Computer-aided sperm analysis: a useful tool to evaluate patient's response to varicocelectomy

OpenAIRE

Ariagno, Julia I; Mendeluk, Gabriela R; Furlan, Mar?a J; Sardi, M; Chenlo, P; Curi, Susana M; Pugliese, Mercedes N; Repetto, Herberto E; Cohen, Mariano

2016-01-01

Preoperative and postoperative sperm parameter values from infertile men with varicocele were analyzed by computer-aided sperm analysis (CASA) to assess if sperm characteristics improved after varicocelectomy. Semen samples of men with proven fertility (n = 38) and men with varicocele-related infertility (n = 61) were also analyzed. Conventional semen analysis was performed according to WHO (2010) criteria and a CASA system was employed to assess kinetic parameters and sperm concentration. Se...

Ultra-differentiation of Sperm Tail of Lesser Egyptian Jerboa, Jaculus jaculus (Family: Dipodidae

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Osama M. Sarhan

2016-01-01

Full Text Available In the present study, events of sperm tail differentiation in Lesser Egyptian Jerboa, Jaculus jaculus were studied for the first time. Generally, stages of sperm tail differentiation are more or less similar to that described by other studies in other rodents. In the present species, special structures were observed. These structures include, first: the formation of a hollow large unit of microtubules that appears to surround the nuclear envelope at its equatorial plane. The manchette microtubules (MMs are re-oriented toward the longitudinal direction and attached along hollow large unit of microtubules. Second, the formation of perinuclear space filled with an electron-translucent substance surrounds the posterior third of the developing nucleus. Third, the nuclear fossa and the connecting piece were inserted in the ventrodorsal region of the nucleus. Fourth, the fibrous sheath (FS is formed of dextral spiral fibrous ribs. Finally, the sperm tail of the present species has a single outer FS, however, other rodents, having additional inner fibrous units, between the outer FS and the inner developing axoneme.

A Systematic Analysis of a Deep Mouse Epididymal Sperm Proteome

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Chauvin, Theodore; Xie, Fang; Liu, Tao; Nicora, Carrie D.; Yang, Feng; Camp, David G.; Smith, Richard D.; Roberts, Kenneth P.

2012-12-21

Spermatozoa are highly specialized cells that, when mature, are capable of navigating the female reproductive tract and fertilizing an oocyte. The sperm cell is thought to be largely quiescent in terms of transcriptional and translational activity. As a result, once it has left the male reproductive tract, the sperm cell is essentially operating with a static population of proteins. It is therefore theoretically possible to understand the protein networks contained in a sperm cell and to deduce its cellular function capabilities. To this end we have performed a proteomic analysis of mouse sperm isolated from the cauda epididymis and have confidently identified 2,850 proteins, which is the most comprehensive sperm proteome for any species reported to date. These proteins comprise many complete cellular pathways, including those for energy production via glycolysis, β-oxidation and oxidative phosphorylation, protein folding and transport, and cell signaling systems. This proteome should prove a useful tool for assembly and testing of protein networks important for sperm function.

Testicular sperm is superior to ejaculated sperm for ICSI in cryptozoospermia: An update systematic review and meta-analysis.

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Kang, Yi-No; Hsiao, Ya-Wen; Chen, Chien-Yu; Wu, Chien-Chih

2018-05-18

Intracytoplasmic sperm injection (ICSI) is well established and provides patients with severely impaired sperm quality with an opportunity to father a child. However, previous studies do not clearly indicate whether male with cryptozoospermia should use testicular sperm or ejaculated sperm for ICSI. The newest systematic review of this topic also gave a controversial conclusion that was based on incorrect pooling result. Moreover, two clinical studies published after the systematic review. In the present update systematic review and meta-analysis, a comprehensive citation search for relevant studies was performed using the Cochrane library databases, Embase, Ovid MEDLINE, PubMed, ScienceDirect, Scopus, and Web of Science up to September 2017. The search returned 313 records, in which six studies were included in quantitative synthesis. These studies involved 578 male infertility patients who had undergone 761 ICSI cycles. The risk ratios favour fresh testicular sperm for good quality embryo rate (1.17, 95% CI 1.05-1.30, P = 0.005), implantation rate (95% CI 1.02-2.26, P = 0.04), and pregnancy rate (RR = 1.74, 95% CI 1.20-2.52, P = 0.004). In conclusion, the existing evidence suggests that testicular sperm is better than ejaculated sperm for ICSI in male with cryptozoospermia.

The effects of male age on sperm analysis by motile sperm organelle morphology examination (MSOME

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Silva Liliane FI

2012-03-01

Full Text Available Abstract Background This study aimed to investigate the influence of age on sperm quality, as analysed by motile sperm organelle morphology examination (MSOME. Methods Semen samples were collected from 975 men undergoing evaluation or treatment for infertility. Sperm cells were evaluated at 8400× magnification using an inverted microscope equipped with Nomarski (differential interference contrast optics. Two forms of spermatozoa were considered: normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV, defined as vacuoles occupying > 50% of the sperm nuclear area. At least 200 spermatozoa per sample were evaluated, and the percentages of normal and LNV spermatozoa were determined. The subjects were divided into three groups according to age: Group I, less than or equal to 35 years; Group II, 36-40 years; and Group III, greater than or equal to 41 years. Results There was no difference in the percentages of normal sperm between the two younger (I and II groups (P >0.05. The percentage of normal sperm in the older group (III was significantly lower than that in the younger (I and II groups (P P >0.05. The percentage of LNV spermatozoa was significantly higher in the older group (III than in the younger (I and II groups (P P P Conclusion The results demonstrated a consistent decline in semen quality, as reflected by morphological evaluation by MSOME, with increased age. Considering the relationship between nuclear vacuoles and DNA damage, these age-related changes predict that increased paternal age should be associated with unsuccessful or abnormal pregnancy as a consequence of fertilisation with damaged spermatozoa. Given that sperm nuclear vacuoles can be evaluated more precisely at high magnification, these results support the routine use of MSOME for ICSI as a criterion for semen analysis.

Automated Analysis of Human Sperm Number and Concentration (Oligospermia) Using Otsu Threshold Method and Labelling

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Susrama, I. G.; Purnama, K. E.; Purnomo, M. H.

2016-01-01

Oligospermia is a male fertility issue defined as a low sperm concentration in the ejaculate. Normally the sperm concentration is 20-120 million/ml, while Oligospermia patients has sperm concentration less than 20 million/ml. Sperm test done in the fertility laboratory to determine oligospermia by checking fresh sperm according to WHO standards in 2010 [9]. The sperm seen in a microscope using a Neubauer improved counting chamber and manually count the number of sperm. In order to be counted automatically, this research made an automation system to analyse and count the sperm concentration called Automated Analysis of Sperm Concentration Counters (A2SC2) using Otsu threshold segmentation process and morphology. Data sperm used is the fresh sperm directly in the analysis in the laboratory from 10 people. The test results using A2SC2 method obtained an accuracy of 91%. Thus in this study, A2SC2 can be used to calculate the amount and concentration of sperm automatically

Microfluidic single sperm entrapment and analysis

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de Wagenaar, B.; Berendsen, Johanna Theodora Wilhelmina; Berendsen, J.T.W.; Bomer, Johan G.; Olthuis, Wouter; van den Berg, Albert; Segerink, Loes Irene

2015-01-01

Selection of healthy spermatozoa is of crucial importance for the success rates of assisted reproduction technologies (ART) such as in vitro fertilization and intra-cytoplasmic sperm injection. Although sperm selection for ART procedures is predominantly based on sperm motility, successful

Sexual selection and the adaptive evolution of PKDREJ protein in primates and rodents.

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Vicens, Alberto; Gómez Montoto, Laura; Couso-Ferrer, Francisco; Sutton, Keith A; Roldan, Eduardo R S

2015-02-01

PKDREJ is a testis-specific protein thought to be located on the sperm surface. Functional studies in the mouse revealed that loss of PKDREJ has effects on sperm transport and the ability to undergo an induced acrosome reaction. Thus, PKDREJ has been considered a potential target of post-copulatory sexual selection in the form of sperm competition. Proteins involved in reproductive processes often show accelerated evolution. In many cases, this rapid divergence is promoted by positive selection which may be driven, at least in part, by post-copulatory sexual selection. We analysed the evolution of the PKDREJ protein in primates and rodents and assessed whether PKDREJ divergence is associated with testes mass relative to body mass, which is a reliable proxy of sperm competition levels. Evidence of an association between the evolutionary rate of the PKDREJ gene and testes mass relative to body mass was not found in primates. Among rodents, evidence of positive selection was detected in the Pkdrej gene in the family Cricetidae but not in Muridae. We then assessed whether Pkdrej divergence is associated with episodes of sperm competition in these families. We detected a positive significant correlation between the evolutionary rates of Pkdrej and testes mass relative to body mass in cricetids. These findings constitute the first evidence of post-copulatory sexual selection influencing the evolution of a protein that participates in the mechanisms regulating sperm transport and the acrosome reaction, strongly suggesting that positive selection may act on these fertilization steps, leading to advantages in situations of sperm competition. © The Author 2014. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Comparison of sperm subpopulation structures in first and second ejaculated semen from Japanese black bulls by a cluster analysis of sperm motility evaluated by a CASA system.

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Kanno, Chihiro; Sakamoto, Kentaro Q; Yanagawa, Yojiro; Takahashi, Yoshiyuki; Katagiri, Seiji; Nagano, Masashi

2017-08-04

In the present study, bull sperm in the first and second ejaculates were divided into subpopulations based on their motility characteristics using a cluster analysis of data from computer-assisted sperm motility analysis (CASA). Semen samples were collected from 4 Japanese black bulls. Data from 9,228 motile sperm were classified into 4 clusters; 1) very rapid and progressively motile sperm, 2) rapid and circularly motile sperm with widely moving heads, 3) moderately motile sperm with heads moving frequently in a short length, and 4) poorly motile sperm. The percentage of cluster 1 varied between bulls. The first ejaculates had a higher proportion of cluster 2 and lower proportion of cluster 3 than the second ejaculates.

Evaluation of the effect of implanted depleted uranium on male reproductive success, sperm concentration, and sperm velocity

International Nuclear Information System (INIS)

Arfsten, Darryl P.; Schaeffer, David J.; Johnson, Eric W.; Robert Cunningham, J.; Still, Kenneth R.; Wilfong, Erin R.

2006-01-01

Depleted uranium (DU) projectiles have been used in battle in Iraq and the Balkans and will continue to be a significant armor-penetrating munition for the US military. As demonstrated in the Persian Gulf War, battle injury from DU projectiles and shrapnel is a possibility, and removal of embedded DU fragments from the body is not always practical because of their location in the body or their small size. Previous studies in rodents have demonstrated that implanted DU mobilizes and translocates to the gonads, and natural uranium may be toxic to spermatazoa and the male reproductive tract. In this study, the effects of implanted DU pellets on sperm concentration, motility, and male reproductive success were evaluated in adult (P1) Sprague-Dawley rats implanted with 0, 12, or 20, DU pellets of 1x2 mm or 12 or 20 tantalum (Ta) steel pellets of 1x2 mm. Twenty DU pellets of 1x2 mm (760 mg) implanted in a 500-g rat are equal to approximately 0.2 pound of DU in a 154-lb (70-kg) person. Urinary analysis found that male rats implanted with DU were excreting uranium at postimplantation days 27 and 117 with the amount dependent on dose. No deaths or evidence of toxicity occurred in P1 males over the 150-day postimplantation study period. When assessed at postimplantation day 150, the concentration, motion, and velocity of sperm isolated from DU-implanted animals were not significantly different from those of sham surgery controls. Velocity and motion of sperm isolated from rats treated with the positive control compound α-chlorohydrin were significantly reduced compared with sham surgery controls. There was no evidence of a detrimental effect of DU implantation on mating success at 30-45 days and 120-145 days postimplantation. The results of this study suggest that implantation of up to 20 DU pellets of 1x2 mm in rats for approximately 21% of their adult lifespan does not have an adverse impact on male reproductive success, sperm concentration, or sperm velocity

Raman micro-spectroscopy analysis of different sperm regions: a species comparison.

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Amaral, S; Da Costa, R; Wübbeling, F; Redmann, K; Schlatt, S

2018-04-01

Is Raman micro-spectroscopy a valid approach to assess the biochemical hallmarks of sperm regions (head, midpiece and tail) in four different species? Non-invasive Raman micro-spectroscopy provides spectral patterns enabling the biochemical characterization of the three sperm regions in the four species, revealing however high similarities for each region among species. Raman micro-spectroscopy has been described as an innovative method to assess sperm features having the potential to be used as a non-invasive selection tool. However, except for nuclear DNA, the identification and assignment of spectral bands in Raman-profiles to the different sperm regions is scarce and controversial. Raman spectra from head, midpiece and tail of four different species were obtained. Sperm samples were collected and smeared on microscope slides. Air dried samples were subjected to Raman analysis using previously standardized procedures. Sperm samples from (i) two donors attending the infertility clinic at the Centre of Reproductive Medicine and Andrology; (ii) two C57BL/6 -TgN (ACTbEGFP) 1Osb adult mice; (iii) two adult Cynomolgus monkeys (Macaca fascicularis) and (iv) two sea urchins (Arbacia punctulata) were used to characterize and compare their spectral profiles. Differences and similarities were confirmed by principal component analysis (PCA). Several novel region-specific peaks were identified. The three regions could be differentiated by distinctive Raman patterns irrespective of the species. However, regardless of the specie, their main spectral pattern remains mostly unchanged. These results were corroborated by the PCA analysis and suggest that the basic constituents of spermatozoa are biochemically similar among species. Further research should be performed in live sperm to validate the detected spectral bands and their use as markers of distinctive regions. Raman peaks that have never been described in the sperm cell were detected. Particularly important are those that

Proteomics analysis of ram sperm by heavy ion radiation

International Nuclear Information System (INIS)

He Yuxuan; Li Hongyan; Zhang Hong

2013-01-01

The objective of this study was to investigate the proteome changes induced by heavy ion radiation using irradiated ram sperm by a two-dimensional electrophoresis (2-DE) analysis. The 2D gels were stained with Coomassie Brilliant Blue. Differentially expressed proteins were detected by PDQuest 8.0 software and subjected to ion trap mass spectrometer equipped with a surveyor HPLC system, and differential protein spots were identified. Results showed there are five differential protein spots in irradiated sperm gels, four up-regulated protein spots and one spot missed. The differentially expressed protein spots were identified to be two up-regulated proteins including enolase, and enolase 1. It was concluded there was proteome changes induced by heavy ion radiation in ram sperm, which may be useful to clarify the physiology state of ram sperm in heavy ion radiation and provide a theoretical basis for radiation ram breeding. (authors)

Cytometric analysis of shape and DNA content in mammalian sperm

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Gledhill, B.L.

1983-10-10

Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. Sperm were analyzed by flow cytometry and slit-scan flow analysis for injury following the exposure of testes to mutagens. The utility of flow cytometry in genotoxin screening and monitoring of occupational exposure was evaluated. The technique proved valuable in separation of X- and Y-chromosome bearing sperm and the potential applicability of this technique in artificial insemination and a solution, of accurately assessing the DNA content of sperm were evaluated-with reference to determination of X- and Y-chromosome bearing sperm.

Cytometric analysis of shape and DNA content in mammalian sperm

International Nuclear Information System (INIS)

Gledhill, B.L.

1983-01-01

Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. Sperm were analyzed by flow cytometry and slit-scan flow analysis for injury following the exposure of testes to mutagens. The utility of flow cytometry in genotoxin screening and monitoring of occupational exposure was evaluated. The technique proved valuable in separation of X- and Y-chromosome bearing sperm and the potential applicability of this technique in artificial insemination and a solution, of accurately assessing the DNA content of sperm were evaluated-with reference to determination of X- and Y-chromosome bearing sperm

Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

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Matsuura, Koji; Huang, Han-Wei; Chen, Ming-Cheng; Chen, Yu; Cheng, Chao-Min

2017-04-01

Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.

Cytometric analysis of mammalian sperm for induced morphologic and DNA content errors

International Nuclear Information System (INIS)

Pinkel, D.

1983-01-01

Some flow-cytometric and image analysis procedures under development for quantitative analysis of sperm morphology are reviewed. The results of flow-cytometric DNA-content measurements on sperm from radiation exposed mice are also summarized, the results related to the available cytological information, and their potential dosimetric sensitivity discussed

Sperm head's birefringence: a new criterion for sperm selection.

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Gianaroli, Luca; Magli, M Cristina; Collodel, Giulia; Moretti, Elena; Ferraretti, Anna P; Baccetti, Baccio

2008-07-01

To investigate the characteristics of birefringence in human sperm heads and apply polarization microscopy for sperm selection at intracytoplasmic sperm injection (ICSI). Prospective randomized study. Reproductive Medicine Unit, Società Italiana Studi Medicina della Riproduzione, Bologna, Italy. A total of 112 male patients had birefringent sperm selected for ICSI (study group). The clinical outcome was compared with that obtained in 119 couples who underwent a conventional ICSI cycle (control group). The proportion of birefringent spermatozoa was evaluated before and after treatment in relation to the sperm sample quality. Embryo development and clinical outcome in the study group were compared with those in the controls. Proportion of birefringent sperm heads, rates of fertilization, cleavage, pregnancy, implantation, and ongoing implantation. The proportion of birefringent spermatozoa was significantly higher in normospermic samples when compared with oligoasthenoteratospermic samples with no progressive motility and testicular sperm extraction samples. Although fertilization and cleavage rates did not differ between the study and control groups, in the most severe male factor condition (oligoasthenoteratospermic with no progressive motility and testicular sperm extraction), the rates of clinical pregnancy, ongoing pregnancy, and implantation were significantly higher in the study group versus the controls. The analysis of birefringence in the sperm head could represent both a diagnostic tool and a novel method for sperm selection.

Sperm structure and motility in the eusocial naked mole-rat, Heterocephalus glaber: a case of degenerative orthogenesis in the absence of sperm competition?

