover motile or structurally-normal spermatozoa, respectively. Despite the importance of sperm energy production to achieving fertilization, there is little knowledge about the influence of cryopreservation or post-thaw processing on felid sperm metabolism. We conducted a comparative study of domestic cat and cheetah sperm metabolism after cryopreservation and post-thaw processing. We hypothesized that freezing/thawing impairs sperm metabolism and that swim-up, but not density gradient centrifugation, recovers metabolically-normal spermatozoa. Ejaculates were cryopreserved, thawed, and processed by swim-up, Accudenz gradient centrifugation, or conventional washing (representing the 'control'). Sperm glucose and pyruvate uptake, lactate production, motility, and acrosomal integrity were assessed. Mitochondrial membrane potential (MMP) was measured in cat spermatozoa. In both species, lactate production, motility, and acrosomal integrity were reduced in post-thaw, washed samples compared to freshly-collected ejaculates. Glucose uptake was minimal pre- and post-cryopreservation, whereas pyruvate uptake was similar between treatments due to high coefficients of variation. In the cat, swim-up, but not Accudenz processing, recovered spermatozoa with increased lactate production, pyruvate uptake, and motility compared to controls. Although confounded by differences in non-specific fluorescence among processing methods, MMP values within treatments were positively correlated to sperm motility and acrosomal integrity. Cheetah spermatozoa isolated by either selection method exhibited improved motility and/or acrosomal integrity, but remained metabolically compromised. Collectively, findings revealed a metabolically-robust subpopulation of cryopreserved cat, but not cheetah, spermatozoa

  16. Psychosocial interventions for premature ejaculation

    Directory of Open Access Journals (Sweden)

    Tamara Melnik

    Full Text Available BACKGROUND: Premature ejaculation (PE is a very common sexual dysfunction among patients, and with varying prevalence estimates ranging from 3% to 20%. Although psychological issues are present in most patients with premature PE, as a cause or as a consequence, research on the effects of psychological approaches for PE has in general not been controlled or randomised and is lacking in long-term follow up. OBJECTIVE: To assess the efficacy of psychosocial interventions for PE. CRITERIA FOR CONSIDERING STUDIES FOR THIS REVIEW: Trials were searched in computerized general and specialized databases, such as: MEDLINE by PubMed (1966 to 2010; PsycINFO (1974 to 2010; EMBASE (1980 to 2010; LILACS (1982 to 2010; the Cochrane Central Register of Controlled Trials (Cochrane Library, 2010; and by checking bibliographies, and contacting manufacturers and researchers. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials evaluating psychosocial interventions compared with different psychosocial interventions, pharmacological interventions, waiting list, or no treatment for PE. DATA COLLECTION AND ANALYSIS: Information on patients, interventions, and outcomes was extracted by at least two independent reviewers using a standard form. The primary outcome measure for comparing the effects of psychosocial interventions to waiting list and standard medications was improvement in IELT (i.e., time from vaginal penetration to ejaculation. The secondary outcome was change in validated PE questionnaires. MAIN RESULTS: In one study behavioral therapy (BT was significantly better than waiting list for duration of intercourse (MD (mean difference 407.90 seconds, 95% CI 302.42 to 513.38, and couples' sexual satisfaction (MD -26.10, CI -50.48 to -1.72. BT was also significantly better for a new functional-sexological treatment (FS (MD 412.00 seconds, 95% CI 305.88 to 518.12, change over time in subjective perception of duration of intercourse (Women: MD 2

  17. Hydroxyflutamide alters the characteristics of live boar spermatozoa.

    Science.gov (United States)

    Zarzycka, Marta; Kotwicka, Malgorzata; Jendraszak, Magdalena; Skibinska, Izabela; Kotula-Balak, Malgorzata; Bilinska, Barbara

    2014-10-15

    Our previous study revealed that in vitro incubation of boar ejaculates with hydroxyflutamide (OH-Flu) causes changes in sperm plasma membrane integrity and its stability and sperm mitochondrial oxidative capability. To broaden the knowledge of cellular physiology of spermatozoa, we investigated direct effects of OH-Flu administered for 2 and 24 hours at concentrations of 5, 50, and 100 μg/mL, on sperm mitochondrial membrane potential and mitochondrial superoxide anion production using JC-1 dye and MitoSOX Red fluorescent probe, respectively. We further measured phosphatidylserine membrane translocation (PST) from the inner to the outer layer of the sperm plasma membrane using an annexin-V binding assay. To provide new information of direct effects of OH-Flu on cell signaling pathway, we measured sperm intracellular calcium ion dynamics using Fluo-3. Finally, we assessed sperm motility using a computer-assisted spermatozoa analysis system. Motile sperm were highlighted using the "C-Ruch" computer program for detailed analysis of the straight line velocity distribution. For each functional test, boar spermatozoa were examined and analyzed by flow cytometry and/or confocal microscopy. The results revealed a significant decrease (Psperm mitochondrial membrane potential and a concomitant increase (Psperm motility. Hydroxyflutamide significantly decreased (Psperm subpopulation percentage after 15 minutes and reduced the straight line velocity distribution (Psperm intracellular calcium ion concentration. Altogether, the altered in vitro characteristics of live boar spermatozoa provide new insight into direct effects of OH-Flu on sperm mitochondrial membrane potential, superoxide anion production, translocation of membrane phosphatidylserine, free calcium ion dynamics, and sperm motility.

  18. Pregnancy rates after artificial insemination with cooled stallion spermatozoa either with or without single layer centrifugation.

    Science.gov (United States)

    Morrell, J M; Richter, J; Martinsson, G; Stuhtmann, G; Hoogewijs, M; Roels, K; Dalin, A-M

    2014-11-01

    A successful outcome after artificial insemination with cooled semen is dependent on many factors, the sperm quality of the ejaculate being one. Previous studies have shown that spermatozoa with good motility, normal morphology, and good chromatin integrity can be selected by means of colloid centrifugation, particularly single layer centrifugation (SLC) using species-specific colloids. The purpose of the present study was to conduct an insemination trial with spermatozoa from "normal" ejaculates, i.e., from stallions with no known fertility problem, to determine whether the improvements in sperm quality seen in SLC-selected sperm samples compared with uncentrifuged controls in laboratory tests are reflected in an increased pregnancy rate after artificial insemination. In a multicentre study, SLC-selected sperm samples and uncentrifuged controls from eight stallions were inseminated into approximately 10 mares per treatment per stallion. Ultrasound examination was carried out approximately 16 days after insemination to detect an embryonic vesicle. The pregnancy rates per cycle were 45% for controls and 69% for SLC-selected sperm samples, which is statistically significant (P < 0.0018). Thus, the improvement in sperm quality reported previously for SLC-selected sperm samples is associated with an increase in pregnancy rate, even for ejaculates from stallions with no known fertility problem.

  19. SPERM HY-LITER™ for the identification of spermatozoa from sexual assault evidence

    DEFF Research Database (Denmark)

    Westring, Christian Gustav; Wiuf, Morten; Nielsen, S Jock

    2014-01-01

    Accurate microscopic identification of human spermatozoa is important in sexual assault cases. We have compared the results of examinations with (1) a fluorescent microscopy method, SPERM HY-LITER™, and (2) Baecchi's method for identification of human spermatozoa. In 35 artificial, forensic type...

  20. Ejaculate collection efficiency and post-thaw semen quality in wild-caught Griffon vultures from the Sardinian population

    Directory of Open Access Journals (Sweden)

    Zinellu Angelo

    2009-02-01

    Full Text Available Abstract This study aimed to test the feasibility of a programme of semen collection and cryopreservation in Griffon vultures. Four wild-caught individuals kept in captivity because of unrecoverable traumas were used. Semen collection attempts were made twice a week during three consecutive reproductive seasons (December – March using the abdominal massage method. Ejaculation was successfully induced between late January and late February. Semen collection efficiency was rather low (27.9% and it did not vary among individuals (p > 0.05. No differences were found in ejaculate volumes (12.5 +/- 9.1 μl, spermatozoa concentration (28.4 +/- 30.9 million cells/ml and viability (61.3 +/- 13.9% among the 4 vultures. ATP values differed among the four vultures (p

  1. Psychosexual therapy for premature ejaculation.

    Science.gov (United States)

    Althof, Stanley E

    2016-08-01

    Premature ejaculation (PE) is a male sexual dysfunction that creates considerable anguish for the man, his partner and their relationship. PE is not one disorder but includes the four subtypes (lifelong, acquired, natural and subjective) each with unique psychological concerns and issues. Psychological treatment for men and couples with PE addresses sexual skills/techniques but also focuses on issues of self-esteem, performance anxiety and interpersonal conflict. The outcome studies for psychotherapy alone are difficult to interpret and compare because of poor methodological design (lack of control groups, small sample size, poor outcome measures and lack of follow-up). However, the few studies that surmount these methodological hurdles suggest that psychological intervention offers men and couples a promising treatment option. Combination pharmaco- and psychotherapy is the most promising intervention for lifelong and acquired PE and offers superior efficacy to drug alone. This is because men and couples learn sexual skills, address the intrapsychic, interpersonal and cognitive issues that precipitate and maintain the dysfunction.

  2. The epidemiology of premature ejaculation.

    Science.gov (United States)

    Saitz, Theodore Robert; Serefoglu, Ege Can

    2016-08-01

    Vast advances have occurred over the past decade with regards to understanding the epidemiology, pathophysiology and management of premature ejaculation (PE); however, we still have much to learn about this common sexual problem. As a standardized evidence-based definition of PE has only recently been established, the reported prevalence rates of PE prior to this definition have been difficult to interpret. As a result, a large range of conflicting prevalence rates have been reported. In addition to the lack of a standardized definition and operational criteria, the method of recruitment for study participation and method of data collection have obviously contributed to the broad range of reported prevalence rates. The new criteria and classification of PE will allow for continued research into the diverse phenomenology, etiology and pathogenesis of the disease to be conducted. While the absolute pathophysiology and true prevalence of PE remains unclear, developing a better understanding of the true prevalence of the disease will allow for the completion of more accurate analysis and treatment of the disease.

  3. [Sexological intervention on premature ejaculation].

    Science.gov (United States)

    San Martín Blanco, C

    2014-07-01

    Strategies, recommendations and techniques proposed by sex therapy for intervention on premature ejaculation, have represented for nearly four decades the most effective model of intervention in this sexual dysfunction, which currently is complemented by the efficacy of dapoxetine drug treatment. Clinical experience and recent studies support that combined intervention offers the best therapeutic results. In addition in sex therapy, etiologic diagnosis is obtained from the analysis of the interrelationship of the couple. Diagnostic and therapeutic intervention has to be always centered in the relationship, so the techniques and resources must be applied with the expectation of being implemented in the sexual interaction. It will therefore be the relationship that receive treatment, even if medication is used for one of the members of the couple. On the other hand, this model of intervention can be implemented by a professional with training, although not necessarily a specialist. Copyright © 2014 Elsevier España, S.L.U. y Sociedad Española de Medicina Rural y Generalista (SEMERGEN). All rights reserved.

  4. The Protective Effects of N-Acetylcysteine on Exogenous Hydrogen Peroxide and Endogenous Superoxide Anion induced DNA Strand Breakage in Human Spermatozoa%`

    Institute of Scientific and Technical Information of China (English)

    徐德祥; 沈汉民; 王俊南

    2001-01-01

    Objective To explore the protective effects of N-Acetylcysteine (NAC) on exogenous hydrogen peroxide and endogenous superoxide anion-induced DNA strand breakage in human spermatozoa by using the single-cell gel electropherosis (SCGE)Methods Sperm cells were exposed to 0. 5 mmol/L of H2O2 or 5. 0 mmol/L of β -NADPH with or without 0. 1, 0. 5, 1. 0 mmol/L of NAC. The percentage of sperm comet cells and the comet tail lengths were measured in the treated sperm cells by using SCGE.Results Both percentage of comet sperm nuclei and mean tail length in sperm cells exposed to 0. 5 mmol/L hydrogen peroxide with different concentrations of NAC decrease significantly in a dose-dependent manner as compared with sperm cells exposed to H2O2 without NAC or catalase. Although mean tail length in sperm cells exposed to 5. 0 mmol/L of β-NADPH with different concentrations of NAC decreases significantly compared with sperm cells exposed to β-NADPH without NAC or SOD,there were no significant differences on the percentage of sperm comet cells between sperm cells exposed to 5. 0 mmol/L of β-NADPH with different concentrations of NAC and sperm cells exposed to 5. 0 mmol/L of β-NADPH without NAC.Conclusion NAC has a protective effect on exogenous hydrogen peroxide-induced DNA damage, while protective effect of NAC against O2- induced DNA strand break age is significant but very weak.

  5. Ejaculate fractioning effect on llama sperm head morphometry as assessed by the ISAS(®) CASA system.

    Science.gov (United States)

    Soler, C; Sancho, M; García, A; Fuentes, Mc; Núñez, J; Cucho, H

    2014-02-01

    South American camelid sperm characteristics are poorly known compared with those of other domestic animals. The long-term duration of ejaculation makes difficult to gather all the seminal fluid, implying possible ejaculation portion losses. Thus, the aim of this research was to evaluate the characteristics of the morphology and morphometry of the spermatozoa change during ejaculation. The morphometric characterization was tested on nine specimens of the Lanuda breed, using a special artificial vagina. In five of the animals, a fractioning of the ejaculate was performed by taking samples every 5 min. for a total of 20 min. Air-dried seminal smears were stained with Hemacolor and mounted permanently with Eukitt. Morphometric analysis was carried out with the morphometry module of the ISAS(®) CASA system. Almost 350 cells were analysed per sample, with a total number of 3207 spermatozoa. Mean values were given as follows: length: 5.51 μm; width: 3.38 μm; area: 17.75 μm(2) ; perimeter: 14.8 μm; ellipticity: 0.24; elongation: 0.56; rugosity: 0.87; regularity: 1.07; and shape factor: 1.41. Different animals showed differences in their morphometric values. When we compared the values from different fractions, only two samples showed differences in morphometric parameter values and four samples showed differences in shape parameters. Multivariate analysis allowed the size classification of the cells into three classes and five classes of shapes. The distribution of classes among fractions showed no differences. Despite the individual morphometric differences observed in some fractions, the characteristics of the sperm head morphometry can be considered constant along the ejaculatory period in the llama.

  6. Epididymosomes transfer epididymal sperm binding protein 1 (ELSPBP1) to dead spermatozoa during epididymal transit in bovine.

    Science.gov (United States)

    D'Amours, Olivier; Frenette, Gilles; Bordeleau, Louis-Jean; Allard, Nancy; Leclerc, Pierre; Blondin, Patrick; Sullivan, Robert

    2012-10-01

    Previously, we showed that epididymal sperm binding protein 1 (ELSPBP1) characterizes spermatozoa already dead before ejaculation in bovine. In this study, we investigated the presence of ELSPBP1 in bull genital tract as well as its acquisition by spermatozoa during epididymal transit. As assessed by real-time RT-PCR, ELSPBP1 was highly expressed in the caput and the corpus epididymis but was present in lower expression levels in the testis and the cauda epididymis. Immunohistochemistry revealed the same expression pattern. However, Western blot on tissue homogenates showed some discrepancies, as ELSPBP1 was found in a comparable concentration all along the epididymis. This difference was due to the presence of ELSPBP1 in the epididymal fluid. In both caput and cauda epididymal fluid, ELSPBP1 was associated with the epididymosomes, small membranous vesicles secreted by epithelial cells of the epididymis and implicated in the transfer of proteins to spermatozoa. As assessed by immunocytometry, ELSPBP1 was found on a subset of dead spermatozoa in caput epididymis but was found on all dead spermatozoa in cauda epididymis. To assess ELSPBP1 acquisition by spermatozoa, caput epididymal spermatozoa were incubated with cauda epididymosomes under various conditions. ELSPBP1 detection by immunocytometry assay revealed that only spermatozoa already dead before incubation were receptive to ELSPBP1 transfer by epididymosomes. This receptivity was enhanced by the presence of zinc in the incubation medium. This specificity for a sperm subpopulation suggests that an underlying mechanism is involved and that ELSPBP1 could be a tag for the recognition of dead spermatozoa during epididymal transit.

  7. Hydrodynamics of insect spermatozoa

    Science.gov (United States)

    Pak, On Shun; Lauga, Eric

    2010-11-01

    Microorganism motility plays important roles in many biological processes including reproduction. Many microorganisms propel themselves by propagating traveling waves along their flagella. Depending on the species, propagation of planar waves (e.g. Ceratium) and helical waves (e.g. Trichomonas) were observed in eukaryotic flagellar motion, and hydrodynamic models for both were proposed in the past. However, the motility of insect spermatozoa remains largely unexplored. An interesting morphological feature of such cells, first observed in Tenebrio molitor and Bacillus rossius, is the double helical deformation pattern along the flagella, which is characterized by the presence of two superimposed helical flagellar waves (one with a large amplitude and low frequency, and the other with a small amplitude and high frequency). Here we present the first hydrodynamic investigation of the locomotion of insect spermatozoa. The swimming kinematics, trajectories and hydrodynamic efficiency of the swimmer are computed based on the prescribed double helical deformation pattern. We then compare our theoretical predictions with experimental measurements, and explore the dependence of the swimming performance on the geometric and dynamical parameters.

  8. Premature Ejaculation and Utilization of Cognitive Techniques

    Directory of Open Access Journals (Sweden)

    Serkan AKKOYUNLU

    2013-03-01

    Full Text Available Introduction: Premature ejaculation is the most common male sexual dysfunction leading to distress in many couples. Master and Johnson emphasized the concept of early learned experiences and Kaplan emphasized lack of sensory awareness. For treatment sex therapists mainly utilize start-stop and squeeze techniques as homework. Couples enter sex therapy with some cognitive distortions and beliefs about sex and sexuality. These beliefs are also named sexual myths. For some couples using techniques to challenge cognitive distortions and maladaptive beliefs about sex and sexuality can be used. In this paper by presenting a case we discussed how cognitive techniques can be used along with behaviour techniques with couples. Case: Presenting clients are five years married couple who are thirty and twenty nine years old respectively. They attended to the outpatient clinic with the request of the female client. Their main complaint was premature ejaculation. They were diagnosed premature ejaculation using clinical interview. In treatment besides start and stop technique, cognitive techniques were utilized to address dysfunctional beliefs about sexuality. Discussion: Premature ejaculation is a male sexual dysfunction that causes distress and intimacy problems between couples. Stop start and squeeze techniques were accepted as the choice of treatment but their effectiveness is questioned recently. Also cognitive distortions and maladaptive beliefs may hamper therapy progress. Besides that, behavioral techniques utilizing cognitive techniques to lessen the degree of dysfunctional beliefs about sex and sexuality may help the couple to overcome premature ejaculation and enhance sexual satisfaction and intimacy.

  9. Total globozoospermia associated with increased frequency of immature spermatozoa with chromatin defects and aneuploidy: a case report.

    Science.gov (United States)

    Vozdova, M; Rybar, R; Kloudova, S; Prinosilova, P; Texl, P; Rubes, J

    2014-10-01

    Globozoospermia, characterised by the presence of round spermatozoa lacking acrosomes in an ejaculate, is a known cause of male infertility. Semen analysis, including sperm chromatin structure assay, toluidine blue, chromomycin A3 and aniline blue staining and fluorescence in situ hybridisation, was performed in an infertile globozoospermic patient to establish to which extent these genetic factors contributed to his infertility. No spermatozoa capable of hyaluronan (HA) binding were detected in the HA binding assay. Increased rates of immature spermatozoa with defective replacement of histones by protamines, DNA breaks and disturbed chromatin integrity and sperm aneuploid for the sex chromosomes were observed. Intracytoplasmic sperm injection (ICSI) was used in three in vitro fertilisation (IVF) cycles, and enough morphologically well-developing embryos were obtained in each cycle. However, no pregnancy was achieved. The infertility of our couple, resistant to IVF/ICSI treatment, was most probably caused by a combination of male and female factors.

  10. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis)

    OpenAIRE

    Kamran Dorostkar; Sayed Mortaza Alavi-Shoushtari; Aram Mokarizadeh

    2012-01-01

    The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis) bulls. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 0.5, 1, 2, 4 and 8 μg mL-1 sodium selenite and the sperm motility and viability were evaluated at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) min after dil...

  11. A Healthy Live Birth Following ICSI with Retrograde Ejaculated Sperm

    African Journals Online (AJOL)

    AJRH Managing Editor

    This case report describes the identification and successful treatment of a couple ... Keywords: Retrograde ejaculation, ICSI, infertility, Africa ... Diagnostic clues to retrograde ejaculation ... normal hormone profile and patent fallopian tubes.

  12. The mystery is solved-CatSper is the principal calcium channel activated by progesterone in human spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Christopher LR Barratt

    2011-01-01

    @@ Aremarkable advance in sperm physiology has recently been published in Nature.Two groups using patch clamping techniques on human sperm have solved a mystery about the sperm cell that has puzzled both andrologists and those involved in non-genomic cellular signalling for over 20 years.In these papers, Lishko1 and Strunker2 independently demonstrate that the universal characteristic effect of progesterone on sperm-a rapid influx of calcium-is via a sperm-specific channel CatSper.

  13. Evaluation of the Efficiency of Two Different Freezing Media and Two Different Protocols to Preserve Human Spermatozoa from Cryoinjury

    Science.gov (United States)

    Starita, Maria Flavia; Rega, Emilia; Piscitelli, Claudio; Giannini, Pierluigi

    2016-01-01

    It is universally recognized that cryopreservation impairs sperm quality. In order to improve postthawing sperm survival and motility, media of different composition and different protocols have been proposed. However, no clear evidence is available to understand which are the most efficient protocol and medium for sperm cryopreservation. The present study evaluates the efficiency of two different cryopreservation protocols and two common freezing media (FM) containing different cryoprotectants (CPs), TEST Yolk Buffer (TYB) and Sperm Freeze (SF), to preserve human sperm quality. Our data suggest that TYB is better than SF both in terms of postthaw viability and in terms of progressive motility, while the direct addition of FM to the sperm sample resulted in the most efficient protocol in terms of postthaw viability but not in terms of progressive motility. PMID:27529078

  14. Comparison of reproductive outcome in oligozoospermic men with high sperm DNA fragmentation undergoing intracytoplasmic sperm injection with ejaculated and testicular sperm.

    Science.gov (United States)

    Esteves, Sandro C; Sánchez-Martín, Fernando; Sánchez-Martín, Pascual; Schneider, Danielle T; Gosálvez, Jaime

    2015-12-01

    To investigate the effectiveness of intracytoplasmic sperm injection (ICSI) using testicular sperm as a strategy to overcome infertility in men with high sperm DNA fragmentation (SDF). Prospective, observational, cohort study. Private IVF centers. A total of 147 couples undergoing IVF-ICSI and day 3 fresh ETs whose male partner has oligozoospermia and high SDF. Sperm injections were carried out with ejaculated sperm (EJA-ICSI) or testicular sperm (TESTI-ICSI) retrieved by either testicular sperm extraction (TESE) or testicular sperm aspiration (TESA). SDF levels were reassessed on the day of oocyte retrieval in both ejaculated and testicular specimens. Percentage of testicular and ejaculated spermatozoa containing fragmented DNA (%DFI) and clinical pregnancy, miscarriage, and live-birth rates. The %DFI in testicular sperm was 8.3%, compared with 40.7% in ejaculated sperm. For the TESTI-ICSI group versus the EJA-ICSI group, respectively, the clinical pregnancy rate was 51.9% and 40.2%, the miscarriage rate was 10.0% and 34.3%, and the live-birth rate was 46.7% and 26.4%. ICSI outcomes were significantly better in the group of men who had testicular sperm used for ICSI compared with those with ejaculated sperm. SDF was significantly lower in testicular specimens compared with ejaculated counterparts. Our results suggest that TESTI-ICSI is an effective option to overcome infertility when applied to selected men with oligozoospermia and high ejaculated SDF levels. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  15. Quantitative assessment of retrograde ejaculation using semen analysis, comparison with a standardized qualitative questionnaire, and investigating the impact of rhBMP-2.

    Science.gov (United States)

    Tepper, Gil; Rabbani, Ramin; Yousefzadeh, Michelle; Prince, Dean

    2013-05-01

    This was a prospective study evaluating the incidence of retrograde ejaculation (RE) after anterior lumbar interbody fusion (ALIF) based on laboratory analysis of semen and urine. The purpose of this study was to investigate the incidence of RE based on quantitative semen analysis and to compare the rate with that identified using a standardized qualitative questionnaire administered by telephone. RE is a specific sexual dysfunction in which semen passes into the bladder during ejaculation. It has long been a known possible complication of ALIF associated with injury to the superior hypogastric plexus occurring during access to the anterior lumbar spine. Although reported in multiple studies, there is no standardized assessment for RE and many studies do not describe the methods by which it was assessed. Recently, there has been increased interest in ALIF-related RE with respect to the use of recombinant human bone morphogenetic protein (rhBMP-2). However, how RE was diagnosed remains loosely defined. One possible assessment of RE is to analyze semen and urine after ejaculation. Forty-one male patients undergoing ALIF with posterior pedicle screw instrumentation and fusion at L4-L5 and/or L5-L1 consented to participate in the study. The subjects went to a cryobank for preoperative semen and postejaculatory urine analysis. They returned to the cryobank 3 to 6 months after surgery for repeat testing. The semen analysis consisted of ejaculate volume, total sperm count, concentration, and motility, whereas the urine test reported postejaculatory voided urine for spermatozoa. Postoperatively, patients were called by the principle investigator who conducted an interview based on a standardized questionnaire (completed for 36 patients). Patients were considered to have RE based on the questionnaire if they reported less or no ejaculate fluid. These results were compared with those from the laboratory testing to evaluate the value of the questionnaire in assessing RE. rh

  16. Vitrified sperm banks: the new aseptic technique for human spermatozoa allows cryopreservation at -86 °C.

    Science.gov (United States)

    Sánchez, R; Risopatrón, J; Schulz, M; Villegas, J V; Isachenko, V; Isachenko, E

    2012-12-01

    The vitrification technique is simple, quick, cost-effective and has showed a significantly stronger cryoprotective effect in contrast to conventional freezing. The method is based on the rapid cooling of the cell by direct immersion in liquid nitrogen (LN (2) ), thereby avoiding the formation of ice crystals, due to the lower risk of water thawing, which impairs cell function. The aim of this study was to evaluate the effect of storage at -86 °C compared to the conventional -196 °C (under LN (2) ) on essential parameters of the functioning of aseptically vitrified human sperm. Sperm motility, integrity of mitochondrial membrane potential and the rate of DNA fragmentation were determined. The comparison of -86 °C and -196 °C demonstrated no statistical difference in sperm progressive motility (73% vs. 77%), integrity of mitochondrial membrane potential (71% vs. 74%) or DNA fragmentation (3.1% vs. 2.9%). In conclusion, aseptically vitrified sperm can be preserved at -86 °C; eliminating the use of LN (2) simplifies and significantly reduces the costs associated with storage in sperm banks by decreasing the time and space needed for storage, the effort in finding stored samples, and by improving safety for the operator. However, for prolonged storage further studies are needed.

  17. Toward Evidence-Based Genetic Research on Lifelong Premature Ejaculation: A Critical Evaluation of Methodology

    Science.gov (United States)

    2011-01-01

    Recently, four premature ejaculation (PE) subtypes have been distinguished on the basis of the duration of the intravaginal ejaculation latency time (IELT). These four PE subtypes have different etiologies and pathogeneses. Genetic research on PE should consider the existence of these PE subtypes and the accurate measurement of the IELT with a stopwatch. Currently, three methods of genetic research on PE have been used. They differ in the investigated population, tool of measurement, study design, and variables of PE. From animal and human research, it is derived that the central serotonergic system "modulates" ejaculation, whereas the ejaculation (reflex) itself is probably not under direct influence of the serotonergic system, but rather under the influence of other neurotransmitter systems in the spinal cord. For genetic research on PE, it is important to take into account that the (serotonergic) modulation of the IELT is variable among men and may even be absent. This means that serotonergic genetic polymorphisms may only be found in men with PE who respond with an ejaculation delay treatment with a selective serotonin reuptake inhibitor. PMID:21344023

  18. Suitability and effectiveness of single layer centrifugation using Androcoll-P in the cryopreservation protocol for boar spermatozoa.

    Science.gov (United States)

    Martinez-Alborcia, Maria J; Morrell, Jane M; Gil, Maria A; Barranco, Isabel; Maside, Carolina; Alkmin, Diego V; Parrilla, Inmaculada; Martinez, Emilio A; Roca, Jordi

    2013-08-01

    The goal of the present experiment was to evaluate the suitability and effectiveness of single layer centrifugation (SLC), using the pig-specific colloid Androcoll-P, as a routine procedure for selecting boar spermatozoa for cryopreservation. The study focuses special attention on the effectiveness of SLC for processing a whole sperm rich ejaculate fraction and the fertilizing ability of frozen-thawed (FT) sperm selected using SLC prior to freezing. Thirteen sperm rich ejaculate fractions (one per boar) were split into three aliquots. Two aliquots of 15 and 150mL were SLC-processed (500×g for 20min) using 15 and 150mL (v/v) of Androcoll-P-Large and Androcoll-P-XL, respectively. The third aliquot remained un-processed as a control. The percentages of spermatozoa that were morphologically normal and showed rapid and progressive motility (assessed by CASA) spermatozoa were higher (Psperm chromatin dispersion test) were lower (Psperm motility (both total motility and rapid progressive motility), viability and intact nuclear DNA were higher (Psperm (679 in vitro matured oocytes inseminated with a viable sperm:oocyte ratio of 300:1 and coincubated for 6h), measured as the percentage of penetrated oocytes and the mean number of swollen sperm heads and/or male pronuclei in penetrated oocytes. However, there was no effect of SLC-processing on the in vitro ability of putative zygotes to develop to blastocysts. Overall these results indicate that SLC-processing of boar ejaculates using Androcoll-P improves the quality and fertilizing ability of cryosurvival boar sperm. However, efforts should be made to ensure continued high recovery yields before considering the inclusion of SLC as a routine procedure in the cryopreservation protocol of boar ejaculates.

  19. Synergetic Effects of K, Ca, Cu and Zn in Human Semen in Relation to Parameters Indicative of Spontaneous Hyperactivation of Spermatozoa

    OpenAIRE

    Ivan Bolanca; Jasmina Obhodas; Dejan Ljiljak; Lidija Matjacic; Krunoslav Kuna

    2016-01-01

    We have observed that sperm quality parameters indicative of spermatozoa hyperactivation such are lower "linearity" and "straightness", and as showed by this research "elongation", were more pronounced in patients with normal spermiogram compared to the group of men with reduced sperm motility who were undergoing routine in vitro fertilisation. The research encompassed 97 men diagnosed with normozoospermia (n = 20), asthenozoospermia (n = 54) and oligoasthenozoospermia (n = 23). The findings ...

  20. The pathophysiology of acquired premature ejaculation.

    Science.gov (United States)

    McMahon, Chris G; Jannini, Emmanuele A; Serefoglu, Ege C; Hellstrom, Wayne J G

    2016-08-01

    The second Ad Hoc International Society for Sexual Medicine (ISSM) Committee for the Definition of Premature Ejaculation defined acquired premature ejaculation (PE) as a male sexual dysfunction characterized by a the development of a clinically significant and bothersome reduction in ejaculation latency time in men with previous normal ejaculatory experiences, often to about 3 minutes or less, the inability to delay ejaculation on all or nearly all vaginal penetrations, and the presence of negative personal consequences, such as distress, bother, frustration and/or the avoidance of sexual intimacy. The literature contains a diverse range of biological and psychological etiological theories. Acquired PE is commonly due to sexual performance anxiety, psychological or relationship problems, erectile dysfunction (ED), and occasionally prostatitis and hyperthyroidism, consistent with the predominant organic etiology of acquired PE, men with this complaint are usually older, have a higher mean BMI and a greater incidence of comorbid disease including hypertension, sexual desire disorder, diabetes mellitus, chronic prostatitis, and ED compared to lifelong, variable and subjective PE.

  1. Pontine Control of Ejaculation and Female Orgasm

    NARCIS (Netherlands)

    Huynh, Hieu K.; Willemsen, Antoon T. M.; Lovick, Thelma A.; Holstege, Gert

    2013-01-01

    IntroductionThe physiological component of ejaculation shows parallels with that of micturition, as both are essentially voiding activities. Both depend on supraspinal influences to orchestrate the characteristic pattern of activity in the pelvic organs. Unlike micturition, little is known about the

  2. Central nervous system control of ejaculation

    NARCIS (Netherlands)

    Holstege, G

    2005-01-01

    An overview is given of the regions in the spinal cord that are active during ejaculation. Motoneurons involved are the preganglionic sympathetic motoneurons in the upper lumbar spinal cord and the motoneurons in the nucleus of Onuf, located in the upper sacral cord. The first group is involved in t

  3. Trehalose in glycerol-free freezing extender enhances post-thaw survival of boar spermatozoa.

    Science.gov (United States)

    Athurupana, Rukmali; Takahashi, Daisen; Ioki, Sumire; Funahashi, Hiroaki

    2015-01-01

    Cryopreservation of boar semen is still considered suboptimal due to lower fertility as compared with fresh samples when glycerol, a permeating cryoprotectant, is used. Trehalose is a non-permeable cryoprotectant and nonreducing disaccharide known to stabilize proteins and biologic membranes. The aim of this study was to evaluate the cryosurvival and in vitro penetrability of boar spermatozoa when glycerol was replaced with trehalose in a freezing extender. Ejaculated Berkshire semen samples were diluted in egg yolk-based freezing extender containing glycerol (100 mM) or trehalose (0, 50, 100, 150, 200 and 250 mM) and cryopreserved using a straw freezing procedure. Thawed samples were analyzed for motility, viability, mitochondrial membrane potential (MMP), and acrosome integrity. In experiment 2, penetrability of spermatozoa cryopreserved with 100 mM glycerol or trehalose was examined. Replacement of cryoprotectant glycerol (100 mM) with trehalose had no effect on sperm viability, but replacing it with 100 mM trehalose improved motility, MMP and acrosome integrity significantly. Sperm motility and MMP were considerably higher in 100 mM trehalose, whereas the acrosome integrity was substantially higher in 100-250 mM trehalose. The in vitro penetration rate was also significantly higher in spermatozoa cryopreserved with trehalose (61.3%) than in those cryopreserved with glycerol (43.6%). In conclusion, 100 mM non-permeable trehalose can be used to replace glycerol, a permeating cryoprotectant, for maintenance of better post-thaw quality of boar spermatozoa.

  4. Laboratory handling of epididymal and testicular spermatozoa: What can be done to improve sperm injections outcome

    Directory of Open Access Journals (Sweden)

    Sandro C Esteves

    2012-01-01

    Full Text Available Spermatozoa from azoospermic males can be retrieved from either the epididymis or the testis, depending on the type of azoospermia, using different surgical methods such as percutaneous epididymal sperm aspiration (PESA, testicular sperm aspiration (TESA, testicular sperm extraction (TESE, and microsurgical testicular sperm extraction (micro- TESE. After collecting the epididymal fluid or testicular tissue, laboratory techniques are used to remove contaminants, cellular debris, noxious microorganisms, and red blood cells. Processed spermatozoa may be used for intracytoplasmic sperm injection or eventually be cryopreserved. However, spermatozoa collected from either the epididymis or the testis are often compromised and more fragile than ejaculated ones. Therefore, sperm processing techniques should be used with great caution to avoid jeopardizing the sperm fertilizing potential in treatment cycles. In this review, we describe the current methods for processing surgically-retrieved specimens, either fresh or frozen- thawed, and provide the tips and pitfalls for facilitating the handling of such specimens. In addition, we present the available laboratory tools to aid in the identification of viable immotile spermatozoa to be used in conjunction with assisted reproductive techniques. Review of the literature was carried out using PubMed and Science Direct search engines.