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van der Horst Gerhard

2011-12-01

Full Text Available Abstract Background We have studied sperm structure and motility in a eusocial rodent where reproduction is typically restricted to a single male and behaviourally dominant queen. Males rarely compete for access to the queen during her estrus cycle, suggesting little or no role for sperm competition. Results Our results revealed an atypical mammalian sperm structure with spermatozoa from breeding, subordinate and disperser males being degenerate and almost completely lacking a "mammalian phylogenetic stamp". Sperm structure is characterized by extreme polymorphism with most spermatozoa classified as abnormal. Sperm head shapes include round, oval, elongated, lobed, asymmetrical and amorphous. At the ultrastructural level, the sperm head contains condensed to granular chromatin with large open spaces between the chromatin. Nuclear chromatin seems disorganized since chromatin condensation is irregular and extremely inconsistent. The acrosome forms a cap (ca 35% over the anterior part of the head. A well defined nuclear fossa and neck with five minor sets of banded protein structures are present. The midpiece is poorly organized and contains only 5 to 7 round to oval mitochondria. The flagellar pattern is 9+9+2. A distinct degenerative feature of the tail principal piece is the absence of the fibrous sheath. Only 7% motile spermatozoa were observed which had exceptionally slow swimming speeds. Conclusion In this species, sperm form has simplified and degenerated in many aspects and represents a specialised form of degenerative orthogenesis at the cellular level.

Analysis of the impact of cryopreservation and theophylline on motility of sperm

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Elaheh Gorji

2018-06-01

Full Text Available Objective: Sperm parameters, particularly motility, decrease during cryopreservation. Theophylline generally enhances sperm motility. We analyzed effects of theophylline and freezing on sperm motility.Design: Experimental study.Setting: Private IVF lab.Setting: IVF lab of Mehrgan Hospital. Method: 22–55 year-old men participated in this study (30 fresh ejaculation and 8 TESE samples. After sperm analysis, we added theophylline (40 mM to half of our samples as case group to compare motility with the remaining samples as control group. Cryopreservation was performed in two groups. After thawing, motility of both groups was recorded. Furthermore, theophylline (40 mM was applied to both groups after thawing again. Result: After adding theophylline, sperm motility improved significantly in all samples. Sperm motility reduced in control group more than the study group after freeze-thaw procedure (P < 0.002, normal morphology <5%. Sperm motility was not enhanced significantly by re-adding of theophylline to the two groups. Interactions between stages and groups were statistically significant in semen and biopsy samples (p < 0.001. Conclusion: Adding theophylline before freezing can preserve motility of sperms in samples with different parameters and even sperms extracted in testicular biopsy. Theophylline may have protective impact on sperms in freezing procedure. Keywords: Sperm motility, Theophylline, Freezing, Morphology, Biopsy

New Perspectives on Rodent Models of Advanced Paternal Age: Relevance to Autism

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Claire J Foldi

2011-06-01

Full Text Available Offspring of older fathers have an increased risk of various adverse health outcomes, including autism and schizophrenia. With respect to biological mechanisms for this association, there are many more germline cell divisions in the life history of a sperm relative to that of an oocyte. This leads to more opportunities for copy error mutations in germ cells from older fathers. Evidence also suggests that epigenetic patterning in the sperm from older men is altered. Rodent models provide an experimental platform to examine the association between paternal age and brain development. Several rodent models of advanced paternal age (APA have been published with relevance to intermediate phenotypes related to autism. All four published APA models vary in key features creating a lack of consistency with respect to behavioural phenotypes. A consideration of common phenotypes that emerge from these APA-related mouse models may be informative in the exploration of the molecular and neurobiological correlates of APA.

Computer-assisted sperm analysis (CASA): capabilities and potential developments.

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Amann, Rupert P; Waberski, Dagmar

2014-01-01

Computer-assisted sperm analysis (CASA) systems have evolved over approximately 40 years, through advances in devices to capture the image from a microscope, huge increases in computational power concurrent with amazing reduction in size of computers, new computer languages, and updated/expanded software algorithms. Remarkably, basic concepts for identifying sperm and their motion patterns are little changed. Older and slower systems remain in use. Most major spermatology laboratories and semen processing facilities have a CASA system, but the extent of reliance thereon ranges widely. This review describes capabilities and limitations of present CASA technology used with boar, bull, and stallion sperm, followed by possible future developments. Each marketed system is different. Modern CASA systems can automatically view multiple fields in a shallow specimen chamber to capture strobe-like images of 500 to >2000 sperm, at 50 or 60 frames per second, in clear or complex extenders, and in information for ≥ 30 frames and provide summary data for each spermatozoon and the population. A few systems evaluate sperm morphology concurrent with motion. CASA cannot accurately predict 'fertility' that will be obtained with a semen sample or subject. However, when carefully validated, current CASA systems provide information important for quality assurance of semen planned for marketing, and for the understanding of the diversity of sperm responses to changes in the microenvironment in research. The four take-home messages from this review are: (1) animal species, extender or medium, specimen chamber, intensity of illumination, imaging hardware and software, instrument settings, technician, etc., all affect accuracy and precision of output values; (2) semen production facilities probably do not need a substantially different CASA system whereas biology laboratories would benefit from systems capable of imaging and tracking sperm in deep chambers for a flexible period of time

Environmental toxicants cause sperm DNA fragmentation as detected by the Sperm Chromatin Structure Assay (SCSA[reg])

International Nuclear Information System (INIS)

Evenson, Donald P.; Wixon, Regina

2005-01-01

Studies over the past two decades have clearly shown that reproductive toxicants cause sperm DNA fragmentation. This DNA fragmentation can usually be detected prior to observing alterations of metaphase chromosomes in embryos. Thus, Sperm Chromatin Structure Assay (SCSA)-detected DNA damage is viewed as the molecular precursor to later gross chromosome damage observed under the light microscope. SCSA measurements of animal or human sperm consist of first obtaining a fresh or flash frozen neat semen sample in LN2 or dry ice. Samples are then sent to a SCSA diagnostic laboratory where the samples are thawed, diluted to ∼1-2 x 106 sperm/ml, treated for 30 s with a pH 1.2 detergent buffer and then stained with acridine orange (AO). The low pH partially denatures DNA at the sites of DNA strand breaks and the AO-ssDNA fluoresces red while the AO-dsDNA fluoresces green. Flow cytometry measurements of 5000 sperm/sample provide statistically robust data on the ratio of red to green sperm, the extent of the DNA fragmentation and the standard deviations of measures. Numerous experiments on rodents treated with reproductive toxicants clearly showed that SCSA measures are highly dose responsive and have a very low CV. Different agents that act on germ cells at various stages of development usually showed sperm DNA fragmentation when that germ cell fraction arrived in the epididymis or ejaculate. Some of these treated samples were capable of successful in vitro fertilization but with frequent embryo failure. A 2-year longitudinal study of men living a valley town with a reported abnormal level of infertility and spontaneous miscarriages and also a seasonal atmospheric smog pollution, showed, for the first time, that SCSA measurements of human sperm DNA fragmentation were detectable and correlated with dosage of air pollution while the classical semen measures were not correlated. Also, young men spraying pesticides without protective gear are at an increased risk for elevated

Association between air pollution and sperm quality: A systematic review and meta-analysis

International Nuclear Information System (INIS)

Deng, Zibing; Chen, Fei; Zhang, Meixia; Lan, Lan; Qiao, Zhijiao; Cui, Yan; An, Jinghuan; Wang, Nan; Fan, Zhiwei; Zhao, Xing; Li, Xiaosong

2016-01-01

Exposure to ambient air pollution has been clearly linked to adverse reproductive outcome and fecundation index, but its effects on male semen quality are still uncertain. In this study, we reviewed information from ten studies to get the qualitative evidence of the influence of the ambient air pollution on sperm quality and collected data from six of the ten studies to conduct meta-analysis. The original studies classified participants into different exposure levels and the highest and lowest expose levels were chosen as high expose and low expose groups, respectively. The random-effect model was used in the meta-analysis with the weight mean difference (WMD) as the measure indicator. The WMDs (95% confidence intervals, CIs) of sperm volume, sperm count, semen concentration, sperm progressive motility, total motility, and normal morphology were 0.09 (−0.04, 0.23), 0.46 (−4.47, 5.39), −8.21 (−20.38, 3.96), −7.76 (−16.26, 0.74), −7.61 (−16.97, 1.74) and −3.40 (−7.42, 0.62), respectively. In conclusion, although the differences are not statistically significant between the two groups, the overall trends and evidence from this review indicate the chronic exposure to ambient pollutants at high level may alter men sperm quality. - Highlights: • Comparing to the low exposure group, the high exposure group had larger WMD of the sperm volume and sperm count. • Comparing to the low exposure group, the high exposure group had smaller WMD of the rest sperm parameters. • There was an obvious alteration of the sperm parameters in the two groups. - The overall trends and evidence from this review indicate the chronic exposure to ambient pollutants at high level may alter men sperm quality.

Morphometric analysis of llama (Lama glama) sperm head.

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Casaretto, C; Lombardo, D M; Giuliano, S; Gambarotta, M; Carretero, M I; Miragaya, M H

2012-05-01

Llama production in Argentina has increased, as the international interest in breeding this type of animals has grown in the last years. Considering the great polymorphism that llama spermatozoa present at evaluation using light microscopy, the aim of this study was to objectively evaluate llama sperm head morphometry using digital morphometric analysis. Five ejaculates from each of eight males were obtained to evaluate morphometric parameters of 8000 sperm heads stained with Tinción 15(®). The following average results were obtained for each parameter: size parameters: area 20.09 μm(2), length 6.60 μm, width 4.14 μm, equivalent circle diameter 5.06 μm, curve length 5.79 μm and curve width 3.48 μm; boundary parameters: perimeter 18.54 μm and convex perimeter 17.34 μm; and shape parameters: roundness 1.28 and elongation 1.59. Morphometric parameters of sperm head were compared between ejaculates of the same male and between males. Significant differences between ejaculates of the same male were found for all parameters evaluated (P < 0.01). Significant differences between males were found for all morphometric parameters (P < 0.01) except for curve length, curve width and perimeter. The differences detected would indicate that there is not a single morphometric pattern for Lama glama sperm head, because parameter values cannot be standardised. © 2011 Blackwell Verlag GmbH.

Sperm dynamics in spiders (Araneae): ultrastructural analysis of the sperm activation process in the garden spider Argiope bruennichi (Scopoli, 1772).

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Vöcking, Oliver; Uhl, Gabriele; Michalik, Peter

2013-01-01

Storage of sperm inside the female genital tract is an integral phase of reproduction in many animal species. The sperm storage site constitutes the arena for sperm activation, sperm competition and female sperm choice. Consequently, to understand animal mating systems information on the processes that occur from sperm transfer to fertilization is required. Here, we focus on sperm activation in spiders. Male spiders produce sperm whose cell components are coiled within the sperm cell and that are surrounded by a proteinaceous sheath. These inactive and encapsulated sperm are transferred to the female spermathecae where they are stored for later fertilization. We analyzed the ultrastructural changes of sperm cells during residency time in the female genital system of the orb-web spider Argiope bruennichi. We found three clearly distinguishable sperm conditions: encapsulated sperm (secretion sheath present), decapsulated (secretion sheath absent) and uncoiled sperm (cell components uncoiled, presumably activated). After insemination, sperm remain in the encapsulated condition for several days and become decapsulated after variable periods of time. A variable portion of the decapsulated sperm transforms rapidly to the uncoiled condition resulting in a simultaneous occurrence of decapsulated and uncoiled sperm. After oviposition, only decapsulated and uncoiled sperm are left in the spermathecae, strongly suggesting that the activation process is not reversible. Furthermore, we found four different types of secretion in the spermathecae which might play a role in the decapsulation and activation process.

Recent advances in bird sperm morphometric analysis and its role in male gamete characterization and reproduction technologies

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Julian Santiago-Moreno

2016-01-01

Full Text Available Postcopulatory sexual selection through sperm competition may be an important evolutionary force affecting many reproductive traits, including sperm morphometrics. Environmental factors such as pollutants, pesticides, and climate change may affect different sperm traits, and thus reproduction, in sensitive bird species. Many sperm-handling processes used in assisted reproductive techniques may also affect the size of sperm cells. The accurately measured dimensions of sperm cell structures (especially the head can thus be used as indicators of environmental influences, in improving our understanding of reproductive and evolutionary strategies, and for optimizing assisted reproductive techniques (e.g., sperm cryopreservation for use with birds. Computer-assisted sperm morphometry analysis (CASA-Morph provides an accurate and reliable method for assessing sperm morphometry, reducing the problem of subjectivity associated with human visual assessment. Computerized systems have been standardized for use with semen from different mammalian species. Avian spermatozoa, however, are filiform, limiting their analysis with such systems, which were developed to examine the approximately spherical heads of mammalian sperm cells. To help overcome this, the standardization of staining techniques to be used in computer-assessed light microscopical methods is a priority. The present review discusses these points and describes the sperm morphometric characteristics of several wild and domestic bird species.

Relationship between sperm parameters and intracytoplasmic sperm injection outcome

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Shahla Chaichian

2015-12-01

Full Text Available Objectives: With the adventure of intracytoplasmic sperm injection (ICSI technique, great progresses have developed in the treatment of infertility. Concentration on the properties of male’s gamete has been encouraged by the increasing concerns about the causes of ICSI failure. We hence conducted this study to investigate the probable association of sperm parameters with ISCI outcome. Methods: A total of 523 couples referred to Isfahan Fertility and Sterility Center from January 2007 to June 2008 for ICSI. Semen analysis was performed before ICSI procedure according to the WHO criteria. Patients were assigned into successful ICSI (case and failed ICSI (control groups. Sperm parameters were then compared between the 2 groups. Results: One hundred and six patients (20% had successful ICSI results (case group compared with 417 couples (80% with undesirable ICSI outcomes (control group. Among evaluated factors, sperm agglutination (p = 0.007, sperm concentration (p = 0.043, leukocytospermia (p = 0.026 and head abnormality of sperm (p = 0.019 showed statistically significant differences between two groups with differing ICSI results. None of the other semen parameters revealed significant differences between these two groups. Conclusion: Our study showed that some sperm parameters are associated with desirable ICSI outcome. However, it is unclear whether these associations are causal.

Validation of a novel computer-assisted sperm analysis (CASA) system using multitarget-tracking algorithms.

Science.gov (United States)

Tomlinson, Mathew James; Pooley, Karen; Simpson, Tracey; Newton, Thomas; Hopkisson, James; Jayaprakasan, Kannamanadias; Jayaprakasan, Rajisha; Naeem, Asad; Pridmore, Tony

2010-04-01

To determine the accuracy and precision of a novel computer-assisted sperm analysis (CASA) system by comparison with existing recommended manual methods. Prospective study using comparative measurements of sperm concentration and motility on latex beads and immotile and motile sperm. Tertiary referral fertility center with strong academic links. Sperm donors and male partners of couples attending for fertility investigations. None. Achievement of Accubead target value for high and low concentration suspensions. Repeatability as demonstrated by coefficients of variation and intraclass correlation coefficients. Correlation and limits of agreement between CASA and manual methods. The CASA measurements of latex beads and sperm concentrations demonstrated a high level of accuracy and repeatability. Repeated Accubead measurements attained the required target value (mean difference from target of 2.61% and 3.71% for high- and low-concentration suspensions, respectively) and were highly reproducible. Limits of agreement analysis suggested that manual and CASA counts compared directly could be deemed to be interchangeable. Manual and CASA motility measurements were highly correlated for grades a, b, and d but could not be deemed to be interchangeable, and manual motility estimates were consistently higher for motile sperm. The novel CASA system was able to provide semen quality measurements for sperm concentration and motility measurements which were at least as reliable as current manual methods. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

EDC IMPACT: Reduced sperm counts in rats exposed to human relevant mixtures of endocrine disrupters

DEFF Research Database (Denmark)

Axelstad Petersen, Marta; Hass, Ulla; Scholze, M.

2018-01-01

and the high doses of the total and the anti-androgenic mixture, compared to controls. In all dose groups, epididymal sperm counts were reduced several months after end of exposure, i.e. at 10 months of age. Interestingly, the same pattern of effects was seen for paracetamol as for mixtures with diverse modes...... of action. Reduced sperm count was seen at a dose level reflecting human therapeutic exposure to paracetamol. Environmental chemical mixtures affected sperm count at the lowest mixture dose indicating an insufficient margin of safety for the most exposed humans. This causes concern for exposure of pregnant......Human semen quality is declining in many parts of the world, but the causes are ill defined. In rodents, impaired sperm production can be seen with early life exposure to certain endocrine-disrupting chemicals, but the effects of combined exposures are not properly investigated. In this study, we...

Factors influencing boar sperm cryosurvival.

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Roca, J; Hernández, M; Carvajal, G; Vázquez, J M; Martínez, E A

2006-10-01

Optimal sperm cryopreservation is a prerequisite for the sustainable commercial application of frozen-thawed boar semen for AI. Three experiments were performed to identify factors influencing variability of postthaw sperm survival among 464 boar ejaculates. Sperm-rich ejaculate fractions were cryopre-served using a standard freezing-thawing procedure for 0.5-mL plastic straws and computer-controlled freezing equipment. Postthaw sperm motility (assessed with a computer-assisted semen analysis system) and viability (simultaneously probed by flow cytometry analysis after triple-fluorescent stain), evaluated 30 and 150 min postthaw, were used to estimate the success of cryopreservation. In the first experiment, 168 unselected ejaculates (1 ejaculate/boar), from boars of 6 breeds with a wide age range (8 to 48 mo), were cryopreserved over a 12-mo period to evaluate the predictive value of boar (breed and age), semen collection, transport variables (season of ejaculate collection, interval between collections, and ejaculate temperature exposure), initial semen traits, and sperm quality before freezing on sperm survival after freezing-thawing. In Exp. 2, 4 ejaculates from each of 29 boars, preselected according to their initial semen traits and sperm quality before freezing, were collected and frozen over a 6-mo period to evaluate the influence of interboar and intraboar ejaculate variability in the survival of sperm after cryopreservation. In Exp. 3, 12 ejaculates preselected as for Exp. 2, from each of 15 boars with known good sperm cryosurvival, were collected and frozen over a 12-mo period to estimate the sustainability of sperm cryosurvival between ejaculates over time. Boar and semen collection and transport variables were not predictive of sperm cryosurvival among ejaculates. Initial semen traits and sperm quality variables observed before freezing explained 23.2 and 10.9%, respectively, of the variation in postthaw sperm motility and viability. However, more that

Spermiogram and sperm head morphometry assessed by multivariate cluster analysis results during adolescence (12-18 years and the effect of varicocele

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Fernando Vásquez

2016-01-01

Full Text Available This work evaluates sperm head morphometric characteristics in adolescents from 12 to 18 years of age, and the effect of varicocele. Volunteers between 150 and 224 months of age (mean 191, n = 87, who had reached oigarche by 12 years old, were recruited in the area of Barranquilla, Colombia. Morphometric analysis of sperm heads was performed with principal component (PC and discriminant analysis. Combining seminal fluid and sperm parameters provided five PCs: two related to sperm morphometry, one to sperm motility, and two to seminal fluid components. Discriminant analysis on the morphometric results of varicocele and nonvaricocele groups did not provide a useful classification matrix. Of the semen-related PCs, the most explanatory (40% was related to sperm motility. Two PCs, including sperm head elongation and size, were sufficient to evaluate sperm morphometric characteristics. Most of the morphometric variables were correlated with age, with an increase in size and decrease in the elongation of the sperm head. For head size, the entire sperm population could be divided into two morphometric subpopulations, SP1 and SP2, which did not change during adolescence. In general, for varicocele individuals, SP1 had larger and more elongated sperm heads than SP2, which had smaller and more elongated heads than in nonvaricocele men. In summary, sperm head morphometry assessed by CASA-Morph and multivariate cluster analysis provides a better comprehension of the ejaculate structure and possibly sperm function. Morphometric analysis provides much more information than data obtained from conventional semen analysis.