  5. Biological Processes that Prepare Mammalian Spermatozoa to Interact with an Egg and Fertilize It

    Directory of Open Access Journals (Sweden)

    Daulat R. P. Tulsiani

    2012-01-01

    Full Text Available In the mouse and other mammals studied, including man, ejaculated spermatozoa cannot immediately fertilize an egg. They require a certain period of residence in the female genital tract to become functionally competent cells. As spermatozoa traverse through the female genital tract, they undergo multiple biochemical and physiological changes collectively referred to as capacitation. Only capacitated spermatozoa interact with the extracellular egg coat, the zona pellucida. The tight irreversible binding of the opposite gametes triggers a Ca2+-dependent signal transduction cascade. The net result is the fusion of the sperm plasma membrane and the underlying outer acrosomal membrane at multiple sites that causes the release of acrosomal contents at the site of sperm-egg adhesion. The hydrolytic action of the acrosomal enzymes released, along with the hyperactivated beat pattern of the bound spermatozoon, is important factor that directs the sperm to penetrate the egg coat and fertilize the egg. The sperm capacitation and the induction of the acrosomal reaction are Ca2+-dependent signaling events that have been of wide interest to reproductive biologists for over half a century. In this paper, we intend to discuss data from this and other laboratories that highlight the biological processes which prepare spermatozoa to interact with an egg and fertilize it.

  6. Liposome encapsulated soy lecithin and cholesterol can efficiently replace chicken egg yolk in human semen cryopreservation medium.

    Science.gov (United States)

    Mutalik, Srinivas; Salian, Sujith Raj; Avadhani, Kiran; Menon, Jyothsna; Joshi, Haritima; Hegde, Aswathi Raju; Kumar, Pratap; Kalthur, Guruprasad; Adiga, Satish Kumar

    2014-06-01

    Cryopreservation of spermatozoa plays a significant role in reproductive medicine and fertility preservation. Chicken egg yolk is used as an extender in cryopreservation of human spermatozoa using glycerol egg yolk citrate (GEYC) buffered medium. Even though 50% survival of spermatozoa is generally achieved with this method, the risk of high levels of endotoxins and transmission pathogens from chicken egg yolk is a matter of concern. In the present study we attempted to establish a chemically defined cryopreservation medium which can replace the chicken egg yolk without affecting sperm survival. Ejaculates from 28 men were cryopreserved with GEYC based freezing medium or liposome encapsulated soy lecithin-cholesterol based freezing medium (LFM). The semen samples were subjected to rapid thawing after 14 days of storage in liquid nitrogen. Post-thaw analysis indicated significantly higher post-thaw motility and sperm survival in spermatozoa cryopreserved with LFM compared to conventional GEYC freezing medium. The soy lecithin and cholesterol at the ratio of 80:20 with sucrose showed the highest percentage of post-thaw motility and survival compared to the other compositions. In conclusion, chemically defined cryopreservation medium with liposome encapsulated soy lecithin and cholesterol can effectively replace the chicken egg yolk from human semen cryopreservation medium without compromising post-thaw outcome.

  7. An investigation of the potential effect of sperm nuclear vacuoles in human spermatozoa on DNA fragmentation using a neutral and alkaline Comet assay.

    Science.gov (United States)

    Pastuszek, E; Kiewisz, J; Skowronska, P; Liss, J; Lukaszuk, M; Bruszczynska, A; Jakiel, G; Lukaszuk, K

    2017-03-01

    Presence of vacuoles and degree of sperm DNA damage are considered to be the basic factors used for the assessment of sperm fertilization capacity. We aimed to investigate the link between these two parameters. According to our knowledge, this is the first study where the Comet assay was used to assess the degree of DNA fragmentation of sperm categorized by Motile Sperm Organelle Morphology Examination (MSOME) Grades. Semen samples from 10 patients were assessed. Spermatozoa were graded into four MSOME groups according to the Vanderzwalmen's criteria. A total of 3930 motile spermatozoa were selected one-by-one using an inverted microscope and transferred onto two different slides. The degree of DNA fragmentation was analyzed by alkaline and neutral Comet assay. Results of the neutral Comet assay showed that Grade I spermatozoa (absence of vacuoles) presented significantly lower dsDNA fragmentation level (mean: 3.13 ± 1.17%) than Grade II (maximum of two small vacuoles; mean: 10.34 ± 2.65%), Grade III (more than two small vacuoles or at least one large vacuole; mean: 23.88 ± 8.37%), and Grade IV (large vacuoles associated with abnormal head shapes or other abnormalities; mean: 36.94 ± 7.78%; p fragmentation level (mean: 8.33 ± 3.62%) than Grade III (mean: 25.64 ± 9.15%) and Grade IV (mean: 40.10 ± 9.10%, p  0.05). Probably, the vacuoles may be responsible for double strand DNA breaks rather than single strand DNA breaks (only 2.39% spermatozoa in MSOME Grade II, 1.76% in III, and 3.16% in IV has single strand breaks). The results demonstrate that lower MSOME grading correlates with lower sperm DNA fragmentation. Therefore, the observation of sperm nuclear vacuoles using real-time optical microscopy without precise DNA fragmentation examination is not sufficient for optimal sperm selection for intracytoplasmic sperm injection. © 2017 American Society of Andrology and European Academy of Andrology.

  8. Detection of invariant natural killer T cells in ejaculates from infertile patients with chronic inflammation of genital tract.

    Science.gov (United States)

    Duan, Yong-Gang; Chen, Shujian; Haidl, Gerhard; Allam, Jean-Pierre

    2017-04-03

    Chronic inflammation of genital tract is thought to play a major role in male fertility disorder. Natural killer (NK) T cells are a heterogeneous group of T cells that share properties of both T cells and NK cells which display immunoregulatory properties. However, little is known regarding the presence and function of NK T cells in ejaculates from patients with chronic inflammation of genital tract. Invariant NK T (iNK T) cells were detected by invariant (Vα24-JαQ) TCR chain in ejaculates from patients suffering from chronic inflammation of genital tract (CIGT) using flow cytometry and immunofluorescence of double staining (n=40). Inflammatory cytokines interleukin (IL)-6, IL-17, and IFN-γ were detected in cell-free seminal plasma using an enzyme-linked immunosorbent assay (ELISA). The correlation between the percentage of iNK T cells and spermatozoa count, motility, vitality, seminal IL-6, IL-17, and IFN-γ was investigated. Significant percentages of iNK T cells above 10% were detected in 50% (CIGT-NKT(+) group). A negative correlation was detected between the percentage of iNK T cells and spermatozoa count (r=-.5957, P=.0056), motility (r=-.6163, P=.0038), and vitality (r=-.8032, P=.0019) in CIGT-NKT(+) group (n=20). Interestingly, a significant correlation of iNK T cells to seminal IL-6 (r=.7083, P=.0005), IFN-γ (r=.9578, PT cells and IL-17 (r=-.1557, P=.5122) in CIGT-NKT(+) group. The proliferative response of iNK T cells could accompany an inflammatory response to spermatozoa and consequently influence sperm quality through secretion of IFN-γ but not IL-17 under chronic inflammatory condition.

  9. The drug treatment of premature ejaculation

    OpenAIRE

    Hisasue, Shin-ichi

    2016-01-01

    The management recommendation for both acquired premature ejaculation (APE) and lifelong PE (LPE) are similar, such as a behavioral/psychotherapy, a pharmacotherapy and a combination of these treatments. For the drug treatment for PE, gold standard is selective serotonin reuptake inhibitors (SSRIs) including dapoxetine or paroxetine. The drug treatment for PE is still developing and some new promising therapeutic options have been proposed. Topical anesthetics, tramadol, and alpha-1 blockers ...

  10. Emerging and investigational drugs for premature ejaculation.

    Science.gov (United States)

    McMahon, Chris G

    2016-08-01

    Over the past 20-30 years, the premature ejaculation (PE) treatment paradigm, previously limited to behavioural psychotherapy, has expanded to include drug treatment. Pharmacotherapy for PE predominantly targets the multiple neurotransmitters and receptors involved in the control of ejaculation which include serotonin, dopamine, oxytocin, norepinephrine, gamma amino-butyric acid (GABA) and nitric oxide (NO). The objective of this article is to review emerging PE interventions contemporary data on the treatment of PE was reviewed and critiqued using the principles of evidence-based medicine. Multiple well-controlled evidence-based studies have demonstrated the efficacy and safety of selective serotonin reuptake inhibitors (SSRIs) in delaying ejaculation, confirming their role as first-line agents for the medical treatment of lifelong and acquired PE. Daily dosing of SSRIs is likely to be associated with superior fold increases in IELT compared to on-demand SSRIs. On-demand SSRIs are less effective but may fulfill the treatment goals of many patients. Integrated pharmacotherapy and CBT may achieve superior treatment outcomes in some patients. PDE-5 inhibitors alone or in combination with SSRIs should be limited to men with acquired PE secondary to co-morbid ED. New on-demand rapid acting SSRIs, oxytocin receptor antagonists, or single agents that target multiple receptors may form the foundation of more effective future on-demand medication. Current evidence confirms the efficacy and safety of dapoxetine, off-label SSRI drugs, tramadol and topical anaesthetics drugs. Treatment with α1-adrenoceptor antagonists cannot be recommended until the results of large well-designed RCTs are published in major international peer-reviewed medical journals. As our understanding of the neurochemical control of ejaculation improves, new therapeutic targets and candidate molecules will be identified which may increase our pharmacotherapeutic armamentarium.

  11. The pathophysiology of lifelong premature ejaculation.

    Science.gov (United States)

    Waldinger, Marcel D

    2016-08-01

    For many decades it has been thought that lifelong premature ejaculation (PE) is only characterized by persistent early ejaculations. Despite enormous progress of in vivo animal research, and neurobiological, genetic and pharmacological research in men with lifelong PE, our current understanding of the mechanisms behind early ejaculations is far from complete. The new classification of PE into four PE subtypes has shown that the symptomatology of lifelong PE strongly differs from acquired PE, subjective PE and variable PE. The phenotype of lifelong PE and therefore also the pathophysiology of lifelong PE is much more complex. A substantial number of men with lifelong PE not only have PE, but also premature erection and premature penile detumescence as part of an acute hypertonic or hypererotic state when engaged in an erotic situation or when making love. As both erectio praecox, ejaculatio praecox, detumescentia praecox, and the hypererotic state are part of the phenotype lifelong PE, it is argued that lifelong PE is not only a disturbance of the timing of ejaculation but also a disturbance of the timing of erection, detumescence and arousal. Since 1998, the pathophysiology of lifelong PE was thought to be mainly mediated by the central serotonergic system in line with genetic polymorphisms of specific serotonergic genes. However, by accepting that lifelong PE is characterized by the reversible hypertonic state the hypothesis of mainly serotonergic dysfunction is no longer tenable. Instead, it has been postulated that the pathophysiology of lifelong PE is mediated by a very complex interplay of central and peripheral serotonergic, dopaminergic, oxytocinergic, endocrinological, genetic and probably also epigenetic factors. Progress in research of lifelong PE can only be accomplished when a stopwatch is used to measure the IELT and the cut-off point of 1 minute for the definition of lifelong PE is maintained. Current use of validated questionnaires, neglect of

  12. The pathophysiology of lifelong premature ejaculation

    Science.gov (United States)

    2016-01-01

    For many decades it has been thought that lifelong premature ejaculation (PE) is only characterized by persistent early ejaculations. Despite enormous progress of in vivo animal research, and neurobiological, genetic and pharmacological research in men with lifelong PE, our current understanding of the mechanisms behind early ejaculations is far from complete. The new classification of PE into four PE subtypes has shown that the symptomatology of lifelong PE strongly differs from acquired PE, subjective PE and variable PE. The phenotype of lifelong PE and therefore also the pathophysiology of lifelong PE is much more complex. A substantial number of men with lifelong PE not only have PE, but also premature erection and premature penile detumescence as part of an acute hypertonic or hypererotic state when engaged in an erotic situation or when making love. As both erectio praecox, ejaculatio praecox, detumescentia praecox, and the hypererotic state are part of the phenotype lifelong PE, it is argued that lifelong PE is not only a disturbance of the timing of ejaculation but also a disturbance of the timing of erection, detumescence and arousal. Since 1998, the pathophysiology of lifelong PE was thought to be mainly mediated by the central serotonergic system in line with genetic polymorphisms of specific serotonergic genes. However, by accepting that lifelong PE is characterized by the reversible hypertonic state the hypothesis of mainly serotonergic dysfunction is no longer tenable. Instead, it has been postulated that the pathophysiology of lifelong PE is mediated by a very complex interplay of central and peripheral serotonergic, dopaminergic, oxytocinergic, endocrinological, genetic and probably also epigenetic factors. Progress in research of lifelong PE can only be accomplished when a stopwatch is used to measure the IELT and the cut-off point of 1 minute for the definition of lifelong PE is maintained. Current use of validated questionnaires, neglect of

  13. Characterization of northern pintail (Anas acuta) ejaculate and the effect of sperm preservation on fertility.

    Science.gov (United States)

    Penfold, L M; Harnal, V; Lynch, W; Bird, D; Derrickson, S R; Wildt, D E

    2001-02-01

    Northern pintail duck semen and sperm traits were characterized, and the fertility of cold-stored spermatozoa was investigated using artificial insemination. Excellent quality ejaculates containing high proportions of motile spermatozoa were collected from drakes within 20 s by a massage technique. Semen was collected in Beltsville poultry semen extender, pooled and cold-stored (4 degrees C) for 0, 24, 48 or 72 h. Hens were inseminated with 100 microl twice a week, and eggs were assessed for fertilization and hatch success. Fertilization success was similar (P > 0.05) for semen cold-stored for 0 (51.6%), 24 (51.5%), 48 (41.1%) and 72 h (22.3%; P > 0.05). Similar (P > 0.05) percentages of fertilized eggs hatched to live offspring (73.1, 71.4, 87.0 and 80.0%, respectively). Fresh semen was also equilibrated with 1 or 4% dimethylsulphoxide or glycerol, and cryopreserved at the following rates: (1) approximately 60 degrees C min(-1) (in liquid nitrogen [LN(2)] vapour) for 10 min; (2) 1 degrees C min(-1) to -20 degrees C, LN(2) vapour for 10 min; and (3) 1 degrees C min(-1) to -35 degrees C, all followed by immersion in LN(2). After thawing for 30 s at 37 degrees C or 20 min at 4 degrees C, sperm motility and viability were assessed. The highest numbers of motile spermatozoa were recovered after slow-fast freezing (2) and thawing at 0 degrees C (P artificial insemination. Nonetheless, cold storage provides an effective means of short-term storage with no loss of fertility in this waterfowl species.

  14. Cryopreservation of spermatozoa from wild-born Namibian cheetahs (Acinonyx jubatus) and influence of glycerol on cryosurvival.

    Science.gov (United States)

    Crosier, Adrienne E; Pukazhenthi, Budhan S; Henghali, Josephine N; Howard, Jogayle; Dickman, Amy J; Marker, Laurie; Wildt, David E

    2006-04-01

    Sperm cryopreservation is a valuable tool for the genetic management of ex situ populations. This study was conducted to assess: (1) semen characteristics of wild-born cheetahs; and (2) the impact of three types of glycerol influence (duration of exposure, temperature, and method of addition) on sperm cryosensitivity. To evaluate the impact of duration of glycerol exposure, spermatozoa were incubated in Test Yolk Buffer (TYB) with 4% glycerol at ambient temperature (approximately 22 degrees C) for 15 vs. 60 min before cryopreservation. To evaluate the influence of temperature and method of glycerol addition, spermatozoa were resuspended at ambient temperature either in TYB with 0% glycerol followed by addition of 8% glycerol (1:1 v/v; at ambient temperature vs. 5 degrees C) or directly in TYB with 4% glycerol. All samples were cryopreserved in straws over liquid nitrogen vapor and evaluated for sperm motility and acrosomal integrity after thawing. Semen samples (n = 23; n = 13 males) contained a high proportion (78%) of pleiomorphic spermatozoa. Ejaculates also contained a high proportion of acrosome-intact (86%) and motile spermatozoa (78%). Immediately after thawing, a significant proportion of spermatozoa retained intact acrosomes (range, 48-67%) and motility (range, 40-49%). After thawing, incubation in glycerol for 60 min at ambient temperature before freezing decreased (p 0.05) impact on sperm cryosurvival. In summary, (1) wild-born cheetahs produce high proportions of pleiomorphic spermatozoa but with a high proportion of intact acrosomes; and (2) resuspension in 4% glycerol, followed by exposure for up to 60 min at ambient temperature, had minimal effect on sperm motility and acrosomal integrity after cryopreservation. Results indicate the feasibility of cryopreserving cheetah spermatozoa under field conditions, providing a user-friendly method to capture and store gametes to enhance genetic management.

  15. Vitrification Increased Vacuolization of Human Spematozoa: Application of MSOME Technology.

    Science.gov (United States)

    Taherzadeh, Sara; Khalili, Mohammad Ali; Agha-Rahimi, Azam; Anbari, Fateme; Ghazali, Shahin; Macchiarelli, Guido

    2017-01-01

    Sperm vitrification is a technique of ice and cryoprotectant free cryopreservation by direct plunging of sperm suspension into liquid nitrogen (LN2). The aim of this study was to investigate the influence of cryoprotectant free-vitrification on human sperm fine structure by MSOME technology and the fertility potential by zona binding assay (ZBA). 20 normo-ejaculates were prepared by swim up technique, and supernatants were divided into two parts of fresh and vitrified groups. For vitrification, sperm was dropped into LN2. Sperm motility, morphology, viability and MSOME were evaluated for each sample. In MSOM morphologically normal sperm (class 1), ≤2 small vacuoles (class 2), and one large vacuole or >2 small vacuoles (class 3) were evaluated. Also, fertility potential was evaluated by zona binding assay. Data was analyzed using paired t-test or Willcoxon's test and p-value <0.05 was considered significant. Vitrification significantly reduced both progressive motility, viability and morphology. Also, normal morphology of spermatozoa decreased significantly after vitrification. In MSOME evaluation, normal motile spermatozoa (Class 1) decreased from 23.00±12.44 to 16.00.56±10.79 after vitrification (p=0.008). Although spermatozoa classes 2 and 3 were increased, the difference was not significant. Moreover, fertility potential of motile spermatozoa was reduced after vitrification (9.0±13.87 vs. 13.40±22.73; p=0.07). Vitrification increased the rate of vacuolization in motile sperm head. Therefore, MSOME technology is recommended for assessment of sperm fine morphology in ICSI program used cryopreserved spermatozoa.

  16. The osmotic tolerance of boar spermatozoa and its usefulness as sperm quality parameter.

    Science.gov (United States)

    Yeste, Marc; Briz, Mailo; Pinart, Elisabeth; Sancho, Sílvia; Bussalleu, Eva; Bonet, Sergi

    2010-06-01

    Predicting the fertility outcome of ejaculates is very important in the field of porcine reproduction. The aims of this study were to determine the effects of different osmotic treatments on boar spermatozoa and to correlate them with fertility and prolificacy, assessed as non-return rates within 60 days (NRR(60d)) of the first inseminations, and litter size (LS), respectively. Sperm samples (n=100) from one hundred healthy Piétrain boars were used to assess 48 treatments combining different osmolalities (ranged between 100 and 4000 mOsm kg(-1)), different compounds used to prepare anisotonic solutions, and two different modalities: return and non-return to isotonic conditions. Sperm quality was evaluated before and after applying the treatments on the basis of analyses of sperm viability, motility, morphology and percentages of acrosome-intact spermatozoa. Statistical analyses were performed using a one-way ANOVA and post hoc Tukey's test, linear regression analyses (Pearson correlation and multiple regression) and Jackknife cross-validation. Although three conventional parameters: sperm viability, sperm morphology and the percentages of acrosome-intact spermatozoa were significantly correlated with NRR(60d) and with LS, their respective osmotic tolerance parameters (defined for each parameter and treatment regarding with negative control) presented a higher Pearson coefficient with both fertility and prolificacy in three treatments (150 mOsm kg(-1) with non-return to isotonic conditions, 200 mOsm kg(-1) with return and 500 mOsm kg(-1) using sodium citrate and non-return to isotonic conditions). We conclude that osmotic resistance in sperm viability, sperm morphology and acrosome-intactness in the treatments mentioned above could be assessed along with classical parameters to better predict the fertilising ability of a given ejaculate.

  17. Characterization of alkaline phosphatase activity in seminal plasma and in fresh and frozen-thawed stallion spermatozoa.

    Science.gov (United States)

    Bucci, Diego; Giaretta, Elisa; Spinaci, Marcella; Rizzato, Giovanni; Isani, Gloria; Mislei, Beatrice; Mari, Gaetano; Tamanini, Carlo; Galeati, Giovanna

    2016-01-15

    Alkaline phosphatase (AP) has been studied in several situations to elucidate its role in reproductive biology of the male from different mammalian species; at present, its role in horse sperm physiology is not clear. The aim of the present work was to measure AP activity in seminal plasma and sperm extracts from freshly ejaculated as well as in frozen-thawed stallion spermatozoa and to verify whether relationship exists between AP activity and sperm quality parameters. Our data on 40 freshly ejaculated samples from 10 different stallions demonstrate that the main source of AP activity is seminal plasma, whereas sperm extracts contribution is very low. In addition, we found that AP activity at physiological pH (7.0) is significantly lower than that observed at pH 8.0, including the optimal AP pH (pH 10.0). Alkaline phosphatase did not exert any effect on sperm-oocyte interaction assessed by heterologous oocyte binding assay. Additionally, we observed a thermal stability of seminal plasma AP, concluding that it is similar to that of bone isoforms. Positive correlations were found between seminal plasma AP activity and sperm concentration, whereas a negative correlation was present between both spermatozoa extracts and seminal plasma AP activity and seminal plasma protein content. A significant decrease in sperm extract AP activity was found in frozen-thawed samples compared with freshly ejaculated ones (n = 21), concomitantly with the decrease in sperm quality parameters. The positive correlation between seminal plasma AP activity measured at pH 10 and viability of frozen-thawed spermatozoa suggests that seminal plasma AP activity could be used as an additional predictive parameter for stallion sperm freezability. In conclusion, we provide some insights into AP activity in both seminal plasma and sperm extracts and describe a decrease in AP after freezing and thawing.

  18. Zinc and iron concentration and SOD activity in human semen and seminal plasma.

    Science.gov (United States)

    Marzec-Wróblewska, Urszula; Kamiński, Piotr; Lakota, Paweł; Szymański, Marek; Wasilow, Karolina; Ludwikowski, Grzegorz; Kuligowska-Prusińska, Magdalena; Odrowąż-Sypniewska, Grażyna; Stuczyński, Tomasz; Michałkiewicz, Jacek

    2011-10-01

    The aim of the present study was to measure zinc (Zn) and iron (Fe) concentration in human semen and superoxide dismutase (SOD) activity in seminal plasma and correlate the results with sperm quality. Semen samples were obtained from men (N = 168) undergoing routine infertility evaluation. The study design included two groups based on the ejaculate parameters. Group I (n = 39) consisted of males with normal ejaculate (normozoospermia), and group II (n = 129) consisted of males with pathological spermiogram. Seminal Zn and Fe were measured in 162 samples (group I, n = 38; group II, n = 124) and SOD activity in 149 samples (group I, n = 37; group II, n = 112). Correlations were found between SOD activity and Fe and Zn concentration, and between Fe and Zn concentration. SOD activity was negatively associated with volume of semen and positively associated with rapid progressive motility, nonprogressive motility, and concentration. Negative correlation was stated between Fe concentration and normal morphology. Mean SOD activity in seminal plasma of semen from men of group I was higher than in seminal plasma of semen from men of group II. Fe concentration was higher in teratozoospermic males than in males with normal morphology of spermatozoa in group II. Our results suggest that Fe may influence spermatozoa morphology.

  19. Premature ejaculation and serotonergic antidepressants-induced delayed ejaculation : the involvement of the serotonergic system

    NARCIS (Netherlands)

    Waldinger, MD; Berendsen, HHG; Blok, BFM; Olivier, B; Holstege, G

    1998-01-01

    Premature ejaculation has generally been considered a psychosexual disorder with psychogenic aetiology. Although still mainly treated by behavioural therapy, in recent years double-blind studies have indicated the beneficial effects of some of the serotonergic antidepressants (SSRIs) in delaying eja

  20. Ejaculate parameters in patients with abdominal obesity

    Directory of Open Access Journals (Sweden)

    E. A. Epanchintseva

    2015-01-01

    Full Text Available Objective: the definition of association of levels of sex steroid hormones and ejaculate parameters with different types of fat distribution in infertile men with overweight and obesity. Materials and methods. A total of 119 somatically healthy Russian men who contacted Novosibirsk Center of Reproductive Medicine in 2012–2014 with the problem of infertility have been examined. Based on the results of anthropometric surveys all the men were divided into 3 groups. The 1st group included men with overweight, obesity, and upper type of fat distribution (the ratio of waist circumference (WC to the hip circumference (HC ≥ 0.95; the 2nd group – men with overweight, obesity and lower type of fat distribution (WC/HC < 0.95; the 3rd group – men with normal body weight. Questionnaires have been completed; determination in serum of concentrations of total testosterone, estradiol, sex hormones binding globulin (SHBG; free testosterone calculated. Special study of ejaculate included semen analysis, sperm morphology assessment by strict criteria of Kruger MAP test, NCA-test, analysis of DNA fragmentation of sperm. Results. In all 3 groups frequency of medical and social risk factors occurrence for infertility were analyzed: sexually transmitted infections, 88 chronic prostatitis, the systematic consumption of alcohol and smoking. It was revealed that these factors occurred with a high, but not significantly different frequency in men of 3 groups: the frequency of sexually transmitted infections in the 1st, 2nd and 3rd groups was 65.8; 61.0 and 63.2 %; systematic consumption of alcohol – 85.4; 78.1 and 63.2 %; systematic smoking – 36.6; 53.7 and 34.21 %; chronic prostatitis – 68.3; 56.1 and 50.0 % respectively. The average concentrations of sex steroid hormones and SHBG in the serum of men of all groups did not go beyond the reference range. Patients of the 1st and 2nd groups had significantly lower concentration of total testosterone in serum

  1. Ejaculate parameters in patients with abdominal obesity

    Directory of Open Access Journals (Sweden)

    E. A. Epanchintseva

    2015-04-01

    Full Text Available Objective: the definition of association of levels of sex steroid hormones and ejaculate parameters with different types of fat distribution in infertile men with overweight and obesity. Materials and methods. A total of 119 somatically healthy Russian men who contacted Novosibirsk Center of Reproductive Medicine in 2012–2014 with the problem of infertility have been examined. Based on the results of anthropometric surveys all the men were divided into 3 groups. The 1st group included men with overweight, obesity, and upper type of fat distribution (the ratio of waist circumference (WC to the hip circumference (HC ≥ 0.95; the 2nd group – men with overweight, obesity and lower type of fat distribution (WC/HC < 0.95; the 3rd group – men with normal body weight. Questionnaires have been completed; determination in serum of concentrations of total testosterone, estradiol, sex hormones binding globulin (SHBG; free testosterone calculated. Special study of ejaculate included semen analysis, sperm morphology assessment by strict criteria of Kruger MAP test, NCA-test, analysis of DNA fragmentation of sperm. Results. In all 3 groups frequency of medical and social risk factors occurrence for infertility were analyzed: sexually transmitted infections, 88 chronic prostatitis, the systematic consumption of alcohol and smoking. It was revealed that these factors occurred with a high, but not significantly different frequency in men of 3 groups: the frequency of sexually transmitted infections in the 1st, 2nd and 3rd groups was 65.8; 61.0 and 63.2 %; systematic consumption of alcohol – 85.4; 78.1 and 63.2 %; systematic smoking – 36.6; 53.7 and 34.21 %; chronic prostatitis – 68.3; 56.1 and 50.0 % respectively. The average concentrations of sex steroid hormones and SHBG in the serum of men of all groups did not go beyond the reference range. Patients of the 1st and 2nd groups had significantly lower concentration of total testosterone in serum

  2. The drug treatment of premature ejaculation.

    Science.gov (United States)

    Hisasue, Shin-Ichi

    2016-08-01

    The management recommendation for both acquired premature ejaculation (APE) and lifelong PE (LPE) are similar, such as a behavioral/psychotherapy, a pharmacotherapy and a combination of these treatments. For the drug treatment for PE, gold standard is selective serotonin reuptake inhibitors (SSRIs) including dapoxetine or paroxetine. The drug treatment for PE is still developing and some new promising therapeutic options have been proposed. Topical anesthetics, tramadol, and alpha-1 blockers will be the next strategies of the drug treatment for PE in the future.

  3. Premature ejaculation: current and future treatments

    Institute of Scientific and Technical Information of China (English)

    Levent Gurkan; Matthew Oommen; Wayne J. G. Hellstrom

    2008-01-01

    Premature ejaculation (PE) is recognized to be the most common male sexual disorder. PE provides difficulties for professionals who treat this condition because there is neither a universally accepted definition nor a medication approved by the Food and Drug Administration (FDA). Despite these shortcomings, physicians continue to diagnose their patients with PE according to major guidelines and treat them with either behavioral therapies or off-label medications. This review focuses on current and emerging treatment options and medications for PE. Advantages and limitations of each treatment option are discussed in the light of current published peer-reviewed literature.

  4. Sperm Hy-Liter™: an effective tool for the detection of spermatozoa in sexual assault exhibits.

    Science.gov (United States)

    De Moors, Anick; Georgalis, Tina; Armstrong, Gail; Modler, Jeff; Frégeau, Chantal J

    2013-05-01

    A fluorescence-based assay specifically targeting human spermatozoa was tested and optimized for best staining results using a variety of mock sexual assault samples. Swab clippings versus whole swabs were evaluated for best sample preparation and to simplify workflow (direct application versus swab extraction). The practicality and sensitivity of Sperm Hy-Liter™ was compared to our current phase contrast microscopy protocol for searching for the presence of spermatozoa. Sperm Hy-Liter™ was more sensitive than phase contrast microscopy and was able to detect spermatozoa more effectively in actual sexual assault samples (recent [N=240] or 24 years old [N=4]) containing few spermatozoa. Correlations were drawn between the Sperm Hy-Liter™ spermatozoa counts and the AmpFlSTR(®) Profiler(®) Plus male profiles generated from the sperm cell DNA fractions of semen containing swabs and swab clippings. In addition, recovered spermatozoa from Sperm Hy-Liter™-stained slides with greater than 40 spermatozoa produced full STR male profiles in 20.3% of slides tested and partial STR male profiles in 52.8% of slides tested. The adoption of Sperm Hy-Liter™ offers a means to standardize and improve the efficiency of the microscopic screening of sexual assault evidence.

  5. Sperm competition dynamics: ejaculate fertilising efficiency changes differentially with time

    Directory of Open Access Journals (Sweden)

    Burke Terry

    2008-12-01

    Full Text Available Abstract Background A fundamental challenge in evolutionary biology is to resolve the mechanisms that maintain paternity a hypervariable fitness component. Because females are often sexually promiscuous, this challenge hinges on establishing the mechanisms through which the ejaculates of different males compete for fertilisation (sperm competition. The competitive quality of an ejaculate is mediated by the relative number of live sperm and their motile performance. The differential rate at which rival ejaculates lose their fertilising efficiency over time is therefore expected to influence the outcome of sperm competition. Results Here, we artificially inseminated into sets of replicate domestic hens, Gallus gallus domesticus, experimentally engineered heterospermic ejaculates containing a large number of low-quality sperm from one male, and a lower number of high-quality sperm from another male. Large, low-quality ejaculates fertilised the first eggs produced after insemination, but small, high-quality ejaculates prevailed in the long run despite their numerical disadvantage. Conclusion Together, these results provide the first experimental demonstration that the relative competitive value of an ejaculate changes drastically over the time during which competing ejaculates are stored within the reproductive tract of a female, resulting in a marked temporal pattern of variation in paternity. A high level of replication makes these results robust. However, our study was restricted to few males of a well characterised study population, and future work should explore the generality of these results.

  6. Male masturbation device for the treatment of premature ejaculation

    Directory of Open Access Journals (Sweden)

    J.E. Rodríguez

    2016-03-01

    Conclusions: The patients who used the device Flip Hole increased of significant form the control grade on the ejaculation, and its interpersonal ejaculation distress and they reduced difficulties. To further evaluate the validity of these claims, randomized controlled trials should occur.

  7. Time Will Tell : Genetic Influences on Ejaculation Time

    NARCIS (Netherlands)

    Janssen, P.K.C.

    2014-01-01

    Lifelong premature ejaculation (PE) is a male sexual disorder characterized by persistent short ejaculation times of less than about 1 minute, in more than 90% of intercourses, with nearly every female partner, since the first sexual activities of a man in puberty or adolescence (Waldinger 2007). In

  8. Painful Ejaculation with Cyclobenzaprine: A Case Report and Literature Review

    Directory of Open Access Journals (Sweden)

    Molly B. Kraus, MD

    2015-12-01

    Conclusion: Painful ejaculation is likely an underreported side effect of tricyclic antidepressants and cyclobenzaprine use. Fortunately, these symptoms are reversible and discontinuation of these medications is typically an effective cure. Kraus MB, Wie CS, Gorlin AW, Wisenbaugh ES, and Rosenfeld DM. Painful ejaculation with cyclobenzaprine: A case report and literature review. Sex Med 2015;3:343–345.

  9. Sperm competition dynamics: ejaculate fertilising efficiency changes differentially with time

    Science.gov (United States)

    2008-01-01

    Background A fundamental challenge in evolutionary biology is to resolve the mechanisms that maintain paternity a hypervariable fitness component. Because females are often sexually promiscuous, this challenge hinges on establishing the mechanisms through which the ejaculates of different males compete for fertilisation (sperm competition). The competitive quality of an ejaculate is mediated by the relative number of live sperm and their motile performance. The differential rate at which rival ejaculates lose their fertilising efficiency over time is therefore expected to influence the outcome of sperm competition. Results Here, we artificially inseminated into sets of replicate domestic hens, Gallus gallus domesticus, experimentally engineered heterospermic ejaculates containing a large number of low-quality sperm from one male, and a lower number of high-quality sperm from another male. Large, low-quality ejaculates fertilised the first eggs produced after insemination, but small, high-quality ejaculates prevailed in the long run despite their numerical disadvantage. Conclusion Together, these results provide the first experimental demonstration that the relative competitive value of an ejaculate changes drastically over the time during which competing ejaculates are stored within the reproductive tract of a female, resulting in a marked temporal pattern of variation in paternity. A high level of replication makes these results robust. However, our study was restricted to few males of a well characterised study population, and future work should explore the generality of these results. PMID:19087292

  10. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia.

    Science.gov (United States)

    Zhou, Jun-Hao; Zhou, Qi-Zhao; Lyu, Xiao-Ming; Zhu, Ting; Chen, Zi-Jian; Chen, Ming-Kun; Xia, Hui; Wang, Chun-Yan; Qi, Tao; Li, Xin; Liu, Cun-Dong

    2015-01-01

    Cysteine-rich secretory protein 2 (CRISP2) is an important sperm protein and plays roles in spermatogenesis, modulation of flagellar motility, acrosome reaction, and gamete fusion. Clinical evidence shows a reduced CRISP2 expression in spermatozoa from asthenozoospermic patients, but the molecular mechanism underlying its reduction remains unknown. Herein, we carried out a study focusing on the CRISP2 reduction and its roles in asthenozoospermia. Initially, through analyzing CRISP2 expression and methylation on CRISP2 promoter activity in sperm, we observed a decreased expression of CRISP2 protein rather than its mRNA in the ejaculated spermatozoa from asthenozoospermic patients and no methylation in the CRISP2 promoter, suggesting CRISP2 expression may be regulated in the sperm at the posttranscriptional level. Subsequently, we found that microRNA 27b (miR-27b), predicted as a candidate regulator of CRISP2 using bioinformatics, was highly expressed in the ejaculated spermatozoa from asthenozoospermic patients. Luciferase reporter assay and transfection experiments disclosed that this microRNA could target CRISP2 by specifically binding its 3' untranslated region, suppressing CRISP2 expression. Extended clinical observation further confirmed a highly expressed miR-27b and its obviously negative correlation with CRISP2 protein expression in ejaculated spermatozoa samples from asthenozoospermic patients. Finally, we conducted a retrospective follow-up study to support that either high miR-27b expression or low CRISP2 protein expression was significantly associated with low sperm progressive motility, abnormal morphology, and infertility. Thus, this study provides the first preliminary insight into the mechanism leading to the reduced CRISP2 expression in asthenozoospermia, offering a potential therapeutic target for treating male infertility or for male contraception.

  11. Premature ejaculation: A clinical review for the general physician.

    Science.gov (United States)

    Chung, Eric; Gilbert, Brent; Perera, Marlon; Roberts, Matthew J

    2015-10-01

    Premature ejaculation is one of the most common sexual dysfunctions in men. Recent epidemiological studies suggest its prevalence in Australia may range from 21-31% This article will discuss the current definition of premature ejaculation from a urological perspective. It will provide an understanding of the pathogenesis of premature ejaculation, as well as assessment and management options. Premature ejaculation can have a significant adverse effect on the quality of life for the patient and his sexual partners. It can potentially lead to psychological distress, diminished self- esteem, anxiety, erectile dysfunction, reduced libido and poor interpersonal relationships. Most men feel reluctant to discuss premature ejaculation with their general practitioner despite its psychological, emotional and relational effects. Effective, evidence-based treatment options are available and physicians should feel confident when exploring ways to improve the quality of life for men with sexual dysfunction.

  12. A review of female ejaculation and the Grafenberg spot.

    Science.gov (United States)

    Belzer, E G

    1984-01-01

    A review of recent works related to the concept of female ejaculation, defined as a partial, infertile homologue to male ejaculation, indicated that we have insufficient evidence to accept or reject the concept on scientific grounds. The claim that "female ejaculate" is similar to semen from a vasectomized male is without foundation. Although recent discussion of female ejaculation has tended to focus on whether or not the fluid sometimes expelled from the urethra upon orgasm is invariably urine, the Bartholin's glands are another potential source of orgasmic expulsions which could properly be called "female ejaculate." "Grafenberg spot" is a neologism referring to an area which some women report is erotically sensitive when massaged via the anterior vaginal wall. "Urethral sponge" is another term which seems to refer to this anatomical/phenomenological reality. The relationship between stimulation of this area and subsequent orgasmic expulsion has yet to be studied systematically.

  13. Freeze-dried stallion spermatozoa: evaluation of two chelating agents and comparative analysis of three sperm DNA damage assays.

    Science.gov (United States)

    Olaciregui, M; Luño, V; Martí, J I; Aramayona, J; Gil, L

    2016-11-01

    During the freeze-drying procedure, sperm DNA might become damaged by both freezing and drying stresses. Sperm DNA status can be detected using well-established assays; however, most techniques are expensive and involve elaborate protocols and equipment. Indirect assessments can provide alternative strategies. The objective of this study was to compare a simple test of DNA status using Diff-Quik (DQ) with two established procedures: acridine orange test (AOT) and sperm chromatin dispersion (SCD) on freeze-dried (FD) stallion spermatozoa. Ejaculated spermatozoa from three stallions were freeze-dried in basic medium supplemented with two different chelating agents: EGTA or EDTA. After rehydration, the spermatozoa were subjected to DNA damage detection using a SCDt, AOT and DQ stain simultaneously. The results showed that the DNA damage levels in the EGTA group were significantly lower than those in the EDTA group. AOT detected a significantly higher proportion of spermatozoa with fragmented DNA than DQ and SCD. The results of the SCD test and DQ stain exhibited a significant positive correlation for DNA fragmentation (r = 0.528), whereas a negative correlation was observed between SCD, DQ and AOT (r = -0.134 and r = -0.332 respectively). The present study shows that both the SCD test and DQ assay are effective methods for detecting FD stallion sperm DNA fragmentation, whereas using of AOT is questionable.

  14. Intra-uterine insemination with prepared sperm vs. unprepared first split ejaculates. A randomized study.

    Science.gov (United States)

    Goldenberg, M; Rabinovici, J; Bider, D; Lunenfeld, B; Blankstein, J; Weissenberg, R

    1992-01-01

    In this randomized prospective study, we determined the conception rate following intra-uterine insemination with washed and prepared sperm, or with the first portion of a split ejaculate, in couples with longstanding male (n = 27, 70 treatment cycles) or cervical infertility (n = 14, 29 treatment cycles). Folliculogenesis and ovulation were induced by human menopausal gonadotropin and human chorionic gonadotropin. Significantly more couples conceived in the male infertility group following intra-uterine insemination with washed sperm, than after intra-uterine insemination with split ejaculate (9 vs. 2; P less than 0.05), while no difference in pregnancy rate (2 vs. 2) was found by the two intra-uterine insemination methods in the cervical infertility group.

  15. Effect of Interventions for Premature Ejaculation in the Treatment of Chronic Prostatitis with Secondary Premature Ejaculation.

    Science.gov (United States)

    Chen, Chang-Qing; Yi, Qing-Tong; Chen, Chu-Hong; Gong, Min

    2016-08-01

    Objective To evaluate the effect of interventions for premature ejaculation (PE) in the management of patients with chronic prostatitis and secondary premature ejaculation. Methods Totally 90 patients diagnosed as chronic prostatitis with PE were randomly divided into control group (n=45) and interventional group (n=45). Control group received a conventional therapy consisted of oral administration of antibiotics,α-receptor blocker,and proprietary Chinese medicine for clearing away heat and promoting diuresis. Interventional group received a conventional therapy combined with treatment for ameliorating the PE symptom (oral dapoxetine on-demand and ejaculation control exercise).National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI),Chinese Index of Sexual Function for Premature Ejaculation (CIPE)-5 questionnaires,intravaginal ejaculatory latency time,and the number of coituses per week were applied for evaluating the treatment outcomes. Results Follow-up was accomplished in 35 and 38 patients in the control and interventional group.The CIPE-5 score,intravaginal ejaculatory latency time,and the number of coituses per week were significantly improved in both two groups but more significantly in interventional group (all P<0.05). The NIH-CPSI pain,urination,and quality of life subscores and total score were improved significantly in both two groups after treatment,but the NIH-CPSI pain and quality of life subscores had been improved more significantly in the interventional group (all P<0.05). The variation of NIH-CPSI was negatively correlated with that of CIPE-5 in both two groups (r=-0.362,P=0.016;r=-0.330,P=0.021). Conclusions For CP with secondary PE patients,the interventions for PE can not only improve the quality of sexual life but also help improve the NIH-CPSI pain and quality of life subscores. PE should be routinely screened and treated during the management of CP.p.

  16. Characteristics of stallion epididymal spermatozoa at collection and effect of two refrigeration protocols on the quality of the frozen/thawed sperm cells.

    Science.gov (United States)

    Guimarães, T; Lopes, G; Ferreira, P; Leal, I; Rocha, A

    2012-12-01

    Cryopreservation of epididymal spermatozoa is a useful tool to preserve genetic material of valuable stallions after emergency castration or unexpected death. For that, testicles and epididymides are generally sent refrigerated to the laboratory. Collection of epididymal spermatozoa is a simple procedure that reduces the volume of the material to be shipped, and may improve the quality of the chilled epididymal sperm cells. In the present study we compared the characteristics of frozen/thawed epididymal spermatozoa after refrigeration of the epididymis or after direct refrigeration of the extended epididymal sperm cells. Ejaculated sperm samples were obtained from 10 healthy stallions with at least 15 days of sexual rest, before routine orchiectomies. Spermatozoa were recovered from the epididymal tail immediately after castration (EPI), after refrigeration of the epididymis for 24h at 4°C (EPI R) and recovered from epididymal tail immediately after castration and stored for 24h at 4°C (EPI RR). Total motility, straight-line velocity, percentage of rapid cells, viability and morphological defects were similar (p>0.05) among different treatments, and post-thaw viability was higher (psperm. The similarity of post-thaw parameters led us to conclude that immediate collection and refrigeration of the epididymal sperm cells or refrigeration of the whole epididymis are equally efficient as a means of transporting material for 24h before cryopreservation of epididymal spermatozoa.

  17. Stallion spermatozoa: putative target of estrogens; presence of the estrogen receptors ESR1, ESR2 and identification of the estrogen-membrane receptor GPER.

    Science.gov (United States)

    Arkoun, Brahim; Gautier, Camille; Delalande, Christelle; Barrier-Battut, Isabelle; Guénon, Isabelle; Goux, Didier; Bouraïma-Lelong, Hélène

    2014-05-01

    Among mammals, the stallion produces the largest amount of testicular estrogens. These steroid hormones are produced mainly by Leydig and Sertoli cells in the testis and also in the epididymis. Their role in horse testicular physiology and their ability to act on spermatozoa are still unknown. In order to determine if spermatozoa are targets for estrogens, the presence of estrogen receptors in mature ejaculated spermatozoa has been investigated. The presence of a single isoform of ESR1 (66kDa) and ESR2 (61kDa) was found by Western-blot analysis in samples from seven stallions. Confocal analysis mainly showed a flagellar localization for both receptors. Immuno-TEM experiments revealed that they are mostly located near the membranes, which are classically associated with rapid, non-genomic, effects. Moreover, we evidenced the expression of the seven transmembrane estradiol binding receptor GPER in colt testis. The protein was also localized at the connecting piece in mature spermatozoa. In conclusion, our results suggest that horse spermatozoa are a target for estrogens, which could act on several receptors either during the epididymal transit and/or in the female genital tract.

  18. Selection of physiological spermatozoa during intracytoplasmic sperm injection.

    Science.gov (United States)

    Torki-Boldaji, B; Tavalaee, M; Bahadorani, M; Nasr-Esfahani, M H

    2017-02-01

    Sperm genomic integrity has a significant effect on intracytoplasmic sperm injection (ICSI) outcomes, especially post-implantation. Spermatozoa selected based on motility and morphology do not guarantee the genomic integrity of spermatozoa. Nearly fifty percentage of spermatozoa in infertile men with normal morphology present different degrees of DNA fragmentation. However, capacitated or hyperactivated spermatozoa show lower degrees of DNA fragmentation. Therefore, selection of hyperactivated spermatozoa may improve ICSI outcome. Routinely, for ICSI, fast-moving spermatozoa with A or B motility pattern are mainly selected for injection. The result of this study shows that in processed semen samples, hyperactivated spermatozoa are mainly observed in B motility pattern while, in viscous medium like polyvinylpyrrolidone (PVP), hyperactivated spermatozoa are mainly present in spermatozoa with C pattern of motility (nonprogressive). Therefore, we propose spermatozoa with C motility pattern which contains the main population of physiological or hyperactivated spermatozoa should be selected for ICSI.

  19. AB035. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator mir-27b in the spermatozoa of patients with asthenozoospermia

    Science.gov (United States)

    Zhou, Jun-Hao; Zhou, Qi-Zhao; Lyu, Xiao-Ming; Zhu, Ting; Chen, Zi-Jian; Chen, Ming-Kun; Xia, Hui; Wang, Chun-Yan; Qi, Tao; Li, Xin; Liu, Cun-Dong

    2016-01-01

    Background Cysteine-rich secretory protein 2 (CRISP2) is an important sperm protein and plays roles in spermatogenesis, modulation of flagellar motility, acrosome reaction, and gamete fusion. Clinical evidence shows a reduced CRISP2 expression in spermatozoa from asthenozoospermic patients, but the molecular mechanism underlying its reduction remains unknown. Herein, we carried out a study focusing on the CRISP2 reduction and its roles in asthenozoospermia Methods Spermatozoa were isolated from 90 study subjects’ ejaculated semen samples. DNA methylation was evaluated using bisulfite-sequencing PCR and methylation-specific PCR. The CRISP2 mRNA and protein expression levels were examined in the ejaculated spermatozoa by qRT-PCR and Western blot respectively. miRNA expression was detected by qRT-PCR. The direct regulatory effect of miR-27b on CRISP2 was predicted computationally and validated via luciferase reporter assay and in vitro experiments in which miR-27b mimic or inhibitor was transfected into 293T cells. Respective correlations of miR-27b and CRISP2 protein expression with clinical features were analyzed using Spearman’s correlation coefficient. Results Initially, low expression of CRISP2 protein rather than its mRNA was observed in the ejaculated spermatozoa from asthenozoospermic patients relative to normozoospermic males. Meanwhile, methylation was not found in CRISP2 promoter. These data suggest a possible post-transcriptional regulation of CRISP2 in asthenozoospermia. Subsequently, bioinformatics prediction, luciferase reporter assay and miR-27b transfection experiments revealed that miR-27b could specifically target CRISP2 by binding its 3’-UTR, suppressing CRISP2 expression post-transcriptionally. Further evidence was provided by the clinical observation of a high miR-27b expression in the ejaculated spermatozoa from asthenozoospermic patients and a negative correlation between miR-27b and CRISP2 protein expression. Finally, a retrospective

  20. Sexual function of premature ejaculation patients assayed with Chinese Index of Premature Ejaculation

    Institute of Scientific and Technical Information of China (English)

    Yi-MingYuan; Zhong-ChengXin; HuiJiang; Yan-JieGuo; Wu-JiangLiu; LongTian; Ji-ChuanZhu

    2004-01-01

    Aim: To assess the psychometric properties of the Chinese Index of Premature Ejaculation (CIPE).Methods: The sexual function of 167 patients with and 114 normal controls without premature ejaculation (PE) were evaluated with CIPE. All subjects were married and had regular sexual activity. The CIPE has 10 questions, focusing on libido, erectile function, ejaculatory latency, sexual satisfaction and difficulty in delaying ejaculation, self-confidence and depression. Each question was responded to on a 5 point Likert-type scale. The individual question score and the total scale score were analyzed between the two groups. Results: There were no significant differences between the age, duration of marriage and educational level (P>0.05) of patients with and without PE and normal controls. The mean latency of patients with PE and normal controls were 1.6±1.2 and 10.2±9.5 minutes,respectively. Significant differences between patients with (26.7±4.6) PE and normal controls (41.9±4.0) were observed on the total score of CIPE (P15 point) 19.8%, moderate (10~14 point) 62.8% and severe (<9point) 16.7%. Conclusion:The CIPE-5 is a useful method for the evaluation of sexual function of patients with PE and can be used as a clinical endpoint for clinical trials studying the efficacy of pharmacological intervention.

  1. Insight on pathogenesis of lifelong premature ejaculation: inverse relationship between lifelong premature ejaculation and obesity.

    Science.gov (United States)

    Gökçe, A; Ekmekcioglu, O

    2010-01-01

    Although both biological and psychological factors are important in the etiology, the exact pathogenesis of lifelong premature ejaculation (PE) remains to be clarified. Obesity is a worldwide epidemic that contributes to many chronic diseases. Obesity is associated with erectile dysfunction, but the relationship between obesity and PE has not yet been specifically investigated. The aim of this study was to evaluate the relationships of these two conditions. Between January 2008 and December 2009, we evaluated consecutive patients with lifelong PE in the urology outpatient clinic. Control cases without lifelong PE were selected randomly among cases attending the department of internal medicine for a checkup procedure. The age and sex of control group were matched with that of the study group. Body mass index (BMI) of each case was calculated using the World Health Organization criteria by the measurements of the physician instead of relying on verbal expressions. The mean (+/-s.d.) age of the premature ejaculators was 31.7+/-5.7 (range 21-51) years and in the control cases it was 32.3+/-6.7 (range 22-54) years. The comparison of the mean (+/-s.d.) weight between the study (74.1+/-11.2 kg) and control groups (81.9+/-6.4 kg) revealed a significant difference (Pobese cases in the control group (n=26, 24.1%) was three times greater than the obese premature ejaculators (Pobesity, and we found that patients with lifelong PE were leaner than the healthy control cases.

  2. Comparison of post-thaw DNA integrity of boar spermatozoa assessed with the neutral comet assay and Sperm-Sus Halomax test kit.

    Science.gov (United States)

    Fraser, L; Parda, A; Filipowicz, K; Strzeżek, J

    2010-10-01

    In this study, we tested the hypothesis whether the neutral Comet assay (NCA) and the Sperm-Sus-Halomax (SSH) test kit could provide similar measurements of post-thaw DNA fragmentation of boar spermatozoa. Whole ejaculates or sperm-rich fractions of boar semen were frozen in an extender containing lactose, lipoprotein fractions isolated from ostrich egg yolk (LPFo), glycerol (lactose-LPFo-G) or in a standard boar semen extender (K3), without the addition of cryoprotective substances. In all boars, both the NCA and SSH test showed similar levels of post-thaw sperm DNA fragmentation in samples of the same ejaculates, regardless of the ejaculate collection procedure and extender. Yet, the levels of post-thaw sperm DNA damage, detected by the NCA and SSH test, were more accentuated in spermatozoa frozen in the absence of cryoprotective substances. Both the NCA and SSH detected variations among individual boars in terms of post-thaw sperm DNA fragmentation. Agreement between the measurements of the NCA and SSH was confirmed by scatter plots of differences, suggesting that the DNA integrity tests could detect the same sperm populations, which were susceptible to cryo-induced DNA damage. The findings of this study indicate that the NCA and the SSH test are effective in detecting similar levels of sperm DNA fragmentation and reinforce their importance in the assessment of frozen-thawed boar semen quality.

  3. SPERM HY-LITER™ for the identification of spermatozoa from sexual assault evidence.

    Science.gov (United States)

    Westring, Christian G; Wiuf, Morten; Nielsen, S Jock; Fogleman, James C; Old, Jennifer B; Lenz, Camilla; Reich, Karl A; Morling, Niels

    2014-09-01

    Accurate microscopic identification of human spermatozoa is important in sexual assault cases. We have compared the results of examinations with (1) a fluorescent microscopy method, SPERM HY-LITER™, and (2) Baecchi's method for identification of human spermatozoa. In 35 artificial, forensic type samples, spermatozoa were identified in 45.7% with SPERM HY-LITER™ in Copenhagen, in 54.3% in the laboratory of the manufacturer of SPERM HY-LITER™, and 40.0% of the samples with Baecchi's staining method. When differences occurred between the two methods, it was significantly more often that SPERM HY-LITER™ detected spermatozoa when Baecchi's method did not (ts=6.567, df=1, P=0.048). This trend was also seen in selected compromised or degraded samples and in selected adjudicative samples. The reactions with spermatozoa from dog, horse, pig and bull were negative with SPERM HY-LITER™, whereas Baecchi's method was non-selective. Data from forensic casework samples in Copenhagen from two years (2008 and 2009) are presented. The samples from 2008 were investigated using Baecchi's method, while those from 2009 were investigated using SPERM HY-LITER™. The frequencies of positive results were similar between the two methods for the two years (27.9% and 32.1% respectively). Analysis of acid phosphatase (ACP) activity for the positive results obtained for these two years does not support the use of a negative ACP result as a prescreen for microscopic analysis for spermatozoa.

  4. Classification and definition of premature ejaculation.

    Science.gov (United States)

    Parnham, Arie; Serefoglu, Ege Can

    2016-08-01

    Premature ejaculation (PE) is a poorly understood condition and is considered as the most common sexual disorder in men. The ambiguity surrounding PE is in part due to the difficulty in conducting and interpreting research in the absence of a standardised definition that adequately encompasses the characteristics of these patients. An enhanced awareness of sexual dysfunctions in the recent decades has lead to an increase in scientific research that has challenged the traditional paradigm regarding PE. This has also enabled to establish a universal definition and classification of the disease. A move to a more evidence based approach has improved the clinicians' ability to define those who need medical treatment, as well as perform further research in this complex condition.

  5. On Chip Spermatozoa and Leucocytes Counter

    NARCIS (Netherlands)

    Segerink, Loes; Sprenkels, Ad; Vermes, István; Berg, van den Albert; Kim, Tae Song; Lee, Yoon-Sik; Chung, Taek-Dong; Jeon, Noo Li; Lee, Sang-Hoon; Suh, Kahp-Yang; Choo, Jaebum; Kim, Yong-Kweon

    2009-01-01

    In this project we aim to develop a semen quality test system based on a lab-on-a-chip. Since an important quality parameter of semen is the concentration of spermatozoa, we focus on counting the spermatozoa by using impedance measurements in a microchannel. However, semen contains also an interferi

  6. Ejaculation increases the serum prostate-specific antigen concentration.

    Science.gov (United States)

    Tchetgen, M B; Song, J T; Strawderman, M; Jacobsen, S J; Oesterling, J E

    1996-04-01

    To determine the effect of ejaculation on the serum prostate-specific antigen (PSA) concentration in men at risk for developing prostate cancer. A prospective, community-based study was conducted in which 64 men, aged 49 to 79 years, underwent a serum PSA determination immediately before ejaculation (baseline) and at 1 hour, 6 hours, and 24 hours following ejaculation. The serum PSA also was measured 48 hours and 1 week after ejaculation if the concentration had not returned to the baseline value by the previous time interval. All subjects abstained from ejaculation for a minimum of 7 days prior to the study and until the PSA concentration returned to the baseline level. Absolute and relative change in serum PSA concentration, as well as the time to return to baseline PSA concentration following ejaculation, were assessed. The serum PSA concentration increased following ejaculation in 87% of the subjects. The mean baseline PSA was 1.8 ng/mL (median, 0.7 ng/mL). The mean absolute PSA change +/- standard deviation 1 hour, 6 hours, 24 hours, and 48 hours after ejaculation was 0.8 +/- 1.32 ng/mL, 0.3 +/- 0.66 ng/mL, 0.2 +/- 0.33 ng/mL, and 0.4 +/- 0.40 ng/mL, respectively. The mean relative PSA change +/- standard error 1 hour, 6 hours, 24 hours, and 48 hours after ejaculation was 41 +/- 4%, 9 +/- 1.5%, 8 +/- 1.3%, and 10 +/- 2.3%, respectively. The absolute and relative changes in PSA concentration noted 1 hour, 6 hours, and 24 hours after ejaculation were statistically significant (P = 0.0001). A strong correlation was observed between absolute change in PSA and baseline serum PSA, at each time interval (1 hour: r = 0.68, 6 hours: r = 0.77, 24 hours: r = 0.70; P returned to baseline by 24 hours (95% confidence interval (Cl) = 83% to 97%), whereas 97% of subjects returned to baseline by 48 hours (95% Cl = 89% to 99%). Ejaculation causes a significant increase in the serum PSA concentration in men between 49 and 79 years of age that may persist for up to 48 hours. This

  7. Electrophoretic and zymographic characterization of proteins isolated by various extraction methods from ejaculated and capacitated boar sperms.

    Science.gov (United States)

    Zigo, Michal; Jonáková, Věra; Maňásková-Postlerová, Pavla

    2011-06-01

    The presented work focuses on electrophoretic and zymographic characterization of boar sperm proteins isolated by various extraction methods and on comparison of the protein profiles obtained from ejaculated and in vitro capacitated spermatozoa. Sperm proteins of ejaculated and in vitro capacitated boar sperms were isolated with the following agents: 1% v/v Triton X-100, 1% v/v Triton X-114, 2% v/v acetic acid, 1% m/v sodium dodecyl sulphate (SDS), 30 mM N-octyl-β-D-glucopyranoside (OBG), rehydration buffer (RHB) for isoelectric focusing and finally by the freezing-thawing approach. The extracts were characterized in terms of 1-DE, 2-DE protein profiles, 1-DE glycoprotein staining and proteinase and hyaluronidase substrate zymographic profiles. The results have shown quantitative and qualitative differences in 1-DE protein and glycoprotein profiles with respect to the employed isolation approach. These differences were seen even more clearly in 2-DE protein profiles, where it was possible to distinguish the presence/absence, changes in relative abundance and pI/M(r) shifts of various protein spots. Proteinase and hyaluronidase zymograms supported the prediction that various isolation protocols result in various profiles of enzymatically active molecules.

  8. Negative effect of combined cysteine and glutathione in soy lecithin-based extender on post-thawed ram spermatozoa.

    Science.gov (United States)

    Zhandi, Mahdi; Sharafi, Mohsen

    2015-09-01

    This study was conducted to evaluate the effect of combined cysteine and glutathione in soy lecithin-based semen extender on post-thawed ram sperm quality. A total of 28 ejaculates were collected twice a week (from four rams) during breeding season. In each replicate, semen samples (n = 4, one ejaculate for each ram) were pooled and divided into three equal parts, and each part was diluted with one of following extender: (1) soy lecithin-based extender containing no cysteine and no glutathione (C0-G0), (2) soy lecithin-based extender containing cysteine (5 mM) and glutathione (5 mM) (C5-G5), and (3) soy lecithin-based extender containing cysteine (10 mM) and glutathione (10 mM) (C10-G10). After freeze-thawing process, motility and velocity parameters, plasma membrane integrity and functionality, mitochondrial activity, and apoptosis features of spermatozoa were evaluated. The obtained results showed that total and progressive motility, plasma membrane integrity and functionality, and live post-thawed spermatozoa was lower in C10-G10 extender compared to C0-G0 and C5-G5 extenders (P 0.05). In conclusion, it seems that high concentration of combined cysteine and glutathione in soy lecithin-based semen extender has a detrimental effect of post-thawed ram sperm quality.

  9. Serotonin transporter promoter region (5-HTTLPR) polymorphism is associated with the intravaginal ejaculation latency time in Dutch men with lifelong premature ejaculation

    NARCIS (Netherlands)

    Janssen, Paddy K C; Bakker, Steven C; Réthelyi, Janos; Zwinderman, Aeilko H; Touw, Daan J; Olivier, Berend; Waldinger, Marcel D

    2009-01-01

    INTRODUCTION: Lifelong premature ejaculation (LPE) is characterized by persistent intravaginal ejaculation latency times (IELTs) of less than 1 minute, and has been postulated as a neurobiological dysfunction with genetic vulnerability for the short IELTs, related to disturbances of central serotoni

  10. Serotonin transporter promoter region (5-HTTLPR) polymorphism is associated with the intravaginal ejaculation latency time in Dutch men with lifelong premature ejaculation

    NARCIS (Netherlands)

    Janssen, Paddy K C; Bakker, Steven C; Réthelyi, Janos; Zwinderman, Aeilko H; Touw, Daan J; Olivier, Berend; Waldinger, Marcel D

    INTRODUCTION: Lifelong premature ejaculation (LPE) is characterized by persistent intravaginal ejaculation latency times (IELTs) of less than 1 minute, and has been postulated as a neurobiological dysfunction with genetic vulnerability for the short IELTs, related to disturbances of central

  11. Serotonin Transporter Promoter Region (5-HTTLPR) Polymorphism is Associated with the Intravaginal Ejaculation Latency Time in Dutch Men with Lifelong Premature Ejaculation

    NARCIS (Netherlands)

    P.K.C. Janssen; S.C. Bakker; J. Réthelyi; A.H. Zwinderman; D.J. Touw; B. Olivier; M.D. Waldinger

    2009-01-01

    Introduction. Lifelong premature ejaculation (LPE) is characterized by persistent intravaginal ejaculation latency times (IELTs) of less than 1 minute, and has been postulated as a neurobiological dysfunction with genetic vulnerability for the short IELTs, related to disturbances of central serotoni

  12. Spontaneous ejaculation with the use of ADHD drugs

    NARCIS (Netherlands)

    Oosterhuis, I.; Heijting, L.W.; Van Puijenbroek, E.P.

    2009-01-01

    Background: Methylphenidate and atomoxetine are indicated for treatment of attention deficit-hyperactivity disorder (ADHD). The Netherlands Pharmacovigilance Centre Lareb received two reports of spontaneous ejaculation associated with the use of these drugs. Objective: To describe two case reports o

  13. Variability in ejaculation rate and libido of boars during reproductive ...

    African Journals Online (AJOL)

    You

    2015-08-29

    Aug 29, 2015 ... Assessment of libido based only on the duration of ejaculation is insufficient, so it is ... A digital timer was used to measure the time intervals in minutes (min). ..... An assessment of the breeding maturity of insemination boars.

  14. IN VITRO EFFECTS OF COPPER ON THE MOTILITY AND VIABILITY OF SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    Zuzana Kňažická

    2012-06-01

    Full Text Available Copper (Cu is an environmental risk factor which has various effects on the animal and human organism. The target of this study was to investigate the effects of Cu on motility and viability of spermatozoa in vitro. Specifically, we examined the dose- and time-dependent effect of copper (II chloride (CuCl2 on the survival of spermatozoa during different time periods (Time 0 h, 1 h, 2 h. The percentage of motile spermatozoa was determined after exposure to concentrations of 3.9; 7.8; 15.6; 31.2; 62.5; 125; 250; 500; 1000 µmol.dm-3 of CuCl2 using the Sperm VisionTM CASA (Computer Assisted Semen Analyzer system. The cell viability was measured by the MTT (metabolic activity assay. The initial spermatozoa motility showed slightly increased values at doses < 31.20 μmol.dm-3 of CuCl2 compared to the control group. In this time, the lowest spermatozoa motility was recorded significantly (P<0.001 in the group A using the highest dose of CuCl2 (1000 μmol.dm-3. After 1 h of cultivation we proved that the average motility values decreased proportionally to the increasing concentration of CuCl2. The low doses (< 7.80 μmol.dm-3 of CuCl2 increased the spermatozoa motility and concurrently of mitochondrial activity (Time 2 h. The obtained data confirm that Cu (in the form CuCl2 at high doses acts as a toxic element on the spermatozoa motility and it has a destructive effect on the mitochondrial complex, which is necessary for their life processes. The low concentrations (< 7.80 μmol.dm-3 of CuCl2 stimulated the mitochondrial activity of cells and maintained of spermatozoa motility during the short-term of cultivation.

  15. Inositol tri-phosphate inhuman and ascidian spermatozoa.

    Science.gov (United States)

    Tosti, E; Palumbo, A; Dale, B

    1993-05-01

    Using a specific protein binding assay we have shown that a spermatozoon of the ascidian Ciona intestinalis contains 1.58 +/- 0.74 x 10(-19) moles of inositol 1,4,5-tri-phosphate (InsP3), while a human spermatozoon contains 6.4 +/- 0.14 x 10(-19) moles. Induction of the acrosome reaction (AR) in both species, by exposure to the calcium ionophore A23187, does not significantly alter levels of InsP3, suggesting that phosphatidylinositol (PI) turnover is not necessary for the calcium ionophore induced AR. Furthermore, PI turnover in ascidian spermatozoa appears to be insensitive to lithium and phorbol ester. The high intracellular concentration of InsP3 in spermatozoa, corresponding to 50-200 microM, suggests it may play a role in egg activation.

  16. Effect of sex-sorting and cryopreservation on the post-thaw sperm quality of Iberian red deer spermatozoa.

    Science.gov (United States)

    Anel-López, L; García-Álvarez, O; Parrilla, I; Del Olmo, D; Maroto-Morales, A; Fernandez-Santos, M R; Ortiz, J A; Soler, A J; Martínez, E M; Vazquez, J M; Garde, J J

    2017-02-01

    This study investigated the effect of sex-sorting and cryopreservation on post-thaw characteristics and fertility of red deer (Cervus elaphus) sperm for the first time. Semen was collected by electroejaculation from 10 mature stags during the breeding season, and each ejaculate split into four experimental groups: Bulk sorted spermatozoa, sorted but not sexed (BSS); sorted high purity X-spermatozoa (XSS); sorted high purity Y-spermatozoa (YSS); and, control non-sorted spermatozoa (NS). Following, all samples were frozen over liquid nitrogen. Two straws per stag and sample type were analyzed immediately post-thaw and following a 2-h incubation period at 37 °C. Post-thaw total motility (TM) as assessed by CASA was not different (P sperm. For XSS, post-thaw TM was lower (39%, P  0.05) to that of YSS (47%) sperm. The percentage of apoptotic spermatozoa as assessed by PI/YO-PRO-1 and flow cytometry analysis, was higher (17%, P ≤ 0.05) for XSS sperm than NS (12%), BSS (13%) and YSS (14%) sperm. Following incubation there were no differences (P > 0.05) in TM or percent apoptosis among treatments. Post-thaw chromatin stability calculated as the DNA fragmentation index (%DFI) was similar among treatments; following incubation %DFI increased in all except YSS, which displayed the lowest value (P sperm, respectively (P sperm displayed acceptable post-thaw sperm evaluation parameters and the expected offspring sex ratio. More studies are needed to understand the source of sperm damage that may compromise the fertility of Y-sorted red deer sperm.

  17. Effect of Selenium, Zinc, Vitamin C and E on Boar Ejaculate Quality at Heat Stress

    Directory of Open Access Journals (Sweden)

    Pavel Horký

    2016-01-01

    Full Text Available The aim of experiment was to test effect of selected antioxidants (selenium, zinc, vitamin C and E to reduce the impact of heat stress at boars. In the experiment, boars of Duroc breed were tested. The first control group (n = 10 was not supplemented with antioxidants. The second experimental group (n = 10 was supplemented with antioxidants in the following quantities of 0.5 mg of selenium (seleno-methionine, 100 mg of zinc (zinc-methionine, 70 mg of vitamin E (alpha‑tocopherol and 350 mg of vitamin C (ascorbic acid per kilogram of their feed. The experiment was carried out for 120 days and took place in summer (June to September. During the experiment, average and maximum daily temperatures, where boars were stabled, were monitored. Average daily temperature ranged from 12 to 28 °C. Maximum temperature during the day was from 13 to 32 °C. The evaluation of the semen quality has revealed increased number of abnormal spermatozoa in the control group of boars by 39 % (P < 0.05. There were observed no significant changes at other monitored parameters (ejaculate volume, total count of produced sperm, motility and sperm concentration. The results show that the addition of selenium, zinc, vitamin C and E may reduce the effect of heat stress to some extent at breeding boars.