Association between air pollution and sperm quality: A systematic review and meta-analysis.

Science.gov (United States)

Deng, Zibing; Chen, Fei; Zhang, Meixia; Lan, Lan; Qiao, Zhijiao; Cui, Yan; An, Jinghuan; Wang, Nan; Fan, Zhiwei; Zhao, Xing; Li, Xiaosong

2016-01-01

Exposure to ambient air pollution has been clearly linked to adverse reproductive outcome and fecundation index, but its effects on male semen quality are still uncertain. In this study, we reviewed information from ten studies to get the qualitative evidence of the influence of the ambient air pollution on sperm quality and collected data from six of the ten studies to conduct meta-analysis. The original studies classified participants into different exposure levels and the highest and lowest expose levels were chosen as high expose and low expose groups, respectively. The random-effect model was used in the meta-analysis with the weight mean difference (WMD) as the measure indicator. The WMDs (95% confidence intervals, CIs) of sperm volume, sperm count, semen concentration, sperm progressive motility, total motility, and normal morphology were 0.09 (-0.04, 0.23), 0.46 (-4.47, 5.39), -8.21 (-20.38, 3.96), -7.76 (-16.26, 0.74), -7.61 (-16.97, 1.74) and -3.40 (-7.42, 0.62), respectively. In conclusion, although the differences are not statistically significant between the two groups, the overall trends and evidence from this review indicate the chronic exposure to ambient pollutants at high level may alter men sperm quality. Copyright © 2015 Elsevier Ltd. All rights reserved.

Temporal trends in sperm count: a systematic review and meta-regression analysis.

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Levine, Hagai; Jørgensen, Niels; Martino-Andrade, Anderson; Mendiola, Jaime; Weksler-Derri, Dan; Mindlis, Irina; Pinotti, Rachel; Swan, Shanna H

2017-11-01

Reported declines in sperm counts remain controversial today and recent trends are unknown. A definitive meta-analysis is critical given the predictive value of sperm count for fertility, morbidity and mortality. To provide a systematic review and meta-regression analysis of recent trends in sperm counts as measured by sperm concentration (SC) and total sperm count (TSC), and their modification by fertility and geographic group. PubMed/MEDLINE and EMBASE were searched for English language studies of human SC published in 1981-2013. Following a predefined protocol 7518 abstracts were screened and 2510 full articles reporting primary data on SC were reviewed. A total of 244 estimates of SC and TSC from 185 studies of 42 935 men who provided semen samples in 1973-2011 were extracted for meta-regression analysis, as well as information on years of sample collection and covariates [fertility group ('Unselected by fertility' versus 'Fertile'), geographic group ('Western', including North America, Europe Australia and New Zealand versus 'Other', including South America, Asia and Africa), age, ejaculation abstinence time, semen collection method, method of measuring SC and semen volume, exclusion criteria and indicators of completeness of covariate data]. The slopes of SC and TSC were estimated as functions of sample collection year using both simple linear regression and weighted meta-regression models and the latter were adjusted for pre-determined covariates and modification by fertility and geographic group. Assumptions were examined using multiple sensitivity analyses and nonlinear models. SC declined significantly between 1973 and 2011 (slope in unadjusted simple regression models -0.70 million/ml/year; 95% CI: -0.72 to -0.69; P regression analysis reports a significant decline in sperm counts (as measured by SC and TSC) between 1973 and 2011, driven by a 50-60% decline among men unselected by fertility from North America, Europe, Australia and New Zealand. Because

Effect of an isotonic lubricant on sperm collection and sperm quality.

Science.gov (United States)

Agarwal, Ashok; Malvezzi, Helena; Sharma, Rakesh

2013-05-01

To assess the influence of an isotonic lubricant used during sperm sample collection on [1] ease of collection and [2] resultant sperm quality. Paired randomized cross-over design. Tertiary hospital. Healthy men over 18 years old with normal semen analysis as per World Health Organization 2010 guidelines. Collection of semen sample from 22 subjects by masturbation with or without the use of Pre-Seed personal lubricant. Qualitative survey results and quantitative sperm function outcomes were measured to determine resultant sperm quality and collection experience with and without Pre-Seed lubricant. The qualitative questionnaire results showed that 73% of donors prefer the semen collection process with the isotonic lubricant and 55% recommended the use of lubricant in their everyday collection. The motility, viability, membrane integrity, levels of reactive oxygen species, total antioxidant capacity, and percentage of DNA damage in collected semen samples were not affected by the use of the lubricant. More donors prefer, and find it easier, to collect semen samples with the use of the lubricant. The isotonic lubricant Pre-Seed did not compromise sperm quality as evaluated in an array of sperm assays, suggesting its safe use in fertility patients as required during sperm collection. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Semen analysis and sperm function tests: How much to test?

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S S Vasan

2011-01-01

Full Text Available Semen analysis as an integral part of infertility investigations is taken as a surrogate measure for male fecundity in clinical andrology, male fertility, and pregnancy risk assessments. Clearly, laboratory seminology is still very much in its infancy. In as much as the creation of a conventional semen profile will always represent the foundations of male fertility evaluation, the 5th edition of the World Health Organization (WHO manual is a definitive statement on how such assessments should be carried out and how the quality should be controlled. A major advance in this new edition of the WHO manual, resolving the most salient critique of previous editions, is the development of the first well-defined reference ranges for semen analysis based on the analysis of over 1900 recent fathers. The methodology used in the assessment of the usual variables in semen analysis is described, as are many of the less common, but very valuable, sperm function tests. Sperm function testing is used to determine if the sperm have the biologic capacity to perform the tasks necessary to reach and fertilize ova and ultimately result in live births. A variety of tests are available to evaluate different aspects of these functions. To accurately use these functional assays, the clinician must understand what the tests measure, what the indications are for the assays, and how to interpret the results to direct further testing or patient management.

Computer-aided sperm analysis (CASA) in the medical laboratory: CASA in diagnostic andrology and assisted conception.

Science.gov (United States)

Tomlinson, Mathew J; Naeem, Asad

2018-03-21

CASA has been used in reproductive medicine and pathology laboratories for over 25 years, yet the 'fertility industry' generally remains sceptical and has avoided automation, despite clear weaknesses in manual semen analysis. Early implementers had difficulty in validating CASA-Mot instruments against recommended manual methods (haemocytometer) due to the interference of seminal debris and non-sperm cells, which also affects the accuracy of grading motility. Both the inability to provide accurate sperm counts and a lack of consensus as to the value of sperm kinematic parameters appear to have continued to have a negative effect on CASA-Mot's reputation. One positive interpretation from earlier work is that at least one or more measures of sperm velocity adds clinical value to the semen analysis, and these are clearly more objective than any manual motility analysis. Moreover, recent CASA-Mot systems offer simple solutions to earlier problems in eliminating artefacts and have been successfully validated for sperm concentration; as a result, they should be viewed with more confidence in relation to motility grading. Sperm morphology and DNA testing both require an evidence-based consensus and a well-validated (reliable, reproducible) assay to be developed before automation of either can be of real clinical benefit.

The Effect of Glyphosate on Human Sperm Motility and Sperm DNA Fragmentation

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George Anifandis

2018-05-01

Full Text Available Glyphosate is the active ingredient of Roundup®, which is one of the most popular herbicides worldwide. Although many studies have focused on the reproductive toxicity of glyphosate or glyphosate-based herbicides, the majority of them have concluded that the effect of the specific herbicide is negligible, while only a few studies indicate the male reproductive toxicity of glyphosate alone. The aim of the present study was to investigate the effect of 0.36 mg/L glyphosate on sperm motility and sperm DNA fragmentation (SDF. Thirty healthy men volunteered to undergo semen analysis for the purpose of the study. Sperm motility was calculated according to WHO 2010 guidelines at collection time (zero time and 1 h post-treatment with glyphosate. Sperm DNA fragmentation was evaluated with Halosperm® G2 kit for both the control and glyphosate-treated sperm samples. Sperm progressive motility of glyphosate-treated samples was significantly reduced after 1 h post-treatment in comparison to the respective controls, in contrast to the SDF of glyphosate-treated samples, which was comparable to the respective controls. Conclusively, under these in vitro conditions, at high concentrations that greatly exceed environmental exposures, glyphosate exerts toxic effects on sperm progressive motility but not on sperm DNA integrity, meaning that the toxic effect is limited only to motility, at least in the first hour.

Improving sperm banking efficiency in endangered species through the use of a sperm selection method in brown bear (Ursus arctos) thawed sperm.

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Anel-Lopez, L; Ortega-Ferrusola, C; Álvarez, M; Borragán, S; Chamorro, C; Peña, F J; Morrell, J; Anel, L; de Paz, P

2017-06-26

Sperm selection methods such as Single Layer Centrifugation (SLC) have been demonstrated to be a useful tool to improve the quality of sperm samples and therefore to increase the efficiency of other artificial reproductive techniques in several species. This procedure could help to improve the quality of genetic resource banks, which is essential for endangered species. In contrast, these sperm selection methods are optimized and focused on farm animals, where the recovery task is not as important as in endangered species because of their higher sperm availability. The aim of this study was to evaluate two centrifugation methods (300 x g/20 min and 600 x g/10 min) and three concentrations of SLC media (Androcoll-Bear -80, 65 and 50%) to optimise the procedure in order to recover as many sperm with the highest quality as possible. Sperm morphology could be important in the hydrodynamic relationship between the cell and centrifugation medium and thus the effect of sperm head morphometry on sperm yield and its hydrodynamic relationship were studied. The samples selected with Androcoll-Bear 65% showed a very good yield (53.1 ± 2.9) although the yield from Androcoll-Bear 80% was lower (19.3 ± 3.3). The latter showed higher values of motility than the control immediately after post-thawing selection. However, both concentrations of colloid (65 and 80%) showed higher values of viable sperm and viable sperm with intact acrosome than the control. After an incubation of 2 h at 37 °C, the samples from Androcoll-Bear 80% had higher kinematics and proportion of viable sperm with intact acrosome. In the morphometric analysis, the sperm selected by the Androcoll-Bear 80% showed a head with a bigger area which was more elongated than the sperm from other treatments. We conclude that sperm selection with Androcoll-Bear at either 65% or 80% is a suitable technique that allows a sperm population with better quality than the initial sample to be obtained. We recommend the

BMI in relation to sperm count

DEFF Research Database (Denmark)

Sermondade, N; Faure, C; Fezeu, L

2013-01-01

BACKGROUND The global obesity epidemic has paralleled a decrease in semen quality. Yet, the association between obesity and sperm parameters remains controversial. The purpose of this report was to update the evidence on the association between BMI and sperm count through a systematic review...... with meta-analysis. METHODS A systematic review of available literature (with no language restriction) was performed to investigate the impact of BMI on sperm count. Relevant studies published until June 2012 were identified from a Pubmed and EMBASE search. We also included unpublished data (n = 717 men...... studies were included in the meta-analysis, resulting in a sample of 13 077 men from the general population and attending fertility clinics. Data were stratified according to the total sperm count as normozoospermia, oligozoospermia and azoospermia. Standardized weighted mean differences in sperm...

Sperm length evolution in the fungus-growing ants

DEFF Research Database (Denmark)

Baer, B.; Dijkstra, M. B.; Mueller, U. G.

2009-01-01

-growing ants, representing 9 of the 12 recognized genera, and mapped these onto the ant phylogeny. We show that average sperm length across species is highly variable and decreases with mature colony size in basal genera with singly mated queens, suggesting that sperm production or storage constraints affect...... the evolution of sperm length. Sperm length does not decrease further in multiply mating leaf-cutting ants, despite substantial further increases in colony size. In a combined analysis, sexual dimorphism explained 63.1% of the variance in sperm length between species. As colony size was not a significant...... predictor in this analysis, we conclude that sperm production trade-offs in males have been the major selective force affecting sperm length across the fungus-growing ants, rather than storage constraints in females. The relationship between sperm length and sexual dimorphism remained robust...

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.

Analysis of sperm quality in recombinant inbred mouse strains: correlation of sperm head shape with sperm abnormalities and with the incidence of supplementary spermatozoa in the perivitelline space.

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Krzanowska, H; Styrna, J; Wabik-Sliz, B

1995-07-01

Recombinant inbred strains were developed from reciprocal crosses between two inbred strains of mice (CBA and KE) differing in sperm head shape, proportion of normal sperm heads (CBA, 95%; KE, 78%) and fertilization efficiency (CBA, 100% of fertilized ova; KE, 72%), to determine whether the indices of sperm morphology and function were correlated. The following parameters were analysed in recombinant inbred and progenitor strains: index of sperm head shape (head width in the middle of its length/head length), percentage of abnormal sperm heads, percentage of spermatozoa with progressive movements, efficiency of penetration of hyaluronic acid polymer (Sperm Select) and percentage of fertilized ova after mating males from the tested strains with females from an outbred stock. For each investigated character, recombinant inbred strains, recombinant inbred EXCB and CBXE, could be divided into at least three categories: KE-like, CBA-like and intermediate, suggesting that in each case a minimum of two genes was involved. Recombinant strains derived from the reciprocal crosses of progenitor strains differed only with respect to the proportion of abnormal sperm heads, showing the involvement of the Y chromosome in determining this character. Penetration into Sperm Select was significantly correlated both with fertilization efficiency and sperm motility, while correlation with the proportion of normal spermatozoa did not reach the level of significance. However, there was a significant negative correlation of both sperm abnormalities and the incidence of supplementary spermatozoa in the perivitelline space with the index of sperm head shape.(ABSTRACT TRUNCATED AT 250 WORDS)

Mobile phones affect multiple sperm quality traits: a meta-analysis [v1; ref status: indexed, http://f1000r.es/ny

Directory of Open Access Journals (Sweden)

Madhukar Shivajirao Dama

2013-02-01

Full Text Available As mobile phone usage is growing rapidly, there is a need for a comprehensive analysis of the literature to inform scientific debates about the adverse effects of mobile phone radiation on sperm quality traits. Therefore, we conducted a meta-analysis of the eligible published research studies on human males of reproductive age. Eleven studies were eligible for this analysis. Based on the meta-analysis, mobile phone use was significantly associated with deterioration in semen quality (Hedges’s g = -0.547; 95% CI: -0.713, -0.382; p < 0.001. The traits particularly affected adversely were sperm concentration, sperm morphology, sperm motility, proportion of non-progressive motile sperm (%, proportion of slow progressive motile sperm (%, and sperm viability. Direct exposure of spermatozoa to mobile phone radiation with in vitro study designs also significantly deteriorated the sperm quality (Hedges’s g = -2.233; 95% CI: -2.758, -1.708; p < 0.001, by reducing straight line velocity, fast progressive motility, Hypo-osmotic swelling (HOS test score, major axis (µm, minor axis (µm, total sperm motility, perimeter (µm, area (µm2, average path velocity, curvilinear velocity, motile spermatozoa, and  acrosome reacted spermatozoa (%. The strength of evidence for the different outcomes varied from very low to very high. The analysis shows that mobile phone use is possibly associated with a number of deleterious effects on the spermatozoa.

Sperm Production Rate, Gonadal and Extragonadal Sperm ...

African Journals Online (AJOL)

Five healthy West African Dwarf (WAD) rams, 1.5 to 2.5 years of age and weighing between 15 kg to 20 kg were used to determine daily sperm production, gonadal and exragonadal sperm reserves. Gonadal and extragonadal sperm reserves were estimated by the haemocytometric method, while the daily sperm production ...

Advanced forensic validation for human spermatozoa identification using SPERM HY-LITER™ Express with quantitative image analysis.

Science.gov (United States)

Takamura, Ayari; Watanabe, Ken; Akutsu, Tomoko

2017-07-01

Identification of human semen is indispensable for the investigation of sexual assaults. Fluorescence staining methods using commercial kits, such as the series of SPERM HY-LITER™ kits, have been useful to detect human sperm via strong fluorescence. These kits have been examined from various forensic aspects. However, because of a lack of evaluation methods, these studies did not provide objective, or quantitative, descriptions of the results nor clear criteria for the decisions reached. In addition, the variety of validations was considerably limited. In this study, we conducted more advanced validations of SPERM HY-LITER™ Express using our established image analysis method. Use of this method enabled objective and specific identification of fluorescent sperm's spots and quantitative comparisons of the sperm detection performance under complex experimental conditions. For body fluid mixtures, we examined interference with the fluorescence staining from other body fluid components. Effects of sample decomposition were simulated in high humidity and high temperature conditions. Semen with quite low sperm concentrations, such as azoospermia and oligospermia samples, represented the most challenging cases in application of the kit. Finally, the tolerance of the kit against various acidic and basic environments was analyzed. The validations herein provide useful information for the practical applications of the SPERM HY-LITER™ Express kit, which were previously unobtainable. Moreover, the versatility of our image analysis method toward various complex cases was demonstrated.