  18. Effect of Three Days Storage of Coated Spermatozoa at Cooling and Adding Seminal Plasma on Ram Fertility

    Directory of Open Access Journals (Sweden)

    Alireza Vaferi

    2016-08-01

    Full Text Available Introduction Artificial insemination (AI has only been used as a supplement to natural mating. AI, when used in conjunction with accurate progeny testing schemes, can substantially increase the rate of genetic progress compared with that of natural service. Moreover, the use of AI causes the limitation of the transmitted diseases. Cervical insemination with frozen-thawed ram semen has not been widely adopted, probably because of the relative poor fertility obtained. Thus using fresh and diluted semen is only approach for performing AI. AI is currently limited by the poor fertility achieved after cervical insemination with the storage of liquid semen at sub-ambient temperature. The success of this procedure in sheep is restricted by the short length of time that ram sperm can be stored in a liquid state. Moreover, the effect of cooling on sperm differs depending on species. It is also well known that ram spermatozoa are more sensitive to cold-shock stress than those of other species. Seminal plasma, as physiological secretion, is a complex mixture of secretions originating from testis, epididymis and accessory sex glands which is mixed with epididymal sperm at ejaculation; it serves as the carrier of sperm to the female genital tract. This mixture contains numerous factors such as organic and nonorganic material which play an important role in the final maturation of the spermatozoa through hormonal, enzymatic and surface-modifying events. During natural mating, a mechanism may be activated to separate spermatozoa from seminal plasma. After being ejaculated into the vagina, sperm swim through cervical mucus and enter the uterus within minutes (>30 min; cervical mucus acts as a barrier for seminal plasma. In the artificial insemination industry, seminal plasma with all the useful and harmful components is not removed from semen and is in contact with sperm throughout cooling, freezing and storage. On the other hand, it was demonstrated that the

  19. Dimethylformamide improves the in vitro characteristics of thawed stallion spermatozoa reducing sublethal damage.

    Science.gov (United States)

    Morillo Rodriguez, A; Balao da Silva, C; Macías-García, B; Gallardo Bolaños, J M; Tapia, J A; Aparicio, I M; Ortega-Ferrusola, C; Peña, F J

    2012-12-01

    A total of 42 ejaculates were used in the experiment; six ejaculates per stallion, obtained from seven Pure Spanish stallions (PRE), were split and frozen in freezing media with different concentrations and combinations of cryoprotectant (CPA): (i) Cáceres (skim milk based extender) containing 2.5% glycerol (2.5GL), (ii) Cáceres containing 1.5% glycerol and 1.5% dimethylformamide (1.5%GL-1.5%DMFA), (iii) Cáceres extender supplemented with 1.5% glycerol and 2.5% dimethylformamide (1.5%GL-2.5%DMFA) and (iv) Cáceres extender supplemented with 4% dimethylformamide (4%DMFA). After at least 4 weeks of storage in liquid nitrogen (LN), straws were thawed and semen analysed by computer-assisted sperm analysis and flow cytometry (membrane lipid architecture (Merocyanine 540), integrity and sublethal damage (YoPro-1) and mitochondrial membrane potential (JC-1)). After thawing, better results were observed in samples frozen in 4%DMFA or in combinations of 1.5%GL-2.5%DMFA, in fact total motility increased by 16% in the 4%DMFA group compared to 2.5%GL (P DFMA had a lower percentage of mitochondria with lower membrane potential. It is concluded that DMFA improves the outcome of cryopreservation of stallion spermatozoa mainly reducing sublethal cryodamage.

  20. Human sperm cells swimming in micro-channels

    CERN Document Server

    Denissenko, Petr; Smith, David; Kirkman-Brown, Jackson

    2012-01-01

    The migratory abilities of motile human spermatozoa in vivo are essential for natural fertility, but it remains a mystery what properties distinguish the tens of cells which find an egg from the millions of cells ejaculated. To reach the site of fertilization, sperm must traverse narrow and convoluted channels, filled with viscous fluids. To elucidate individual and group behaviors that may occur in the complex three-dimensional female tract environment, we examine the behavior of migrating sperm in assorted micro-channel geometries. Cells rarely swim in the central part of the channel cross-section, instead traveling along the intersection of the channel walls (`channel corners'). When the channel turns sharply, cells leave the corner, continuing ahead until hitting the opposite wall of the channel, with a distribution of departure angles, the latter being modulated by fluid viscosity. If the channel bend is smooth, cells depart from the inner wall when the curvature radius is less than a threshold value clo...

  1. Selenium in human male reproductive organs.

    Science.gov (United States)

    Oldereid, N B; Thomassen, Y; Purvis, K

    1998-08-01

    The objective of the study was to obtain information on the concentration and distribution of selenium throughout the human male reproductive tract. Material was removed at autopsy from 41 men who had died suddenly and unexpectedly. Semen samples were also provided from 184 men attending an andrology clinic for fertility investigation and from 32 healthy volunteers. Significant positive correlations in the selenium concentration were demonstrated between the different reproductive organs, the testis having the highest concentrations. No correlation was found between the concentration of selenium in the genital organs and liver, kidney or blood, suggesting that its uptake and/or biochemical activity in the reproductive organs may be controlled by similar mechanisms not shared by the other organs. No significant age-dependent changes could be detected in tissue selenium concentrations. In a group of men under fertility investigation, a significant positive correlation was obtained between seminal plasma concentrations of selenium and concentrations of spermatozoa in the same ejaculate. A significant positive correlation between concentrations of zinc and selenium in the same ejaculates indicated that selenium may arise largely from the prostate gland.

  2. Efficacy of Dapoxetine in the Treatment of Premature Ejaculation

    Science.gov (United States)

    McMahon, Chris G.

    2011-01-01

    Introduction Premature ejaculation (PE) is a common male sexual disorder which is associated with substantial personal and interpersonal negative psychological factors. Pharmacotherapy of PE with off-label antidepressant SSRI drugs is common. Development and regulatory approval of drugs specifically for the treatment of PE will reduce reliance on off-label treatments and serve to fill a unmet treatment need. Aim To review evidence supporting the efficacy and safety of dapoxetine in the treatment of PE. Methods MEDLINE and the proceedings of major international and regional scientific meetings during the period 1994–2010 were searched for publications or abstracts using the word dapoxetine in the title, abstract or keywords. This search was then manually cross-referenced for all papers. This review encompasses studies of dapoxetine pharmacokinetics, animal studies, human phase 1, 2 and 3 efficacy and safety studies and drug-interaction studies. Results Dapoxetine is a potent selective serotonin re-uptake inhibitor, which is administered on-demand 1–3 hours prior to planned sexual contact. Dapoxetine is rapidly absorbed and eliminated, resulting in minimal accumulation and has dose-proportional pharmacokinetics, which are unaffected by multiple dosing. Dapoxetine 30 mg and 60 mg has been evaluated in 5 randomized, double-blind, placebo-controlled studies in 6081 men aged ≥18 years. Outcome measures included stopwatch-measured intravaginal ejaculatory latency time (IELT), Premature Ejaculation Profile (PEP) inventory items, clinical global impression of change (CGIC) in PE, and adverse events. Mean IELT, all PEP items and CGIC improved significantly with both doses of dapoxetine vs. placebo (P < 0.001 for all). The most common treatment related adverse effects included nausea (11.0% for 30 mg, 22.2% for 60 mg), dizziness (586% for 30 mg, 10.9% for 60 mg), and headache (5.6% for 30 mg, 8.8% for 60 mg), and evaluation of validated rated scales demonstrated no SSRI

  3. Current and emerging therapies in premature ejaculation: Where we are coming from, where we are going.

    Science.gov (United States)

    Martin, Christopher; Nolen, Hunter; Podolnick, Jason; Wang, Run

    2017-01-01

    Premature ejaculation is the most common form of sexual dysfunction among men. The pathophysiology of premature ejaculation appears to be multifactorial, implicating the need for multimodal therapeutic regimens to successfully treat premature ejaculation. Multiple treatment regimens have been shown to be effective in extending the time between penetration and ejaculation. These treatment modalities include everything from behavioral modifications and medications to diet alterations and major surgery. The goal of the present article was to review the commonly used treatment regimens used in the treatment of premature ejaculation, as well as to introduce and discuss the newest treatment routines under study for the treatment of premature ejaculation.

  4. Function of ram spermatozoa frozen in diluents supplemented with casein and vegetable oils.

    Science.gov (United States)

    Del Valle, I; Souter, A; Maxwell, W M C; Muiño-Blanco, T; Cebrián-Pérez, J A

    2013-05-01

    The aim of this study was to assess biologically safer components as alternatives to egg yolk for the frozen storage of ram semen using casein, coconut or palm oil in either Salamon's diluent (S) or a swim-up medium (SU). Ejaculates were frozen as pellets and sperm motility (subjectively) and acrosome integrity (FITC-PNA/PI) by flow cytometry were assessed at 0, 3 and 6h after thawing and incubation at 37°C. Three experiments were done: different concentrations of palm oil (5%, 10% and 20%); casein added as emulsifier and protective agent; and differences between egg yolk, coconut and palm oil in S and SU. 20% of oil added to SU accounted for a lesser percentage (Poil levels on viable cells. When casein was added to diluents containing 5% of palm oil, no differences were found between palm or casein (P>0.05). No differences were found when S and SU were compared neither as groups nor between S alone and containing coconut or palm oil; however, SU alone yielded less motility than SU 5% coconut. However, in both groups, S and SU, egg yolk accounted for the greatest values in both bases. These results indicate that none of biologically safer media components (casein, palm or coconut oil) used in this study maintained the function of ram spermatozoa after freeze-thawing better than S-containing egg yolk. The application of vegetable oils as substitutes for egg yolk in diluents for the cryopreservation of ram spermatozoa requires further research.

  5. EFFECT OF NON-ENZYMATIC ANTIOXIDANTS IN EXTENDER ON POST-THAW QUALITY OF BUFFALO (BUBALUS BUBALIS BULL SPERMATOZOA

    Directory of Open Access Journals (Sweden)

    S. M. H. ANDRABI, M. S. ANSARI, N. ULLAH AND M. AFZAL

    2008-12-01

    Full Text Available The objective of the present study was to determine the effect of non-enzymatic antioxidants (vitamin C or E in tris-citric acid buffer (TCA on post-thaw quality of buffalo (Bubalus bubalis bull spermatozoa. Split pooled buffalo bull ejaculates were diluted in TCA egg yolk glycerol extender containing either vitamin C (TCAC, vitamin E (TCAE or without antioxidant (TCAN at 37°C. Extended semen was cooled to 4C in 2 h and equilibrated for 4 h at 4C. Cooled semen was then filled in 0.5 ml straws at 4C and frozen in programmable cell freezer. Thawing of semen was performed at 37°C for 30 seconds. Sperm motility, plasma membrane integrity and sperm morphology (acrosome integrity, head, mid-piece and tail abnormalities of each semen sample were evaluated. Percentage of post-thaw spermatozoal motility assessed visually at 0 and 6 h and the post-thaw percentage of spermatozoa with intact plasma membranes at 0 h were higher (P0.05. In conclusion, non-enzymatic antioxidants, particularly vitamin E, in the tris citric acid extender may improve the quality of frozen-thawed buffalo bull spermatozoa.

  6. How to select immotile but viable spermatozoa on the day of intracytoplasmic sperm injection? An embryologist's view.

    Science.gov (United States)

    Nordhoff, V

    2015-03-01

    A viable spermatozoon is the prerequisite for initiating fertilization in intracytoplasmic sperm injection. Usually motility is the primary sign used to determine a sperm's viability. However, in every in vitro fertilization (IVF) laboratory cases are observed in which none or only few spermatozoa are motile. This can occur in treatment cycles involving ejaculated samples but is most common in cases where surgically extracted testicular spermatozoa are used. To aid in selection, several techniques have been developed to identify viable spermatozoa from the immotile fraction. Amongst the more commonly used approaches are (i) the hypo-osmotic swelling test (ii) chemical substances for induction of tail movements (iii) the sperm tail flexibility test and (iv) laser-assisted immotile sperm selection. All can be used routinely in an IVF laboratory with each having both strengths and weaknesses. It is the purpose of this short review to focus on the technical issues involved in the performance of each of these techniques and to highlight the advantages and disadvantages of each approach.

  7. Acrosin activity in turkey spermatozoa: assay by clinical method and effect of zinc and benzamidine on the activity.

    Science.gov (United States)

    Glogowski, J; Jankowski, J; Faruga, A; Ottobre, J S; Ciereszko, A

    2001-09-15

    We optimized a clinical assay developed for measuring total acrosin activity for mammalian and fish semen for use in turkey spermatozoa. The main modifications included dilution of semen to a final concentration of 25 to 1000 x 10(3) spermatozoa, an increase of Triton X-100 concentration to 0.05% and 1 hr preincubation without substrate, Acrosin activity in turkey spermatozoa was much higher than in human spermatozoa (about 100-times) but similar to that of boar sperm. To optimize this assay for turkey spermatozoa, it was necessary to use higher Triton X-100 concentrations in the reaction mixture. There was a better catalytic efficiency at higher temperatures and a special requirement for a preincubation period for proacrosin activation. We observed high inhibition of acrosin activity by zinc added during preincubation (90% at 0.01 mM of zinc chloride). Benzamidine also inhibited turkey acrosin, and the extent of inhibition was similar for the incubation or preincubation period. When zinc ions were added during incubation, this inhibition was lower (24%). The results suggest that zinc influences proacrosin activation of turkey spermatozoa. This influence may be important for successful long-term storage of spermatozoa in the hen's oviduct.

  8. SPERM HY-LITER™ for the identification of spermatozoa from sexual assault evidence

    DEFF Research Database (Denmark)

    Westring, Christian Gustav; Wiuf, Morten; Nielsen, S Jock;

    2014-01-01

    Accurate microscopic identification of human spermatozoa is important in sexual assault cases. We have compared the results of examinations with (1) a fluorescent microscopy method, SPERM HY-LITER™, and (2) Baecchi's method for identification of human spermatozoa. In 35 artificial, forensic type...... was non-selective. Data from forensic casework samples in Copenhagen from two years (2008 and 2009) are presented. The samples from 2008 were investigated using Baecchi's method, while those from 2009 were investigated using SPERM HY-LITER™. The frequencies of positive results were similar between the two...

  9. Effect of seminal plasma zinc–binding proteins on motility and membrane integrity of canine spermatozoa stored at 5°C

    Directory of Open Access Journals (Sweden)

    Mogielnicka-Brzozowska Marzena

    2014-03-01

    Full Text Available Sperm surface binding sites for non-zinc-binding proteins (nZnBPs and zinc-binding proteins (ZnBPs were studied by the fluorescence technique with biotin-labelled proteins. The nZnBPs binding pattern was unspecific, no characteristic sites on plasmalemma were found. ZnBPs were attached mainly to the acrosomal region of sperm head and to the sperm flagellum. ZnBPs added to the incubation mixture of the canine spermatozoa allowed the preservation of higher values for total motility, progressive motility, curvilinear line velocity, straight line velocity, and beat cross frequency (P < 0.05, both at time 0 and after 1 h incubation at 5ºC. The addition of nZnBPs to the incubation mixture caused only weak positive effects when compared with control sample (PBS. A higher percentage of canine-ejaculated spermatozoa with intact membranes were observed when ejaculate was incubated with ZnBPs in comparison to control sample stored with PBS (P < 0.05 or nZnBPs (P < 0.05. Spermatozoa diluted with ZnBPs and nZnBPs exhibited a higher percentage of cells with active mitochondria when compared with control, both at time 0 and after 1 h; however, no statistical differences were observed. Our results emphasise the role of seminal plasma protein in securing the correct quantity and availability of zinc ions as a component regulating the motility of canine spermatozoa. The protective effect of ZnBPs against the cooling effect may be due to their ability of preventing sperm membrane damage.

  10. Sperm FISH and chromatin integrity in spermatozoa from a t(6;10;11) carrier.

    Science.gov (United States)

    Olszewska, Marta; Huleyuk, Nataliya; Fraczek, Monika; Zastavna, Danuta; Wiland, Ewa; Kurpisz, Maciej

    2014-05-01

    Complex chromosome rearrangements (CCRs) are structurally balanced or unbalanced aberrations involving more than two breakpoints on two or more chromosomes. CCRs can be a potential reason for genomic imbalance in gametes, which leads to a drastic reduction in fertility. In this study, the meiotic segregation pattern, aneuploidy of seven chromosomes uninvolved in the CCR and chromatin integrity were analysed in the ejaculated spermatozoa of a 46,XY,t(6;10;11)(q25.1;q24.3;q23.1)mat carrier with asthenozoospermia and a lack of conception. The frequency of genetically unbalanced spermatozoa was 78.8% with a prevalence of 4:2 segregants of 38.2%, while the prevalence of the adjacent 3:3 mode was 35.3%. Analysis of the aneuploidy of chromosomes 13, 15, 18, 21, 22, X and Y revealed an approximately fivefold increased level in comparison with that of the control group, indicating the presence of an interchromosomal effect. Sperm chromatin integrity status was evaluated using chromomycin A3 and aniline blue staining (deprotamination), acridine orange test and TUNEL assay (sperm DNA fragmentation). No differences were found when comparisons were made with a control group. We suggest that the accumulation of genetically unbalanced spermatozoa, significantly increased sperm aneuploidy level and decreased sperm motility (20%, progressive) were not responsible for the observed lack of reproductive success in the analysed infertile t(6;10;11) carrier. Interestingly, in the case described herein, a high level of sperm chromosomal imbalance appears not to be linked to sperm chromatin integrity status.

  11. Relationship between premature ejaculation and depression

    Science.gov (United States)

    Xia, Yue; Li, Juanjuan; Shan, Guang; Qian, Huijun; Wang, Tao; Wu, Wei; Chen, Jun; Liu, Luhao

    2016-01-01

    Abstract Background: Premature ejaculation (PE) is the most prevalent male sexual dysfunction. Epidemiologic findings are inconsistent concerning the risk for depression associated with PE. Objective: The aim of this study was to investigate the potential association between between depression and risk of PE. Data sources: We conducted a literature search of PubMed, Embase, and the Cochrane Library from these databases’ inception through June 2014 for observational epidemiological studies examining the association between depression on risk of PE. Study eligibility criteria: Studies were selected if they reported the risk estimates for PE associated with depression. Participants: patients>18 years of age suffering from PE. Interventions: a history of depressive disorder. Study appraisal and synthesis methods: These odds ratios (ORs) were pooled using a random or fixed effects model and were tested for heterogeneity. Subgroup analysis was employed to explore heterogeneity. Results: Eight trials involving 18,035 patients were included in the meta-analysis. Depression were statistically significantly associated with the risk of PE (OR = 1.63, 95% CI:1.42–1.87). There was no evidence of between-study heterogeneity (P = 0.623, I2 = 0.0%). The association was similar when stratified by mean age, geographical area, study design, sample size, publication year, and controlling key confounders. Limitations: The severity of depression and PE could not be identified due to unavailable data of trials. No evidence of publication bias was observed. Conclusions: These findings provide evidence that depression is associated with a significantly increased risk of PE. In addition, more prospective studies are necessary to evaluate the association and identify the ideal treatment. Systematic review registration number: CRD42016041272 PMID:27583879

  12. Dorsal penile nerves and primary premature ejaculation

    Institute of Scientific and Technical Information of China (English)

    ZHANG Hai-feng; ZHANG Chun-ying; LI Xing-hua; FU Zhong-ze; CHEN Zhao-yan

    2009-01-01

    Background Based on our clinical experience, the number of dorsal penile nerves in patients with primary premature ejaculation (PPE) is not consistent with the average number (2 branches). In this study, we evaluated the number and distribution of dorsal penile nerves among healthy Chinese adults and patients with PPE.Methods The dorsal nerve of the penis, the deep dorsal vein of the penis, and the dorsal artery of the penis between the deep fascia of the penis and the albuginea penis were carefully educed, observed, and counted in 38 adult autopsy specimens. The number and distribution of the dorsal penile nerve in 128 surgical patients with PPE were determined. Results The numbers of dorsal penile nerves of the 38 cases were as follows:7 branches in 1 case; 6 branches in 1 case; 5 branches in 6 cases; 4 branches in 9 cases; 3 branches in 14 cases; and 2 branches in 7 cases. Most of the dorsal nerves were parallel to each other and in the dorsum of the penis. In only 8 cases, the branches were connected by some communicating branches. In 4 cases, 1 or 2 thin dorsal nerves continued their pathway over the ventral aspect of the penis. The average number of branches of the dorsal penile nerve in patients with PPE was 7.16. Conclusions Based on the study of 38 cases, the average number of dorsal penile nerves was 3.55 branches and that of patients with PPE was greater. These preliminary results suggest that the excessive dorsal penile nerves may have an impact on PPE via increased sensitivity and provide topographic data for the possible treatment of PPE.

  13. Genetic dosage and position effect of small supernumerary marker chromosome (sSMC) in human sperm nuclei in infertile male patient.

    Science.gov (United States)

    Olszewska, Marta; Wanowska, Elzbieta; Kishore, Archana; Huleyuk, Nataliya; Georgiadis, Andrew P; Yatsenko, Alexander N; Mikula, Mariya; Zastavna, Danuta; Wiland, Ewa; Kurpisz, Maciej

    2015-11-30

    Chromosomes occupy specific distinct areas in the nucleus of the sperm cell that may be altered in males with disrupted spermatogenesis. Here, we present alterations in the positioning of the human chromosomes 15, 18, X and Y between spermatozoa with the small supernumerary marker chromosome (sSMC; sSMC(+)) and spermatozoa with normal chromosome complement (sSMC(-)), for the first time described in the same ejaculate of an infertile, phenotypically normal male patient. Using classical and confocal fluorescent microscopy, the nuclear colocalization of chromosomes 15 and sSMC was analyzed. The molecular cytogenetic characteristics of sSMC delineated the karyotype as 47,XY,+der(15)(pter->p11.2::q11.1->q11.2::p11.2->pter)mat. Analysis of meiotic segregation showed a 1:1 ratio of sSMC(+) to sSMC(-) spermatozoa, while evaluation of sperm aneuploidy status indicated an increased level of chromosome 13, 18, 21 and 22 disomy, up to 7 × (2.7 - 15.1). Sperm chromatin integrity assessment did not reveal any increase in deprotamination in the patient's sperm chromatin. Importantly, we found significant repositioning of chromosomes X and Y towards the nuclear periphery, where both chromosomes were localized in close proximity to the sSMC. This suggests the possible influence of sSMC/XY colocalization on meiotic chromosome division, resulting in abnormal chromosome segregation, and leading to male infertility in the patient.

  14. Chloride channels are involved in sperm motility and are downregulated in spermatozoa from patients with asthenozoospermia.

    Science.gov (United States)

    Liu, Shan-Wen; Li, Yuan; Zou, Li-Li; Guan, Yu-Tao; Peng, Shuang; Zheng, Li-Xin; Deng, Shun-Mei; Zhu, Lin-Yan; Wang, Li-Wei; Chen, Li-Xin

    2016-06-03

    Human spermatozoa encounter an osmotic decrease from 330 to 290 mOsm l-1 when passing through the female reproductive tract. We aimed to evaluate the role of chloride channels in volume regulation and sperm motility from patients with asthenozoospermia. Spermatozoa were purified using Percoll density gradients. Sperm volume was measured as the forward scatter signal using flow cytometry. Sperm motility was analyzed using computer-aided sperm analysis (CASA). When transferred from an isotonic solution (330 mOsm l-1 ) to a hypotonic solution (290 mOsm l-1 ), cell volume was not changed in spermatozoa from normozoospermic men; but increased in those from asthenozoospermic samples. The addition of the chloride channel blockers, 4,4'-diisothiocyanatostilbene-2,2'- isulfonic acid (DIDS) or 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) to the hypotonic solution caused the normal spermatozoa to swell but did not increase the volume of those from the asthenozoospermic semen. DIDS and NPPB decreased sperm motility in both sets of semen samples. The inhibitory effect of NPPB on normal sperm motility was much stronger than on spermatozoa from the asthenozoospermic samples. Both sperm types expressed ClC-3 chloride channels, but the expression levels in the asthenozoospermic samples were much lower, especially in the neck and mid-piece areas. Spermatozoa from men with asthenozoospermia demonstrated lower volume regulating capacity, mobility, and ClC-3 expression levels (especially in the neck) than did normal spermatozoa. Thus, chloride channels play important roles in the regulation of sperm volume and motility and are downregulated in cases of asthenozoospermia.

  15. Effects of ejaculation by penile vibratory stimulation on bladder capacity in men with spinal cord lesions

    DEFF Research Database (Denmark)

    Laessøe, Line; Sønksen, Jens; Bagi, Per

    2003-01-01

    We examined the effects of ejaculation by penile vibratory stimulation on bladder capacity in men with spinal cord lesions.......We examined the effects of ejaculation by penile vibratory stimulation on bladder capacity in men with spinal cord lesions....

  16. Morphological characterisation of vesicular structures in the canine ejaculate

    DEFF Research Database (Denmark)

    Goericke-Pesch, Sandra Kathrin; Hauck, S; Bergmann, M

    2015-01-01

    Membrane vesicles (MV) have been identified in seminal plasma from various species and they are thought to have a significant impact on semen quality and fertilisation. Although recently presence of MV has been also described in the canine ejaculate, detailed knowledge on their morphology is miss....... Furthermore, the presence of MV in the castrated azoospermic male confirms an at least partly prostatic origin of canine MV.......Membrane vesicles (MV) have been identified in seminal plasma from various species and they are thought to have a significant impact on semen quality and fertilisation. Although recently presence of MV has been also described in the canine ejaculate, detailed knowledge on their morphology...... normospermic dogs (n=15), hypokinozoospermic dogs (n=2, h) and one castrated azoospermic dog (a). For TEM, a new preparation protocol was used resulting in a higher MV retrieval rate. Using fractionated semen samples, most MV were identified in the second (sperm-rich) fraction in LM. Using pooled ejaculates...

  17. Interrelationships among measures of premature ejaculation: the central role of perceived control.

    Science.gov (United States)

    Patrick, Donald L; Rowland, David; Rothman, Margaret

    2007-05-01

    Domains of premature ejaculation (PE) include short intravaginal ejaculatory latency time (IELT), poor perceived control over ejaculation, decreased satisfaction with sexual intercourse, and personal distress and interpersonal difficulty related to ejaculation. How these measures interrelate is unknown. Here, we evaluated the interrelationships between these PE-specific variables, applying cross-sectional data from a large U.S. observational study of men with PE. We analyzed data from men with PE identified in a previously reported observational study. PE was diagnosed by experienced clinicians using the criteria specified in the Diagnostic and Statistical Manual of Mental Disorders, fourth edition, text revision. Subjects reported their stopwatch-measured IELT, perceived control over ejaculation, satisfaction with sexual intercourse, personal distress related to ejaculation, and interpersonal difficulty related to ejaculation. Relationships between variables were assessed using bivariate correlations, and the strength and significance of direct or indirect effects between variables were evaluated using a form of regression analysis known as path analysis. Bivariate Pearson correlation coefficients for all relationships were significant at the P interpersonal difficulty related to ejaculation. When all variables were included in the model, IELT showed a significant direct effect on perceived control over ejaculation but did not show a significant direct effect on ejaculation-related personal distress or satisfaction with sexual intercourse. Perceived control over ejaculation showed a significant direct effect on both ejaculation-related personal distress and satisfaction with sexual intercourse, which each showed direct effects on interpersonal difficulty related to ejaculation. The patient's perception of control over ejaculation is central to understanding how PE is associated with satisfaction with sexual intercourse and ejaculation-related distress. In contrast

  18. Male masturbation device for the treatment of premature ejaculation

    OpenAIRE

    Rodríguez, J E; López, A.

    2016-01-01

    Objective: To determine the efficiency as a treatment of the first line masturbator aid device for patients with premature ejaculation (PE). Methods: A whole of 18 cases with lifelong PE used a masturbator TENGA, 5 times per week for 6 weeks and a minimum of 5 min for every use. Premature Ejaculation Profile (PEP) was used to measure the effects of its use. As a main outcome measure was used the proportion of patients who achieved criteria for clinical benefit, defined as achieving a two-c...

  19. Healthy offspring from freeze-dried mouse spermatozoa held on the International Space Station for 9 months.

    Science.gov (United States)

    Wakayama, Sayaka; Kamada, Yuko; Yamanaka, Kaori; Kohda, Takashi; Suzuki, Hiromi; Shimazu, Toru; Tada, Motoki N; Osada, Ikuko; Nagamatsu, Aiko; Kamimura, Satoshi; Nagatomo, Hiroaki; Mizutani, Eiji; Ishino, Fumitoshi; Yano, Sachiko; Wakayama, Teruhiko

    2017-06-06

    If humans ever start to live permanently in space, assisted reproductive technology using preserved spermatozoa will be important for producing offspring; however, radiation on the International Space Station (ISS) is more than 100 times stronger than that on Earth, and irradiation causes DNA damage in cells and gametes. Here we examined the effect of space radiation on freeze-dried mouse spermatozoa held on the ISS for 9 mo at -95 °C, with launch and recovery at room temperature. DNA damage to the spermatozoa and male pronuclei was slightly increased, but the fertilization and birth rates were similar to those of controls. Next-generation sequencing showed only minor genomic differences between offspring derived from space-preserved spermatozoa and controls, and all offspring grew to adulthood and had normal fertility. Thus, we demonstrate that although space radiation can damage sperm DNA, it does not affect the production of viable offspring after at least 9 mo of storage on the ISS.

  20. Expression and Subcellular Localization of Retinoic Acid Receptor-α (RARα) in Healthy and Varicocele Human Spermatozoa: Its Possible Regulatory Role in Capacitation and Survival.

    Science.gov (United States)

    Perrotta, Ida; Perri, Mariarita; Santoro, Marta; Panza, Salvatore; Caroleo, Maria C; Guido, Carmela; Mete, Annamaria; Cione, Erika; Aquila, Saveria

    2015-01-01

    Varicocele, an abnormal tortuosity and dilation of veins of the pampiniform plexus, is the most common identifiable and correctable cause of male infertility. It is now becoming apparent that signaling through vitamin A metabolites, such as all-trans retinoic acid (ATRA), is indispensable for spermatogenesis and disruption of retinoic acid receptor-α (RARα) function may result in male sterility and aberrant spermatogenesis. Herein, we investigated by Western blot and immunogold electron microscopy the expression profiles and subcellular localization of RARα in healthy and varicocele human sperm; in addition, we analyzed the effects of ATRA on cholesterol efflux and sperm survival utilizing enzymatic colorimetric CHOD-PAP method and Eosin Y technique, respectively. In varicocele samples, a strong reduction of RARα expression was observed. Immunogold labeling evidenced cellular location of RARα also confirming its reduced expression in "varicocele" samples. Sperm responsiveness to ATRA treatment was reduced in varicocele sperm. Our study showed that RARα is expressed in human sperm probably with a dual role in promoting both cholesterol efflux and survival. RARα might be involved in the pathogenesis of varicocele as its expression is reduced in pathologic samples. Thus, ATRA administration in procedures for artificial insemination or dietary vitamin A supplementation might represent a promising therapeutic approach for the management of male infertility.

  1. Bidirectional swimming in spermatozoa of Tephritid flies.

    Science.gov (United States)

    Baccetti, B; Gibbons, B H; Gibbons, I R

    1989-10-01

    Our observations show that spermatozoa of the Mediterranean fruit fly Ceratitis capitata and of Dacus oleae and Dacus dorsalis are capable of swimming backwards as well as forwards, and that they can change direction abruptly. The preferred direction is backwards, observed in spermatozoa obtained from the male genitalia. Forwards swimming spermatozoa were frequently seen in the spermatheca and close to the eggs. The change in swimming direction appears to be effected solely by a change in the direction of bend propagation, with no significant change in other waveform parameters. In vitro reactivated spermatozoa swim forwards only and require a minimum free Ca++ concentration of about 10(-6) M for movement. A switching of wave propagation from one direction to the other under control of intracellular free Ca++ concentration is suggested. Perhaps the backwards movement allows easier delivery of spermatozoa from the common envelope embedding the heads in the male apparatus, and assures a more efficient movement of the sperm towards the egg, especially given the enormous relative length of the head. The forwards movement is favoured in order to orient the sperm for penetration of the micropile.

  2. AB087. The expression of cysteine-rich secretory protein 2 (CRISP2) and its specific regulator miR-27b in the spermatozoa of patients with asthenozoospermia

    Science.gov (United States)

    Zhou, Junhao; Zhou, Qizhao; Lyu, Xiaoming; Zhu, Ting; Chen, Zijian; Chen, Mingkun; Xia, Hui; Wang, Chunyan; Qi, Tao; Li, Xin; Liu, Cundong

    2016-01-01

    Objective Cysteine-rich secretory protein 2 (CRISP2) is an important sperm protein and plays roles in spermatogenesis, modulation of flagellar motility, acrosome reaction, and gamete fusion. Clinical evidence shows a reduced CRISP2 expression in spermatozoa from asthenozoospermic patients, but the molecular mechanism underlying its reduction remains unknown. Herein, we carried out a study focusing on the CRISP2 reduction and its roles in asthenozoospermia. Methods Spermatozoa were isolated from 90 study subjects’ ejaculated semen samples. DNA methylation was evaluated using bisulfite-sequencing PCR and methylation-specific PCR. The CRISP2 mRNA and protein expression levels were examined in the ejaculated spermatozoa by qRT-PCR and Western blot respectively. MiRNA expression was detected by qRT-PCR. The direct regulatory effect of miR-27b on CRISP2 was predicted computationally and validated via luciferase reporter assay and in vitro experiments in which miR-27b mimic or inhibitor was transfected into 293T cells. Respective correlations of miR-27b and CRISP2 protein expression with clinical features were analyzed using Spearman’s correlation coefficient. Results Initially, low expression of CRISP2 protein rather than its mRNA was observed in the ejaculated spermatozoa from asthenozoospermic patients relative to normozoospermic males. Meanwhile, methylation was not found in CRISP2 promoter. These data suggest a possible post-transcriptional regulation of CRISP2 in asthenozoospermia. Subsequently, bioinformatics prediction, luciferase reporter assay and miR-27b transfection experiments revealed that miR-27b could specifically target CRISP2 by binding its 3’-UTR, suppressing CRISP2 expression post-transcriptionally. Further evidence was provided by the clinical observation of a high miR-27b expression in the ejaculated spermatozoa from asthenozoospermic patients and a negative correlation between miR-27b and CRISP2 protein expression. Finally, a retrospective

  3. Incorporation of nanoparticles within mammalian spermatozoa using in vitro capacitation

    Science.gov (United States)

    There is still much unknown about the journey of spermatozoa within the female genital tract. Recent studies have investigated mammalian spermatozoa labeling with fluorescent quantum dot nanoparticles (QD) for non-invasive imaging. Furthermore, the incorporation of these QD within the spermatozoa ma...