BMI in relation to sperm count: an updated systematic review and collaborative meta-analysis.

Science.gov (United States)

Sermondade, N; Faure, C; Fezeu, L; Shayeb, A G; Bonde, J P; Jensen, T K; Van Wely, M; Cao, J; Martini, A C; Eskandar, M; Chavarro, J E; Koloszar, S; Twigt, J M; Ramlau-Hansen, C H; Borges, E; Lotti, F; Steegers-Theunissen, R P M; Zorn, B; Polotsky, A J; La Vignera, S; Eskenazi, B; Tremellen, K; Magnusdottir, E V; Fejes, I; Hercberg, S; Lévy, R; Czernichow, S

2013-01-01

BACKGROUND The global obesity epidemic has paralleled a decrease in semen quality. Yet, the association between obesity and sperm parameters remains controversial. The purpose of this report was to update the evidence on the association between BMI and sperm count through a systematic review with meta-analysis. METHODS A systematic review of available literature (with no language restriction) was performed to investigate the impact of BMI on sperm count. Relevant studies published until June 2012 were identified from a Pubmed and EMBASE search. We also included unpublished data (n = 717 men) obtained from the Infertility Center of Bondy, France. Abstracts of relevant articles were examined and studies that could be included in this review were retrieved. Authors of relevant studies for the meta-analysis were contacted by email and asked to provide standardized data. RESULTS A total of 21 studies were included in the meta-analysis, resulting in a sample of 13 077 men from the general population and attending fertility clinics. Data were stratified according to the total sperm count as normozoospermia, oligozoospermia and azoospermia. Standardized weighted mean differences in sperm concentration did not differ significantly across BMI categories. There was a J-shaped relationship between BMI categories and risk of oligozoospermia or azoospermia. Compared with men of normal weight, the odds ratio (95% confidence interval) for oligozoospermia or azoospermia was 1.15 (0.93-1.43) for underweight, 1.11 (1.01-1.21) for overweight, 1.28 (1.06-1.55) for obese and 2.04 (1.59-2.62) for morbidly obese men. CONCLUSIONS Overweight and obesity were associated with an increased prevalence of azoospermia or oligozoospermia. The main limitation of this report is that studied populations varied, with men recruited from both the general population and infertile couples. Whether weight normalization could improve sperm parameters should be evaluated further.

Sperm quality analysis in XX, XY and YY males of the Nile tilapia (Oreochromis niloticus).

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Gennotte, V; François, E; Rougeot, C; Ponthier, J; Deleuze, S; Mélard, C

2012-07-01

In Nile tilapia (Oreochromis niloticus), individuals with atypical sexual genotype are commonly used in farming (use of YY males to produce all-male offspring), but they also constitute major tools to study sex determinism mechanisms. In other species, sexual genotype and sex reversal procedures affect different aspects of biology, such as growth, behavior and reproductive success. The aim of this study was to assess the influence of sexual genotype on sperm quality in Nile tilapia. Milt characteristics were compared in XX (sex-reversed), XY and YY males in terms of gonadosomatic index, sperm count, sperm motility and duration of sperm motility. Sperm motility was measured by computer-assisted sperm analysis (CASA) quantifying several parameters: total motility, progressive motility, curvilinear velocity, straight line velocity, average path velocity and linearity. None of the sperm traits measured significantly differed between the three genotypes. Mean values of gonadosomatic index, sperm concentration and sperm motility duration of XX, XY and YY males, respectively ranged from 0.92 to 1.33%, from 1.69 to 2.22 ×10(9) cells mL(-1) and from 18'04″ to 27'32″. Mean values of total motility and curvilinear velocity 1 min after sperm activation, respectively ranged from 53 to 58% and from 71 to 76 μm s(-1) for the three genotypes. After 3 min of activity, all the sperm motility and velocity parameters dropped by half and continued to slowly decrease thereafter. Seven min after activation, only 9 to 13% of spermatozoa were still progressive. Our results prove that neither sexual genotype nor hormonal sex reversal treatments affect sperm quality in male Nile tilapias with atypical sexual genotype. Copyright © 2012 Elsevier Inc. All rights reserved.

Immature germ cells in semen - correlation with total sperm count and sperm motility.

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Patil, Priya S; Humbarwadi, Rajendra S; Patil, Ashalata D; Gune, Anita R

2013-07-01

Current data regarding infertility suggests that male factor contributes up to 30% of the total cases of infertility. Semen analysis reveals the presence of spermatozoa as well as a number of non-sperm cells, presently being mentioned in routine semen report as "round cells" without further differentiating them into leucocytes or immature germ cells. The aim of this work was to study a simple, cost-effective, and convenient method for differentiating the round cells in semen into immature germ cells and leucocytes and correlating them with total sperm counts and motility. Semen samples from 120 males, who had come for investigation for infertility, were collected, semen parameters recorded, and stained smears studied for different round cells. Statistical analysis of the data was done to correlate total sperm counts and sperm motility with the occurrence of immature germ cells and leucocytes. The average shedding of immature germ cells in different groups with normal and low sperm counts was compared. The clinical significance of "round cells" in semen and their differentiation into leucocytes and immature germ cells are discussed. Round cells in semen can be differentiated into immature germ cells and leucocytes using simple staining methods. The differential counts mentioned in a semen report give valuable and clinically relevant information. In this study, we observed a negative correlation between total count and immature germ cells, as well as sperm motility and shedding of immature germ cells. The latter was statistically significant with a P value 0.000.

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...... spermatozoa and whether VD serum levels are associated with semen quality. METHODS Cross-sectional association study of semen quality and VD serum level in 300 men from the general population, and in vitro studies on spermatozoa from 40 men to investigate the effects of VD on intracellular calcium, sperm......M). 1,25(OH)(2)D(3) increased intracellular calcium concentration in human spermatozoa through VDR-mediated calcium release from an intracellular calcium storage, increased sperm motility and induced the acrosome reaction in vitro. CONCLUSIONS 1,25(OH)(2)D(3) increased intracellular calcium...

Influence of Post-Mortem Sperm Recovery Method and Extender on Unstored and Refrigerated Rooster Sperm Variables.

Science.gov (United States)

Villaverde-Morcillo, S; Esteso, M C; Castaño, C; Santiago-Moreno, J

2016-02-01

Many post-mortem sperm collection techniques have been described for mammalian species, but their use in birds is scarce. This paper compares the efficacy of two post-mortem sperm retrieval techniques - the flushing and float-out methods - in the collection of rooster sperm, in conjunction with the use of two extenders, i.e., L&R-84 medium and Lake 7.1 medium. To determine whether the protective effects of these extenders against refrigeration are different for post-mortem and ejaculated sperm, pooled ejaculated samples (procured via the massage technique) were also diluted in the above extenders. Post-mortem and ejaculated sperm variables were assessed immediately at room temperature (0 h), and after refrigeration at 5°C for 24 and 48 h. The flushing method retrieved more sperm than the float-out method (596.5 ± 75.4 million sperm vs 341.0 ± 87.6 million sperm; p < 0.05); indeed, the number retrieved by the former method was similar to that obtained by massage-induced ejaculation (630.3 ± 78.2 million sperm). For sperm collected by all methods, the L&R-84 medium provided an advantage in terms of sperm motility variables at 0 h. In the refrigerated sperm samples, however, the Lake 7.1 medium was associated with higher percentages of viable sperm, and had a greater protective effect (p < 0.05) with respect to most motility variables. In conclusion, the flushing method is recommended for collecting sperm from dead birds. If this sperm needs to be refrigerated at 5°C until analysis, Lake 7.1 medium is recommended as an extender. © 2015 Blackwell Verlag GmbH.

Hypericum perforatum as a cognitive enhancer in rodents: A meta-analysis

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Ben-Eliezer, Daniel; Yechiam, Eldad

2016-01-01

Considered an antidepressant and anti-anxiety agent, Hypericum perforatum affects multiple neurotransmitters in a non-competitive synergistic manner, and may have nootropic potential. We quantitatively reviewed the pre-clinical literature to examine if there is a cognitive-enhancing effect of H. perforatum in healthy rodents. Additionally, within these studies, we compared the effects observed in intact rodents versus those whose performance has been impaired, mostly through stress manipulations. The meta-analysis incorporated studies that examined the effect of H. perforatum versus placebo on memory indices of task performance. All analyses were based on weighting different studies according to their inverse variance. Thirteen independent studies (published 2000–2014) involving 20 experimental comparisons met our inclusion criteria. The results showed a large positive effect of H. perforatum on cognitive performance for intact, healthy rodents (d = 1.11), though a larger effect emerged for stress-impaired rodents (d = 3.10 for restraint stress). The positive effect on intact rodents was observed in tasks assessing reference memory as well as working memory, and was not moderated by the type of memory or motivation (appetitive versus aversive). Thus, while primarily considered as a medication for depression, H. perforatum shows considerable nootropic potential in rodents. PMID:27762349

Effect of transfection and co-incubation of bovine sperm with exogenous DNA on sperm quality and functional parameters for its use in sperm-mediated gene transfer.

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Arias, María Elena; Sánchez-Villalba, Esther; Delgado, Andrea; Felmer, Ricardo

2017-02-01

Sperm-mediated gene transfer (SMGT) is based on the capacity of sperm to bind exogenous DNA and transfer it into the oocyte during fertilization. In bovines, the progress of this technology has been slow due to the poor reproducibility and efficiency of the production of transgenic embryos. The aim of the present study was to evaluate the effects of different sperm transfection systems on the quality and functional parameters of sperm. Additionally, the ability of sperm to bind and incorporate exogenous DNA was assessed. These analyses were carried out by flow cytometry and confocal fluorescence microscopy, and motility parameters were also evaluated by computer-assisted sperm analysis (CASA). Transfection was carried out using complexes of plasmid DNA with Lipofectamine, SuperFect and TurboFect for 0.5, 1, 2 or 4 h. The results showed that all of the transfection treatments promoted sperm binding and incorporation of exogenous DNA, similar to sperm incorporation of DNA alone, without affecting the viability. Nevertheless, the treatments and incubation times significantly affected the motility parameters, although no effect on the integrity of DNA or the levels of reactive oxygen species (ROS) was observed. Additionally, we observed that transfection using SuperFect and TurboFect negatively affected the acrosome integrity, and TurboFect affected the mitochondrial membrane potential of sperm. In conclusion, we demonstrated binding and incorporation of exogenous DNA by sperm after transfection and confirmed the capacity of sperm to spontaneously incorporate exogenous DNA. These findings will allow the establishment of the most appropriate method [intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF)] of generating transgenic embryos via SMGT based on the fertilization capacity of transfected sperm.

Confocal Microscopy and Image Analysis Indicates a Region-Specific Relation between Active Caspases and Cytoplasm in Ejaculated and Epididymal Sperm

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García Vazquez, Susana; Aragón Martínez, Andrés; Flores-Alonso, Juan Carlos

2012-01-01

Previously, it was suggested a relation between the presence of apoptosis markers with cytoplasm in mammalian sperm. In this work, flow cytometry, confocal microscopy and image analysis were used to analyze the relationship between active caspase-3 and -7 and intracellular esterases expression in ejaculated or epididymal ram sperm. Sperm obtained from ejaculates from the caput, corpus, or cauda of the epididymis were treated with an inhibitor of active caspase-3 and -7 and a marker of cytoplasmic esterases. Additionally, ejaculated sperm were incubated for one, two, or three hours before evaluation for active caspases. Sperm subpopulations positive for active caspases and/or intracellular esterases were detected by flow cytometry and confocal microscopy; however, image analysis of confocal images showed that the correlation between active caspases and cytoplasmic esterases in sperm is region-specific. Lower values of Spearman correlation coefficients were found when whole sperm or head sperm was analyzed; however, a high correlation was observed for midpiece sperm. Incubation of sperm for two or three hours promoted the autoactivation of caspases. It has been suggested that the presence of apoptotic markers in sperm are related with a process of abortive apoptosis and with errors during spermiogenesis. Our results permit us suggest that the origin of the relationship between active caspases and cytoplasmic esterases is due to differentiation errors occurring during spermiogenesis because the percentages of sperm with active caspases are not different in the caput, corpus, or cauda of the epididymis. In this study we demonstrate that existing sperm subpopulations can express active caspases and intracellular esterases and that the correlation between these molecules is high in midpiece sperm. PMID:22530029

Individual adjustment of sperm expenditure accords with sperm competition theory.

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Pilastro, Andrea; Scaggiante, Marta; Rasotto, Maria B

2002-07-23

Sperm competition theory predicts that males should strategically allocate their sperm reserves according to the level of sperm competition, defined as the probability that the sperm of two males compete for fertilizing a given set of ova. Substantial evidence from numerous animal taxa suggests that, at the individual level, sperm expenditure increases when the risk of sperm competition is greater. In contrast, according to the "intensity model" of sperm competition [Parker, G. A., Ball, M. A., Stockley, P. & Gage, M. J. G. (1996) Proc. R. Soc. London Ser. B 263, 1291-1297], when more than two ejaculates compete during a given mating event, sperm expenditure should decrease as the number of competing males increases. Empirical evidence supporting this prediction, however, is still lacking. Here we measured sperm expenditure in two gobiid fishes, the grass (Zosterisessor ophiocephalus) and black goby (Gobius niger), in which up to six sneakers can congregate around the nest of territorial males and release their sperm when females spawn. We show that, in accordance with theory, sneaker males of both species release fewer sperm as the number of competitors increases.

Implementing an open-access CASA software for the assessment of stallion sperm motility: Relationship with other sperm quality parameters.

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Giaretta, Elisa; Munerato, Mauro; Yeste, Marc; Galeati, Giovanna; Spinaci, Marcella; Tamanini, Carlo; Mari, Gaetano; Bucci, Diego

2017-01-01

Setting an open-access computer assisted sperm analysis (CASA) may benefit the evaluation of motility in mammalian sperm, especially when economic constraints do not allow the use of a commercial system. There have been successful attempts to develop such a device in Zebra fish sperm and the system has been used in very few studies on mammalian spermatozoa. Against this background, the present study aimed at developing an open-access CASA system for mammalian sperm using the horse as a model and based upon the Image J software previously established for Zebra fish sperm. Along with determining the sperm progressive motility and other kinetic parameters (such as amplitude of lateral head displacement), the "results" window was adjusted to simplify subsequent statistical analyses. The path window was enriched with colored sperm trajectories on the basis of the subpopulation they belong to and a number that allowed the sperm track to be associated to the sperm motility data shown in the "results" window. Data obtained from the novel plugin (named as CASA_bgm) were compared with those of the commercial CASA Hamilton-Thorn IVOS Vers.12, through Bland Altman's plots. While the percentage of total and progressive motile sperm, VCL, VAP, VSL, LIN and STR and ALH were in agreement with those obtained with the commercial system, BCF significantly differed between the two systems probably due to their settings. Interestingly, a positive and significant correlation between the percentages of total motile sperm evaluated through CASA_bgm and those showing high mitochondrial membrane potential evaluated by JC-1 staining was found. In conclusion, CASA_bgm ImageJ plugin could be useful and reliable for stallion sperm motility analysis and it is our aim to apply this system to other mammalian species. Copyright © 2016 Elsevier B.V. All rights reserved.

Pulmonary exposure to carbonaceous nanomaterials and sperm quality

DEFF Research Database (Denmark)

Skovmand, Astrid; Lauvas, Anna Jacobsen; Christensen, Preben

2018-01-01

. Pulmonary inflammation was determined by differential cell count in bronchoalveolar lavage fluid. Epididymal sperm concentration and motility were measured by computer-assisted sperm analysis. Epididymal sperm viability and morphological abnormalities were assessed manually using Hoechst 33,342/PI...... inflammation is a potential modulator of endocrine function. The aim of this study was to investigate the effects of pulmonary exposure to carbonaceous nanomaterials on sperm quality parameters in an experimental mouse model.Methods: Effects on sperm quality after pulmonary inflammation induced by carbonaceous...... flourescent and Spermac staining, respectively. Epididymal sperm were assessed with regard to sperm DNA integrity (damage). Daily sperm production was measured in the testis, and testosterone levels were measured in blood plasma by ELISA.Results: Neutrophil numbers in the bronchoalveolar fluid showed...

Sperm competition selects for sperm quantity and quality in the Australian Maluridae.

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Rowe, Melissah; Pruett-Jones, Stephen

2011-01-25

When ejaculates from rival males compete for fertilization, there is strong selection for sperm traits that enhance fertilization success. Sperm quantity is one such trait, and numerous studies have demonstrated a positive association between sperm competition and both testes size and the number of sperm available for copulations. Sperm competition is also thought to favor increases in sperm quality and changes in testicular morphology that lead to increased sperm production. However, in contrast to sperm quantity, these hypotheses have received considerably less empirical support and remain somewhat controversial. In a comparative study using the Australian Maluridae (fairy-wrens, emu-wrens, grasswrens), we tested whether increasing levels of sperm competition were associated with increases in both sperm quantity and quality, as well as an increase in the relative amount of seminiferous tubule tissue contained within the testes. After controlling for phylogeny, we found positive associations between sperm competition and sperm numbers, both in sperm reserves and in ejaculate samples. Additionally, as sperm competition level increased, the proportion of testicular spermatogenic tissue also increased, suggesting that sperm competition selects for greater sperm production per unit of testicular tissue. Finally, we also found that sperm competition level was positively associated with multiple sperm quality traits, including the proportion of motile sperm in ejaculates and the proportion of both viable and morphologically normal sperm in sperm reserves. These results suggest multiple ejaculate traits, as well as aspects of testicular morphology, have evolved in response to sperm competition in the Australian Maluridae. Furthermore, our findings emphasize the importance of post-copulatory sexual selection as an evolutionary force shaping macroevolutionary differences in sperm phenotype.