  4. Identification of the inorganic pyrophosphate metabolizing, ATP substituting pathway in mammalian spermatozoa.

    Directory of Open Access Journals (Sweden)

    Young-Joo Yi

    Full Text Available Inorganic pyrophosphate (PPi is generated by ATP hydrolysis in the cells and also present in extracellular matrix, cartilage and bodily fluids. Fueling an alternative pathway for energy production in cells, PPi is hydrolyzed by inorganic pyrophosphatase (PPA1 in a highly exergonic reaction that can under certain conditions substitute for ATP-derived energy. Recombinant PPA1 is used for energy-regeneration in the cell-free systems used to study the zymology of ATP-dependent ubiquitin-proteasome system, including the role of sperm-borne proteasomes in mammalian fertilization. Inspired by an observation of reduced in vitro fertilization (IVF rates in the presence of external, recombinant PPA1, this study reveals, for the first time, the presence of PPi, PPA1 and PPi transporter, progressive ankylosis protein ANKH in mammalian spermatozoa. Addition of PPi during porcine IVF increased fertilization rates significantly and in a dose-dependent manner. Fluorometric assay detected high levels of PPi in porcine seminal plasma, oviductal fluid and spermatozoa. Immunofluorescence detected PPA1 in the postacrosomal sheath (PAS and connecting piece of boar spermatozoa; ANKH was present in the sperm head PAS and equatorial segment. Both ANKH and PPA1 were also detected in human and mouse spermatozoa, and in porcine spermatids. Higher proteasomal-proteolytic activity, indispensable for fertilization, was measured in spermatozoa preserved with PPi. The identification of an alternative, PPi dependent pathway for ATP production in spermatozoa elevates our understanding of sperm physiology and sets the stage for the improvement of semen extenders, storage media and IVF media for animal biotechnology and human assisted reproductive therapies.

  5. Fusion of artificial membranes with mammalian spermatozoa. Specific involvement of the equatorial segment after acrosome reaction.

    Science.gov (United States)

    Arts, E G; Kuiken, J; Jager, S; Hoekstra, D

    1993-11-01

    The fusogenic properties of bovine and human spermatozoa membranes were investigated, using phospholipid bilayers (liposomes) as target membranes. Fusion was monitored by following lipid mixing, as revealed by an assay based on resonance-energy transfer. In addition, fusion was visualized by fluorescence microscopy, using fluorescent lipid vesicles. Cryopreserved bovine sperm fused with liposomes before induction of the acrosome reaction, fluorescence being located in essentially all spermatozoa membrane domains. Fresh bovine and human spermatozoa fused with liposomes only after the induction of the acrosome reaction, as triggered by calcium ionophore A23187 or zonae pellucidae (proteins), while the fluorescence distribution was mainly restricted to the equatorial segment (ES). However, with spermatozoa that had undergone a freeze/thawing cycle, domains other than ES also became labeled. Hence, the redistribution of the lipid probes over the entire membrane occurring during lipid mixing with cryopreserved bovine sperm is probably related to membrane perturbations caused by long-term cryopreservation. Fusion with liposomes was governed by spermatozoa factors and required the presence of acidic phospholipids like cardiolipin and phosphatidylserine in the liposomal bilayer. Incorporation of the zwitterionic lipid phosphatidylcholine in the vesicles inhibited the fusion reaction. Fusion was pH dependent. The results indicate that the ES is the primary domain of spermatozoa membranes that harbours the fusogenic capacity of sperm. Liposomes appear a valuable tool in further characterizing the properties of this domain, which has been claimed [Yanagimachi, R. (1988) in The physiology of reproduction (Knobil, E. & Neill, J., eds) pp. 135-185, Raven Press, New York] to represent the putative, initial fusion site for the oocyte.

  6. Role of C-type natriuretic peptide in the function of normal human sperm

    Directory of Open Access Journals (Sweden)

    Hui Xia

    2016-01-01

    Full Text Available C-type natriuretic peptide (CNP is a newly discovered type of local regulatory factor that mediates its biological effects through the specific, membrane-bound natriuretic peptide receptor-B (NPR-B. Recent studies have established that CNP is closely related to male reproductive function. The aims of this study were to determine the distribution of CNP/NPR-B in human ejaculated spermatozoa through different methods (such as immunolocalization, real time polymerase chain reaction and Western Blot, and then to evaluate the influence of CNP on sperm function i n vitro, such as motility and acrosome reaction. Human semen samples were collected from consenting donors who met the criteria of the World Health Organization for normozoospermia. Our results show that the specific receptor NPR-B of CNP is localized in the acrosomal region of the head and the membrane of the front-end tail of the sperm, and there is no signal of CNP in human sperm. Compared with the control, CNP can induce a significant dose-dependent increase in spermatozoa motility and acrosome reaction. In summary, CNP/NPR-B can affect sperm motility and acrosome reaction, thus regulating the reproductive function of males. CNP may be a new key factor in regulating sperm function.

  7. Influence of chemical elements on mammalian spermatozoa.

    Science.gov (United States)

    Marzec-Wróblewska, U; Kamiński, P; Lakota, P

    2012-01-01

    Exposure to heavy metals is the most important risk factor in the assessment of spermatogenesis. About 30-40 % cases of infertility are caused by the male factor, and most of them are due to the small quantity of spermatozoa or to inferior spermatozoa quality. The negative impact on sperm motility, morphology and concentration of such chemical elements as Al, Cr, Cd, Pb or Fe was observed, while positive influence was noticed for Zn, Mg, and Ca. The influence of Mn, Cu, Ni or Se on spermatozoa is ambiguous. Chemical elements known as necessary for capacitation and acrosome reaction are Zn, Mg and Ca, while Cd and Pb disturb initiation and progress of the acrosome reaction. The positive effect of chemical elements Al, Cd, Cr, Cu, Ni, Pb, Se, and Zn, lies in their protection against oxidative stress. On the other hand, Al, Cu and Ni induce structural changes in the testes and epididymis or influence interactions with other chemical elements.

  8. The nitric oxide synthase of mouse spermatozoa.

    Science.gov (United States)

    Herrero, M B; Goin, J C; Boquet, M; Canteros, M G; Franchi, A M; Perez Martinez, S; Polak, J M; Viggiano, J M; Gimeno, M A

    1997-07-01

    Nitric oxide synthase (NOS) was evidenced in mature mouse spermatozoa by means of biochemical techniques and Western blot. During 120 min of incubation, 10(7) spermatozoa synthesized 7 +/- 2 pmol of L-[14C]citrulline. Besides, L-citrulline formation depended on the incubation time and on the concentration of L-arginine present in the incubation medium. Different concentrations of N(G)-nitro-L-arginine methyl ester (L-NAME) but not aminoguanidine, inhibited L-[14C]citrulline formation. Western-blot analysis of solubilized sperm proteins revealed a unique band of M(r)=140 kDa with the neural, endothelial and inducible NOS antisera tested. These results provide evidence that mature mouse sperm contains a NOS isoform and that spermatozoa have the potential ability to synthesize NO, suggesting a role for endogenous NO on mammalian sperm function.

  9. Patient reported outcomes in the assessment of premature ejaculation.

    Science.gov (United States)

    Althof, Stanley E

    2016-08-01

    The term 'Patient Reported Outcome', abbreviated as PRO, was introduced by the US Food and Drug Administration (FDA) which proposed guidance on the development and validation of PROs. Previously PROs were known as self-report diaries, event-logs, self-administered questionnaires, and clinician administered rating scales. PROs seek to capture the subjective perceptions of patients and/or partner's related to their specific symptoms, degree of bother, efficacy of a medication or psychotherapy intervention, and quality of life issues related to a specific condition. This article reviews the essential psychometric and regulatory agency requirements in the development of PROs. The constructs of reliability, various forms of validity, sensitivity, and specificity as well as concerns with translating a PRO into a different language are reviewed. Three PROs, the Premature Ejaculation Profile (PEP), the Index of Premature Ejaculation (IPE) and the Premature Ejaculation Diagnostic Tool (PEDT) all used in the assessment of premature ejaculation (PE) are discussed. These questionnaires meet or exceed all the psychometric requirements and have been employed in clinical trials and observational studies of men with PE. The article concludes on discussing some of the limitations of PRO use and recommendations for the future.

  10. Retrograde ejaculation and sexual dysfunction in men with diabetes mellitus

    DEFF Research Database (Denmark)

    Fedder, J; Kaspersen, Maja Døvling; Brandslund, I;

    2013-01-01

    Retrograde ejaculation (RE) and erectile dysfunction may be caused by diabetes mellitus (DM), but the prevalence of RE among DM patients is unknown. A prospective, blinded case-control study comparing men with DM with matched controls according to RE and erectile dysfunction was performed. Twenty...

  11. A NEW METHOD TO DETECT ACROSOME-REACTED SPERMATOZOA USING BIOTINYLATED SOYBEAN TRYPSIN-INHIBITOR

    NARCIS (Netherlands)

    ARTS, EGJM; KUIKEN, J; JAGER, S

    1994-01-01

    Objective: To develop a method to detect acrosome-reacted spermatozoa on human zonae pellucidae using only commercially available reagents and without need for sperm fixation. Design: Sperm head labeling with biotinylated soybean trypsin inhibitor (SBTI-biotin) was compared with results of a known m

  12. FUSION OF ARTIFICIAL MEMBRANES WITH MAMMALIAN SPERMATOZOA - SPECIFIC INVOLVEMENT OF THE EQUATORIAL SEGMENT AFTER ACROSOME REACTION

    NARCIS (Netherlands)

    ARTS, EGJM; KUIKEN, J; JAGER, S; HOEKSTRA, D

    1993-01-01

    The fusogenic properties of bovine and human spermatozoa membranes were investigated, using phospholipid bilayers (liposomes) as target membranes. Fusion was monitored by following lipid mixing, as revealed by an assay based on resonance-energy transfer. In addition, fusion was visualized by fluores

  13. Cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa in a chemically defined extender

    Institute of Scientific and Technical Information of China (English)

    Ya-Hui Li; Ke-Jun Cai; Lei Su; Mo Guan; Xie-Chao He; Hong Wang; Andras Kovacs; Wei-Zhi Ji

    2005-01-01

    Aim: To establish a method for cynomolgus monkey sperm cryopreservation in a chemically defined extender.Methods: Semen samples were collected by electro-ejaculation from four sexually mature male cynomolgus monkeys.The spermatozoa were frozen in straws by liquid nitrogen vapor using egg-yolk-free Tes-Tris (mTTE) synthetic extender and glycerol as cryoprotectant. The effects of glycerol concentration (1%, 3%, 5%, 10% and 15% [v/v])and its equilibration time (10 min, 30 min, 60 min and 90 min) on post-thaw spermatozoa were examined by sperm motility and sperm head membrane integrity. Results: The post-thaw motility and head membrane integrity of spermatozoa were significantly higher (P < 0.05) for 5% glycerol (42.95 ± 2.55 and 50.39 ± 2.42, respectively) than those of the other groups (1%: 19.19 ± 3.22 and 24.84 ± 3.64; 3%: 34.23 ± 3.43 and 41.37 ± 3.42; 10%:15.68 ± 2.36 and 21.39 ± 3.14; 15%: 7.47 ± 1.44 and 12.90 ± 2.18). The parameters for 30 min equilibration (42.95 ± 2.55 and 50.39 ± 2.42) were better (P < 0.05) than those of the other groups (10 min: 31.33 ± 3.06 and 38.98 ± 3.31; 60 min: 32.49 ± 3.86 and 40.01 ± 4.18; 90 min: 31.16 ± 3.66 and 38.30 ± 3.78). Five percent glycerol and 30 min equilibration yielded the highest post-thaw sperm motility and head membrane integrity. Conclusion:Cynomolgus monkey spermatozoa can be successfully cryopreserved in a chemically defined extender, which is related to the concentration and the equilibration time of glycerol.

  14. Premature Ejaculation, Selective Serotonin Reuptake Inhibitor (ssri) Induced Delayed Ejaculation In The Framework Of The Emotional Motor System

    NARCIS (Netherlands)

    Waldinger, Marcel D; Berendsen, Hemmie H.G.; Blok, Bertil F.M.; Olivier, Berend; Holstege, Gert

    1998-01-01

    Premature ejaculation has generally been considered a psychosexual disorder with psychogenic aetiology. Although still mainly treated by behavioural therapy, in recent years double-blind studies have indicated the beneficial effects of some of the serotonergic anti depressants (SSRIs) in delaying ej

  15. Fecal endocrine profiles and ejaculate traits in black-footed cats (Felis nigripes) and sand cats (Felis margarita).

    Science.gov (United States)

    Herrick, J R; Bond, J B; Campbell, M; Levens, G; Moore, T; Benson, K; D'Agostino, J; West, G; Okeson, D M; Coke, R; Portacio, S C; Leiske, K; Kreider, C; Polumbo, P J; Swanson, W F

    2010-01-15

    Information regarding the reproductive biology of black-footed cats (BFC) and sand cats (SC) is extremely limited. Our objectives were to: (1) validate fecal hormone analysis (estrogens, E; progestagens, P; androgens, T) for noninvasive monitoring of gonadal activity; (2) characterize estrous cyclicity, ovulatory mechanisms, gestation, and seasonality; and (3) evaluate male reproductive activity via fecal androgen metabolites and ejaculate traits. In both species, the estrous cycle averaged 11-12 days. In BFC (n=8), estrus lasted 2.2+/-0.2 days with peak concentrations of E (2962.8+/-166.3 ng/g feces) increasing 2.7-fold above basal concentrations. In SC (n=6), peak concentrations of E (1669.9+/-83.5 ng/g feces) during estrus (2.9+/-0.2 days) were 4.0-fold higher than basal concentrations. Nonpregnant luteal phases occurred in 26.5% (26 of 98) of BFC estrous cycles, but were not observed in SC (0 of 109 cycles). In both species, P concentrations during pregnancy were elevated (32.3+/-3.0 microg/g feces BFC; 8.5+/-0.7 microg/g feces SC) approximately 10-fold above basal concentrations. Fecal T concentrations in males averaged 3.1+/-0.1 microg/g feces in BFC and 2.3+/-0.0 microg/g feces in SC. Following electroejaculation, 200 to 250 microl of semen was collected containing 29.9 (BFC) to 36.5 (SC)x10(6) spermatozoa with 40.4 (SC) to 46.8 (BFC)% normal morphology. All females exhibited estrous cycles during the study and spermatozoa were recovered from all males on every collection attempt, suggesting poor reproductive success in these species may not be due to physiological infertility.

  16. Oral Sex, Semen Displacement, and Sexual Arousal: Testing the Ejaculate Adjustment Hypothesis

    Directory of Open Access Journals (Sweden)

    Michael N. Pham

    2013-07-01

    Full Text Available Male Indian Flying Foxes (Pteropus giganteus that spend more time performing oral sex on a female also spend more time copulating with her. In humans, men who spend more time copulating with their regular partner also perform more “semen-displacing” copulatory behaviors (e.g., deeper, more vigorous penile thrusting. We investigated whether men who spend more time performing oral sex on their regular partner also spend more time copulating with her and perform more semen-displacing copulatory behaviors. We proposed and tested the ejaculate adjustment hypothesis for men's copulatory behaviors: Men adjust their copulatory behaviors to increase their sexual arousal and consequent ejaculate quality, thereby increasing their chances of success in sperm competition. Two hundred and thirty-three men in a committed, heterosexual relationship responded to questions about their copulatory behavior and sexual arousal during their most recent sexual encounter with their long-term partner. The results indicated that men who spend more time performing oral sex on their partner also spend more time copulating with her, perform more semen-displacing copulatory behaviors, and report greater sexual arousal. We discuss limitations to the current research and highlight the heuristic value of sperm competition theory for understanding human sexual behaviors.

  17. In vitro effects of nonylphenol on motility, mitochondrial, acrosomal and chromatin integrity of ram and boar spermatozoa.

    Science.gov (United States)

    Uguz, C; Varisli, O; Agca, C; Evans, T; Agca, Y

    2015-10-01

    The objective of this study was to determine the effects of nonylphenol (NP) on viability of ram and boar sperm in vitro. Ram or boar spermatozoa were exposed to 1, 10, 100, 250 and 500 μg NP ml(-1) for 1, 2, 3 or 4 h. Computer-assisted sperm motility analysis (CASA) system was used to evaluate sperm motility characteristics. Flow cytometry was used to determine mitochondrial membrane potential (MMP) and chromatin integrity, while epifluorescent microscopy was used to determine sperm acrosomal status. Exposure of both species spermatozoa to 250 and 500 μg NP ml(-1) was detrimental to progressive motility (P boar spermatozoa with high MMP declined drastically after exposures to ≥250 μg ml(-1) NP (P boar spermatozoa and 10 μg ml(-1) NP for ram spermatozoa. These data show adverse effects of NP on ram and boar spermatozoa and thus its potential harmful effects on male reproduction as NP is found in fruits, vegetables, human milk, fish and livestock products.

  18. How is a giant sperm ejaculated? Anatomy and function of the sperm pump, or "Zenker organ," in Pseudocandona marchica (Crustacea, Ostracoda, Candonidae)

    Science.gov (United States)

    Yamada, Shinnosuke; Matzke-Karasz, Renate

    2012-07-01

    `Giant sperm', in terms of exceptionally long spermatozoa, occur in a variety of taxa in the animal kingdom, predominantly in arthropod groups, but also in flatworms, mollusks, and others. In some freshwater ostracods (Cypridoidea), filamentous sperm cells reach up to ten times the animal's body length; nonetheless, during a single copulation several dozen sperm cells can be transferred to the female's seminal receptacle. This highly effective ejaculation has traditionally been credited to a chitinous-muscular structure within the seminal duct, which has been interpreted as a sperm pump. We investigated this organ, also known as the Zenker organ, of a cypridoid ostracod, Pseudocandona marchica, utilizing light and electron microscope techniques and produced a three-dimensional reconstruction based on serial semi-thin histological sections. This paper shows that numerous muscle fibers surround the central tube of the Zenker organ, running in parallel with the central tube and that a thin cellular layer underlies the muscular layer. A cellular inner tube exists inside the central tube. A chitinous-cellular structure at the entrance of the organ has been recognized as an ejaculatory valve. In male specimens during copulation, we confirmed a small hole derived from the passage of a single spermatozoon through the valve. The new data allowed for proposing a detailed course of operation of the Zenker organ during giant sperm ejaculation.

  19. Seasonal variations of cauda epididymal spermatozoa of bucks

    Directory of Open Access Journals (Sweden)

    Dilip Kumar Swain

    2016-09-01

    Full Text Available Objective: The study was conducted to evaluate the influence of season on cauda epididymal spermatozoa isolated from bucks. Materials and methods: Testes of 30 mature bucks were collected from local slaughter house, and were processed for the retrieval of cauda epididymal spermatozoa for evaluation. Testes were collected in three seasons (winter, summer and rainy, and each season was having 10 pairs of testicles. Recovered spermatozoa from the cauda epididymis were processed immediately for evaluation of semen attributes (Spermatozoa motility, viability, plasma membrane integrity, acrosomal status and DNA integrity. Results: Physiological effect of season was observed on progressive motility, percent of live spermatozoa, spermatozoal membrane integrity (HOST, acrosomal integrity, capacitation status and DNA integrity. Progressive motility, percent live spermatozoa, HOST positive spermatozoa, were found significantly (P<0.05 high in summer season, whereas, significantly (P<0.05 lower comet positive spermatozoa were found in summer season as compared to rainy and winter. Compromised acrosomal status was seen in winter and rainy seasons as compared to summer. Conclusion: Compromised acrosome along with plasma membrane and higher percentage of spermatozoa with damaged DNA in cauda spermatozoa were observed during winter and rainy seasons as compared to summer season. Summer season was found to be the most suitable season for collection of cauda epididymal spermatozoa and can effectively be used for assisted reproduction with further investigations of associated mechanisms. [J Adv Vet Anim Res 2016; 3(3.000: 263-267

  20. PERUBAHAN VIABILITAS DAN STRUKTUR SUBSELULER SPERMATOZOA DOMBA SETELAH PENGERINGBEKUAN

    Directory of Open Access Journals (Sweden)

    Takdir Saili

    2009-12-01

    Full Text Available Several methods i.e. cooling, freezing, and freeze-drying have been widely used to preserve spermatozoa with various degree of success. Freeze-drying appears to provide a method to preserve spermatozoa in a dry state without requiring liquid nitrogen for storing frozen spermatozoa. Freeze-drying procedures can have a detrimental effect on plasma membrane and acrosomal cap of the spermatozoa. In this experiment study, the viability and subcellular changes of freeze-dried ram spermatozoa were evaluated using staining method and scanning electron microscopy. The results revealed that all freeze-dried spermatozoa were dead following evaluation using eosin staining and Hoechst-propidium iodide staining methods. Morover, plasma membrane and acrosomal cap of freeze-dried ram spermatozoa was disrupted observed using scanning electron microscope.

  1. The 5-HT₁A receptor CG polymorphism influences the intravaginal ejaculation latency time in Dutch Caucasian men with lifelong premature ejaculation

    NARCIS (Netherlands)

    Janssen, Paddy K C; van Schaik, R; Zwinderman, Aeilko H; Olivier, Berend; Waldinger, Marcel D

    2014-01-01

    INTRODUCTION: Lifelong premature ejaculation (LPE) is characterized by persistent intravaginal ejaculation latency times (IELTs) of less than 1 min, and has been postulated as a neurobiological dysfunction related to diminished serotonergic neurotransmission with 5-HT₁A receptor hyperfunction and 5-

  2. The 5-HT2C receptor gene Cys23Ser polymorphism influences the intravaginal ejaculation latency time in Dutch Caucasian men with lifelong premature ejaculation

    NARCIS (Netherlands)

    Janssen, Paddy Kc; Schaik, Ron van; Olivier, Berend; Waldinger, Marcel D

    2014-01-01

    It has been postulated that the persistent short intravaginal ejaculation latency time (IELT) of men with lifelong premature ejaculation (LPE) is related to 5-hydroxytryptamine (HT)2C receptor functioning. The aim of this study was to investigate the relationship of Cys23Ser 5-HT2C receptor gene pol

  3. Serotonin Transporter Promoter Region (5-HTTLPR) Polymorphism Is Not Associated With Paroxetine-Induced Ejaculation Delay in Dutch Men With Lifelong Premature Ejaculation

    NARCIS (Netherlands)

    Janssen, Paddy K C; Zwinderman, Aeilko H; Olivier, Berend; Waldinger, Marcel D

    2014-01-01

    PURPOSE: To investigate the association between the 5-HT-transporter-gene-linked promoter region (5-HTTLPR) polymorphism and 20-mg paroxetine-induced ejaculation delay in men with lifelong premature ejaculation (LPE). MATERIALS AND METHODS: This was a prospective study of 10 weeks of paroxetine trea

  4. Relationship between premature ejaculation and depression in Korean males.

    Science.gov (United States)

    Son, Hwancheol; Song, Sang Hoon; Lee, Jun-Young; Paick, Jae-Seung

    2011-07-01

    The psychological impacts of premature ejaculation (PE), which include guilt, anxiety, and distress, have been well established in Western countries. However, in Asia, although a substantial number of epidemiological studies have surveyed the prevalence of PE, researchers have not thoroughly investigated the relationship between PE and depression, or have defined PE properly. We studied the association between PE and depression and other psychological disturbances, in a Korean cohort by applying an appropriate definition for PE and validated outcome measures of depression. METHODS.  A total of 956 males (≥20 years) were initially approached via an Internet survey company. Participants were asked to complete a questionnaire requesting detailed medical and sexual histories, which included questions from the Erectile Function Domain score in the International Index of Erectile Function (IIEF-EF) and the Beck Depression Inventory (BDI). The prevalence of PE was evaluated using two different definitions-self-assessed PE and presumed PE. Presumed PE was defined as a short ejaculation time (an estimated intravaginal ejaculatory latency time ≤5 minutes), an inability to control ejaculation, and the presence of distress resulting from PE. Ejaculation-related questionnaire, the IIEF-EF, and BDI. A total of 334 men were evaluated. The prevalence of PE was 10.5% according to the Presumed PE definition, whereas by self-assessment, it was 25.4%. Self-assessed PE patients suffered from various psychological problems, such as depression, low self-esteem, bother, and low sexual satisfaction. Even after excluding erectile dysfunction (ED) subjects, a significant relationship was found between self-assessed PE and depression. Moreover, after further classification of the Self-assessed PE group, we found that subjects included in this group, but not in the Presumed PE group, suffered more from psychological burden than any other members of the cohort. Korean men with subjective

  5. Behavioral Therapies for Management of Premature Ejaculation: A Systematic Review

    Directory of Open Access Journals (Sweden)

    Katy Cooper, PhD

    2015-09-01

    Conclusions: There is limited evidence that physical behavioral techniques for PE improve IELT and other outcomes over waitlist and that behavioral therapies combined with drug treatments give better outcomes than drug treatments alone. Further RCTs are required to assess psychotherapeutic approaches to PE. Cooper K, Martyn‐St James M, Kaltenthaler E, Dickinson K, Cantrell A, Wylie K, Frodsham L, and Hood C. Behavioral therapies for management of premature ejaculation: A systematic review. Sex Med 2015;3:174–188.

  6. Selective resection of dorsal nerves of penis for premature ejaculation.

    Science.gov (United States)

    Zhang, G-X; Yu, L-P; Bai, W-J; Wang, X-F

    2012-12-01

    Premature ejaculation (PE) is one of the most prevalent male sexual dysfunctions. Selective resection of the dorsal nerve (SRDN) of penis has recently been used for the treatment of PE and has shown some efficacy. To further clarify the efficacy and safety of SRDN on PE, we performed a preliminary, randomized, placebo-controlled clinical observational study. Persons with the complaints of rapid ejaculation, asking for circumcision because of redundant foreskin, intravaginal ejaculation latency time (IELT) within 2 min, not responding to antidepressant medication or disliking oral medication were randomly enrolled in two groups. From April 2007 to August 2010, a total of 101 eligible persons were enrolled, 40 of them received SRDN which dorsal nerves of the penis were selectively resected, and those (n = 61) enrolled in the control group were circumcised only. IELT and the Brief Male Sexual Function Inventory (BMSFI) questionnaire were implemented pre- and post-operatively for the evaluation of the effect and safety of the surgery. There are no statistically significant differences in the baseline data including mean ages, mean IELTs, perceived control abilities and the BMSFI mean scores between the two groups. With regard to the post-operative data of the surgery, both IELTs and perceived control abilities were significantly increased after SRDN (1.1 ± 0.9 min vs. 3.8 ± 3.1 min for pre- and post-operative IELT, respectively, p 0.05). Also, there were no statistically significant differences both in BMSFI composite and subscale scores between the two groups after surgery. Hence, we conclude that SRDN is effective in delaying ejaculation and improving ejaculatory control, whereas erectile function is not affected. The results imply that SRDN may be an alternative method for the treatment of PE for some patients.

  7. Oxidative phosphorylation is essential for felid sperm function, but is substantially lower in cheetah (Acinonyx jubatus) compared to domestic cat (Felis catus) ejaculate.

    Science.gov (United States)

    Terrell, Kimberly A; Wildt, David E; Anthony, Nicola M; Bavister, Barry D; Leibo, S P; Penfold, Linda M; Marker, Laurie L; Crosier, Adrienne E

    2011-09-01

    Compared with the normospermic domestic cat, sperm metabolic function is compromised in the teratospermic cat and cheetah, but the pathway(s) involved in this deficiency are unknown. Glycolysis is essential for sperm motility, yet it appears to function normally in spermatozoa of either species regardless of structural morphology. We conducted a comparative study to further understand the mechanisms of energy production in felid spermatozoa, with the hypothesis that oxidative phosphorylation is required for normal sperm function and is impaired in teratospermic ejaculates. Electroejaculates from both species were stained with MitoTracker to quantify mitochondrial membrane potential (MMP) or were incubated to assess changes in sperm function (motility, acrosomal integrity, and lactate production) after mitochondrial inhibition with myxothiazol. Sperm midpiece dimensions also were quantified. Sperm mitochondrial fluorescence (directly proportional to MMP) was ~95% lower in the cheetah compared with the normospermic and teratospermic cat, despite the cheetah having a 10% longer midpiece. In both species, MMP was increased 5-fold in spermatozoa with retained cytoplasm compared with structurally normal cells. Inhibition of oxidative phosphorylation impaired sperm function in both species, but a 100-fold higher inhibitor concentration was required in the cat compared with the cheetah. Collectively, findings revealed that oxidative phosphorylation was required for sperm function in the domestic cat and cheetah. This pathway of energy production appeared markedly less active in the cheetah, indicating a species-specific vulnerability to mitochondrial dysfunction. The unexpected, cross-species linkage between retained cytoplasmic droplets and elevated MMP may reflect increased concentrations of metabolic enzymes or substrates in these structures.

  8. Apoptotic-like changes of boar spermatozoa in freezing media supplemented with different antioxidants.

    Science.gov (United States)

    Trzcińska, M; Bryła, M

    2015-01-01

    This study evaluated the effect of supplementing the freezing extender with exogenous anti-oxidants on apoptotic-like changes in post-thaw boar spermatozoa. A total of 36 ejaculates were resuspended in standard lactose-egg yolk-glycerol extender supplemented with antioxidant to final concentrations of 0 (as control), 2.5mM GSH (group I), 5.0 mM GSH (group II), 150 IU/mL SOD (group III), 300 IU/mL SOD (group IV), 200 IU/mL CAT (group V), 400 IU/mL CAT (group VI), 150 IU/mL SOD+200 IU/mL CAT (group VII), 300 IU/mL SOD+400 IU/mL CAT (group VIII). Sperm motility and apoptotic-like changes were determined before and after freeze-thawing. The various markers of apoptotic-like changes were measured: plasma membrane permeability by YO-PRO-1/PI assay, phosphatidylserine (PS) translocation across the plasma membrane using fluorescein-labeled Annexin-V, mitochondrial transmembrane potential detected by JC-1, and DNA fragmentation evaluated by TUNEL assay. The highest percentage of progressive motile sperm was noticed in group II (PM% 64.2±15.4) compared with control (PM% 36.8±5.5). The supplementation of 400 IU/mL CAT (group VI) revealed significant (Psperm survival compared with the control. Evaluation by TUNEL assay revealed that cryopreservation and thawing did not induce DNA fragmentation in boar spermatozoa.

  9. Localization of CD9 Molecule on Bull Spermatozoa: Its Involvement in the Sperm-Egg Interaction.

    Science.gov (United States)

    Antalíková, J; Jankovičová, J; Simon, M; Cupperová, P; Michalková, K; Horovská, Ľ

    2015-06-01

    Tetraspanin CD9 is one of the egg membrane proteins known to be essential in fertilization process. The presence and localization of CD9 molecule in spermatozoa and its possible function in reproduction are still unclear. In our study, we describe the localization of CD9 on bull spermatozoa. In the immunofluorescence assay, the positive signal has been observed in the high proportion of sperm cells as a fine grains either on the apical part or through the entire anterior region of sperm head. CD9 recognized by monoclonal antibody IVA-50 was detected on freshly ejaculated (83.4 ± 3.7%) and frozen-thawed (84.3 ± 2.3%) sperm. The same reaction pattern was observed on sperm capacitated for 1 h, 2 h, 3 h and 4 h (83.6 ± 2.0%; 84.0 ± 1.5%; 85.7 ± 0.8%; 77.5 ± 10.8%). The presence of CD9 exclusively on plasma membrane of the bovine sperm has been detected by Western blot analysis of the protein fractions after the discontinuous sucrose gradient fractionation of the bull sperm. Moreover, probable role of the sperm CD9 molecule in fertilization process of cattle has been suggested as sperm treatment with anti-CD9 antibody significantly reduced (by 25%, p ≤ 0.001) the number of fertilized oocytes compared to control group in fertilization assay in vitro.

  10. Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination.

    Science.gov (United States)

    Ogata, Kazuko; Sasaki, Aiko; Kato, Yuka; Takeda, Arisa; Wakabayashi, Mikio; Sarentonglaga, Borjigin; Yamaguchi, Mio; Hara, Asuka; Fukumori, Rika; Nagao, Yoshikazu

    2015-01-01

    The present study was conducted to evaluate whether supplementation of semen extender with glutathione (GSH) can maintain the quality of frozen-thawed canine spermatozoa. Eighteen ejaculates were obtained from 5 dogs and placed in extender (20% egg yolk, Tris, citric acid, lactose, raffinose, antibiotics and 6.5% glycerol) containing 0 (control), 2.5, 5, 7.5 or 10 mM GSH. The samples were cooled to 4 C and then frozen in liquid nitrogen vapor. Motility parameters of the sperm were evaluated at 0, 1, 2, 3, 4, 12 and 24 h after thawing. Sperm motility was higher in the 5 mM GSH group than in the control or 2.5 and 10 mM GSH groups; this effect was observed at 1 to 24 h after thawing (P control. Lipid peroxidation (LP) levels immediately after thawing were lower in the 5 and 10 mM GSH groups compared with the control, while those at 12 h after thawing did not differ significantly. Frozen-thawed semen in the 5 mM GSH group was used for transcervical insemination of 4 bitches, resulting in delivery of 5 puppies from 2 bitches. These results indicate that supplementation of semen extender with 5 mM GSH was effective in improving motility, longevity and acrosomal integrity and inhibiting LP levels in post-thaw canine spermatozoa, without any adverse impacts on full-term development after transcervical insemination.

  11. Aneuploidy rates for chromosomes X/Y and 18 among preselected spermatozoa in men with severe teratospermia.

    Science.gov (United States)

    Levron, Jacob; Aviram-Goldring, Ayala; Rienstien, Shlomit; Bider, David; Dor, Jehoshua; Raviv, Gil

    2013-09-01

    Eight infertile men with various degrees of oligoasthenoteratozoospermia and repeated implantation failure were selected for this study due to exceptionally high rates of sperm aneupoidy in their ejaculates. All subjects had normal physical examination, karyotype and serum FSH concentration. Prior to IVF treatment, spermatozoa was collected, processed, micromanipulated and tested for chromosomes X, Y and 18 using fluorescence in-situ hybridization. Aneupoidy rates for chromosomes X, Y and 18 were determined among sperm population selected for normal morphology using high-order magnification light microscopy. A second group of fast motile spermatozoa were collected using an intracytoplasmic sperm injection pipette from the medium-oil interface from microdroplets. The average aneuploidy rates for the three chromosomes were 7.6% (395/5182) in the sperm specimen before selection, 8.7% (116/1326) in the normal morphology selected group and 4.3% (59/1388; Pchromosomal abnormalities (P<0.001). Preselection of the most rapid sperm subpopulation for intracytoplasmic sperm injection may improve the qualities of the fertilizing spermatozoon. Copyright © 2013 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  12. Low long non-coding RNA HOTAIR expression is associated with down-regulation of Nrf2 in the spermatozoa of patients with asthenozoospermia or oligoasthenozoospermia.

    Science.gov (United States)

    Zhang, Lixin; Liu, Zhineng; Li, Xiaokang; Zhang, Ping; Wang, Jia; Zhu, Dandan; Chen, Xinping; Ye, Lihua

    2015-01-01

    HOTAIR, a long noncoding RNA, regulates development and progression of tumor cells and function of normal stem cells. However, the role and the molecular mechanism of HOTAIR in the spermatozoa of patients with asthenozoospermia and oligoasthenozoospermia are still unclear. Herein, 45 healthy control, 45 asthenozoospermic patients and 45 oligoasthenozoospermic patients were enrolled. Initially, through analyzing HOTAIR expression, we observed a decreased level of HOTAIR expression in patients. Subsequently, we found that there was a positive correlation between HOTAIR expression and Nrf2 expression in patients. The low expression of HOTAIR was also observed to be associated with specific sperm function parameters, including motility and vitality. In the ejaculated spermatozoa from patients, low level of histone H4 acetylation of the Nrf2 gene promoter was observed. Finally, we found that downregulation of HOTAIR expression reduced histone H4 acetylation in Nrf2 promoter and Nrf2 expression. Therefore, this study demonstrated that HOTAIR expression was low in the spermatozoa of patients with asthenozoospermia and oligoasthenozoospermia, which resulted in down-regulation of Nrf2 expression. Our data suggested the decrease of HOTAIR expression led to ROS related defects in sperm function.

  13. Successful intracytoplasmic sperm injection with testicular spermatozoa from a man with multiple morphological abnormalities of the sperm flagella: a case report.

    Science.gov (United States)

    Yang, Shenmin; Gao, Liang; Wang, Wei; Ding, Jie; Xu, Yongle; Li, Hong

    2017-10-02

    The purpose of this study is to analyze the sperm morphology of a Chinese man affected with multiple morphological abnormalities of the sperm flagella (MMAF) and observe the intracytoplasmic sperm injection (ICSI) outcome. A Chinese man was diagnosed with multiple morphological abnormalities of the sperm flagella by semen analysis and electron microscopy. Testicular spermatozoa were injected intracytoplasmically, and the following ICSI results were observed. All the spermatozoa from his ejaculate were immotile and morphologically abnormal in the flagellum. In transmission electron microscopy assays, most spermatozoa showed disorganized fibrous sheath, accompanied by distortion of various cytoskeletal components, and missing of the central pair microtubules. Testicular sperm was injected to the oocytes in two ICSI cycles, with fertilization rates of 45.5 and 40.0%. Finally, a healthy female infant was delivered at the second ICSI cycle. Fertilization and pregnancy could be achieved by intracytoplasmic sperm injection, regardless of severe flagellar defects. ICSI is effective for MMAF-affected man, and testicular sperm is an alternative when no motile sperm is available.

  14. A study of aneuploidy and DNA fragmentation in spermatozoa of three men with sex chromosome mosaicism including a 45,X cell line.

    Science.gov (United States)

    Nguyen, Minh Huong; Morel, Frederic; Bujan, Louis; May-Panloup, Pascale; De Braekeleer, Marc; Perrin, Aurore

    2015-06-01

    Meiotic segregation of mosaic males with a 45,X cell line has been little examined. In this study, we evaluated the risk of aneuploid gametes using fluorescence in situ hybridization (FISH) and DNA fragmentation in ejaculated spermatozoa of three men with sex chromosome mosaicism including a 45,X cell line. Triple- and dual-color FISH were performed. Sperm DNA fragmentation was detected using the TUNEL assay. A significantly increased frequency of XY disomic spermatozoa was observed for patients (P)1 and P2. A significant increase in diploidy and autosomal aneuploidy was found in P2 and P3, respectively. The rate of DNA fragmentation was not different from that observed in a control group. Data from the literature are scarce (only 3 cases reported), making comparison of the present data difficult, especially as the frequencies of the cell lines comprising the mosaicism differed between patients. Furthermore, the proportion of the different cell lines can differ from one tissue to another in the same patient. Whether the relative levels of the several cell lines present in the mosaicism can influence the rate of aneuploid spermatozoa remains unknown.

  15. Sperm economy between female mating frequency and male ejaculate allocation.

    Science.gov (United States)

    Abe, Jun; Kamimura, Yoshitaka

    2015-03-01

    Why females of many species mate multiply is a major question in evolutionary biology. Furthermore, if females accept matings more than once, ejaculates from different males compete for fertilization (sperm competition), which confronts males with the decision of how to allocate their reproductive resources to each mating event. Although most existing models have examined either female mating frequency or male ejaculate allocation while assuming fixed levels of the opposite sex's strategies, these strategies are likely to coevolve. To investigate how the interaction of the two sexes' strategies is influenced by the level of sperm limitation in the population, we developed models in which females adjust their number of allowable matings and males allocate their ejaculate in each mating. Our model predicts that females mate only once or less than once at an even sex ratio or in an extremely female-biased condition, because of female resistance and sperm limitation in the population, respectively. However, in a moderately female-biased condition, males favor partitioning their reproductive budgets across many females, whereas females favor multiple matings to obtain sufficient sperm, which contradicts the predictions of most existing models. We discuss our model's predictions and relationships with the existing models and demonstrate applications for empirical findings.

  16. The Ability of Honey Bee Drones to Ejaculate

    Directory of Open Access Journals (Sweden)

    Czekońska Krystyna

    2015-12-01

    Full Text Available The effectiveness of two methods of collecting semen from honeybee Apis mellifera drones was compared, and the reasons for problems with ejaculating semen were analysed. Among 275 drones, 100 were stimulated to release semen using a manual method, 100 with the use of chloroform, and from 75 drones the reproductive organs were dissected for analysis and evaluation. It was found that the principal causes of problems that drones had with ejaculating their semen were anatomical changes or a delay in the development of the mucus glands. It was also found that the method employing chloroform was less efficient in the first phase of eversion of the endophallus, compared with the manual method. The method with the use of chloroform allows the determination of the proportion of drones, which do not evert the endophallus because of poor or delayed development of mucus glands, as well as the proportion of drones which evert the organ, but do not ejaculate semen because of the absence of semen in the seminal vesicles.

  17. Evaluation of Sperm Parameters of Infertile Men with Retrograde Ejaculation

    Institute of Scientific and Technical Information of China (English)

    Hong-xing ZHONG; Wei-jie ZHU; Jing LI

    2006-01-01

    Objective To investigate sperm parameters of infertile men with retrograde ejaculation.Methods Twelve infertile men with retrograde ejaculation (group A) were enrolled into this study. Sperm samples were obtained from the postejaculation urine. After sperm recovery and washing procedure, sperm parameters were assessed. Twelve semen samples from normospermic donors were used as the control (group B).Results In all retrograde cases, motile sperm with forward movement were observed in the medium. Motility of group A was significantly lower than that of group B (P<0. 01).In group A, sperm motility ranged from 11% to 56%, sperm with intact both head and tail membranes was 42.2 ± 12.3%, sperm count ranged (13-85)×106/ml, and the sperm survival time was highly shortened. Sperm with normal morphology and intact acrosome were observed in retrograde specimens.Conclusion Sperm parameters recovered from retrograde specimens were highly variable between subjects. The toxicity of urine caused deleterious to sperm functions.Motile sperm could be collected by sperm recovery procedure. Sperm parameters could meet the requirement for the use of assisted reproductive techniques for treating infertile men with retrograde ejaculation.

  18. Changes in family functions in patients with secondary premature ejaculation

    Directory of Open Access Journals (Sweden)

    Mustafa Arı

    2011-06-01

    Full Text Available We aimed to inverstigate changes in family functions in patients with premature ejaculation.Materials and methods: In the present study, study group were randomly selected from Mustafa Kemal University Medical School Research and Training Hospital Urology Department outpatients clinic. Control group were selected among healthy volunteers. Totally 30 patients were included in the PE group and 30 healthy volunteers were included in the control group. Subjects were examined by the same psychiatrist. Beck Anxiety Inventory and Family Assessment Scale were applied to both groupsResults: Compared with the control group, premature ejaculation patients had significantly higher anxiety scores (p=0.001 and more deterioration in problem solving (p=0.001, communication (p=0.022, affective responsiveness (p=0.011, behavior control (p=0.032, and affective involvement in their families (p=0.011. There were no difference in terms of roles and general functions scores (p>0.05.Conclusion: It can be concluded that there is deterioration in family functions in patients with premature ejaculation, Therefore, approaches targeting family functions may be beneficial in the treatment of these patients.

  19. Recent advances in the treatment of premature ejaculation

    Directory of Open Access Journals (Sweden)

    Kate D Linton

    2010-01-01

    Full Text Available Kate D Linton, Kevan R WylieSheffield Teaching Hospitals, Sheffield, UKAbstract: Premature ejaculation (PE is the most common sexual problem affecting men. It can affect men at all ages and has a serious impact on the quality of life for men and their partners. Currently there are no pharmaceutical agents approved for use in the UK, and so all drugs used for this condition are off label. Behavioral therapy has been used to treat PE, but the results are not durable once therapy has been concluded. Several topical therapies have been used including severance-secret (SS cream, lignocaine spray, lidocaine-prilocaine cream and lidocaine-prilocaine spray (TEMPE. There has been recent interest in the selective serotonin reuptake inhibitors (SSRIs for the treatment of PE, due to the fact that one of their common side effects is delayed ejaculation. Currently used SSRIs have several non-sexual side effects and long half lives, therefore there has been interest in developing a short acting, efficacious SSRI that can be used on-demand for PE. Dapoxetine has been recently evaluated for the treatment of PE by several groups, and results so far appear promising.Keywords: premature ejaculation, topical therapies, selective serotonin re-uptake inhibitor, dapoxetine

  20. Current pharmacological agents for the treatment of premature ejaculation

    Directory of Open Access Journals (Sweden)

    Onur Dede

    2014-06-01

    Full Text Available This study was aimed to review and assess the update studies regarding medical treatment for premature ejaculation (PE. It is the most common sexual problem affecting men. It can affect men at all ages and has a serious impact on the quality of life for men and their partners. A wide variety of therapeutic modalities have been tried for treatment of premature ejaculation. Psychological therapies may be helpful for patients with complaint PE. Several topical therapies have been used including lidocaine cream, lidocaine-prilocaine cream. There has been recent interest in the selective serotonin reuptake inhibitors (SSRI for the treatment of PE, due to the fact that one of their common side effects is delayed ejaculation. Currently used SSRIs have several non-sexual side effects and long half lives, therefore there has been interest in developing a short acting, and efficacious SSRI that can be used on-demand for PE. Dapoxetine has been recently evaluated for the treatment of PE by several groups, and results so far appear promising.

  1. Role of Frenular Web Preservation on Ejaculation Latency Time

    Directory of Open Access Journals (Sweden)

    Alborz Salavati

    2012-10-01

    Full Text Available Premature ejaculation (PE is one of prevalent male sexual dysfunctions worldwide. Despite many psychiatric backgrounds, yet there are speculations about organic etiologies considering both anatomic and physiologic points of view. This survey assesses effect of frenular web preservation on premature ejaculation. One thousand and forty otherwise healthy men being visited for urolithiasis (asymptomatic patients were asked for PE according to the International Society of Sexual Medicine definition criteria as intravaginal ejaculation latency time (IELT less than a minute according to stop watch checked by patients' partner and were examined for presence of frenular web. Frenular web defined as a residual of frenulum after a circumcision. Overall prevalence of PE was 18.2% (n=102. We found the presence of frenulum at physical examination in 255 out of 560 (45.5%. Prevalence of PE was 20.7% (n=53 and 16% (n=49 in patients with frenular web preserved and without it, respectively. PE was higher among the men with frenulum preserved; but no statistically significant differences were seen (P=0.70. We did not find any relationship between frenular web and PE, and concerns about this, during circumcision, may not be justified. PE is a not only a problem of local anatomical condition but many psychological and neurological factors could interact with it.

  2. Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen.

    Science.gov (United States)

    Miró, J; Taberner, E; Rivera, M; Peña, A; Medrano, A; Rigau, T; Peñalba, A

    2009-11-01

    The aim of this work was to study the effects of dilution and centrifugation (i.e., two methods of reducing the influence of the seminal plasma) on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey (Equus asinus) semen. Fifty ejaculates from nine Catalonian jackasses were collected. Gel-free semen was diluted 1:1, 1:5 or 1:10 with Kenney extender. Another sample of semen was diluted 1:5, centrifuged, and then resuspended with Kenney extender until a final dilution of 25x10(6) sperm/ml was achieved (C). After 24 h, 48 h or 72 h of refrigerated storage at 5 degrees C, aliquots of these semen samples were incubated at 37 degrees C for 5 min. The percentage of viable sperm was determined by staining with eosin-nigrosin. The motility characteristics of the spermatozoa were examined using the CASA system (Microptic, Barcelona, Spain). At 24h, more surviving spermatozoa were seen in the more diluted and in the centrifuged semen samples (1:1 48.71%; 1:5 56.58%, 1:10 62.65%; C 72.40%). These differences were maintained at 48 h (1:1 34.31%, 1:5 40.56%, 1:10 48.52%, C 66.30%). After 72 h, only the C samples showed a survival rate of above 25%. The four known donkey motile sperm subpopulations were maintained by refrigeration. However, the percentage of motile sperms in each subpopulation changed with dilution. Only the centrifuged samples, and only at 24h, showed exactly the same motile sperm subpopulation proportions as recorded for fresh sperm. However, the 1:10 dilutions at 24 and 48 h, and the centrifuged semen at 48 h, showed few variations compared to fresh sperm. These results show that the elimination of seminal plasma increases the survival of spermatozoa and the maintenance of motility patterns. The initial sperm concentration had a significant (P<0.05) influence on centrifugation efficacy, but did not influence the number of spermatozoa damaged by centrifugation. In contrast, the percentage of live

  3. Spontaneous ejaculation in a wild Indo-Pacific bottlenose dolphin (Tursiops aduncus).

    Science.gov (United States)

    Morisaka, Tadamichi; Sakai, Mai; Kogi, Kazunobu; Nakasuji, Akane; Sakakibara, Kasumi; Kasanuki, Yuria; Yoshioka, Motoi

    2013-01-01

    Spontaneous ejaculation, which is defined as the release of seminal fluids without apparent sexual stimulation, has been documented in boreoeutherian mammals. Here we report spontaneous ejaculation in a wild Indo-Pacific bottlenose dolphin (Tursiops aduncus), and present a video of this rare behavior. This is the first report of spontaneous ejaculation by an aquatic mammal, and the first video of this behavior in animals to be published in a scientific journal.

  4. Spontaneous ejaculation in a wild Indo-Pacific bottlenose dolphin (Tursiops aduncus.

    Directory of Open Access Journals (Sweden)

    Tadamichi Morisaka

    Full Text Available Spontaneous ejaculation, which is defined as the release of seminal fluids without apparent sexual stimulation, has been documented in boreoeutherian mammals. Here we report spontaneous ejaculation in a wild Indo-Pacific bottlenose dolphin (Tursiops aduncus, and present a video of this rare behavior. This is the first report of spontaneous ejaculation by an aquatic mammal, and the first video of this behavior in animals to be published in a scientific journal.

  5. Percentage of motile spermatozoa at 22 hours after swim-up procedure: An indicator for intracytoplasmic sperm injection?

    Science.gov (United States)

    Inoue, Taketo; Yonezawa, Yukiko; Sugimoto, Hironobu; Uemura, Mikiko; Ono, Yuri; Kishi, Junji; Emi, Nobuyuki; Ono, Yoshiyuki

    2016-09-01

    The decision to use in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), or split insemination (IVF-ICSI) in the first cycle is based on the number of motile sperm. Hence, total fertilization failure (TFF) often occurs during IVF cycles, despite normozoospermia. To investigate whether the cumulative motile swim-up spermatozoa percentage at 22 hours post-insemination (MSPPI) is an indicator for ICSI, we analyzed TFF, fertilization, blastocyst development, chemical pregnancy, clinical pregnancy, and live birth rates. This prospective study was performed using data obtained from 260 IVF cycles. At 22 hours after insemination, the remaining swim-up spermatozoa were observed and divided into six groups according to MSPPI (<10%, 10% to <30%, 30% to <50%, 50% to <70%, 70% to <90%, and 90% to 100%). Regardless of the ejaculated motile sperm concentration (0.6-280×10(6)/mL motile spermatozoa), the incidence of TFF significantly increased when MSPPI was <10%, and the fertilization rate significantly decreased when MSPPI was <30%. We found that cumulative MSPPI correlated with the cumulative fertilization rate (Spearman correlation, 0.508, p<0.001). Regarding embryo development, we observed no significant differences in the rates of blastocyst development, chemical pregnancy, clinical pregnancy, or live birth among all groups. Our findings suggest that MSPPI is a viable indicator for split IVF-ICSI and ICSI. Taken together, by employing the MSPPI test in advance before IVF, ICSI, or split IVF-ICSI cycles, unnecessary split IVF-ICSI and ICSI may be avoided.

  6. Zinc and magnesium in bull and boar spermatozoa.

    Science.gov (United States)

    Arver, S; Eliasson, R

    1980-11-01

    Mean +/- s.e.m. concentrations (nmol/10(8) cells) of zinc and magnesium in bull spermatozoa were 30.6 +/- 6.6 and 119 +/- 28.8, respectively. Corresponding values for boar spermatozoa were 16.9 +/- 1.98 and 57.1 +/- 4.3. Bull spermatozoa washed twice in a standard buffered salt solution, pH 7.75, lost 72.6% of their zinc and 46.5% of their magnesium. Boar spermatozoa lost 40% of Zn and 18% of Mg, respectively. Addition of albumin (4% final concentration) to the washing solution did not increase the loss of ions from bull spermatozoa but increased the loss of zinc and magnesium from boar spermatozoa to 52% and 41%, respectively.

  7. Stallion spermatozoa selected by single layer centrifugation are capable of fertilization after storage for up to 96 h at 6°C prior to artificial insemination

    Directory of Open Access Journals (Sweden)

    Lindahl Johanna

    2012-07-01

    Full Text Available Abstract Background One of the challenges faced by equine breeders is ensuring delivery of good quality semen doses for artificial insemination when the mare is due to ovulate. Single Layer Centrifugation (SLC has been shown to select morphologically normal spermatozoa with intact chromatin and good progressive motility from the rest of the ejaculate, and to prolong the life of these selected spermatozoa in vitro. The objective of the present study was a proof of concept, to determine whether fertilizing ability was retained in SLC-selected spermatozoa during prolonged storage. Findings Sixteen mares were inseminated with SLC-selected sperm doses that had been cooled and stored at 6°C for 48 h, 72 h or 96 h. Embryos were identified in 11 mares by ultrasound examination 16–18 days after presumed ovulation. Conclusion SLC-selected stallion spermatozoa stored for up to 96 h are capable of fertilization.

  8. Ejaculate of sneaker males is pheromonally inconspicuous in the black goby, Gobius niger (Teleostei, Gobiidae).

    Science.gov (United States)

    Locatello, L; Mazzoldi, C; Rasotto, M B

    2002-11-01

    The black goby, Gobius niger, shows alternative male mating tactics, i.e., parental and sneaker males. Males release a sexual pheromone that attracts females and stimulates aggressive displays in males. This pheromone is produced by the mesorchial gland, a structure well developed in parental males but markedly undeveloped in sneakers. We measured the behavioral response of parental males to the ejaculates of males performing different reproductive tactics. Parental males reacted to the ejaculate of other parental males, with stereotypic aggressive behaviors, but not to the ejaculate of sneakers; consequently sneaker male ejaculate appears to be pheromonally inconspicuous.

  9. Non-motile tetraploid spermatozoa of Misgurnus loach hybrids

    OpenAIRE

    Zhao, Yan; Fujimoto, Takafumi; Psenicka, Martin; Saito, Taiju; Arai, Katsutoshi

    2016-01-01

    We have compared various properties of spermatozoa from the wild diploid male pond loach Misgurnus anguillicaudatus to those from the interspecific male hybrid of the cross between a female M. anguillicaudatus and a male mud loach M. mizolepis. Our results show that spermatozoa from this interspecific hybrid had poor motility, low viability, abnormal morphology, a larger volume of mitochondrial mass per cell and higher ATP content of spermatozoa with tetraploid DNA content, and they were pres...

  10. Zinc content in epididymal spermatozoa of metoclopramide-treated rats.

    Science.gov (United States)

    Kaminska, B; Rozewicka, L; Dominiak, B; Mielnicka, M; Mikulska, D

    1987-01-01

    Zinc content was determined separately in spermatozoa taken from epididymal caput and cauda in rats. It was revealed that spermatozoa transported from the epididymal caput to the cauda reduce about 54% of zinc. This reduction is significantly inhibited in spermatozoa of rats receiving metoclopramide. That is also accompanied by a fall of testosterone level in blood serum and of delta 5, 3 beta-HSD activity in Leydig cells. It was found out that the reduction of zinc in spermatozoa at the time of their passage through the epididymis is the process that depends on androgens.

  11. Acrosome reaction is impaired in spermatozoa of obese men: a preliminary study.

    Science.gov (United States)

    Samavat, Jinous; Natali, Ilaria; Degl'Innocenti, Selene; Filimberti, Erminio; Cantini, Giulia; Di Franco, Alessandra; Danza, Giovanna; Seghieri, Giuseppe; Lucchese, Marcello; Baldi, Elisabetta; Forti, Gianni; Luconi, Michaela

    2014-11-01

    To compare spontaneous (Sp-AR) and P-induced acrosome reaction (AR) in spermatozoa of obese and lean subjects. Bariatric unit at a university hospital. Prospective, observational study. Twenty-three obese (mean±SD body mass index [BMI], 44.3±5.9 kg/m2) and 25 age-matched lean (BMI, 24.2±3.0 kg/m2) subjects. None. Spontaneous and P-induced AR in spermatozoa of obese and lean subjects. A statistically significant difference was found between obese and lean cohorts in total T and calculated free T, E2, glycated hemoglobin, and high-density lipoproteins, whereas among the routine semen parameters analyzed, only immotile sperm percentage and ejaculate volume differed significantly. Spermatozoa of obese (n=13) vs. lean men (n=19) showed a higher Sp-AR (17.9%±7.2% vs. 8.3%±4.2%), which resulted in a reduced ability to respond to P evaluated as the AR-after-P-challenge parameter (3.5%±3.2% vs. 17.6%±9.2%). Multivariate analysis adjusted for age revealed a significant correlation between BMI, waist, E2, and glycated hemoglobin with both Sp-AR (age-adjusted r=0.654, r=0.711, r=0.369, and r=0.644, respectively) and AR-after-P-challenge (age-adjusted r=-0.570, r=-0.635, r=-0.507, and r=-0.563, respectively). A significant difference in sperm cholesterol content was reported between obese and lean men (29.8±19.5 vs. 19.1±14.6 ng/μg of proteins). Sperm AR is impaired in obese men, showing reduced response to P and elevated Sp-AR, associated with altered circulating levels of E2 and sperm cholesterol content. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  12. Assessment of bovine spermatozoa viability using different cooling protocols prior to cryopreservation

    Directory of Open Access Journals (Sweden)

    Paulo B.D. Gonçalves

    2010-11-01

    Full Text Available The aim of our study was to evaluate the effect of different cooling rates on the post-thawing quality of bovine spermatozoa. Ejaculated semen from a 24-month-old Jersey bull was collected using an artificial vagina and diluted in a commercial extender to evaluate spermatozoan concentration and motility subjectively before cooling and freezing and after thawing. Straws were allocated to four cooling curves: rapid (RD, semi-rapid (SRD, semi-slow (SSLW and slow (SLW. The temperature was decreased from 25ºC to 4ºC in 10, 50, 110 and 135 min, which represents a cooling rate of 2.06, 0.40, 0.18 and 0.15ºC/min, respectively. Then straws were frozen and stored at -196ºC. After thawing, one aliquot of each straw was used for evaluation. Spermatozoan integrity and mitochondrial function were evaluated using a combination of fluorescent probes containing 100 mg/mL FITC-PSA, 0.5 μg/mL PI and 153 μM JC-1. At the end of cooling, spermatozoan motility did not differ among RD (63.3%, SRD (66.7%, SSLW (66.7% and SLW (80.0%. However, normal spermatozoan morphology was lower in SRD (84.8% compared to RD (91.7%, SSLW (91.7% and SLW (90.3% (P<0.05. In thawed semen, spermatozoan motility and normal morphology did not differ among RD (40.0%; 88.8%, SRD (43.3%; 82.5%, SSLW (40.0%; 87.2% and SLW (36.7%; 88.0%. The percentage of damaged spermatozoa, including plasma and acrosome membrane damage and low mitochondrial potential, was higher in RD compared to the others (P<0.05. In conclusion, a rapid cooling curve is detrimental to the spermatozoa and affects the post-thaw spermatozoan integrity of bovine frozen semen.

  13. Correlation between aneuploidy, apoptotic markers and DNA fragmentation in spermatozoa from normozoospermic patients.

    Science.gov (United States)

    Vendrell, Xavier; Ferrer, Minerva; García-Mengual, Elena; Muñoz, Patricia; Triviño, Juan Carlos; Calatayud, Carmen; Rawe, Vanesa Y; Ruiz-Jorro, Miguel

    2014-04-01

    Genetic and biochemical sperm integrity is essential to ensure the reproductive competence. However, spermatogenesis involves physiological changes that could endanger sperm integrity. DNA protamination and apoptosis have been studied extensively. Furthermore, elevated rates of aneuploidy and DNA injury correlate with reproductive failures. Consequently, this study applied the conventional spermiogram method in combination with molecular tests to assess genetic integrity in ejaculate from normozoospermic patients with implantation failure by retrospectively analysing aneuploidy (chromosomes 18, X, Y), DNA fragmentation, externalization of phosphatidylserine and mitochondrial membrane potential status before and after magnetic activated cell sorting (MACS). Aneuploid, apoptotic and DNA-injured spermatozoa decreased significantly after MACS. A positive correlation was detected between reduction of aneuploidy and decreased DNA damage, but no correlation was determined with apoptotic markers. The interactions between apoptotic markers, DNA integrity and aneuploidy, and the effect of MACS on these parameters, remain unknown. In conclusion, use of MACS reduced aneuploidy, DNA fragmentation and apoptosis. A postulated mechanism relating aneuploidy and DNA injury is discussed; on the contrary, cell death markers could not be related. An 'apoptotic-like' route could explain this situation. Copyright © 2013 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  14. An evidence-based unified definition of lifelong and acquired premature ejaculation : report of the second international society for sexual medicine ad hoc committee for the definition of premature ejaculation

    National Research Council Canada - National Science Library

    Serefoglu, Ege Can; McMahon, Chris G; Waldinger, Marcel D; Althof, Stanley E; Shindel, Alan; Adaikan, Ganesh; Becher, Edgardo F; Dean, John; Giuliano, Francois; Hellstrom, Wayne Jg; Giraldi, Annamaria; Glina, Sidney; Incrocci, Luca; Jannini, Emmanuele; McCabe, Marita; Parish, Sharon; Rowland, David; Segraves, R Taylor; Sharlip, Ira; Torres, Luiz Otavio

    2014-01-01

    INTRODUCTION: The International Society for Sexual Medicine (ISSM) Ad Hoc Committee for the Definition of Premature Ejaculation developed the first evidence-based definition for lifelong premature ejaculation (PE...

  15. Emerging treatments for premature ejaculation: focus on dapoxetine

    Directory of Open Access Journals (Sweden)

    Wayne JG Hellstrom

    2008-12-01

    Full Text Available Wayne JG HellstromChief, Section of Andrology, Department of Urology, Tulane University School of Medicine, New Orleans, LA, USAAbstract: Premature ejaculation (PE is a common problem in men worldwide. It has a significant impact on affected men and their partners in terms of self-esteem, dissatisfaction with their sexual relationships, personal distress, and interpersonal difficulty. Psychological therapies may achieve short-term improvements, but there are limited data on the long-term success of these methods. Oral therapy with long-acting selective serotonin reuptake inhibitors (SSRIs improves intravaginal ejaculatory latency time (IELT, but these agents are designed to be administered daily and may be associated with unwanted sexual side effects and withdrawal symptoms upon abrupt discontinuation. Dapoxetine is a short-acting SSRI that can be taken as needed (prn by men with PE. It has been studied in five separate multicenter, randomized, double-blind, placebo-controlled trials involving more than 6000 men with PE. In four studies that evaluated IELT as an endpoint (N = 4843, dapoxetine 30 and 60 mg prn achieved statistically significant increases in IELT versus placebo. Dapoxetine also showed statistically significant improvements in perceived control over ejaculation, PE-related personal distress, and other patient-reported outcomes in all five trials. Dapoxetine treatment is generally well-tolerated, with low incidences of discontinuation syndrome, sexual dysfunction, and treatment-emergent mood symptoms. The most common adverse events with dapoxetine included nausea, diarrhea, headache, dizziness, and somnolence.Keywords: dapoxetine, discontinuation syndrome, premature ejaculation, selective serotonin reuptake inhibitor

  16. Selective serotonin reuptake inhibitors in the treatment of premature ejaculation

    Institute of Scientific and Technical Information of China (English)

    WANG Wei-fu; CHANG Le; Suks Minhas; David J Ralph

    2007-01-01

    Objective To review and assess the update studies regarding se lective serotonin reuptake inhibitors (SSRIs) in the treatment of premature ejaculation (PE) and then provide practical recommendations and possible mechanisms concerning state of the art knowledge for the use of SSRIs in alleviating PE.Data sources Using the Medline, 48 articles published from January 1st, 1996 to August 1st, 2006 concerning the use of SSRIs and their possible mechanisms in alleviating PE were found and reviewed.Study selection PE, rapid ejaculation, early ejaculation and SSRIs were employed as the keywords, and relevant articles about the use of SSRIs and their possible mechanisms in the treatment of PE were selected.Results Many kinds of SSRIs, such as fluoxetine, sertraline, paroxetine and citalopram, have widely been employed to treat PE. However, their effects are moderate and there is no a universal agreement about the kind, dose, protocol and duration. Dapoxetine, as the first prescription treatment of PE, may change this bottle-neck situation. SSRIs are suggested to be used in young men with lifelong PE, and acquired PE when etiological factors are removed but PE still exists. Phosphodiesterase 5 inhibitors (PDE5-Is) are suggested to be employed alone or combined with SSRIs when SSRIs fail to treat PE or sexual dysfunction associated with SSRIs occurs. The protocol of taking drugs on demand based on taking them daily for a suitable period is proposed to be chosen firstly. The possible mechanisms include increasing serotonergic neurotransmission and activating 5-hydroxytryptamine 2C (5-HT2C) receptors, then switching the ejaculatory threshold to a higher level, decreasing the penile sensitivity and their own effect of antidepression.Conclusion The efficacies of the current SSRIs are moderate in the treatment of PE and they have not been approved by the FDA, therefore new SSRI like dapoxetine needs to be further evaluated.

  17. Changes in the structures of motile sperm subpopulations in dog spermatozoa after both cryopreservation and centrifugation on PureSperm(®) gradient.

    Science.gov (United States)

    Dorado, J; Alcaráz, L; Duarte, N; Portero, J M; Acha, D; Hidalgo, M

    2011-05-01

    The aims of the present study were to: (1) determine if discrete motile sperm subpopulations exist and their incidence in fresh dog ejaculates, (2) evaluate the effects of cryopreservation on the distribution of spermatozoa within the different subpopulations, and (3) determine the effect of the discontinuous PureSperm(®) gradient on the sperm subpopulation structure of frozen-thawed dog spermatozoa. Semen from 5 dogs were collected and cryopreserved following a standard protocol. After thawing, semen samples were selected by centrifugation on PureSperm(®). Sperm motility (assessed by computerized-assisted semen analysis, CASA) was assessed before freezing, just after thawing and after preparation on the PureSperm(®) gradients. Cryopreservation had a significant (Psperm suspensions with improved motility (Psperm subpopulation structure was perfectly maintained after freezing and thawing. The selected sperm samples was enrich in Subpopulation 4, reaching a proportion of 31.9% of the present spermatozoa, in contrast with the unselected sperm samples, where this sperm subpopulation accounted for 24.9% of the total. From these results, we concluded that four well-defined motile sperm subpopulations were present either in fresh semen, in unselected sperm samples or in selected preparations from dogs. The discontinuous PureSperm(®) gradient is a simple method to improve the quality of canine frozen-thawed semen samples, since Subpopulation 4 (high-speed and progressive spermatozoa) was more frequently observed after preparation on the gradient. Finally, this study also demonstrated that the general motile sperm structure present in dog remains constant despite the effect caused by either cryopreservation or separation on PureSperm(®) gradient.

  18. Activity of antioxidative enzymes in fresh and frozen thawed buffalo (Bubalus bubalis) spermatozoa in relation to lipid peroxidation and semen quality

    Institute of Scientific and Technical Information of China (English)

    G Kadirvel; SatishKumar; S K Ghosh; P Perumal

    2014-01-01

    Objective:To investigate the activity of antioxidative enzymes in fresh and frozen thawed spermatozoa in relation to lipid peroxidation and semen quality in buffalo(Bubalus bubalis) bulls.Methods:Forty two semen ejaculates from seven buffalo bulls were collected by artificial vagina method and were used for the study.Sperm motility, livability, plasma membrane and acrosomal integrity, buffalo cervical mucous penetration test were assessed in fresh and frozen thawed semen.Intracellular antioxidative enzymatic activity such as super oxide dismutase (SOD), catalase(CAT), glutathione peroxidase(GSHPx) and reduced glutathione(GSH), reactive oxygen species(ROS) and lipid peroxidation(LPO) were estimated in fresh and frozen thawed semen.Results:A significant(P<0.01) reduction in activity of antioxidative enzymes(SOD by 47.7%,GSHPx by62.7% andGSH by58.6%) in frozen thawed spermatozoa as compared to fresh spermatozoa was found.Although the catalase activity was varied from0 to3.8IU/109sperm in fresh semen, but after freezing and thawing this activity was not detectable.These enzyme activities had a strong positive association with sperm motility, membrane integrity and distance traveled by vanguard spermatozoa in buffalo cervical mucus and negative correlation withLPO andROS. However, no significant correlation with acrosomal integrity was found.Conclusion:It was concluded that loss of activity of intracellular antioxidative enzymes was evident after freezing and thawing and there was a strong association between the antioxidative enzyme activities,ROS, lipid peroxidation and sperm function in buffalo semen.

  19. Efficacy of Clomipramine, Sertraline and Terazosin Treatments in Premature Ejaculation

    OpenAIRE

    Tuncel, Altuğ; Aslan, Yılmaz; Başar, M. Murad; Atan, Ali

    2014-01-01

    Aim: To compare the efficacy of oral clomipramine, sertraline and terazosin to placebo in premature ejaculation. Materials and Methods: A total of 90 patients aged from 20 to 58 years were enrolled in this study. Patients were randomized into 4 groups. Group 1 (n: 22) took placebo and served as controls. Group 2 (n: 23) patients took 25 mg clomipramine HCl nightly; Group 3 (n: 20) 50 mg sertraline nightly; and Group 4 (n: 25) 5 mg terazosin nightly. The medications were used for two month...

  20. Tramadol use in premature ejaculation: Daily versus sporadic treatment

    Directory of Open Access Journals (Sweden)

    Amil H Khan

    2013-01-01

    Full Text Available Aim: Premature ejaculation (PME is defined as ejaculation with the minimal sexual stimulation before, on or shortly after penetration and or before a person wishes it. It is a function of the time between intra-vaginal penetration and intra-vaginal ejaculation. Tramadol has shown efficacy in PME when used as sporadic basis. In this study, we compared the use of 100 mg of tramadol as sporadic treatment (administered 6-8 h before coitus versus continued treatment with the objective of evaluating the therapeutic results of both modalities. We assumed our alternative hypothesis that they have similar effects. Materials and Methods: A prospective study was carried out on 60 patients divided into two groups of 30 patients each. Intra-vaginal ejaculation latency time (IELT and coital frequency were measured both prior to and after the treatment. Group A received tramadol 100 mg daily for 4 weeks and on request (sporadically for 4 weeks more. Group B was given placebo in the same manner. Results were statistically analyzed using the Student t-test. Results : Mean IELT prior to treatment was 59.2 s in Group A and 58.7 s in Group B. Mean pre-treatment coital frequency was 2.44 times/week for Group A and 2.13 times/week for Group B. Mean IELT was 202.5 s after continued tramadol treatment and 238.2 s after sporadic treatment in Group A. Mean IELT with daily placebo was 94.8 s and with sporadic placebo was 96.6 s. Coital frequency increased to 4.32 times/week with daily tramadol treatment and 4.86 times with sporadic treatment. Coital frequency increased to 2.88 times/week with daily placebo treatment and 3.23 times with sporadic treatment. Conclusions: The results of PME treatment with tramadol are similar with both continued and sporadic administration. The sex life of patients improved and they reported greater satisfaction with the sporadic treatment.