Boundaries of Ethics, Sperm on the Border –The Globalization of Danish Sperm

DEFF Research Database (Denmark)

Willum Adrian, Stine

as sperm donation in parliamentary debates, and in the media, often raises ethical questions. This presentation inquires into how the bending of boundaries by “inappropriate parents”, fertility travelers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation......’s situational analysis based on Anselm Strauss’ social world arena theory. Furthermore, I draw theoretically on Karen Barad and Donna Haraway’s notion of diffractive readings. I use their understanding of accountability to question and intervene in the doing of ethics in practice......., relatedness and family by politicians and bioethicists, in the parliament and in the media. I will argue that the globalization of Danish sperm is a story of combined subversive acts by women and men using the technologies, sperm banks and private fertility clinics. I will explore what the labeling...

Sperm competition promotes diversity of sperm bundles in Ohomopterus ground beetles

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Takami, Yasuoki; Sota, Teiji

2007-07-01

Diversification of sperm morphology has been investigated in the context of sperm competition, but the adaptive significance of sperm bundles is still unclear. In analyzing 10 taxa of the genus Carabus subgenus Ohomopterus and one related Carabus ground beetles, we found that dimorphic sperm bundles occurred in most species with varied degrees of bimodality, whereas sperm were generally monomorphic. Comparative analyses with phylogenetically independent contrasts revealed that the sizes of large and small sperm bundles evolved more rapidly than, and were not correlated with, the length of sperm, suggesting more intense selection on sperm bundle sizes and their independent responses to different evolutionary forces. The size of large sperm bundles was positively correlated with male genital morphology (pertinent to displacement of rival spermatophores) and postcopulatory guarding duration as well as male body length, suggesting that larger sperm bundles have been favored when the risk of spermatophore displacement is high. Larger sperm bundles may be advantageous because of their ability to migrate more rapidly into the spermatheca. In contrast, no clear association was detected between the small sperm bundle size and mating traits despite its rapid diversification. The present study provides the first record of heteromorphic sperm bundles, the diversity of which may be promoted by sperm competition.

A comparative analysis of the morphology and evolution of permanent sperm depletion in spiders.

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Peter Michalik

2011-01-01

Full Text Available Once thought to be energetically cheap and easy to produce, empirical work has shown that sperm is a costly and limited resource for males. In some spider species, there is behavioral evidence that sperm are permanently depleted after a single mating. This extreme degree of mating investment appears to co-occur with other reproductive strategies common to spiders, e.g. genital mutilation and sexual cannibalism. Here we corroborate that sperm depletion in the golden orb-web spider Nephila clavipes is permanent by uncovering its mechanistic basis using light and electron microscopy. In addition, we use a phylogeny-based statistical analysis to test the evolutionary relationships between permanent sperm depletion (PSD and other reproductive strategies in spiders. Male testes do not produce sperm during adulthood, which is unusual in spiders. Instead, spermatogenesis is nearly synchronous and ends before the maturation molt. Testis size decreases as males approach their maturation molt and reaches its lowest point after sperm is transferred into the male copulatory organs (pedipalps. As a consequence, the amount of sperm available to males for mating is limited to the sperm contained in the pedipalps, and once it is used, males lose their ability to fertilize eggs. Our data suggest that PSD has evolved independently at least three times within web-building spiders and is significantly correlated with the evolution of other mating strategies that limit males to monogamy, including genital mutilation and sexual cannibalism. We conclude that PSD may be an energy-saving adaptation in species where males are limited to monogamy. This could be particularly important in web-building spiders where extreme sexual size dimorphism results in large, sedentary females and small, searching males who rarely feed as adults and are vulnerable to starvation. Future work will explore possible energetic benefits and the evolutionary lability of PSD relative to other

Temporal trends in sperm count

DEFF Research Database (Denmark)

Levine, Hagai; Jørgensen, Niels; Martino-Andrade, Anderson

2017-01-01

a predefined protocol 7518 abstracts were screened and 2510 full articles reporting primary data on SC were reviewed. A total of 244 estimates of SC and TSC from 185 studies of 42 935 men who provided semen samples in 1973-2011 were extracted for meta-regression analysis, as well as information on years.......006, respectively). WIDER IMPLICATIONS: This comprehensive meta-regression analysis reports a significant decline in sperm counts (as measured by SC and TSC) between 1973 and 2011, driven by a 50-60% decline among men unselected by fertility from North America, Europe, Australia and New Zealand. Because......BACKGROUND: Reported declines in sperm counts remain controversial today and recent trends are unknown. A definitive meta-analysis is critical given the predictive value of sperm count for fertility, morbidity and mortality. OBJECTIVE AND RATIONALE: To provide a systematic review and meta-regression...

Comparative proteomics analysis of sheep sperm under two doses of heavy ion to irradiation

International Nuclear Information System (INIS)

Li Hongyan; Zhao Xingxu; He Yuxuan; Zhang Yong; Zhang Hong; Wang Yanling; Li Fadi; Ma Youji

2011-01-01

The object of this study was to investigate differential proteomic expressions in sheep sperm protein under two doses (0.5 and 0.3 kGy) heavy ion radiation. The current research presented the protein changes using two-dimensional gel electrophoresis (2-DE) after staining with silver nitrate, differential expression proteins were detected by PDQuest 8.0 software and subjected to ion trap mass spectrometer equipped with a Surveyor HPLC system, and differential spots of protein were identified. Results showed that eight common different expressed protein spots in two doses 2D gels were identified to be three up-regulated proteins (glutaredoxin -1, transcription factor AP -2-alpha and enolase). It was concluded that there was significant difference at protein level in sheep sperm after heavy ion radiation and differential proteome expression analysis may be useful to clarify the physiology state of sheep sperm in heavy ion radiation, which laid a foundation for the further studies on heavy ion radiation of sheep sperm proteomics. (authors)

A systematic review and meta-analysis to determine the effect of sperm DNA damage on in vitro fertilization and intracytoplasmic sperm injection outcome

Science.gov (United States)

Simon, Luke; Zini, Armand; Dyachenko, Alina; Ciampi, Antonio; Carrell, Douglas T

2017-01-01

Sperm DNA damage is prevalent among infertile men and is known to influence natural reproduction. However, the impact of sperm DNA damage on assisted reproduction outcomes remains controversial. Here, we conducted a meta-analysis of studies on sperm DNA damage (assessed by SCSA, TUNEL, SCD, or Comet assay) and clinical pregnancy after IVF and/or ICSI treatment from MEDLINE, EMBASE, and PUBMED database searches for this analysis. We identified 41 articles (with a total of 56 studies) including 16 IVF studies, 24 ICSI studies, and 16 mixed (IVF + ICSI) studies. These studies measured DNA damage (by one of four assays: 23 SCSA, 18 TUNEL, 8 SCD, and 7 Comet) and included a total of 8068 treatment cycles (3734 IVF, 2282 ICSI, and 2052 mixed IVF + ICSI). The combined OR of 1.68 (95% CI: 1.49–1.89; P IVF and/or ICSI treatment. In addition, the combined OR estimates of IVF (16 estimates, OR = 1.65; 95% CI: 1.34–2.04; P IVF + ICSI studies (16 estimates, OR = 2.37; 95% CI: 1.89–2.97; P IVF and/or ICSI treatment. PMID:27345006

Contrasting patterns of evolutionary constraint and novelty revealed by comparative sperm proteomic analysis in Lepidoptera.

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Whittington, Emma; Forsythe, Desiree; Borziak, Kirill; Karr, Timothy L; Walters, James R; Dorus, Steve

2017-12-02

Rapid evolution is a hallmark of reproductive genetic systems and arises through the combined processes of sequence divergence, gene gain and loss, and changes in gene and protein expression. While studies aiming to disentangle the molecular ramifications of these processes are progressing, we still know little about the genetic basis of evolutionary transitions in reproductive systems. Here we conduct the first comparative analysis of sperm proteomes in Lepidoptera, a group that exhibits dichotomous spermatogenesis, in which males produce a functional fertilization-competent sperm (eupyrene) and an incompetent sperm morph lacking nuclear DNA (apyrene). Through the integrated application of evolutionary proteomics and genomics, we characterize the genomic patterns potentially associated with the origination and evolution of this unique spermatogenic process and assess the importance of genetic novelty in Lepidopteran sperm biology. Comparison of the newly characterized Monarch butterfly (Danaus plexippus) sperm proteome to those of the Carolina sphinx moth (Manduca sexta) and the fruit fly (Drosophila melanogaster) demonstrated conservation at the level of protein abundance and post-translational modification within Lepidoptera. In contrast, comparative genomic analyses across insects reveals significant divergence at two levels that differentiate the genetic architecture of sperm in Lepidoptera from other insects. First, a significant reduction in orthology among Monarch sperm genes relative to the remainder of the genome in non-Lepidopteran insect species was observed. Second, a substantial number of sperm proteins were found to be specific to Lepidoptera, in that they lack detectable homology to the genomes of more distantly related insects. Lastly, the functional importance of Lepidoptera specific sperm proteins is broadly supported by their increased abundance relative to proteins conserved across insects. Our results identify a burst of genetic novelty

Hyaluronic acid binding ability of human sperm reflects cellular maturity and fertilizing potential: selection of sperm for intracytoplasmic sperm injection.

Science.gov (United States)

Huszar, Gabor; Ozkavukcu, Sinan; Jakab, Attila; Celik-Ozenci, Ciler; Sati, G Leyla; Cayli, Sevil

2006-06-01

The current concepts of sperm biochemical markers and the central role of the HspA2 chaperone protein, a measure of sperm cellular maturity and fertilizing potential, are reviewed. Because HspA2 is a component of the synaptonemal complex, low HspA2 levels and increased frequency of chromosomal aneuploidies are related in diminished maturity sperm. We also suggest a relationship between HspA2 expression in elongating spermatids and events of late spermiogenesis, such as cytoplasmic extrusion and plasma membrane remodeling that aid the formation of the zona pellucida binding and hyaluronic acid binding sites. The presence of hyaluronic acid receptor on the plasma membrane of mature sperm, coupled with hyaluronic acid coated glass or plastic surfaces, facilitates testing of sperm function and selection of single mature sperm for intracytoplasmic sperm injection. The frequencies of sperm with chromosomal disomy are reduced approximately fourfold to fivefold in hyaluronic acid selected sperm compared with semen sperm, comparable to the increase in such abnormalities in intracytoplasmic sperm injection offspring. Hyaluronic acid binding also excludes immature sperm with cytoplasmic extrusion, persistent histones, and DNA chain breaks. Hyaluronic acid mediated sperm selection is a novel technique that is comparable to sperm zona pellucida binding. Hyaluronic acid selected sperm will also alleviate the risks related to intracytoplasmic sperm injection fertilization with sperm of diminished maturity that currently cause worldwide concern.

Isolating Sperm from Cell Mixtures Using Magnetic Beads Coupled with an Anti-PH-20 Antibody for Forensic DNA Analysis.

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Xing-Chun Zhao

Full Text Available Vaginal swabs taken in rape cases usually contain epithelial cells from the victim and sperm from the assailant and forensic DNA analysis requires separation of sperm from these cell mixtures. PH-20, which is a glycosylphosphatidylinositol-anchored hyaluronidase located on the head of sperm, has important functions in fertilization. Here we describe a newly developed method for sperm isolation using anti-PH-20 antibody-coupled immunomagnetic beads (anti-PH-20 IMBs. Optical microscopy and scanning electron microscopy showed the IMBs recognized the head of sperm specifically and exhibited a great capacity to capture sperm cells. However, we found it necessary to incubate the IMB-sperm complex with DNase I before sperm lysis in order to remove any female DNA completely. We compared the sensitivity of anti-PH-20 IMBs in sperm and epithelial cell discrimination to those coated with a different anti-sperm antibody (anti-SP-10, anti-ADAM2 or anti-JLP. Only the anti-PH-20 IMBs succeeded in isolating sperm from cell mixtures at a sperm/epithelial cell ratio of 103:105. Further, our method exhibited greater power and better stability for sperm isolation compared to the traditional differential lysis strategy. Taken together, the anti-PH-20 IMB method described here could be effective for the isolation of sperm needed to obtain a single-sourced DNA profile as an aid to identifying the perpetrator in sexual assault cases.

Appraisal and standardization of curvilinear velocity (VCL) cut-off values for CASA analysis of Japanese quail (Coturnix japonica) sperm.

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Farooq, U; Malecki, I A; Mahmood, M; Martin, G B

2017-06-01

One of the basic steps in objective analysis of sperm motility is the subdivision of a motile sperm population into slow, medium and rapid categories based on their velocity. However, for CASA analysis of quail sperm, the velocity values for categorization of slow, medium and rapid sperm have not yet been standardized. To identify the cut-off values of "velocity curvilinear" (VCL) for quail sperm categorization, we captured and analysed 22,300 tracks of quail sperm using SCA ® -CASA. The median and mean VCL values were 85 and 97 μm/s. To define the VCL cut-off values, we used two methods. In the first, we identified the upper (rapid sperm) and lower (slow sperm) cut-off values using: (i) median VCL ± 25% or ± 50% or ± 75% of median VCL value; (ii) first and third quartile values of VCL data (i.e. 25% cut-off setting); and (iii) 33% and 66% of VCL data. Among these settings, sperm categories and their corresponding motility characteristics recorded using the "25%" setting (i.e. slow ≤36 ≤ medium ≤154 ≤ rapid) were found the most realistic and coherent with male ranking by fertility. In the second method, we calculated heteroscedasticity in the total VCL data using PCA and the two-step clustering method. With this approach, the mean of the high and low clusters was 165 and 51 μm/s, respectively. Together, the mean from two methods suggested that, for SCA ® -CASA categorization of quail sperm, sperm should be classed as "rapid" at VCL ≥160 μm/s and "slow" at VCL ≤45 μm/s. © 2017 Blackwell Verlag GmbH.

Effects of diluting medium and holding time on sperm motility analysis by CASA in ram.

Science.gov (United States)

Mostafapor, Somayeh; Farrokhi Ardebili, Farhad

2014-01-01

The aim of this study was to evaluate the effects of dilution rate and holding time on various motility parameters using computer-assisted sperm analysis (CASA). The semen samples were collected from three Ghezel rams. Samples were diluted in seminal plasma (SP), phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA) and Bioexcell. The motility parameters that computed and recorded by CASA include curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), straightness (STR), linearity (LIN), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF). In all diluters, there was a decrease in the average of all three parameters of sperms movement velocity as the time passed, but density of this decrease was more intensive in SP. The average of ALH between diluters indicated a significant difference, as it was more in Bioexcell in comparison with the similar amount in SP and PBS. The average of LIN in the diluted sperms in Bioexcell was less than two other diluters in all three times. The motility parameters of the diluted sperms in Bioexcell and PBS indicated an important and considerable difference with the diluted sperms in SP. According to the gained results, the Bioexcell has greater ability in preserving motility of sperm in comparison with the other diluters but as SP is considered as physiological environment for sperm. It seems that the evaluation of the motility parameters in Bioexcell and PBS cannot be an accurate and comparable evaluation with SP.

Effects of diluting medium and holding time on sperm motility analysis by CASA in ram

Directory of Open Access Journals (Sweden)

Somayeh Mostafapor

2014-06-01

Full Text Available The aim of this study was to evaluate the effects of dilution rate and holding time on various motility parameters using computer-assisted sperm analysis (CASA. The semen samples were collected from three Ghezel rams. Samples were diluted in seminal plasma (SP, phosphate-buffered saline (PBS containing 1% bovine serum albumin (BSA and Bioexcell. The motility parameters that computed and recorded by CASA include curvilinear velocity (VCL, straight line velocity (VSL, average path velocity (VAP, straightness (STR, linearity (LIN, amplitude of lateral head displacement (ALH, and beat cross frequency (BCF. In all diluters, there was a decrease in the average of all three parameters of sperms movement velocity as the time passed, but density of this decrease was more intensive in SP. The average of ALH between diluters indicated a significant difference, as it was more in Bioexcell in comparison with the similar amount in SP and PBS. The average of LIN in the diluted sperms in Bioexcell was less than two other diluters in all three times. The motility parameters of the diluted sperms in Bioexcell and PBS indicated an important and considerable difference with the diluted sperms in SP. According to the gained results, the Bioexcell has greater ability in preserving motility of sperm in comparison with the other diluters but as SP is considered as physiological environment for sperm. It seems that the evaluation of the motility parameters in Bioexcell and PBS cannot be an accurate and comparable evaluation with SP.

Sperm-macrophage interaction in the mouse: a quantitative assay in vitro using 111indium oxine-labeled sperm

International Nuclear Information System (INIS)

Olive, D.L.; Weinberg, J.B.; Haney, A.F.

1987-01-01

The role of reproductive tract macrophages in contraception and reproductive failure has become widely recognized. However, in vitro analysis of sperm phagocytosis by macrophages has relied upon a semi-quantitative method of sperm counting that is of limited accuracy and reproducibility. We have developed an assay using murine sperm labeled with 111 indium oxine, and results indicate the labeling to be rapid and efficient. Incorporation of 111 indium into sperm increased the dose and sperm concentration and reached 90% maximal uptake after 15 min incubation, with maximal uptake occurring at 30 min. No decrease in sperm motility was noted with levels of oxine in excess of those required for significant labeling. Maximal labeling efficiency occurred in phosphate-buffered saline (PBS), with Dulbecco's modified Eagle's medium (DMEM) + 10% adult bovine serum (ABS) producing significantly less uptake. Label dissociation was detectable in PBS at room temperature, but at 37 degrees C in DMEM + 10% ABS, loss of label occurred at a rate of 23.5%/h. Addition of labeled sperm to murine macrophage monolayers under optimal conditions resulted in uptake of 111 indium by macrophages, while free label was unincorporated. Results indicated assay specificity for macrophage-limited uptake, with insignificant label uptake by nonphagocytic murine fibroblasts and better sensitivity than sperm counting. Macrophages from Bacillus Calmette-Guerin (BCG)-infected mice resulted in a decrease in sperm uptake. Female macrophages showed greater capacity for sperm uptake than those of the male mouse. These initial studies demonstrated the utility of this model system in enhancing the understanding of sperm-macrophage interaction in the female reproductive tract

Analysis of the functional aspects and seminal plasma proteomic profile of sperm from smokers.

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Antoniassi, Mariana Pereira; Intasqui, Paula; Camargo, Mariana; Zylbersztejn, Daniel Suslik; Carvalho, Valdemir Melechco; Cardozo, Karina H M; Bertolla, Ricardo Pimenta

2016-11-01

To evaluate the effect of smoking on sperm functional quality and seminal plasma proteomic profile. Sperm functional tests were performed in 20 non-smoking men with normal semen quality, according to the World Health Organization (2010) and in 20 smoking patients. These included: evaluation of DNA fragmentation by alkaline Comet assay; analysis of mitochondrial activity using DAB staining; and acrosomal integrity evaluation by PNA binding. The remaining semen was centrifuged and seminal plasma was used for proteomic analysis (liquid chromatography-tandem mass spectrometry). The quantified proteins were used for Venn diagram construction in Cytoscape 3.2.1 software, using the PINA4MS plug-in. Then, differentially expressed proteins were used for functional enrichment analysis of Gene Ontology categories, Kyoto Encyclopedia of Genes and Genomes and Reactome, using Cytoscape software and the ClueGO 2.2.0 plug-in. Smokers had a higher percentage of sperm DNA damage (Comet classes III and IV; P analysis, 422 proteins were identified and quantified, of which one protein was absent, 27 proteins were under-represented and six proteins were over-represented in smokers. Functional enrichment analysis showed the enrichment of antigen processing and presentation, positive regulation of prostaglandin secretion involved in immune response, protein kinase A signalling and arachidonic acid secretion, complement activation, regulation of the cytokine-mediated signalling pathway and regulation of acute inflammatory response in the study group (smokers). In conclusion, cigarette smoking was associated with an inflammatory state in the accessory glands and in the testis, as shown by enriched proteomic pathways. This state causes an alteration in sperm functional quality, which is characterized by decreased acrosome integrity and mitochondrial activity, as well as by increased nuclear DNA fragmentation. © 2016 The Authors BJU International © 2016 BJU International Published by John

A technical assessment of the porcine ejaculated spermatozoa for a sperm-specific RNA-seq analysis.

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Gòdia, Marta; Mayer, Fabiana Quoos; Nafissi, Julieta; Castelló, Anna; Rodríguez-Gil, Joan Enric; Sánchez, Armand; Clop, Alex

2018-04-26

The study of the boar sperm transcriptome by RNA-seq can provide relevant information on sperm quality and fertility and might contribute to animal breeding strategies. However, the analysis of the spermatozoa RNA is challenging as these cells harbor very low amounts of highly fragmented RNA, and the ejaculates also contain other cell types with larger amounts of non-fragmented RNA. Here, we describe a strategy for a successful boar sperm purification, RNA extraction and RNA-seq library preparation. Using these approaches our objectives were: (i) to evaluate the sperm recovery rate (SRR) after boar spermatozoa purification by density centrifugation using the non-porcine-specific commercial reagent BoviPure TM ; (ii) to assess the correlation between SRR and sperm quality characteristics; (iii) to evaluate the relationship between sperm cell RNA load and sperm quality traits and (iv) to compare different library preparation kits for both total RNA-seq (SMARTer Universal Low Input RNA and TruSeq RNA Library Prep kit) and small RNA-seq (NEBNext Small RNA and TailorMix miRNA Sample Prep v2) for high-throughput sequencing. Our results show that pig SRR (~22%) is lower than in other mammalian species and that it is not significantly dependent of the sperm quality parameters analyzed in our study. Moreover, no relationship between the RNA yield per sperm cell and sperm phenotypes was found. We compared a RNA-seq library preparation kit optimized for low amounts of fragmented RNA with a standard kit designed for high amount and quality of input RNA and found that for sperm, a protocol designed to work on low-quality RNA is essential. We also compared two small RNA-seq kits and did not find substantial differences in their performance. We propose the methodological workflow described for the RNA-seq screening of the boar spermatozoa transcriptome. FPKM: fragments per kilobase of transcript per million mapped reads; KRT1: keratin 1; miRNA: micro-RNA; miscRNA: miscellaneous

Sperm competition and reproductive mode influence sperm dimensions and structure among snakes.

Science.gov (United States)

Tourmente, Maximiliano; Gomendio, Montserrat; Roldan, Eduardo R S; Giojalas, Laura C; Chiaraviglio, Margarita

2009-10-01

The role of sperm competition in increasing sperm length is a controversial issue, because findings from different taxa seem contradictory. We present a comparative study of 25 species of snakes with different levels of sperm competition to test whether it influences the size and structure of different sperm components. We show that, as levels of sperm competition increase, so does sperm length, and that this elongation is largely explained by increases in midpiece length. In snakes, the midpiece is comparatively large and it contains structures, which in other taxa are present in the rest of the flagellum, suggesting that it may integrate some of its functions. Thus, increases in sperm midpiece size would result in more energy as well as greater propulsion force. Sperm competition also increases the area occupied by the fibrous sheath and outer dense fibers within the sperm midpiece, revealing for the first time an effect upon structural elements within the sperm. Finally, differences in male-male encounter rates between oviparous and viviparous species seem to lead to differences in levels of sperm competition. We conclude that the influence of sperm competition upon different sperm components varies between taxa, because their structure and function is different.

Association of sperm apoptosis and DNA ploidy with sperm chromatin quality in human spermatozoa.

Science.gov (United States)

Mahfouz, Reda Z; Sharma, Rakesh K; Said, Tamer M; Erenpreiss, Juris; Agarwal, Ashok

2009-04-01

To examine the relationship among sperm apoptosis, sperm chromatin status, and DNA ploidy in different sperm fractions. Prospective study. Reproductive research center in a tertiary care hospital. Sperm prepared by density gradient were evaluated for sperm count, motility, apoptosis, and sperm chromatin assessment. Sperm count, sperm motility, toluidine blue (TB) results, DNA fragmentation index (%DFI), high DNA stainability, DNA cytometry, and early and late apoptosis. Sperm motility was related to late apoptotic and subhaploid apoptotic sperm (r = -0.56 and -0.53, respectively). The sperm %DFI showed significant correlation with late apoptotic and subhaploid sperm (r = 0.62 and 0.68). TB-stained sperm were significantly correlated with late apoptotic sperm (r = 0.51). Significantly higher proportions of haploid sperm and light blue TB-stained sperm were seen in mature compared with immature fractions. Even in semen samples with low %DFI, semen processing results in a lower incidence of nuclear immaturity and subhaploidy, but the incidence of late apoptotic sperm remains unchanged. Therefore, simultaneous evaluation of apoptosis and sperm chromatin status is important for processing sperm in assisted reproductive procedures.

Subversive practices of sperm donation - globalizing Danish sperm

DEFF Research Database (Denmark)

Willum Adrian, Stine

as the use of donated sperm continuously has been debated as an ethical issue, and increasingly been regulated. In this presentation I will discuss how Denmark became a destination for fertility travelling (sperm donation) as a result of various subversive strategies of family making. The article inquires......-sited ethnography drawing on ethnographic research including observations and interviews from fertility clinics and sperm banks in Denmark during 2002/2003 and 2011- 2013, legislative documents and websites of fertility clinics and sperm banks. The presentation is methodologically inspired by Adele Clarke...

Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples.

Science.gov (United States)

Roque, M; Valle, M; Marques, F; Sampaio, M; Geber, S

2016-04-01

Intracytoplasmic sperm injection (ICSI) may be performed with testicular frozen-thawed spermatozoa in patients with nonobstructive azoospermia (NOA). Sperm retrieval can be performed in advance of oocyte aspiration, as it may avoid the possibility of no recovery of spermatozoa on the day of oocyte pickup. There are few studies available in the literature concerning the use of frozen-thawed spermatozoa obtained from testicular sperm aspiration (TESA). To evaluate the effects and the outcomes of ICSI with frozen-thawed spermatozoa obtained by TESA, we performed a retrospective analysis of 43 ICSI cycles using frozen-thawed TESA. We obtained acceptable results with a fertilisation rate of 67.9%, an implantation rate (IR) of 17.1%, and clinical and ongoing pregnancy rates of 41.9% and 37.2% respectively. The results of this study suggest that performing ICSI using cryopreserved frozen-thawed testicular spermatozoa with TESA as a first option is a viable, safe, economic and effective method for patients with NOA. © 2015 Blackwell Verlag GmbH.

Post-Translational Modifications of Histones in Human Sperm.

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Krejčí, Jana; Stixová, Lenka; Pagáčová, Eva; Legartová, Soňa; Kozubek, Stanislav; Lochmanová, Gabriela; Zdráhal, Zbyněk; Sehnalová, Petra; Dabravolski, Siarhei; Hejátko, Jan; Bártová, Eva

2015-10-01

We examined the levels and distribution of post-translationally modified histones and protamines in human sperm. Using western blot immunoassay, immunofluorescence, mass spectrometry (MS), and FLIM-FRET approaches, we analyzed the status of histone modifications and the protamine P2. Among individual samples, we observed variability in the levels of H3K9me1, H3K9me2, H3K27me3, H3K36me3, and H3K79me1, but the level of acetylated (ac) histones H4 was relatively stable in the sperm head fractions, as demonstrated by western blot analysis. Sperm heads with lower levels of P2 exhibited lower levels of H3K9ac, H3K9me1, H3K27me3, H3K36me3, and H3K79me1. A very strong correlation was observed between the levels of P2 and H3K9me2. FLIM-FRET analysis additionally revealed that acetylated histones H4 are not only parts of sperm chromatin but also appear in a non-integrated form. Intriguingly, H4ac and H3K27me3 were detected in sperm tail fractions via western blot analysis. An appearance of specific histone H3 and H4 acetylation and H3 methylation in sperm tail fractions was also confirmed by both LC-MS/MS and MALDI-TOF MS analysis. Taken together, these data indicate that particular post-translational modifications of histones are uniquely distributed in human sperm, and this distribution varies among individuals and among the sperm of a single individual. © 2015 Wiley Periodicals, Inc.

Trigonellae Semen Enhances Sperm Motility and the Expression of the Cation Sperm Channel Proteins in Mouse Testes

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Do Rim Kim

2015-01-01

Full Text Available Genetic defects during spermatogenesis can lead to a reduction in sperm motility and cause male infertility. The cation channels of sperm (CatSper play a role in the regulation of hyperactivated sperm motility in mouse testes. The effect of Trigonellae Semen (TS on the male reproductive system and CatSper protein in mouse testes during spermatogenesis was examined. C57BL/c mice were divided into the following five groups: normal, cyclophosphamide- (CP- only treated (control group, and three groups treated with varying concentrations of TS with CP (100, 500, and 1000 mg/kg TS and 100 mg/kg CP. Real-time PCR, western blot analysis, and a testosterone immunoassay were performed to assess CatSper protein levels in the five groups. Additionally, sperm cell counts and motility were examined. Results indicate that sperm motility and sperm counts increased in the TS treated groups in a dose-dependent manner (p<0.01. CatSper levels were also significantly higher in the TS treated groups compared to that of the control group (p<0.001. Therefore, TS treatment could enhance sperm function by promoting spermatogenesis and the expression of CatSper proteins in mouse testes.

Variation in sperm morphology among Afrotropical sunbirds

Czech Academy of Sciences Publication Activity Database

Omotoriogun, T. C.; Laskemoen, T.; Rowe, M.; Albrecht, Tomáš; Bowie, R. C. K.; Sedláček, O.; Hořák, D.; Ottosson, U.; Lifjeld, J. T.

2016-01-01

Roč. 158, č. 1 (2016), s. 155-166 ISSN 0019-1019 Institutional support: RVO:68081766 Keywords : comparative analysis * Nectariniidae * phylogenetic signal * sperm competition * sperm size Subject RIV: EG - Zoology Impact factor: 2.279, year: 2016

Viscoelasticity promotes collective swimming of sperm

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Tung, Chih-Kuan; Harvey, Benedict B.; Fiore, Alyssa G.; Ardon, Florencia; Suarez, Susan S.; Wu, Mingming

From flocking birds to swarming insects, interactions of organisms large and small lead to the emergence of collective dynamics. Here, we report striking collective swimming of bovine sperm, with sperm orienting in the same direction within each cluster, enabled by the viscoelasticity of the fluid. A long-chain polyacrylamide solution was used as a model viscoelastic fluid such that its rheology can be fine-tuned to mimic that of bovine cervical mucus. In viscoelastic fluid, sperm formed dynamic clusters, and the cluster size increased with elasticity of the polyacrylamide solution. In contrast, sperm swam randomly and individually in Newtonian fluids of similar viscosity. Analysis of the fluid motion surrounding individual swimming sperm indicated that sperm-fluid interaction is facilitated by the elastic component of the fluid. We note that almost all biological fluids (e.g. mucus and blood) are viscoelastic in nature, this finding highlights the importance of fluid elasticity in biological function. We will discuss what the orientation fluctuation within a cluster reveals about the interaction strength. Supported by NIH Grant 1R01HD070038.

Sperm quality after swim up and density gradient centrifugation sperm preparation with supplementation of alpha lipoic acid (ALA): A preliminary study

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Lestari, Silvia W.; Lestari, Sarah H.; Pujianto, Dwi A.

2018-02-01

Intra uterine insemination (IUI) as one of the treatment for infertility, persists low success rate. A factor that contributes to the unsuccessful of IUI is sperm preparation, performed through Swim-up (SU) and Density Gradient Centrifugation (DGC) methods. Furthermore, studies have shown that Alpha Lipoic Acid (ALA) is a potent antioxidant that could enhance the sperm motility and protect the DNA integrity of the sperm [1]. This study is aimed to re-evaluate the efficiency of the DGC and SU methods in selecting sperm before being transferred for IUI by the supplementation of ALA based on the sperm DNA integrity. Semen samples were obtained from 13 men from partners of women who are infertile (normozoospermia) and underwent IUI. Semen analysis based on the guideline of World Health Organization (WHO) 2010 was performed to measure the sperm motility and velocity, before and after sperm preparation. Then, samples were incubated with Alpha Lipoic Acid (ALA) in 0.625 mg (ALA 1), 1.25 mg (ALA 2) and 2.5 mg (ALA 3). The Sperm Chromatin Dispersion (SCD) test was performed to evaluate the sperm DNA Fragmentation Index (DFI). The percentage of motile sperm was higher in prepared sperm (post-DGC and post-SU) than in whole semen. Furthermore, the percentage of motile sperm was higher in post-DGC compared to post-SU. The level of DFI after the supplementation of ALA was decreased in prepared sperm compared to the whole semen. ALA was proved capable to select the better sperm quality with decreased sperm DNA fragmentation of prepared sperm in the all of DFI category.

der(4)t(Y;4): Three-generation transmission and sperm meiotic segregation analysis.

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Vozdova, Miluse; Horinova, Vera; Wernerova, Vendula; Skalikova, Romana; Rybar, Roman; Prinosilova, Petra; Oracova, Eva; Rubes, Jiri

2011-05-01

We present a family where five members (three males and two females) are carriers of der(4)t(Y;4)(q11.23;p16.3). The adult carriers are phenotypicaly normal and fertile; the boy shows macrocephaly, psychomotor retardation, and atypical autism. The FISH on cultured lymphocytes confirmed that the redundant Yq heterochromatin was attached to the 4p-subtelomeric region maintained on the der(4). Sperm FISH analysis performed in a normospermic der(4) carrier showed a significant distortion of the expected 1:1 ratio of the X- and Y-bearing spermatozoa in favor of the X chromosome and significant lack of Y,der(4)spermatozoa. The overall lack of Y spermatozoa was not balanced even by a relative excess of Y,4 sperm. The analysis of X, Y, 7, 8, 18, and 21 sperm disomy and diploidy did not indicate any interchromosomal effect. The chromosome 4 disomy was significantly increased but still very low to be of considerable reproductive significance. The neurodevelomental phenotype of the boy was probably caused by a gene mutation. The coincidental occurrence of such chromosomal aberration and boy's phenotype might lead to misinterpretation of the causal relationship between these findings. It is necessary to consider the results of chromosomal analysis and clinical records of relatives for provide genetic counseling in such families. Copyright © 2011 Wiley-Liss, Inc.

What use is an infertile sperm? A comparative study of sperm-heteromorphic Drosophila

DEFF Research Database (Denmark)

Holman, Luke; Freckleton, Robert P; Snook, Rhonda R

2007-01-01

Sperm size and number are important determinants of male reproductive success. The genus Drosophila exhibits a remarkable diversity of sperm production strategies, including the production of multiple sperm morphs by individual males, a phenomenon called sperm heteromorphism. Sperm-heteromorphic ......Sperm size and number are important determinants of male reproductive success. The genus Drosophila exhibits a remarkable diversity of sperm production strategies, including the production of multiple sperm morphs by individual males, a phenomenon called sperm heteromorphism. Sperm......-heteromorphic Drosophila species in the obscura group produce large numbers of infertile "parasperm" in addition to fertile eusperm. Parasperm have been hypothesized to perform a number of roles in place of fertilization, predominantly focused on their potential function in postcopulatory sexual selection. However...

Variation in apical hook length reflects the intensity of sperm competition in murine rodents

Czech Academy of Sciences Publication Activity Database

Šandera, M.; Albrecht, Tomáš; Stopka, P.

2013-01-01

Roč. 8, č. 7 (2013), e68427 E-ISSN 1932-6203 Institutional support: RVO:68081766 Keywords : postcopulatory sexual selection * field mice Apodemus * female promiscuity * acrosome reaction * Passerine birds * Muroid rodents * genus Apodemus * cytochrome-b * testes size * house mice Subject RIV: EG - Zoology Impact factor: 3.534, year: 2013

Prevalence of Leishmania species in rodents: A systematic review and meta-analysis in Iran.