  1. Tramadol Use in Premature Ejaculation: Daily Versus Sporadic Treatment

    Science.gov (United States)

    Khan, Amil H.; Rasaily, Deepa

    2013-01-01

    Aim: Premature ejaculation (PME) is defined as ejaculation with the minimal sexual stimulation before, on or shortly after penetration and or before a person wishes it. It is a function of the time between intra-vaginal penetration and intra-vaginal ejaculation. Tramadol has shown efficacy in PME when used as sporadic basis. In this study, we compared the use of 100 mg of tramadol as sporadic treatment (administered 6-8 h before coitus) versus continued treatment with the objective of evaluating the therapeutic results of both modalities. We assumed our alternative hypothesis that they have similar effects. Materials and Methods: A prospective study was carried out on 60 patients divided into two groups of 30 patients each. Intra-vaginal ejaculation latency time (IELT) and coital frequency were measured both prior to and after the treatment. Group A received tramadol 100 mg daily for 4 weeks and on request (sporadically) for 4 weeks more. Group B was given placebo in the same manner. Results were statistically analyzed using the Student t-test. Results: Mean IELT prior to treatment was 59.2 s in Group A and 58.7 s in Group B. Mean pre-treatment coital frequency was 2.44 times/week for Group A and 2.13 times/week for Group B. Mean IELT was 202.5 s after continued tramadol treatment and 238.2 s after sporadic treatment in Group A. Mean IELT with daily placebo was 94.8 s and with sporadic placebo was 96.6 s. Coital frequency increased to 4.32 times/week with daily tramadol treatment and 4.86 times with sporadic treatment. Coital frequency increased to 2.88 times/week with daily placebo treatment and 3.23 times with sporadic treatment. Conclusions: The results of PME treatment with tramadol are similar with both continued and sporadic administration. The sex life of patients improved and they reported greater satisfaction with the sporadic treatment. PMID:24249927

  2. Virtual azoospermia and cryptozoospermia--fresh/frozen testicular or ejaculate sperm for better IVF outcome?

    Science.gov (United States)

    Hauser, Ron; Bibi, Guy; Yogev, Leah; Carmon, Ariella; Azem, Foad; Botchan, Amnon; Yavetz, Haim; Klieman, Sandra E; Lehavi, Ofer; Amit, Ami; Ben-Yosef, Dalit

    2011-01-01

    Men diagnosed as having azoospermia occasionally have a few mature sperm cells in other ejaculates. Other men may have constant, yet very low quality and quantity of sperm cells in their ejaculates, resulting in poor intracytoplasmic sperm injection (ICSI) outcome. It has not been conclusively established which source of sperm cells is preferable for ICSI when both ejaculate and testicular (fresh or frozen) sperm cells are available. It is also unclear whether there is any advantage of fresh over frozen sperm if testicular sperm is to be used. We used ejaculate, testicular (fresh or frozen) sperm cells, or both for ICSI in 13 couples. Five of these couples initially underwent ICSI by testicular sperm extraction, because the males had total azoospermia, and in later cycles with ejaculate sperm cells. Ejaculate sperm cells were initially used for ICSI in the other 8 patients, and later with testicular sperm cells. The fertilization rate was significantly higher when fresh or frozen-thawed testicular sperm cells were used than when ejaculated sperm cells were used. Likewise, the quality of the embryos from testicular (fresh and frozen) sperm was higher than from ejaculated sperm (65.3% vs 53.2%, respectively, P cryptozoospermia because of their superior fertility.

  3. Comparison Between Tadalafil Plus Paroxetine and Paroxetine Alone in the Treatment of Premature Ejaculation

    OpenAIRE

    Moudi; Kasaeeyan

    2016-01-01

    Background Several recent studies have investigated the therapeutic role of phosphodiesterase type 5 (PDE5) inhibitors in premature ejaculation (PE) used in the treatment of erectile dysfunction. Objectives In the present research, the efficacy of paroxetine alone and paroxetine plus tadalafil was compared in patients referred because of premature ejaculation. Patients and Methods ...

  4. Morphological characteristics of spermatozoa before and after renal transplantation

    Institute of Scientific and Technical Information of China (English)

    Long-Gen Xu; Shi-Fang Shi; Xiao-Ping Qi; Xiao-Feng Huang; Hui-Ming Xu; Qi-Zhe Song; Xing-Hong Wang; Zong-Fu Shao; Jun-Rong Zhang

    2005-01-01

    Aim: To investigate the changes of the spermatozoa ultrastructures before and after renal transplantation in uremic patients. Methods: The sperm of five uremic patients before and after transplantation and four healthy volunteers were collected and examined by scanning electron microscopy. Results: Abnormal spermatozoa were found in patients pre-transplantation; abnormalities included deletion of the acrosome, absence of the postacrosomal and postnuclear ring, dumbbell-like changes of the head, tail curling, and absence of the mitochondrial sheath in the midsegment. After renal transplantation, most of the spermatozoa became normal. Conclusion: There are many abnormalities with regard to the appearance and structure of the head, acrosome, mitochondria and tail of the spermatozoa in uremic patients. The majority of the spermatozoa returned to normal after renal transplantation, but a few still presented some abnormalities possibly relating to the administration of immunosuppressants.

  5. The use of comet assay to assess DNA integrity of boar spermatozoa following liquid preservation at 5 degrees C and 16 degrees C.

    Directory of Open Access Journals (Sweden)

    J Strzezek

    2004-03-01

    Full Text Available The comet assay, under neutral conditions, allows the assessment of DNA integrity influenced by sperm ageing, which is manifested in DNA double-strand breaks. Here, we attempted to use a modified neutral comet assay test (single-cell gel electrophoresis, to our knowledge for the first time, to assess DNA integrity of boar spermatozoa during liquid storage for 96 h at 5 degrees C and 16 degrees C. In this comet assay protocol we used 2% beta-mercaptoethanol prior to the lysis procedure, to aid in removing nuclear proteins. Ejaculates from 3 boars (designated A, C and G were diluted with a standard semen extender, Kortowo-3 (K-3, which was supplemented with lipoprotein fractions extracted from hen egg yolk (LPFh or ostrich egg yolk (LPFo. Irrespective of the extender type, the percentage of comet-detected spermatozoa with damaged DNA increased gradually during prolonged storage at 5 degrees C and 16 degrees C. Spermatozoa stored in K-3 extender exhibited elevated levels of DNA damage at both storage temperatures. Significant differences in DNA damage among the boars were more pronounced during storage in LPF-based extenders at 5 degrees C: spermatozoa of boars A and G were less susceptible to DNA damage. The percent of tail DNA in comets was lower in LPF-based extenders, and there were individual variations among the boars. We observed that changes in DNA integrity were dependent on the extender type and storage temperature. A higher level of DNA instability was observed in K-3 extended semen compared with K-3/LPFh or K-3/LPFo extended semen during storage at 5 degrees C. No significant difference in the level of DNA damage between K-3/LPFh and K-3/LPFo was observed. It seems that a long-term storage can affect genomic integrity of boar spermatozoa. The modified neutral comet assay can be used to detect low levels of DNA damage in boar spermatozoa during liquid preservation. Therefore, screening for sperm DNA damage may be used as an additional

  6. DNA fragmentation in spermatozoa: a historical review.

    Science.gov (United States)

    Rex, A S; Aagaard, J; Fedder, J

    2017-07-01

    Sperm DNA Fragmentation has been extensively studied for more than a decade. In the 1940s the uniqueness of the spermatozoa protein complex which stabilizes the DNA was discovered. In the fifties and sixties, the association between unstable chromatin structure and subfertility was investigated. In the seventies, the impact of induced DNA damage was investigated. In the 1980s the concept of sperm DNA fragmentation as related to infertility was introduced as well as the first DNA fragmentation test: the Sperm Chromatin Structure Assay (SCSA). The terminal deoxynucleotidyl transferase nick end labelling (TUNEL) test followed by others was introduced in the nineties. The association between DNA fragmentation in spermatozoa and pregnancy loss has been extensively investigated spurring the need for a therapeutic tool for these patients. This gave rise to an increased interest in the aetiology of DNA damage. The present decade continues within this research area. Some of the more novel methods recently submerging are sorting of cells with increased DNA fragmentation and hyaluronic acid (HA) binding techniques. The clinical value of these tests remains to be elucidated. In spite of half a century of research within the area, this analysis is not routinely implemented into the fertility clinics. The underlying causes are multiple. The abundance of methods has impeded the need for a clinical significant threshold. One of the most promising methods was commercialized in 2005 and has been reserved for larger licensed laboratories. Myriads of reviews and meta-analyses on studies using different assays for analysis of DNA fragmentation, different clinical Artificial Reproductive Treatments (ART), different definitions of successful ART outcome and small patient cohorts have been published. Although the area of DNA fragmentation in spermatozoa is highly relevant in the fertility clinics, the need for further studies focusing on standardization of the methods and clinical

  7. The prevalence of premature ejaculation in young Turkish men.

    Science.gov (United States)

    Karabakan, M; Bozkurt, A; Hirik, E; Celebi, B; Akdemir, S; Guzel, O; Nuhoglu, B

    2016-11-01

    This study was conducted to investigate the prevalence of premature ejaculation (PE) in young Turkish men and to evaluate PE in a population having good physical and mental health. A total of 1230 healthy university graduates aged between 24 and 30 attending the police academy having no physical or mental problems were included in the study. To identify the presence of PE, the participants were asked to complete the premature ejaculation diagnostic tool (PEDT). The mean ages in the PE and non-PE group were 27.3 and 26.7 respectively. There was no statistically significant difference between the two groups concerning age, body mass index (BMI), smoking status and alcohol consumption (P > 0.05). The PE prevalence was found to be 9.2%. The mean PEDT score was calculated as 6.3. Of the participants, 92 scored 11 and higher (9.2%), 66 scored 9 and 10 (6.6%), and the remaining 842 obtained a score equal to or lower than 8 (84.2%). The lower prevalence of PE in young Turkish men compared to the results of studies in the literature can be attributed to the physical and mental well-being of the participants. This study showed that the prevalence of PE in young men with good physical and mental health is lower than that found in the literature.

  8. A single nucleotide polymorphism within the novel sex-linked testis-specific retrotransposed PGAM4 gene influences human male fertility.

    Directory of Open Access Journals (Sweden)

    Hidenobu Okuda

    Full Text Available BACKGROUND: The development of novel fertilization treatments, including in vitro fertilization and intracytoplasmic injection, has made pregnancy possible regardless of the level of activity of the spermatozoa; however, the etiology of male-factor infertility is poorly understood. Multiple studies, primarily through the use of transgenic animals, have contributed to a list of candidate genes that may affect male infertility in humans. We examined single nucleotide polymorphisms (SNPs as a cause of male infertility in an analysis of spermatogenesis-specific genes. METHODS AND FINDING: We carried out the prevalence of SNPs in the coding region of phosphoglycerate mutase 4 (PGAM4 on the X chromosome by the direct sequencing of PCR-amplified DNA from male patients. Using RT-PCR and western blot analyses, we identified that PGAM4 is a functional retrogene that is expressed predominantly in the testes and is associated with male infertility. PGAM4 is expressed in post-meiotic stages, including spermatids and spermatozoa in the testes, and the principal piece of the flagellum and acrosome in ejaculated spermatozoa. A case-control study revealed that 4.5% of infertile patients carry the G75C polymorphism, which causes an amino acid substitution in the encoded protein. Furthermore, an assay for enzymatic activity demonstrated that this polymorphism decreases the enzyme's activity both in vitro and in vivo. CONCLUSION: These results suggest that PGAM4, an X-linked retrogene, is a fundamental gene in human male reproduction and may escape meiotic sex chromosome inactivation. These findings provide fresh insight into elucidating the mechanisms of male infertility.

  9. The importance of transmission electron microscopy analysis of spermatozoa: Diagnostic applications and basic research.

    Science.gov (United States)

    Moretti, Elena; Sutera, Gaetano; Collodel, Giulia

    2016-06-01

    This review is aimed at discussing the role of ultrastructural studies on human spermatozoa and evaluating transmission electron microscopy as a diagnostic tool that can complete andrology protocols. It is clear that morphological sperm defects may explain decreased fertilizing potential and acquire particular value in the field of male infertility. Electron microscopy is the best method to identify systematic or monomorphic and non-systematic or polymorphic sperm defects. The systematic defects are characterized by a particular anomaly that affects the vast majority of spermatozoa in a semen sample, whereas a heterogeneous combination of head and tail defects found in variable percentages are typically non-systematic or polymorphic sperm defects. A correct diagnosis of these specific sperm alterations is important for choosing the male infertility's therapy and for deciding to turn to assisted reproduction techniques. Transmission electron microscopy (TEM) also represents a valuable method to explore the in vitro effects of different compounds (for example drugs with potential spermicidal activity) on the morphology of human spermatozoa. Finally, TEM used in combination with immunohistochemical techniques, integrates structural and functional aspects that provide a wide horizon in the understanding of sperm physiology and pathology. transmission electron microscopy: TEM; World Health Organization: WHO; light microscopy: LM; motile sperm organelle morphology examination: MSOME; intracytoplasmic morphologically selected sperm injection: IMSI; intracytoplasmic sperm injection: ICSI; dysplasia of fibrous sheath: DFS; primary ciliary dyskinesia: PCD; outer dense fibers: ODF; assisted reproduction technologies: ART; scanning electron microscopy: SEM; polyvinylpirrolidone: PVP; tert-butylhydroperoxide: TBHP.

  10. Identification of MicroRNAs in Zebrafish Spermatozoa.

    Science.gov (United States)

    Jia, Kun-Tong; Zhang, Jing; Jia, Peng; Zeng, Lin; Jin, Yilin; Yuan, Yongming; Chen, Jieying; Hong, Yunhan; Yi, Meisheng

    2015-12-01

    MicroRNAs (miRNAs) participate in almost all biological processes. Plenty of evidences show that some testis- or spermatozoa-specific miRNAs play crucial roles in the process of gonad and germ cell development. In this study, the spermatozoa miRNA profiles were investigated through a combination of illumina deep sequencing and bioinformatics analysis in zebrafish. Deep sequencing of small RNAs yielded 11,820,680 clean reads. By mapping to the zebrafish genome, we identified 400 novel and 204 known miRNAs that could be grouped into 104 families. Furthermore, we selected the six highest expressions of known miRNAs to detect their expression patterns in different tissues by stem-loop quantitative real-time polymerase chain reaction. We found that among the six miRNAs, dre-miR-202-5p displayed specific and high expression in zebrafish spermatozoa and testis. Fluorescence in situ hybridization analysis indicated that dre-miR-202-5p was predominantly expressed in all kind of germ cells at different spermatogenetic stages, including spermatogonia and spermatozoa, but barely expressed in the germ cells in the ovary. This sex-biased expression pattern suggests that dre-miR-202-5p might be related to spermatogenesis and the functioning of spermatozoa. The identification of miRNAs in zebrafish spermatozoa and germ cells offers new insights into the spermatogenesis and spermatozoa in the teleost and other vertebrates.

  11. SSRIs and ejaculation: a double-blind, randomized, fixed-dose study with paroxetine and citalopram.

    Science.gov (United States)

    Waldinger, M D; Zwinderman, A H; Olivier, B

    2001-12-01

    Selective serotonin reuptake inhibitors (SSRIs) are known to induce delayed orgasm and ejaculation. However, different SSRIs may differentially delay ejaculation. A double-blind, fixed-dose study in healthy men with lifelong rapid ejaculation was performed to evaluate potential differences between clinically relevant doses of two selective serotonin reuptake inhibitors, paroxetine and citalopram, in their effects on ejaculation. Thirty men with an intravaginal ejaculation latency time (IELT) less than 1 minute were randomly assigned to receive paroxetine (20 mg/day) and citalopram (20 mg/day) for 5 weeks, after taking half the dosage in the first week. During the 1-month baseline and 6-week treatment period, IELTs were measured at home by using a stopwatch procedure. The trial was completed by 23 men. Analysis of variance revealed a between-group difference in the evolution of IELT delay over time (p = 0.0004); the IELT after paroxetine and citalopram gradually increased from 18 and 21 seconds to approximately 170 and 44 seconds, respectively. Paroxetine 20 mg/day exerted a strong delay (8.9-fold increase), whereas citalopram 20 mg/day mildly delayed ejaculation (1.8-fold increase). These results indicate that paroxetine leads to a significant delay in orgasm and ejaculation, whereas citalopram seems to have less of an effect on it.

  12. Influence of Macrophages on the Rooster Spermatozoa Quality.

    Science.gov (United States)

    Kuzelova, L; Vasicek, J; Chrenek, P

    2015-08-01

    The goal of this study was to evaluate the occurrence of macrophages in rooster semen and to investigate their impact on the spermatozoa quality. Ross 308 breeder males (n = 30) with no evidence of genital tract infections were used to determine the concentration of macrophages using fluorescently conjugated acetylated low-density lipoprotein (AcLDL). Subsequently, the roosters were divided into two groups on the basis of semen macrophage concentration, and semen quality was compared in two heterospermic samples. We applied computer-assisted semen analysis (CASA) system to determine motility parameters. Fluorescence microscopy and flow cytometry were used to evaluate occurrence of apoptotic and dead spermatozoa. Spermatozoa fertility potential was examined after intravaginal artificial insemination of hens. Eighteen roosters (control group) contained 0.2-3% of macrophages within spermatozoa population and ten roosters (macrophage group) had 10-15% of macrophages. Males from macrophage group had lower (p < 0.05) motility parameters (total and progressive movement, velocity curved line) and increased concentration of dead spermatozoa detected by flow cytometry and fluorescence microscopy (p < 0.001 and p ˂ 0.05, respectively). Differences (p < 0.05) between fluorescent microscopy and flow cytometry in results on spermatozoa apoptosis and viability were observed. No significant difference was found between groups in fertility of spermatozoa. In conclusion, the higher presence of macrophages in rooster semen may have a negative effect on some parameters of rooster spermatozoa evaluated in vitro. Furthermore, our study suggests that flow cytometry allows more precise examination of spermatozoa viability and apoptosis in a very short time compared with the fluorescent microscopy.

  13. The Efficacy of Cryopreservation and Intrauterine Insemination of Retragrade Ejaculation Sperm%逆行射精精子冷冻保存在宫腔内人工授精技术中的应用

    Institute of Scientific and Technical Information of China (English)

    田晓华; 陈冬丽; 刘华; 王超云; 赵邦霞

    2012-01-01

    目的:探讨逆行射精患者尿液中回收精子的冷冻保存及其在宫腔内人工授精(IUI)周期中的应用效果.方法:2008年12月至2011年1月逆行射精患者共计7例,嘱其按要求留取射精后的尿液,充分洗涤后实施液氮蒸汽法超低温精子冷冻,共冻存精液标本14人份.IUI周期时将冻存精子解冻、优化后行宫腔内人工授精.结果:冷冻前、解冻后前向运动精子总数分别为(19.9±10.4)× 106和(6.9±4.2)× 106,存在显著性差异(P<0.05),解冻优化后前向运动精子总数为(2.6± 1.7)× 106,实施IUI 8个周期,临床妊娠1例.结论:超低温冷冻会明显降低前向运动精子总数,但多次冷冻后一次复苏行IUI可能是治疗逆行射精所致不育的一种较为理想的方法.%Objective: To investigated the effectiveness of cryopreservation for spermatozoa from the urine of patients with retrograde ejaculation and application in intrauterine insemination (IUI) cycles. Methods: Seven patients with retrograde ejaculation were drinken sodium bicarbonate to alkaline urine before masturbating ejaculation. Collected the urine and fully washing,then cryopreservated the sperm by ultra-low temperature of liquid nitrogen vapor. 17straws of frozen spermatozoa were prepared for later IUI cycle. The total number of forward movement sperm were calculated before and after cryopreservation. Results: There is significant difference in total number of forward movement spermatozoa between before and after cryopreservation [(19.9± 10.4)× lCvs (6.9± 4.2)×106, P<0.05].The total number of forward movement spermatozoa were (2.6± 1.7)× 106 after optimization. 17straws of frozen-thawed sperm used in 8 IUI cycles,and one gained clinical pregnancy. Conclusion: Cryopreservation probarbly decline the the total number of forward movement spermatozoa, but cryopreservated sperm several times for one IUI cycle is the feasibility way in the treatment of retragrade ejaculatived infertility.

  14. Microphotokinesigraphic analysis of buffalo spermatozoa I. Characteristic features and significance of the immobile buffalo spermatozoa

    Directory of Open Access Journals (Sweden)

    M. Boichev

    2010-02-01

    Full Text Available The article discusses the results of a study of immobile spermatozoa of buffalo bulls from Bulgarian murrah breed, based on photo images registered in various displays in conditions of dark background photokinesis. The analysis showed the way developer affects the brightness of kinesigrams. The study is based on samples of semen of six buffalos analysed by means of the microphotokinetic method developed by Tchakarov and Natchev. Brightness parameters of the cells and their photokinetic images have been established photometrically; their metric parameters have been defined by means of an eyepiece and object micrometer. It has been found out that the optimal exposure time for a photokinetic analysis of buffalo spermatozoa was(is 5 sec, and the recommended developer for treating of negative registration material is the pyrocatechin developer by Windisch.

  15. Surgery is not indicated for the treatment of premature ejaculation.

    Science.gov (United States)

    Anaissie, James; Yafi, Faysal A; Hellstrom, Wayne J G

    2016-08-01

    Premature ejaculation (PE) is considered the most common male sexual disorder, affecting up to 75% of men at some point in their lives. While medical management is the mainstay of therapy, surgical options such as dorsal nerve neurectomy (DNN), hyaluronic acid (HA) gel glans penis augmentation, and circumcision have been studied as alternative forms of treatment. Preliminary studies have suggested that DNN and HA gel glans penis augmentation are relatively safe and effective, but due to a lack of large, multicenter, randomized-control trials with long-term follow-up, the International Society of Sexual Medicine (ISSM) has been unable to endorse DNN or HA gel glans penis augmentation as options in the treatment of PE. Conflicting data regarding the efficacy and safety of circumcision has similarly led to its exclusion from ISSM recommendations for the treatment of PE. Ethical concerns, particularly the fundamental concept of non-maleficence, are also barriers to the implementation of surgery for PE.

  16. Evaluation of Tris-citric acid, skim milk and sodium citrate extenders for liquid storage of Punjab Urial (Ovis vignei punjabiensis) spermatozoa.

    Science.gov (United States)

    Rakha, Bushra A; Hussain, Iftikhar; Akhter, Shamim; Ullah, Nemat; Andrabi, Syed M H; Ansari, Muhammad S

    2013-09-01

    The Punjab Urial (Ovis vignei punjabiensis) is an endangered subspecie of ovidae, distributed as small scattered populations in the forest belt of the Himalayan foothills of Pakistan and in the areas enclosed by the Indus and the Jhelum rivers. The present study was conducted to evaluate the liquid storage of Punjab Urial spermatozoa in different extenders for use in future in situ conservation activities. Semen was collected by electro-ejaculation from three captive Punjab Urial rams. Suitable ejaculates of individual animals were pooled and divided into three aliquots for dilution with the experimental extenders (Tris-citric acid, skim milk and sodium citrate) at 37°C. Extended semen was cooled from 37°C to 5°C in 2h, and stored for three days at 5°C. Sperm motility (%), viability (%; live/dead), acrosome integrity (%) and plasma membrane integrity (%) were assessed on days 1, 2 and 3 of storage. On day 1, sperm motility, viability as well as acrosome and plasma membrane integrity were similar (p>0.05) in all three experimental extenders. On day 2, sperm motility, viability, acrosome and plasma membrane integrity were higher (psodium citrate based extender. On day 3 of storage, the values of motility, viability and acrosome integrity were higher (psodium citrate based extenders. In conclusion, Tris-citric acid extender appears to be a better option compared with skim milk and sodium citrate extenders for liquid storage of Punjab Urial semen.

  17. Probability to retrieve testicular spermatozoa in azoospermic patients

    Institute of Scientific and Technical Information of China (English)

    H.-J.Glander; L.-C.Horn; W.Dorschner; U.Paasch; J.Kratzsch

    2000-01-01

    Aim: The degree of probability to retrieve spermatozoa from testicular tissue for intracytoplasmic sperm injection into oocytes is of interest for counselling of infertility patients. We investigated the relation of sperm retrieval to clinical data and histological pattern in testicular biopsies from azoospermic patients. Methods: In 264 testicular biopsies from 142 azoospermic patients, the testicular tissue was shredded to separate the spermatozoa, histological semi - thin sections of which were then evaluated using Johnsen score. Results: The retrieval of spermatozoa correlated significantly ( P 18 U/L, testicular volume < 5 mL, mean Johnsen score<5, and maximum Johnsen score < 7.

  18. Impact of prostate volume on erectile dysfunction and premature ejaculation.

    Science.gov (United States)

    Lee, Jun Ho; Lee, Sung Won

    2016-06-01

    We evaluated the impact of total prostate volume (TPV) on the international index of erectile function-5 (IIEF) and the premature ejaculation diagnostic tool (PEDT). A cross-sectional study was conducted that included 8336 men who had participated in a health examination. PEDT, IIEF and transrectal ultrasonography were used. A full metabolic work-up and serum testosterone level checks were also performed. The median age of participants was 51.0 years. In total, 40.1% had IIEF scores ≤16. Additionally, 24.7% were classified as demonstrating premature ejaculation (PE) (PEDT > 10). The severity of erectile dysfunction (ED) significantly increased with the TPV (p trend < 0.001). After adjusting for potential confounding factors, the odds ratio (OR) for IIEF scores ≤ 16 significantly increased in the group with TPVs of 30-39 cm(3) and the group with TPVs ≥ 40 cm(3) compared with the group with TPVs ≤ 19 cm(3) (TPV 30-39 cm(3), OR: 1.204, 95% confidence interval: 1.034-1.403; TPV ≥ 40 cm(3), OR: 1.326: 95% confidence interval: 1.051-1.733) and this relationship was maintained after adjusting for propensity score (TPV ≥ 30 cm(3), OR: 1.138: 95% confidence interval: 1.012-1.280). However, neither PEDT nor PE was correlated with TPV. In conclusion, TPV is significantly and independently correlated with IIEF but not with PEDT. Future investigations should explore the temporal relationship between TPV and ED.

  19. Evidence that nitric oxide synthase is involved in progesterone-induced acrosomal exocytosis in mouse spermatozoa.

    Science.gov (United States)

    Herrero, M B; Viggiano, J M; Pérez Martínez, S; de Gimeno, M F

    1997-01-01

    In a recent work, we detected nitric oxide synthase (NO synthase) in the acrosome and tail of mouse and human spermatozoa by an immunofluorescence technique. Also, NO-synthase inhibitors added during sperm capacitation in vitro reduced the percentage of oocytes fertilized in vitro, suggesting a role for NO synthase in sperm function. Therefore, in the present study the effect of three NO-synthase inhibitors, NG-nitro-L-arginine methyl ester (L-NAME), NG-nitro-D-arginine methyl ester (D-NAME) and L-NG-nitro-arginine (NO2-arg), and of a nitric oxide donor, spermine-NONOate, on the progesterone-induced acrosome reaction of mouse sperm was examined. NO-synthase inhibitors were added at 0, 60 or 90 min during capacitation; at 120 min, mouse epididymal spermatozoa were exposed to 15 microM progesterone for another 15 min. In another set of experiments, different concentrations of spermine-NONOate were added to capacitated spermatozoa for 15 min; in these experiments, progesterone was not included. NO2-arg and L-NAME blocked progesterone-induced exocytosis regardless of the time at which these inhibitors were added. Moreover, D-NAME did not inhibit exocytosis. In contrast, spermine-NONOate stimulated the acrosomal exocytosis in vitro directly. These results provide evidence that mouse sperm NO synthase participates in the progesterone-induced acrosome reaction in vitro and that nitric oxide induces this event.

  20. Phospholipid hydroperoxide glutathione peroxidase: expression pattern during testicular development in mouse and evolutionary conservation in spermatozoa.

    Science.gov (United States)

    Nayernia, Karim; Diaconu, Mihaela; Aumüller, Gerhard; Wennemuth, Gunther; Schwandt, Iris; Kleene, Kenneth; Kuehn, Hartmut; Engel, Wolfgang

    2004-04-01

    Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a selenoprotein belonging to the family of glutathione peroxidases and has been implicated in antioxidative defense and spermatogenesis. PHGPx accounts for almost the entire selenium content of mammalian testis. In an attempt to verify the expression pattern of PHGPx, testes of mouse mutants with arrest at different stages of germ cell development and testes of mice at different ages were subjected to immunostaining with a monoclonal anti-PHGPx antibody. PHGPx was detected in Leydig cells of testes in all developmental stages. In the seminiferous tubuli, the PHGPx staining was first observed in testes of 21-day-old mice which is correlated with the appearance of the first spermatids. This result was confirmed when the testes of mutant mice with defined arrest of germ cell development were used. An immunostaining was observed in the seminiferous tubuli of olt/olt and qk/qk mice which show an arrest at spermatid differentiation. In Western blot analysis of proteins extracted from testes of mutant mice and from developing testes, two signals at 19- and 22-kDa were observed which confirm the existence of two PHGPx forms in testicular cells. In mouse spermatozoa, a subcellular localization of PHGPx and sperm mitochondria-associated cysteine-rich protein (SMCP) was demonstrated, indicating the localization of PHGPx in mitochondria of spermatozoa midpiece. For verifying the midpiece localization of PHGPx in other species, spermatozoa of Drosophila melanogaster, frog, fish, cock, mouse, rat, pig, bull, and human were used in immunostaining using anti-PHGPx antibody. A localization of PHGPx was found in the midpiece of spermatozoa in all species examined. In electronmicroscopical analysis, PHGPx signals were found in the mitochondria of midpiece. These results indicate a conserved crucial role of PHGPx during sperm function and male fertility.

  1. The expression variations of IL-1 beta and TNF-alpha mRNA in human spermatozoa%细胞因子IL-1β和TNF-α mRNA在人精子表达的变化及意义

    Institute of Scientific and Technical Information of China (English)

    白文俊; 朱积川; 邓庆平; 闫征; 何培英

    2001-01-01

    目的 了解IL-1β和TNF-α在人精子表达的变化及意义。方法 应用逆转录聚合酶链式反应(RT-PCR)检测13例正常生育者、30例不育者及30例慢性前列腺炎患者精子IL-1β和TNF-αmRNA表达。结果 不育者精子中IL-1β和TNF-αmRNA表达均显著低于正常生育者(P=0.018和P=0.046)。慢性前列腺炎患者精子中TNF-αmRNA的表达显著低于正常生育者(P=0.018),而IL-1βmRNA表达与正常生育者相比差别无显著性意义(P=0.42)。IL-1β和TNF-αmRNA在不育者与慢性前列腺炎患者精子中的表达差别无显著性意义(P=0.49和P=0.62)。结论 某些因素可能干扰不育症患者精子IL-1β和TNF-α的合成,导致精液质量下降及不育。生殖道炎症也可能影响精子TNF-α表达,其机制和意义有待进一步研究证实。%Objective To investigate the expression variations of IL-1 betaand TNF-alpha in human spermatozoa.Methods IL-1β and TNF-α mRNA expressions were studied by means of reverse transcription polymerase reaction (RT-PCR)in 13 fertile men,in 30 infertiles and in 30 chronic prostatitis patients.Results The expressions of IL-1β and TNF-α mRNA in infertile men were significantly lower than those in fertile men(P=0.018 and P=0.050),and the expressions TNF-α mRNA in chronic prostatitis patients were significantly lower than those in fertile men(P=0.018 ),but the expressions of IL-1β in these patients were not significantly lower than in fertile men(P=0.420). The expression differences of IL-1β and TNF-α mRNA between the infertile men and chronic prostatitis patients were not significantly(P=0.488 and P=0.616).Conclusions Some unknown factors might impair the production of IL-1β and TNF-α in infertile spermatozoa, and that may be responsible for the poor quality of semen and the infertile status in these patients. Furthermore, urogenital inflammation could also disturb the expression of TNF-α in human spermatozoa.

  2. Premature ejaculation results from partners' mismatch: development and validation of index of intra-vaginal ejaculation latency time.

    Science.gov (United States)

    Cai, L; Wen, Y; Jiang, M; Zeng, M; Zhang, B

    2016-05-01

    Mismatch of partners in premature ejaculation (PE) regarding intra-vaginal ejaculation latency time (IELT) is usually neglected. Here we proposed the concept and evaluated the use of index of IELT (IIELT) as an objective diagnostic tool for PE. Data from 103 self-reporting PE patients and 59 normal controls were collected. The expected IELTs of both the male and female partners were provided by each participating couple in two questionnaires. IIELT=stopwatch IELT/(1/2 the male's expected IELT+1/2 the female's expected IELT). The stopwatch IELTs were 1.74±1.4 min (PE group) and 14.45±11.0 min (control group), P<0.05. The expected IELTs were 15.65±8.7 min (men) and 14.16±6.9 min (women) in the PE group, and 21.3±16.1 min (men) and 20.04±13.47 min (women) in the control group, P<0.05. The calculated IIELTs were 0.14±0.12 (PE group) and 0.83±0.60 (control group), P<0.05. The best cut-off point was 0.658, the Youden index was 0.652, sensitivity was 0.991, specificity was 0.661, positive predictive probability was 83.46% and negative predictive probability was 97.6%. We concluded that IIELT was an integrated measurement of the couples' sexual equilibrium and demonstrated that it provided a simple and objective screening indicator for diagnosing self-reported PE.

  3. 18,X,Y aneuploidies and transmission electron microscopy studies in spermatozoa from five carriers of different reciprocal translocations

    Institute of Scientific and Technical Information of China (English)

    Elena Moretti; Nicola Antonio Pascarelli; Valentina Giannerini; Michela Geminiani; Cecilia Anichini; Oiulia Collodel

    2009-01-01

    We analysed ejaculated spermatozoa from five infertile men with different balanced reciprocal translocations to contribute to the study of meiotic segregation of chromosomes 18, X and Y and also to evaluate sperm morphology by transmission electron microscopy (TEM) analysis. Conventional lymphocyte karyotype analyses highlighted dif-ferent reciprocal balanced translocations: t(12; 13), t(4;9), t(X;8), t(8; 10) and t(3; 16). Semen analysis was performed by light and TEM. Fluorescence in situ hybridization was performed directly on sperm nuclei using centromeric probes for chromosomes 18, X and Y. The carriers of the balanced reciprocal translocations considered in the pres-ent study showed a very similar pattern of sperm pathologies: diffused presence of apoptosis and immaturity. All patients showed meiotic segregation derangements, highlighted by the presence of sperm diploidies and sex chro-mosome disomies particularly related to the failure of the first meiotic division. However, an increased incidence of chromosome 18 aneuploidy was detected in spermatozoa from t(X;8) and t(8;10) carriers. We have also reported values from sex chromosomes such as t(X;8), although the X chromosome was involved in translocation. Since pa-tients with reciprocal translocations and spermatogenetic impairment are candidates for intracytoplasmic sperm in-jection cycles, the study of sperm parameters, and particularly of the level of aneuploidy rates, would provide better information for couples at risk and would contribute to the data in the literature for a better understanding of the ef-fects of chromosomal rearrangement on the whole meiotic process and, in particular, on chromosomes not involved in translocation.