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Foroutan, Masoud; Khademvatan, Shahram; Majidiani, Hamidreza; Khalkhali, Hamidreza; Hedayati-Rad, Faezeh; Khashaveh, Shahla; Mohammadzadeh, Habib

2017-08-01

Leishmaniasis are diverse group of diseases caused by numerous species of genus Leishmania. Herein we have contrived a systematic review and meta-analysis on the prevalence of Leishmania species in rodents of Iran. For this purpose, following the general methodology recommended for systematic reviews and meta-analysis, six English databases (PubMed, Science Direct, Scopus, Ovid, Web of Science and Google Scholar) and four Persian databases (Magiran, SID, Iran Doc and Iran Medex) were explored during January 1995 till June 2015. Papers were selected based on 8 pre-defined inclusion criteria. During the years, a total number of 4485 different rodents were captured; among which 1291 cases were Leishmania positive. The calculated weighted prevalence of Leishmania species in rodents was 23% (95% CI=18-28). Given geographical zones of Iran, the highest and lowest prevalence rate was belonged to North 50% (95% CI=40-61) and West 11% (95% CI=5-17), respectively. Rhombomys opimus (1766), Meriones lybicus (1258) and Tatera indica (488) were the three most abundant captured rodents, while the highest prevalence of Leishmania species was observed in Nesokia indica 48% (95% CI=42-54) and followed by R. opimus 39% (95% CI=30-47). Egger's regression test was performed to detect publication bias, which revealed it may not have a significant influence on overall weighted prevalence estimate (P=0.317). Meta-regression analysis demonstrated that there is no significant relationship between overall prevalence with sample size (P=0.1) and year of publication (P=0.7). The results showed remarkable prevalence of Leishmania species in rodent reservoirs. In future, adopting a suitable strategy for control and combat with rodents is necessary. Copyright © 2017 Elsevier B.V. All rights reserved.

Towards microfluidic sperm refinement : impedance-based analysis and sorting of sperm cells

NARCIS (Netherlands)

de Wagenaar, B.; Dekker, Stefan; de Boer, Hans L.; Bomer, Johan G.; Olthuis, Wouter; van den Berg, Albert; Segerink, Loes Irene

2016-01-01

The use of high quality semen for artificial insemination in the livestock industry is essential for successful outcome. Insemination using semen with a high number of sperm cells containing morphological defects has a negative impact on fertilization outcome. Therefore, semen with a high number of

ANALYSIS OR THE POTENTIAL SPERM BIOMARKER, SP22, IN HUMAN SEMEN

Science.gov (United States)

ANALYSIS OF THE POTENTIAL SPERM BIOMARKER SP22 IN HUMAN SEMEN Rebecca A. Morris Ph.D.1, Gary R. Klinefelter Ph.D.1, Naomi L. Roberts 1, Juan D. Suarez 1, Lillian F. Strader 1, Susan C. Jeffay 1 and Sally D. Perreault Ph.D.1 1 U.S. EPA / ORD / National Health a...

Role of WNT signaling in epididymal sperm maturation.

Science.gov (United States)

Cheng, Jin-Mei; Tang, Ji-Xin; Li, Jian; Wang, Yu-Qian; Wang, Xiu-Xia; Zhang, Yan; Chen, Su-Ren; Liu, Yi-Xun

2018-02-01

Spermatozoa maturation, a process required for spermatozoa to acquire progressive motility and the ability to fertilize ova, primarily occurs in the caput and corpus of the epididymis. Despite considerable efforts, the factor(s) promoting epididymal sperm maturation remains unclear. Recently, WNT signaling has been implicated in epididymal sperm maturation. To further investigate WNT signaling function in epididymal sperm maturation, we generated Wntless conditional knockout mice (Wls cKO), Wls flox/flox ; Lcn5-Cre. In these mice, WNTLESS (WLS), a conserved membrane protein required for all WNT protein secretion, was specifically disrupted in the principal cells of the caput epididymidis. Immunoblot analysis showed that WLS was significantly reduced in the caput epididymidis of Wls cKO mice. In the caput epididymidis of Wls cKO mice, WNT 10A and WNT 2b, which are typically secreted by the principal cells of the caput epididymis, were not secreted. Interestingly, sperm motility analysis showed that the WLS deficiency in the caput epididymidis had no effect on sperm motility. Moreover, fertility tests showed that Wls cKO male mice had normal fertility. These results indicate that the disruption of WLS in principal cells of the caput epididymidis inhibits WNT protein secretion but has no effect on sperm motility and male fertility, suggesting that WNT signaling in the caput epididymidis may be dispensable for epididymal sperm maturation in mice.

Analysis of sperm antigens by sodium dodecyl sulfate gel/protein blot radioimmunobinding method

International Nuclear Information System (INIS)

Lee, C.Y.G.; Huang, Y.S.; Hu, P.C.; Gomel, V.; Menge, A.C.

1982-01-01

A radioimmunobinding method based on the blotting of renatured proteins from sodium dodecyl sulfate gels on to nitrocellulose filter papers was developed to analyze the sperm antigens that elicit serum anti-sperm antibodies. In rabbits, serum anti-sperm antibodies were raised by immunization with homologous epididymal spermatozoa mixed with complete Freund's adjuvant. The raised antisera from either male or female rabbits were shown to react with three major sperm protein bands on sodium dodecyl sulfate gels with the corresponding molecular weights of about 70,000 +/- 5000, 14,000, and 13,000, respectively. In humans, the monoclonal antibodies against human sperm were raised by a hybridoma technique. Out of six independent hybrid cell lines that were generated, three of them were shown to secrete immunoglobulins that react with the same two protein bands on sodium dodecyl sulfate gels, which have the approximate molecular weight of 10,000. The same procedure was also used to analyze human serum samples that were shown to contain anti-sperm antibodies by the known techniques. Unique sperm antigens that elicit anti-sperm antibodies in humans were identified and correlated. The results of this study suggest that sodium dodecyl sulfate gel/protein blot radioimmunobinding method may be a sensitive and useful tool for the study of sperm antigens that elicit autoimmune responses and their association with human infertility

Comparison of cryopreserved human sperm in vapor and liquid phases of liquid nitrogen: effect on motility parameters, morphology, and sperm function.

Science.gov (United States)

Punyatanasakchai, Piyaphan; Sophonsritsuk, Areephan; Weerakiet, Sawaek; Wansumrit, Surapee; Chompurat, Deonthip

2008-11-01

To compare the effects of cryopreserved sperm in vapor and liquid phases of liquid nitrogen on sperm motility, morphology, and sperm function. Experimental study. Andrology laboratory at Ramathibodi Hospital, Thailand. Thirty-eight semen samples with normal motility and sperm count were collected from 38 men who were either patients of an infertility clinic or had donated sperm for research. Each semen sample was divided into two aliquots. Samples were frozen with static-phase vapor cooling. One aliquot was plunged into liquid nitrogen (-196 degrees C), and the other was stored in vapor-phase nitrogen (-179 degrees C) for 3 days. Thawing was performed at room temperature. Motility was determined by using computer-assisted semen analysis, sperm morphology was determined by using eosin-methylene blue staining, and sperm function was determined by using a hemizona binding test. Most of the motility parameters of sperm stored in the vapor phase were not significantly different from those stored in the liquid phase of liquid nitrogen, except in amplitude of lateral head displacement. The percentages of normal sperm morphology in both vapor and liquid phases also were not significantly different. There was no significant difference in the number of bound sperm in hemizona between sperm cryopreserved in both vapor and liquid phases of liquid nitrogen. Cryopreservation of human sperm in a vapor phase of liquid nitrogen was comparable to cryopreservation in a liquid phase of liquid nitrogen.

Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine progeny sex

Directory of Open Access Journals (Sweden)

Pilar Santolaria

2016-01-01

Full Text Available This study was designed to determine the ability of computer-assisted sperm morphometry analysis (CASA-Morph with fluorescence to discriminate between spermatozoa carrying different sex chromosomes from the nuclear morphometrics generated and different statistical procedures in the bovine species. The study was divided into two experiments. The first was to study the morphometric differences between X- and Y-chromosome-bearing spermatozoa (SX and SY, respectively. Spermatozoa from eight bulls were processed to assess simultaneously the sex chromosome by FISH and sperm morphometry by fluorescence-based CASA-Morph. SX cells were larger than SY cells on average (P < 0.001 although with important differences between bulls. A simultaneous evaluation of all the measured features by discriminant analysis revealed that nuclear area and average fluorescence intensity were the variables selected by stepwise discriminant function analysis as the best discriminators between SX and SY. In the second experiment, the sperm nuclear morphometric results from CASA-Morph in nonsexed (mixed SX and SY and sexed (SX semen samples from four bulls were compared. FISH allowed a successful classification of spermatozoa according to their sex chromosome content. X-sexed spermatozoa displayed a larger size and fluorescence intensity than nonsexed spermatozoa (P < 0.05. We conclude that the CASA-Morph fluorescence-based method has the potential to find differences between X- and Y-chromosome-bearing spermatozoa in bovine species although more studies are needed to increase the precision of sex determination by this technique.

Why small males have big sperm: dimorphic squid sperm linked to alternative mating behaviours.

Science.gov (United States)

Iwata, Yoko; Shaw, Paul; Fujiwara, Eiji; Shiba, Kogiku; Kakiuchi, Yasutaka; Hirohashi, Noritaka

2011-08-10

Sperm cells are the target of strong sexual selection that may drive changes in sperm structure and function to maximize fertilisation success. Sperm evolution is regarded to be one of the major consequences of sperm competition in polyandrous species, however it can also be driven by adaptation to the environmental conditions at the site of fertilization. Strong stabilizing selection limits intra-specific variation, and therefore polymorphism, among fertile sperm (eusperm). Here we analyzed reproductive morphology differences among males employing characteristic alternative mating behaviours, and so potentially different conditions of sperm competition and fertilization environment, in the squid Loligo bleekeri. Large consort males transfer smaller (average total length = 73 μm) sperm to a female's internal sperm storage location, inside the oviduct; whereas small sneaker males transfer larger (99 μm) sperm to an external location around the seminal receptacle near the mouth. No significant difference in swimming speed was observed between consort and sneaker sperm. Furthermore, sperm precedence in the seminal receptacle was not biased toward longer sperm, suggesting no evidence for large sperm being favoured in competition for space in the sperm storage organ among sneaker males. Here we report the first case, in the squid Loligo bleekeri, where distinctly dimorphic eusperm are produced by different sized males that employ alternative mating behaviours. Our results found no evidence that the distinct sperm dimorphism was driven by between- and within-tactic sperm competition. We propose that presence of alternative fertilization environments with distinct characteristics (i.e. internal or external), whether or not in combination with the effects of sperm competition, can drive the disruptive evolution of sperm size.

Why small males have big sperm: dimorphic squid sperm linked to alternative mating behaviours

Directory of Open Access Journals (Sweden)

Shiba Kogiku

2011-08-01

Full Text Available Abstract Background Sperm cells are the target of strong sexual selection that may drive changes in sperm structure and function to maximize fertilisation success. Sperm evolution is regarded to be one of the major consequences of sperm competition in polyandrous species, however it can also be driven by adaptation to the environmental conditions at the site of fertilization. Strong stabilizing selection limits intra-specific variation, and therefore polymorphism, among fertile sperm (eusperm. Here we analyzed reproductive morphology differences among males employing characteristic alternative mating behaviours, and so potentially different conditions of sperm competition and fertilization environment, in the squid Loligo bleekeri. Results Large consort males transfer smaller (average total length = 73 μm sperm to a female's internal sperm storage location, inside the oviduct; whereas small sneaker males transfer larger (99 μm sperm to an external location around the seminal receptacle near the mouth. No significant difference in swimming speed was observed between consort and sneaker sperm. Furthermore, sperm precedence in the seminal receptacle was not biased toward longer sperm, suggesting no evidence for large sperm being favoured in competition for space in the sperm storage organ among sneaker males. Conclusions Here we report the first case, in the squid Loligo bleekeri, where distinctly dimorphic eusperm are produced by different sized males that employ alternative mating behaviours. Our results found no evidence that the distinct sperm dimorphism was driven by between- and within-tactic sperm competition. We propose that presence of alternative fertilization environments with distinct characteristics (i.e. internal or external, whether or not in combination with the effects of sperm competition, can drive the disruptive evolution of sperm size.

Outdoor air pollution and sperm quality.

Science.gov (United States)

Lafuente, Rafael; García-Blàquez, Núria; Jacquemin, Bénédicte; Checa, Miguel Angel

2016-09-15

Exposure to air pollution has been clearly associated with a range of adverse health effects, including reproductive toxicity, but its effects on male semen quality are still unclear. We performed a systematic review (up to June 2016) to assess the impact of air pollutants on sperm quality. We included 17 semi-ecological, panel, and cohort studies, assessing outdoor air pollutants, such as PM2.5, PM10, NOx, SO2, and O3, and their effects on DNA fragmentation, sperm count, sperm motility, and sperm morphology. Thirteen studies assessed air pollution exposure measured environmentally, and six used biomarkers of air pollution exposure (two did both). We rated the studies using the Newcastle-Ottawa Scale and assessed with the exposure method. Taking into account these factors and the number of studies finding significant results (positive or negative), the evidence supporting an effect of air pollution on DNA fragmentation is weak but suggestive, on sperm motility is limited and probably inexistent, on lower sperm count is inconclusive, and on sperm morphology is very suggestive. Because of the diversity of air pollutants and sperm parameters, and the studies' designs, we were unable to perform a meta-analysis. In summary, most studies concluded that outdoor air pollution affects at least one of the four semen quality parameters included in the review. However, results lack consistency, and furthermore, studies were not comparable. Studies using standardized air pollution and semen measures are required to obtain more reliable conclusions. CRD42015007175. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Slimmer or fertile? Pharmacological mechanisms involved in reduced sperm quality and fertility in rats exposed to the anorexigen sibutramine.

Directory of Open Access Journals (Sweden)

Cibele S Borges

Full Text Available Sperm acquire motility and fertility capacity during epididymal transit, under the control of androgens and sympathetic innervations. It is already known that the acceleration of epididymal sperm transit time can lead to lower sperm quality. In a previous work we showed that rats exposed to the anorexigen sibutramine, a non-selective serotonin-norepinephrine reuptake inhibitor, presented faster sperm transit time, lower epididymal sperm reserves and potentiation of the tension of epididymal duct to norepinephrine exposed acutely in vitro to sibutramine. In the present work we aimed to further investigate pharmacological mechanisms involved in these alterations and the impact on rat sperm quality. For this, adult male Wistar rats were treated with sibutramine (10 mg/kg/day or vehicle for 30 days. Sibutramine decreased final body, seminal vesicle, ventral prostate and epididymal weights, as well as sperm transit time in the epididymal cauda. On the contrary of the in vitro pharmacological assays, in which sibutramine was added directly to the bath containing strips of distal epididymal cauda, the ductal tension was not altered after in vivo sub-chronic exposure to sibutramine. However, there is pharmacological evidence that the endogenous epididymal norepinephrine reserves were reduced in these animals. It was also shown that the decrease in prostate weight can be related to increased tension developed of the gland, due to sibutramine sympathomimetic effects. In addition, our results showed reduced sperm quality after in utero artificial insemination, a more sensitive procedure to assess fertility in rodents. The epididymal norepinephrine depletion exerted by sibutramine, associated with decreases in sperm transit time, quantity and quality, leading to reduced fertility in this experimental model, reinforces the concerns about the possible impact on fertility of man taking sibutramine as well as other non-selective serotonin

Slimmer or fertile? Pharmacological mechanisms involved in reduced sperm quality and fertility in rats exposed to the anorexigen sibutramine.

Science.gov (United States)

Borges, Cibele S; Missassi, Gabriela; Pacini, Enio S A; Kiguti, Luiz Ricardo A; Sanabria, Marciana; Silva, Raquel F; Banzato, Thais P; Perobelli, Juliana E; Pupo, André S; Kempinas, Wilma G

2013-01-01

Sperm acquire motility and fertility capacity during epididymal transit, under the control of androgens and sympathetic innervations. It is already known that the acceleration of epididymal sperm transit time can lead to lower sperm quality. In a previous work we showed that rats exposed to the anorexigen sibutramine, a non-selective serotonin-norepinephrine reuptake inhibitor, presented faster sperm transit time, lower epididymal sperm reserves and potentiation of the tension of epididymal duct to norepinephrine exposed acutely in vitro to sibutramine. In the present work we aimed to further investigate pharmacological mechanisms involved in these alterations and the impact on rat sperm quality. For this, adult male Wistar rats were treated with sibutramine (10 mg/kg/day) or vehicle for 30 days. Sibutramine decreased final body, seminal vesicle, ventral prostate and epididymal weights, as well as sperm transit time in the epididymal cauda. On the contrary of the in vitro pharmacological assays, in which sibutramine was added directly to the bath containing strips of distal epididymal cauda, the ductal tension was not altered after in vivo sub-chronic exposure to sibutramine. However, there is pharmacological evidence that the endogenous epididymal norepinephrine reserves were reduced in these animals. It was also shown that the decrease in prostate weight can be related to increased tension developed of the gland, due to sibutramine sympathomimetic effects. In addition, our results showed reduced sperm quality after in utero artificial insemination, a more sensitive procedure to assess fertility in rodents. The epididymal norepinephrine depletion exerted by sibutramine, associated with decreases in sperm transit time, quantity and quality, leading to reduced fertility in this experimental model, reinforces the concerns about the possible impact on fertility of man taking sibutramine as well as other non-selective serotonin-norepinephrine reuptake inhibitors

Quantitation of sperm bindable IgA and IgG in seminal fluid.

Science.gov (United States)

Howe, S E; Lynch, D M

1986-05-01

Seminal fluid and serum from 95 infertile males were assayed for sperm bindable immunoglobulins using an indirect ELISA with whole target sperm. The ELISA method was compared to seminal fluid and serum immobilization and agglutination assays (functional assays). In this infertile group, the ELISA assay was positive in 22% of seminal fluids (greater than 1.2 fg IgA/sperm and greater than 0.3 fg IgG/sperm). The seminal fluid antibodies were IgA and had an accompanying elevated IgG component in 78% of patients. There was a 96% correlation between negative seminal fluid functional assays and negative ELISA, and a 95% correlation between positive seminal fluid functional assays and positive ELISA. Positive serum sperm antibody tests were found in 71% of the infertile males with positive seminal fluid sperm antibodies, but 29% of the infertile males with strongly positive IgA seminal fluid sperm antibodies showed normal levels of serum sperm antibodies by either ELISA or functional assays. The ELISA method gives reproducible quantitation of sperm antibodies in seminal fluid and correlates well with accepted functional assays. Comparisons with serum sperm antibody assays suggests that seminal fluid sperm antibody analysis complements the serum analysis of sperm antibodies.

Diisopropyl fluorophosphate labeling of sperm-associated proteinases

International Nuclear Information System (INIS)

Odem, R.R.; Willand, J.L.; Polakoski, K.L.