  4. Boar spermatozoa successfully predict mitochondrial modes of toxicity: implications for drug toxicity testing and the 3R principles.

    Science.gov (United States)

    Vicente-Carrillo, A; Edebert, I; Garside, H; Cotgreave, I; Rigler, R; Loitto, V; Magnusson, K E; Rodríguez-Martínez, H

    2015-04-01

    Replacement of animal testing by in vitro methods (3-R principles) requires validation of suitable cell models, preferably obtained non-invasively, defying traditional use of explants. Ejaculated spermatozoa are highly dependent on mitochondrial production and consumption of ATP for their metabolism, including motility display, thus becoming a suitable model for capturing multiple modes of action of drugs and other chemicals acting via mitochondrial disturbance. In this study, a hypothesis was tested that the boar spermatozoon is a suitable cell type for toxicity assessment, providing a protocol for 3R-replacement of animals for research and drug-testing. Boar sperm kinetics was challenged with a wide variety of known frank mito-toxic chemicals with previously shown mitochondrial effects, using a semi-automated motility analyser allied with real-time fluorescent probing of mitochondrial potential (MitoTracker & JC-1). Output of this sperm assay (obtained after 30 min) was compared to cell viability (ATP-content, data obtained after 24-48 h) of a hepatome-cell line (HepG2). Results of compound effects significantly correlated (Psperm variables and for most variables in (HepG2). Dose-dependent decreases of relative ATP content in HepG2 cells correlated to sperm speed (r=0.559) and proportions of motile (r=0.55) or progressively motile (r=0.53) spermatozoa. The significance of the study relies on the objectivity of computerized testing of sperm motility inhibition which is comparable albeit of faster output than somatic cell culture models. Sperm suspensions, easily and painlessly obtained from breeding boars, are confirmed as suitable biosensors for preclinical toxicology screening and ranking of lead compounds in the drug development processes.

  5. Prostasomes of canine seminal plasma - zinc-binding ability and effects on motility characteristics and plasma membrane integrity of spermatozoa.

    Science.gov (United States)

    Mogielnicka-Brzozowska, M; Strzeżek, R; Wasilewska, K; Kordan, W

    2015-06-01

    Prostasomes are small lipid membrane-confined vesicles that are involved in various fertilization-related processes. The aim of this study was to demonstrate canine seminal plasma prostasomes' ability to bind zinc ions, as well as examining their effects on sperm motility characteristics and plasma membrane integrity during cold storage. Ejaculates, collected from five cross-bred dogs (n = 50), were subjected to ultracentrifugation followed by gel filtration (GF) on a Superose 6 column. Prostasomes appeared as a single fraction in the elution profile. Transmission electron microscopy (TEM) analysis of canine prostasomes revealed the presence of membrane vesicles with diameters ranging from 20.3 to 301 nm. The zinc-affinity chromatography on a Chelating Sepharose Fast Flow - Zn(2 +) showed that from 93 to 100% of the prostasome proteins bind zinc ions (P(+) Zn). SDS-PAGE revealed that canine P(+) Zn comprised four protein bands, with low molecular weights (10.2-12 kDa). We have also shown a positive effect of prostasomes (p spermatozoa with intact plasma membrane (SYBR/PI dual staining) and intact acrosome (Giemsa stained), after 2 h storage at 5°C, was showed, in variant A (1.5% of total seminal plasma protein) and B, when compared with Control and variant C (2.5% of total seminal plasma protein). The prostasomes' effect on motility and plasma membrane integrity of canine cold-stored spermatozoa may be related to their ability to bind zinc ions and regulate their availability to the sperm.

  6. The voltage-gated sodium channel nav1.8 is expressed in human sperm.

    Directory of Open Access Journals (Sweden)

    Antonio Cejudo-Roman

    Full Text Available The role of Na(+ fluxes through voltage-gated sodium channels in the regulation of sperm cell function remains poorly understood. Previously, we reported that several genes encoding voltage-gated Na(+ channels were expressed in human testis and mature spermatozoa. In this study, we analyzed the presence and function of the TTX-resistant VGSC α subunit Nav1.8 in human capacitated sperm cells. Using an RT-PCR assay, we found that the mRNA of the gene SCN10A, that encode Na v1.8, was abundantly and specifically expressed in human testis and ejaculated spermatozoa. The Na v1.8 protein was detected in capacitated sperm cells using three different specific antibodies against this channel. Positive immunoreactivity was mainly located in the neck and the principal piece of the flagellum. The presence of Na v1.8 in sperm cells was confirmed by Western blot. Functional studies demonstrated that the increases in progressive motility produced by veratridine, a voltage-gated sodium channel activator, were reduced in sperm cells preincubated with TTX (10 μM, the Na v1.8 antagonist A-803467, or a specific Na v1.8 antibody. Veratridine elicited similar percentage increases in progressive motility in sperm cells maintained in Ca(2+-containing or Ca(2+-free solution and did not induce hyperactivation or the acrosome reaction. Veratridine caused a rise in sperm intracellular Na(+, [Na(+]i, and the sustained phase of the response was inhibited in the presence of A-803467. These results verify that the Na(+ channel Na v1.8 is present in human sperm cells and demonstrate that this channel participates in the regulation of sperm function.

  7. Sperm flagellum volume determines freezability in red deer spermatozoa

    National Research Council Canada - National Science Library

    Ros-Santaella, José Luis; Domínguez-Rebolledo, Alvaro Efrén; Garde, José Julián

    2014-01-01

    ... provides sperm motility. Here, for the first time, we determined the volumes of the flagellum structures in fresh epididymal red deer spermatozoa using a stereological method under phase contrast microscopy...

  8. Lifestyle influences human sperm functional quality

    Institute of Scientific and Technical Information of China (English)

    Mnica Ferreira; Joana Vieira Silva; Vladimiro Silva; Antnio Barros; Margarida Fardilha

    2012-01-01

    Objective:To investigate the impact of acute lifestyle changes on human sperm functional quality.Methods:In the academic festivities week, young and apparently healthy male students who voluntarily submit themselves to acute lifestyle alterations(among the potentially important variations are increase in alcohol, caffeine, and tobacco consumption and circadian rhythm shifts) were used as a model system.Sperm samples were obtained before and after the academic week and compared by traditional semen analysis(n=54) and also tested for cleavedPolyADP-ribose polymerase(PARP) protein, an apoptotic marker(n=35).Results:Acute lifestyle changes that occurred during the academic week festivities(the study model) resulted both in a significant reduction in sperm quality, assessed by basic semen analysis(decrease in sperm concentration, total number of spermatozoa, progressive and non-progressive motility and increase in sperm morphological abnormalities) and by an increase in the expression of the apoptotic marker, cleavedPARP, in the ejaculate.Conclusions:Acute lifestyle changes have clear deleterious effects on sperm quality.We propose cleavedPARP as a novel molecular marker, valuable for assessing spermquality in parallel with the basic semen analysis method.

  9. Sperm variation within a single ejaculate affects offspring development in Atlantic salmon

    Science.gov (United States)

    Immler, Simone; Hotzy, Cosima; Alavioon, Ghazal; Petersson, Erik; Arnqvist, Göran

    2014-01-01

    It is generally believed that variation in sperm phenotype within a single ejaculate has no consequences for offspring performance, because sperm phenotypes are thought not to reflect sperm genotypes. We show that variation in individual sperm function within an ejaculate affects the performance of the resulting offspring in the Atlantic salmon Salmo salar. We experimentally manipulated the time between sperm activation and fertilization in order to select for sperm cohorts differing in longevity within single ejaculates of wild caught male salmon. We found that within-ejaculate variation in sperm longevity significantly affected offspring development and hence time until hatching. Whether these effects have a genetic or epigenetic basis needs to be further evaluated. However, our results provide experimental evidence for transgenerational effects of individual sperm function. PMID:24522632

  10. Storage of sexed boar spermatozoa: Limits and perspectives.

    Science.gov (United States)

    Spinaci, M; Perteghella, S; Chlapanidas, T; Galeati, G; Vigo, D; Tamanini, C; Bucci, D

    2016-01-01

    Despite the great potential application of sex-sorted spermatozoa in swine, the technology is not practiced in the pig industry because of technical factors and species-specific issues. The susceptibility of boar spermatozoa to stresses induced by the sorting procedure, the relative slowness of the sex-sorting process together with the high sperm numbers required for routine artificial insemination in pig are some of the main factors limiting the commercial application of this technology in pigs. This review briefly describes the damage to spermatozoa during sex sorting, focusing on an additional limiting factor: increased susceptibility of sexed boar spermatozoa to injuries induced by liquid storage and cryopreservation that, in turn, impairs sperm quality leading to unsatisfactory results in vivo. Strategies to extend the lifespan of sex-sorted boar spermatozoa and to improve their fertilizing ability after liquid storage or cryopreservation need to be implemented before this technology can be used in pig farms. In this regard, encapsulation in barium alginate membranes could be a promising technique to optimize the in vivo use of sexed boar spermatozoa, by protecting, targeting, and controlling the release of sperm into the female genital tract.

  11. Spermatozoa characteristics in six psittacine species using light microscopy.

    Science.gov (United States)

    Stelzer, G; Schmidt, V; Sobiraj, A; Krautwald-Junghanns, M-E

    2009-12-01

    Even though breeding of companion birds has increased continuously for years, the fecundity assessment of birds has hardly been acknowledged. Knowledge of the structure of spermatozoa is crucial for evaluation of the basic reproductive biology of any species as well as for phylogenetic research and cladistic analyses of internal relationships. Spermatozoa of six different psittacine species (Nymphicus hollandicus, Myiopsitta monachus, Agapornis roseicollis, Melopsittacus undulatus, Tanygnathus lucionensis, Guarouba guarouba) were examined using light microscopy. Head length (nucleus including acrosome), head width, midpiece length and tail length were measured and documented. Significant differences were obvious among almost all of the species for almost all four parameters. However, in all the six species a significant moderate correlation between spermatozoa midpiece lengths and tail lengths (r=0.535, pspermatozoa head lengths and midpiece lengths (r=0.508, pspermatozoa of cockatiels have shorter heads and midpieces than the other psittacines examined in this study. Spermatozoa characteristics in the monk parakeet, blue-naped parrrot and golden conure are previously unreleased and documented for the first time.

  12. Fertilization after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa.

    Science.gov (United States)

    Romero, J; Remohí, J; Mínguez, Y; Rubio, C; Pellicer, A; Gil-Salom, M

    1996-04-01

    To assess the possibility of cryopreserving testicular tissue extracted sperm for intracytoplasmic sperm injection (ICSI). A report of two cases. Our study was approved by the Ethical Committee at the Instituto Valenciano de Infertilidad. In vitro fertilization program at the Instituto Valenciano de Infertilidad. Two azoospermic patients with severe spermatogenic failure but with focal spermatogenesis on testicular biopsies. In both cases, a first ICSI attempt with fresh testicular biopsy extracted sperm was unsuccessful. Cryopreservation of testicular spermatozoa in 100-micro L "pills." Intracytoplasmic sperm injection with thawed testicular spermatozoa. Fertilization rate, cleavage rate, embryo quality, clinical pregnancy. Fertilization rates were 36 percent and 100 percent after ICSI with fresh testicular spermatozoa, and 63 percent and 57 percent after ICSI with cryopreserved testicular sperm. In both cases, cleavage rates and embryo quality were similar when using fresh and cryopreserved testicular spermatozoa. No clinical pregnancies were achieved. High fertilization rates can be obtained after ICSI with frozen-thawed testicular tissue extracted spermatozoa. Cryopreservation of testicular sperm may avoid repetition of testicular biopsies in azoospermic patients in whom the only source of spermatozoa is the testicle.

  13. EFFECTS OF A NEW ANTIBIOTIC COMBINATION ON POST -THAW MOTION CHRACTERISTICS AND MEMBRANE INTEGRITY OF BUFFALO AND SAHIWAL BULL SPERMATOZOA AND ON THE BACTERIOLOGICAL QUALITY OF THEIR SEMEN

    Directory of Open Access Journals (Sweden)

    S. Hasan, S. M. H. Andrabi, R. Muneer, M. Anzar and N. Ahmad

    2001-01-01

    Full Text Available In this study the effects of a new antibiotic combination, i.e., gentamycin, tylosin and linco-spectin (STLS on post-thaw motion characteristics, plasma membrane integrity, sperm morphology and the total aerobic bacterial counts (TABC in buffalo and Sahiwal bull semen were investigated. Ten ejaculates, five each from a buffalo and a Sahiwal bull, possessing more than 60% sperm motility were used. These ejaculates were diluted in Tris-citric acid (TCA extender (at 37 °C; 50 X 106 spermatozoa/mi, containing either GTLS (gentamycin 500 g/ml, tylosin 100 g/ml and linco-spectin 300/600 g/ml, streptomycin 1000 g/ml and penicillin 1000 IU/ml (SP, or negative control with no antibiotics (NCON. Samples were cooled to 4°C in 2 hours, equilibrated at 4°C for 4 hours, filled in 0.5 ml straws, frozen in a controlled rate cell freezer and plunged into liquid nitrogen. Frozen semen was thawed at 37°C for 15 seconds. Post-thaw sperm motion characteristics, plasma membrane integrity and sperm morphology were determined. Total aerobic bacterial counts and the frequency of appearance of bacterial genera were determined in neat semen, after dilution, and after freezing and thawing. Mean motilities (visual; computer-assisted, linear and circular, velocities (straight-line, average path and curvilinear and lateral head displacement (LHD in post- thaw semen samples did not differ due to antibiotics or species. Same was true for sperm plasma membrane integrity. Morphologically abnormal spermatozoa were lower (P<0.05 in GTLS and SP than in NCON. Sperm cells possessing normal acrosomes were higher (P<0.01 in GTLS and SP than in NCON. Total aerobic bacterial counts in post-thaw samples were lower (P<0.05 in GTLS than in SP or NCON. Staphylococcus and micrococcus were lower in samples treated with GTLS than that of SP or NCON. Pseudomonas and E.coli were more frequent in buffaloes than Sahiwal bull samples. Proteus and corynebacteria were scarcely present

  14. Effects of spermatozoa-oviductal cell coincubation time and oviductal cell age on spermatozoa-oviduct interactions.

    Science.gov (United States)

    Aldarmahi, Ahmed; Elliott, Sarah; Russell, Jean; Fazeli, Alireza

    2014-01-01

    The oviduct plays a crucial role in sperm storage, maintenance of sperm viability and sperm transport to the site of fertilisation. The aim of the present study was to investigate the effects of oviductal cell culture passage number, oviductal cell age and spermatozoa-oviduct coincubation times on gene expression in oviductal cells. Immortalised oviductal epithelial cells (OPEC) obtained from two different cell passages (36 and 57) were subcultured three times with and without spermatozoa for 24 h (control group). In a second study, OPEC were cocultured with spermatozoa for different time intervals (0, 4, 12 and 24 h). Expression of adrenomedullin (ADM), heat shock 70 kDa protein 8 (HSPA8) and prostaglandin E synthase (PGES) in OPEC was measured by quantitative polymerase chain reaction. The expression of ADM and HSPA8 was decreased significantly in OPEC cells from Passage 57, particularly in the later subculture group. These effects on HSPA8, but not ADM, expression in OPEC were further altered after coculture with spermatozoa for 24 h. We also demonstrated that spermatozoa-oviduct coculture for 12 and 24 h resulted in significantly higher expression of ADM, HSPA8 and PGES in OPEC. Overall, the data suggest that the OPEC lose some of their properties as a result of oviductal cell aging and that there are spermatozoa-oviduct interactions leading to increased oviductal cell gene expression.

  15. The design and methodology of premature ejaculation interventional studies.

    Science.gov (United States)

    McMahon, Chris G

    2016-08-01

    Large well-designed clinical efficacy and safety randomized clinical trials (RCTs) are required to achieve regulatory approval of new drug treatments. The objective of this article is to make recommendations for the criteria for defining and selecting the clinical trial study population, design and efficacy outcomes measures which comprise ideal premature ejaculation (PE) interventional trial methodology. Data on clinical trial design, epidemiology, definitions, dimensions and psychological impact of PE was reviewed, critiqued and incorporated into a series of recommendations for standardisation of PE clinical trial design, outcome measures and reporting using the principles of evidence based medicine. Data from PE interventional studies are only reliable, interpretable and capable of being generalised to patients with PE, when study populations are defined by the International Society for Sexual Medicine (ISSM) multivariate definition of PE. PE intervention trials should employ a double-blind RCT methodology and include placebo control, active standard drug control, and/or dose comparison trials. Ejaculatory latency time (ELT) and subject/partner outcome measures of control, personal/partner/relationship distress and other study-specific outcome measures should be used as outcome measures. There is currently no published literature which identifies a clinically significant threshold response to intervention. The ISSM definition of PE reflects the contemporary understanding of PE and represents the state-of-the-art multi-dimensional definition of PE and is recommended as the basis of diagnosis of PE for all PE clinical trials.

  16. Level of Sexual Myths Level in Premature Ejaculation Cases

    Directory of Open Access Journals (Sweden)

    Mehmet Gunes

    2016-06-01

    Full Text Available Objective: The aim of this study is to determine level of belief in sexual myths in the cases of premature ejacula­tion (PE which is the most common sexual dysfunction in men. Methods: This study included 100 cases who applied Di­cle University Faculty of Medicine hospitals meet prema­ture ejaculation criteria of DSM-5 and 70 healthy controls. Sociodemographic data form, Hamilton Depression Rat­ing Scale (HDS, Hamilton Anxiety Rating Scale (HAS, Arizona Sexual Experience Scale (ASES-Men form and Sexual Myths Evaluation Form were applied to partici­pants. Results: In the study, rate of belief in sexual myths in PE cases was found significantly higher than healthy controls. In the PE cases, education time less than 10 years, the presence of comorbid sexual dysfunction were found to be statistically significant factors that increase the level of belief in sexual myths in the PE cases, HDS (p=0.0002, HAS (p=0.0001, ASES (p=0.0004 scores were statisti­cally significantly higher than the control group. In the loss of sexual desire in men with comorbid ASES (p=0.0001, with ED, ASES (p=0.001 and HDS (p=0.040 scores were found statistically significantly higher. Conclusions: Sexual information should be given in the appropriate age by educated person in educational insti­tutions.

  17. Seminal fluid mediates ejaculate competition in social insects

    DEFF Research Database (Denmark)

    Den Boer, Susanne Petronella A; Baer, Boris; Boomsma, Jacobus Jan

    2010-01-01

    Queens of ants and bees normally obtain a lifetime supply of sperm on a single day of sexual activity, and sperm competition is expected to occur in lineages where queens receive sperm from multiple males. We compared singly mated (monandrous) and multiply mated (polyandrous) sister groups of ant...... and ants. In Atta leafcutter ants, the negative effect of the seminal fluid of other males was negated by secretion from the queen sperm-storage organ, suggesting that queens may control ejaculate competition after sperm storage.......Queens of ants and bees normally obtain a lifetime supply of sperm on a single day of sexual activity, and sperm competition is expected to occur in lineages where queens receive sperm from multiple males. We compared singly mated (monandrous) and multiply mated (polyandrous) sister groups of ants...... and bees and show that seminal fluid of polyandrous species has a more positive effect on the survival of a male's own sperm than on other males' sperm. This difference was not observed in the monandrous species, suggesting that incapacitation of competing sperm may have independently evolved in both bees...

  18. New agents in the treatment of premature ejaculation.

    Science.gov (United States)

    McMahon, Chris G; McMahon, Chelsea N; Leow, Liang Joo

    2006-12-01

    Premature ejaculation (PE) is a common male sexual disorder. Recent normative data suggest that men with an intravaginal ejaculatory latency time (IELT) of less than 1 minute have "definite" PE, while men with IELTs between 1 and 1.5 minutes have "probable" PE. Although there is insufficient empirical evidence to identify the etiology of PE, there is limited correlational evidence to suggest that men with PE have high levels of sexual anxiety and inherited altered sensitivity of central 5-HT (serotonin) receptors. Pharmacological modulation of the ejaculatory threshold using off-label daily or on-demand selective serotonin re-uptake inhibitors (SSRIs) offers patients a high likelihood of achieving improved ejaculatory control within a few days of initiating treatment, consequential improvements in sexual desire and other sexual domains and is well tolerated. Investigational drugs such as the ejaculo-selective serotonin transport inhibitors (ESSTIs) such as dapoxetine and UK-390,957 represent a major development in sexual medicine. These drugs offer patients the convenience of on-demand dosing, significant improvements in IELT, ejaculatory control, and sexual satisfaction with minimal adverse effects.

  19. Dapoxetine: a new option in the medical management of premature ejaculation

    Science.gov (United States)

    2012-01-01

    Premature ejaculation (PE) is a common male sexual disorder which is associated with substantial personal and interpersonal negative psychological consequences. Pharmacotherapy of PE with off-label antidepressant selective serotonin reuptake inhibitors (SSRIs) is common, effective and safe. Development and regulatory approval of drugs specifically for the treatment of PE will reduce reliance on off-label treatments and serve to fill an unmet treatment need. The objective of this article is to review evidence supporting the efficacy and safety of dapoxetine in the treatment of PE. MEDLINE, Web of Science, PICA, EMBASE and the proceedings of major international and regional scientific meetings were searched for publications or abstracts published during the period 1993–2012 that used the word ‘dapoxetine’ in the title, abstract or keywords. This search was then manually cross referenced for all papers. This review encompasses studies of dapoxetine pharmacokinetics, animal studies, human phase I, II and III studies, independent postmarketing and pharmacovigilance efficacy and safety studies and drug-interaction studies. Dapoxetine is a potent SSRI which is administered on demand 1–3 h prior to planned sexual contact. It is rapidly absorbed and eliminated, resulting in minimal accumulation, and has dose-proportional pharmacokinetics which are unaffected by multiple dosing. Dapoxetine 30 mg and 60 mg has been evaluated in five industry-sponsored randomized, double-blind, placebo-controlled studies in 6081 men aged at least 18 years. Outcome measures included stopwatch-measured intravaginal ejaculatory latency time (IELT), Premature Ejaculation Profile (PEP) inventory items, Clinical Global Impression of Change (CGIC) in PE, and adverse events. Mean IELT, all PEP items and CGIC improved significantly with both doses of dapoxetine versus placebo (all p <0.001). The most common treatment-related adverse effects included nausea (11.0% for 30 mg, 22.2% for 60 mg

  20. Impact of proximal cytoplasmic droplets on quality traits and in-vitro embryo production efficiency of cryopreserved bull spermatozoa

    Directory of Open Access Journals (Sweden)

    Carreira Janaina T

    2012-01-01

    Full Text Available Abstract Background Proximal cytoplasmic droplets (PCDs, a remnant of germ cell cytoplasm, are common non-specific morphological defects in bovine semen. This study evaluated the effect of higher percentages of PCDs on the quality of frozen-thawed bovine semen, embryo production and early embryo development. Methods Three ejaculates from each of five (group 1: PCD ≤ 1%, control and eight adult Bos indicus bulls (group 2: PCD ≥ 24% were analysed. Semen samples were examined for: post-thaw motility, vigour of movement, concentration, sperm morphology, slow thermoresistance test (STT, membrane integrity, acrosome status, mitochondrial function using fluorescent probes association (FITC-PSA, PI and JC-1 and sperm chromatin integrity using acridine orange assay. Two bulls from group 2, with 28.5% and 48.5% PCD, respectively, and three bulls from the control group, each with 0% PCD, were selected for IVF (in vitro fertilisation. Results Semen analyses revealed a significant correlation (P Conclusion Higher PCD levels influenced spermatozoa quality traits. IVF and embryo development data showed that cleavage, blastocyst formation and blastocyst hatching may have been influenced by the interaction of morphology traits and individual bull effects.

  1. Cryopreservation of Spermatozoa in Veterinary Medicine

    Directory of Open Access Journals (Sweden)

    Pesch S

    2007-01-01

    Full Text Available Semen cryopreservation and artificial insemination (AI are the most important biotechnological techniques presently applied in animal breeding. Its large-scale introduction in cattle breeding some 60 years ago aimed at the prevention of genital infections transmittable via natural mating as well as breeding progress and success by rapid spreading of valuable genes. Relating to non-food (pet animals, the avoidance of travel and quarantine restrictions and the conservation of genetic resources are additional advantages. A strict legal framework guarantees sire identity and health innocuousness. Similarly strict guidelines regulate the quality of fresh semen, its processing for cryopreservation and its quality after freezing and thawing. Successful application of AI, particularly when using frozen and thawed semen, requires a proven breeding soundness of both, the semen donor and the semen recipient. Satisfactory results matching those of natural mating can then be obtained. It can be expected that the combined use of AI and sexed spermatozoa in distinct breeding programs will further boost breeding progress.

  2. Larger testes are associated with a higher level of polyandry, but a smaller ejaculate volume, across bushcricket species (Tettigoniidae).

    Science.gov (United States)

    Vahed, Karim; Parker, Darren J; Gilbert, James D J

    2011-04-23

    While early models of ejaculate allocation predicted that both relative testes and ejaculate size should increase with sperm competition intensity across species, recent models predict that ejaculate size may actually decrease as testes size and sperm competition intensity increase, owing to the confounding effect of potential male mating rate. A recent study demonstrated that ejaculate volume decreased in relation to increased polyandry across bushcricket species, but testes mass was not measured. Here, we recorded testis mass for 21 bushcricket species, while ejaculate (ampulla) mass, nuptial gift mass, sperm number and polyandry data were largely obtained from the literature. Using phylogenetic-comparative analyses, we found that testis mass increased with the degree of polyandry, but decreased with increasing ejaculate mass. We found no significant relationship between testis mass and either sperm number or nuptial gift mass. While these results are consistent with recent models of ejaculate allocation, they could alternatively be driven by substances in the ejaculate that affect the degree of polyandry and/or by a trade-off between resources spent on testes mass versus non-sperm components of the ejaculate.

  3. On-Demand Treatment of Premature Ejaculation with Citalopram: A Randomized Double-Blind Study

    Directory of Open Access Journals (Sweden)

    Ghafuri Zahra

    2009-10-01

    Full Text Available "nAs the most common male sexual disorder premature ejaculation (PE, also referred to as early ejaculation (EE or rapid ejaculation (RE, affects 30%-40% of sexually active men. Despite the limited number of available studies comparing the efficacy of selective serotonin re-uptake inhibitors (SSRI they have been thought to have beneficial effects for the treatment of patients with PE. In the present study, we assessed the efficacy of on-demand use of citalopram, in the treatment of premature ejaculation. A randomized double blind study of fixed dose on-demand use of citalopram was performed in Roozbeh Psychiatry Hospital, Tehran University of Medical Sciences. The sample was consisted of 80 married patients diagnosed with PE according to Diagnostic and Statistical Manual of Mental Disorders. The patients were randomly assigned to two groups: group 1 consisting of 42 patients received 20mg citalopram, and group 2 consisting of 38 patients received placebo four hours before intercourse for a 4-week treatment course. The effects of drug on the ejaculatory function in each group were assessed by the intravaginal ejaculation latency time (IELT, and the Chinese Index of Premature Ejaculation (CIPE before and at the end of treatment course. The mean IELT increased from 66.78±36.94 to 80.85±43.05 seconds in group 1 and from 63.44±33.16 to 65.71±34.26 seconds in group 2 (P = 0.000. Mean CIPE score increased 1.14±1.04 and 0.52±0.50 in group 1 and 2 respectively (P = 0.002. The patients treated with on demand citalopram showed significantly greater improvement in IELT and CIPE score compared to the patients receiving placebo. It seems that citalopram may be an effective treatment of premature ejaculation with on-demand usage. However further studies are warranted.

  4. Larger ejaculate volumes are associated with a lower degree of polyandry across bushcricket taxa.

    Science.gov (United States)

    Vahed, Karim

    2006-09-22

    In numerous insects, including bushcrickets (Tettigoniidae), males are known to transfer substances in the ejaculate that inhibit the receptivity of females to further matings, but it has not yet been established whether these substances reduce the lifetime degree of polyandry of the female. The aim of this study was to test the hypothesis that larger ejaculate volumes should be associated with a lower degree of polyandry across tettigoniid taxa, controlling for male body mass and phylogeny. Data on ejaculate mass, sperm number, nuptial gift mass and male mass were taken primarily from the literature. The degree of polyandry for 14 species of European bushcrickets was estimated by counting the number of spermatodoses within the spermathecae of field-caught females towards the end of their adult lifespans. Data for four further species were obtained from the literature. Data were analysed by using both species regression and independent contrasts to control for phylogeny. Multiple regression analysis revealed that, as predicted, there was a significant negative association between the degree of polyandry and ejaculate mass, relative to male body mass, across bushcricket taxa. Nuptial gift size and sperm number, however, did not contribute further to interspecific variation in the degree of polyandry. A positive relationship was found, across bushcricket taxa, between relative nuptial gift size and relative ejaculate mass, indicating that larger nuptial gifts allow the male to overcome female resistance to accepting large ejaculates. This appears to be the first comparative evidence that males can manipulate the lifetime degree of polyandry of their mates through the transfer of large ejaculates.

  5. Serotonin Transporter Promoter Region (5-HTTLPR) Polymorphism Is Not Associated With Paroxetine-Induced Ejaculation Delay in Dutch Men With Lifelong Premature Ejaculation

    Science.gov (United States)

    Janssen, Paddy K.C.; Zwinderman, Aeilko H.; Olivier, Berend

    2014-01-01

    Purpose To investigate the association between the 5-HT-transporter-gene-linked promoter region (5-HTTLPR) polymorphism and 20-mg paroxetine-induced ejaculation delay in men with lifelong premature ejaculation (LPE). Materials and Methods This was a prospective study of 10 weeks of paroxetine treatment in 54 men with LPE. Intravaginal ejaculation latency time (IELT) was measured by stopwatch. Controls consisted of 92 Caucasian men. All men with LPE were genotyped for the 5-HTTLPR polymorphism. Allele frequencies and genotypes of short (S) and long (L) variants of the polymorphism were compared between patients and controls. Associations between the LL, SL, and SS genotypes and fold increase of mean IELT were investigated. Results Of the 54 patients, 43 (79.6%) responded to 20-mg paroxetine treatment with an ejaculation delay, whereas 11 patients (20.4%) did not respond; 44%, 18%, and 18% of the patients showed a fold increase in mean IELT of 2-10, 10-20, and more than 20, respectively. Of the 54 men, 14 (25.9%) had the LL genotype, 29 (53.7%) had the SL genotype, and 11 (20.4%) had the SS genotype. In the 92 controls, the LL, SL, and SS genotypes were present in 27 (29.3%), 41 (44.6%), and 24 (26.1%), respectively. No statistically significant differences were found in 5-HTTLPR allelic variations or in 5-HTTLPR gene variations. In all men treated with 20 mg paroxetine, analysis of variance of the natural logarithm of fold increase in the IELT showed no statistically significant difference according to genotype (p=0.83). Conclusions The 5-HTTLPR polymorphism is not associated with daily 20-mg paroxetine treatment-induced ejaculation delay in men with LPE. PMID:24578810

  6. Nuclear status of immature and mature stallion spermatozoa.

    Science.gov (United States)

    Dias, G M; Retamal, C A; Tobella, L; Arnholdt, A C V; López, M L

    2006-07-15

    'The highly packed chromatin of mature spermatozoa results from replacement of somatic-like histones by highly basic arginine- and cysteine-rich protamines during spermatogenesis, with additional conformational changes in chromatin structure during epididymal transit. The objective of the present study was to compare the nuclear characteristics of immature and mature epididymal stallion spermatozoa, using a variety of experimental approaches. Resistance to in vitro decondensation of chromatin, following exposure to SDS-DTT and alkaline thioglycolate, increased significantly in mature spermatozoa. Evaluation of the thiol-disulfide status (monobromobimane labeling) demonstrated that immature cells obtained from ductulli efferentes contained mostly thiol groups, whereas these groups were oxidized in mature cells collected from the cauda epididymidis. Based on atomic absorption spectrophotometry, maturation of stallion spermatozoa was accompanied by a 60% reduction in the Zn(2+) content of sperm cells, concomitant with increased concentrations of this ion in epididymal fluid. Furthermore, the degree of disulfide bonding was inversely correlated with susceptibility of chromatin to acid denaturation (SCSA). Collectively, these data were consistent with the hypothesis that maturation of stallion spermatozoa involves oxidation of sulphydryl groups to form intra- and intermolecular disulfide links between adjacent protamines, with loss of zinc as an integral feature. These changes endow mechanical and chemical resistance to the nucleus, ensuring efficient transmission of the paternal genome at fertilization.

  7. Morphology, morphometry and ultrastructure of the Amazonian manatee (Sirenia: Trichechidae) spermatozoa

    OpenAIRE

    Amaral,Rodrigo S.; LUCCI, Carolina M.; Fernando C.W. Rosas; Vera M. F. da Silva; Báo,Sônia N.

    2010-01-01

    This study describes the morphological, morphometric and ultrastructural characteristics of the Amazonian manatee Trichechus inunguis (Natterer, 1883) spermatozoon. The spermatozoa were obtained from a urine sample of an adult T. inunguis kept in captivity. The spermatozoa were analyzed by light and transmission electron microscopy. The head of Amazonian manatee spermatozoa had a flat oval shape and a well distinguishable midpiece. The mean dimensions of the spermatozoa were: head length, 7.4...

  8. Effect of low-density lipoproteins,spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 ℃%低密度脂蛋白、精子浓度以及甘油对4℃下存储的公牛精子功能和活力的影响

    Institute of Scientific and Technical Information of China (English)

    Oscar Vera-Munoz; Lamia Amirat-Briand; Djemil Bencharif; Marc Anton; Serge Desherces; Eric Shmitt; Chantal Thorin; Daniel Tainturier

    2011-01-01

    An extender has been developed with low-density lipoproteins(LDLs)that eliminates the microbial risks associated with the use of whole egg yolk.The objective of this study was to assess the effects of substituting egg yolk with LDLs for use as an extender in sperm preservation at 4 ℃,as well as on spermatozoa motility,plasma membrane and acrosome integrity,at two different concentrations(80×106 and 240×106 sperm per ml)for 8 days and to evaluate glycerol toxicity in both extenders.A total of 12 ejaculates were collected from three bulls.Spermatozoa motility was examined using computer-assisted semen analysis.Plasma membrane integrity was determined using the hypo-osmotic swelling test and acrosome integrity with the fluorescein isothiocyanate-Pisum sativum agglutinin test.The semen was subsequently divided into four aliquots and diluted with Tris-egg yolk-glycerol(TEG),Tris-egg yolk without glycerol(TE),LDL with glycerol(LDL+)and LDL without glycerol(LDL-),at 80×106 and 240×106 sperm per ml.This study showed that the LDL+ and