1990-01-01

Proteinase inhibitors have been shown to be capable of preventing various aspects of fertilization. Diisopropyl fluorophosphate (DFP) is an irreversible inhibitor of trypsin-like enzymes that is commercially available in a radiolabeled form. The experiments described herein were designed to determine if DFP would prevent sperm function in live, motile sperm and to identify the sperm proteins bound with DFP. DFP at 5 mM concentrations had no observable effect on sperm motility, but inhibited the penetration of zona-free hamster ova by human sperm (5.5%) compared to controls (33.5%). Acid extracts of motile sperm that had been incubated with radiolabeled DFP and collected by the swim-up procedure demonstrated the presence of radiolabeled DFP, and the autoradiography of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of these extracts localized the uptake of radiolabeled DFP to proteins in the molecular weight region of the proacrosin-acrosin system. Acid-extracted proteinases from semen samples incubated with DFP demonstrated a concentration-dependent inhibition of both esterolytic hydrolysis of benzoyl-arginine ethyl ester on spectrophotometric analysis and proteolytic activity on gelatin SDS-PAGE zymography. DFP-labeled proteins were precipitated by highly specific antibodies to proacrosin. These results demonstrated that DFP is capable of inhibiting sperm function, and that it associates with the proacrosin-acrosin system in live motile sperm

Diisopropyl fluorophosphate labeling of sperm-associated proteinases

Energy Technology Data Exchange (ETDEWEB)

Odem, R.R.; Willand, J.L.; Polakoski, K.L. (Washington Univ. School of Medicine, St. Louis, MO (USA))

1990-02-01

Proteinase inhibitors have been shown to be capable of preventing various aspects of fertilization. Diisopropyl fluorophosphate (DFP) is an irreversible inhibitor of trypsin-like enzymes that is commercially available in a radiolabeled form. The experiments described herein were designed to determine if DFP would prevent sperm function in live, motile sperm and to identify the sperm proteins bound with DFP. DFP at 5 mM concentrations had no observable effect on sperm motility, but inhibited the penetration of zona-free hamster ova by human sperm (5.5%) compared to controls (33.5%). Acid extracts of motile sperm that had been incubated with radiolabeled DFP and collected by the swim-up procedure demonstrated the presence of radiolabeled DFP, and the autoradiography of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of these extracts localized the uptake of radiolabeled DFP to proteins in the molecular weight region of the proacrosin-acrosin system. Acid-extracted proteinases from semen samples incubated with DFP demonstrated a concentration-dependent inhibition of both esterolytic hydrolysis of benzoyl-arginine ethyl ester on spectrophotometric analysis and proteolytic activity on gelatin SDS-PAGE zymography. DFP-labeled proteins were precipitated by highly specific antibodies to proacrosin. These results demonstrated that DFP is capable of inhibiting sperm function, and that it associates with the proacrosin-acrosin system in live motile sperm.

Sperm length, sperm storage and mating system characteristics in bumblebees

DEFF Research Database (Denmark)

Baer, Boris; Schmid-Hempel, Paul; Høeg, Jens Thorvald

2003-01-01

-term storage of sperm, using three bumblebee species with different mating systems as models. We show that individual males produce only one size-class of sperm, but that sperm length is highly variable among brothers, among unrelated conspecific males, and among males of different species. Males of Bombus...

Evidence for nuclear internalization of exogenous DNA into mammalian sperm cells

International Nuclear Information System (INIS)

Francolini, M.; Lavitrano, M.; Lamia, C.L.; French, D.; Frati, L.; Cotelli, F.; Spadafora, C.

1993-01-01

Mature sperm cells have the spontaneous capacity to take up exogenous DNA. Such DNA specifically interacts with the subacrosomal segment of the sperm head corresponding to the nuclear area. Part of the sperm-bound foreign DNA is further internalized into nuclei. Using end-labelled plasmid DNA we have found that 15-22% of the total sperm bound DNA is associated with nuclei as determined on isolated nuclei. On the basis of autoradiographic analysis, nuclear permeability to exogenous DNA seems to be a wide phenomenon involving the majority of the sperm nuclei. In fact, the foreign DNA, incubated with sperm cells for different lengths of time, is found in 45% (10 min) to 65% (2 hr) of the sperm nuclei. Ultrastructural autoradiography on thin sections of mammalian spermatozoa, preincubated with end-labelled plasmid DNA, shows that the exogenous DNA is internalized into the nucleus. This conclusion is further supported by ultrastructural autoradiographic analysis on thin sections of nuclei isolated from spermatozoa preincubated with end-labelled DNA

Full field optical coherence tomography can identify spermatogenesis in a rodent sertoli-cell only model.

Science.gov (United States)

Ramasamy, Ranjith; Sterling, Joshua; Manzoor, Maryem; Salamoon, Bekheit; Jain, Manu; Fisher, Erik; Li, Phillip S; Schlegel, Peter N; Mukherjee, Sushmita

2012-01-01

Microdissection testicular sperm extraction (micro-TESE) has replaced conventional testis biopsies as a method of choice for obtaining sperm for in vitro fertilization for men with nonobstructive azoospermia. A technical challenge of micro-TESE is that the low magnification inspection of the tubules with a surgical microscope is insufficient to definitively identify sperm-containing tubules, necessitating tissue removal and cytologic assessment. Full field optical coherence tomography (FFOCT) uses white light interference microscopy to generate quick high-resolution tomographic images of fresh (unprocessed and unstained) tissue. Furthermore, by using a nonlaser safe light source (150 W halogen lamp) for tissue illumination, it ensures that the sperm extracted for in vitro fertilization are not photo-damaged or mutagenized. A focal Sertoli-cell only rodent model was created with busulfan injection in adult rats. Ex vivo testicular tissues from both normal and busulfan-treated rats were imaged with a commercial modified FFOCT system, Light-CT™, and the images were correlated with gold standard hematoxylin and eosin staining. Light-CT™ identified spermatogenesis within the seminiferous tubules in freshly excised testicular tissue, without the use of exogenous contrast or fixation. Normal adult rats exhibited tubules with uniform size and shape (diameter 328 ±11 μm). The busulfan-treated animals showed marked heterogeneity in tubular size and shape (diameter 178 ± 35 μm) and only 10% contained sperm within the lumen. FFOCT has the potential to facilitate real-time visualization of spermatogenesis in humans, and aid in micro-TESE for men with infertility.

No evidence of trade-offs in the evolution of sperm numbers and sperm size in mammals.

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Tourmente, M; Delbarco Trillo, J; Roldan, E R S

2015-10-01

Post-copulatory sexual selection, in the form sperm competition, has influenced the evolution of several male reproductive traits. However, theory predicts that sperm competition would lead to trade-offs between numbers and size of spermatozoa because increased costs per cell would result in a reduction of sperm number if both traits share the same energetic budget. Theoretical models have proposed that, in large animals, increased sperm size would have minimal fitness advantage compared with increased sperm numbers. Thus, sperm numbers would evolve more rapidly than sperm size under sperm competition pressure. We tested in mammals whether sperm competition maximizes sperm numbers and size, and whether there is a trade-off between these traits. Our results showed that sperm competition maximizes sperm numbers in eutherian and metatherian mammals. There was no evidence of a trade-off between sperm numbers and sperm size in any of the two mammalian clades as we did not observe any significant relationship between sperm numbers and sperm size once the effect of sperm competition was taken into account. Maximization of both numbers and size in mammals may occur because each trait is crucial at different stages in sperm's life; for example size-determined sperm velocity is a key determinant of fertilization success. In addition, numbers and size may also be influenced by diverse energetic budgets required at different stages of sperm formation. © 2015 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2015 European Society For Evolutionary Biology.

Daily Sperm Production, Gonadal and Extra-Gonadal Sperm ...

African Journals Online (AJOL)

SH

animals fed diets 2 and 3 were similar to the control animals but they were significantly (P<0.05) lower than those fed ... Keywords: Prebiotics, probiotics, rabbits, sperm reserves, sperm production. ... Materials and methods .... In: Handbook of.

The impact of BMI on sperm parameters and the metabolite changes of seminal plasma concomitantly.

Science.gov (United States)

Guo, Dan; Wu, Wei; Tang, Qiuqin; Qiao, Shanlei; Chen, Yiqiu; Chen, Minjian; Teng, Mengying; Lu, Chuncheng; Ding, Hongjuan; Xia, Yankai; Hu, Lingqing; Chen, Daozhen; Sha, Jiahao; Wang, Xinru

2017-07-25

The development of male infertility increased rapidly worldwide, which coinciding with the epidemic of obesity. However, the impact of weight abnormalities on sperm quality is still contestable. To assess the correlation between BMI and sperm parameters, we searched relevant articles in PubMed, Embase, Web of science, and Wanfang database published until June 2015 without language restriction. Otherwise, we also recruited some participants who attended fertility clinic as well as some general populations in this report. We performed a systematic review and meta-analysis about BMI and sperm parameters containing total sperm count, concentration, semen volume and sperm motility (overall and progressive). Metabolomic analysis of seminal plasma was performed to explore the mechanism from a new perspective. This study found standardized weighted mean differences (SMD) in sperm parameters (total sperm count, sperm concentration, and semen volume) of abnormal weight groups decreased to different degree compared to normal weight. Dose-response analysis found SMD of sperm count, sperm concentration and semen volume respectively fell 2.4%, 1.3% and 2.0% compared with normal weight for every 5-unit increase in BMI. Metabolomic analysis of seminal plasma showed that spermidine and spermine were likely to play a vital role in the spermatogenesis progress. This systematic review with meta-analysis has confirmed there was a relationship between BMI and sperm quality, suggesting obesity may be a detrimental factor of male infertility.

Cystic fibrosis transmembrane conductance regulator is correlated closely with sperm progressive motility and normal morphology in healthy and fertile men with normal sperm parameters.

Science.gov (United States)

Jiang, L-Y; Shan, J-J; Tong, X-M; Zhu, H-Y; Yang, L-Y; Zheng, Q; Luo, Y; Shi, Q-X; Zhang, S-Y

2014-10-01

Cystic fibrosis transmembrane conductance regulator (CFTR) has been demonstrated to be expressed in mature spermatozoa and correlated with sperm quality. Sperm CFTR expression in fertile men is higher than that in infertile men suffering from teratospermia, asthenoteratospermia, asthenospermia and oligospermia, but it is unknown whether CFTR is correlated with sperm parameters when sperm parameters are normal. In this study, 282 healthy and fertile men with normal semen parameters were classified into three age groups, group (I): age group of 20-29 years (98 cases, 27.1 ± 6.2), group (II): age group of 30-39 years (142 cases, 33.7 ± 2.6) and group (III): age group of more than or equal to 40 years (42 cases, 44.1 ± 4.6). Sperm concentration, total count and progressive motility were analysed by computer-assisted sperm analysis. Sperm morphology was analysed by modified Papanicolaou staining. Sperm CFTR expression was conducted by indirect immunofluorescence staining. There was a significant positive correlation (P sperm progressive motility (r = 0.221) and normal morphology (r = 0.202), but there were no correlations between sperm CFTR expression and semen volume, sperm concentration, sperm total count as well as male age (P > 0.05). Our findings show that CFTR expression is associated with sperm progressive motility and normal morphology in healthy and fertile men with normal sperm parameters, but not associated with the number of spermatozoa and male age. © 2013 Blackwell Verlag GmbH.

Sperm count. Do we need a new reference value?

Science.gov (United States)

Cardona Maya, Walter

2010-03-01

To evaluate the sperm count in fertile men, general population, and infertile men in different regions of the world. Sperm counts were recorded according to their fertility status, proven fertility, men recruited from an andrology/infertility clinic, or healthy men. The average of sperm count in the different studies is lower in infertile men that in fertile men (p>0.001) and in the general population (p>0.001). Based on this analysis the normal sperm count is about 65 million per mL. Using these reference value, only the 25% of the studies in infertile men are above this value, and the 75% studies with fertile men (>65 x 106 sperm/mL).

[Eosin Y-water test for sperm function examination].

Science.gov (United States)

Zha, Shu-wei; Lü, Nian-qing; Xu, Hao-qin

2015-06-01

Based on the principles of the in vitro staining technique, hypotonic swelling test, and water test, the Eosin Y-water test method was developed to simultaneously detect the integrity of the sperm head and tail and sperm membrane structure and function. As a widely used method in clinical laboratories in China, the Eosin Y-water test is methodologically characterized by three advantages. Firstly, both the sperm head and tail can be detected at the same time, which allows easy and comprehensive assessment of membrane damage in different parts of sperm. Secondly, distilled water is used instead of the usual formula solution to simplify and standardize the test by eliminating any potential effects on the water molecules through the sperm membrane due to different osmotic pressure or different sugar proportions and electrolyte solutions. Thirdly, the test takes less time and thus can be repeated before and after treatment. This article focuses on the fundamental principles and modification of the Eosin Y-water test and its application in sperm function examination and routine semen analysis for male infertility, assessment of the quality of sperm retrieved by testicular fine needle aspiration, semen cryopreservation program development, and evaluation of sperm membrane integrity after microwave radiation.

Cytometry of mammalian sperm

Energy Technology Data Exchange (ETDEWEB)

Gledhill, B.L.

1983-10-11

Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples.

Cytometry of mammalian sperm

International Nuclear Information System (INIS)

Gledhill, B.L.

1983-01-01

Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples

The stochastic dance of circling sperm cells: sperm chemotaxis in the plane

Energy Technology Data Exchange (ETDEWEB)

Friedrich, B M; Juelicher, F [Max Planck Institute for the Physics of Complex Systems, Noethnitzer Strasse 38, 01187 Dresden (Germany)], E-mail: ben@pks.mpg.de, E-mail: julicher@pks.mpg.de

2008-12-15

Biological systems such as single cells must function in the presence of fluctuations. It has been shown in a two-dimensional experimental setup that sea urchin sperm cells move toward a source of chemoattractant along planar trochoidal swimming paths, i.e. drifting circles. In these experiments, a pronounced variability of the swimming paths is observed. We present a theoretical description of sperm chemotaxis in two dimensions which takes fluctuations into account. We derive a coarse-grained theory of stochastic sperm swimming paths in a concentration field of chemoattractant. Fluctuations enter as multiplicative noise in the equations for the sperm swimming path. We discuss the stochastic properties of sperm swimming and predict a concentration-dependence of the effective diffusion constant of sperm swimming which could be tested in experiments.

The stochastic dance of circling sperm cells: sperm chemotaxis in the plane

International Nuclear Information System (INIS)

Friedrich, B M; Juelicher, F

2008-01-01

Biological systems such as single cells must function in the presence of fluctuations. It has been shown in a two-dimensional experimental setup that sea urchin sperm cells move toward a source of chemoattractant along planar trochoidal swimming paths, i.e. drifting circles. In these experiments, a pronounced variability of the swimming paths is observed. We present a theoretical description of sperm chemotaxis in two dimensions which takes fluctuations into account. We derive a coarse-grained theory of stochastic sperm swimming paths in a concentration field of chemoattractant. Fluctuations enter as multiplicative noise in the equations for the sperm swimming path. We discuss the stochastic properties of sperm swimming and predict a concentration-dependence of the effective diffusion constant of sperm swimming which could be tested in experiments.

Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine progeny sex.

Science.gov (United States)

Santolaria, Pilar; Pauciullo, Alfredo; Silvestre, Miguel A; Vicente-Fiel, Sandra; Villanova, Leyre; Pinton, Alain; Viruel, Juan; Sales, Ester; Yániz, Jesús L

2016-01-01

This study was designed to determine the ability of computer-assisted sperm morphometry analysis (CASA-Morph) with fluorescence to discriminate between spermatozoa carrying different sex chromosomes from the nuclear morphometrics generated and different statistical procedures in the bovine species. The study was divided into two experiments. The first was to study the morphometric differences between X- and Y-chromosome-bearing spermatozoa (SX and SY, respectively). Spermatozoa from eight bulls were processed to assess simultaneously the sex chromosome by FISH and sperm morphometry by fluorescence-based CASA-Morph. SX cells were larger than SY cells on average (P potential to find differences between X- and Y-chromosome-bearing spermatozoa in bovine species although more studies are needed to increase the precision of sex determination by this technique.

OBJECTIVE EVALUATION OF HYPERACTIVATED MOTILITY IN RAT SPERMATOZA USING COMPUTER-ASSISTED SPERM ANALYSIS (CASA)

Science.gov (United States)

Objective evaluation of hyperactivated motility in rat spermatozoa using computer-assisted sperm analysis.Cancel AM, Lobdell D, Mendola P, Perreault SD.Toxicology Program, University of North Carolina, Chapel Hill, NC 27599, USA.The aim of this study was t...

Sperm competition leads to functional adaptations in avian testes to maximize sperm quantity and quality.

Science.gov (United States)

Lüpold, Stefan; Wistuba, Joachim; Damm, Oliver S; Rivers, James W; Birkhead, Tim R

2011-05-01

The outcome of sperm competition (i.e. competition for fertilization between ejaculates from different males) is primarily determined by the relative number and quality of rival sperm. Therefore, the testes are under strong selection to maximize both sperm number and quality, which are likely to result in trade-offs in the process of spermatogenesis (e.g. between the rate of spermatogenesis and sperm length or sperm energetics). Comparative studies have shown positive associations between the level of sperm competition and both relative testis size and the proportion of seminiferous (sperm-producing) tissue within the testes. However, it is unknown how the seminiferous tissue itself or the process of spermatogenesis might evolve in response to sperm competition. Therefore, we quantified the different germ cell types and Sertoli cells (SC) in testes to assess the efficiency of sperm production and its associations with sperm length and mating system across 10 species of New World Blackbirds (Icteridae) that show marked variation in sperm length and sperm competition level. We found that species under strong sperm competition generate more round spermatids (RS)/spermatogonium and have SC that support a greater number of germ cells, both of which are likely to increase the maximum sperm output. However, fewer of the RS appeared to elongate to mature spermatozoa in these species, which might be the result of selection for discarding spermatids with undesirable characteristics as they develop. Our results suggest that, in addition to overall size and gross morphology, testes have also evolved functional adaptations to maximize sperm quantity and quality.

The Semen pH Affects Sperm Motility and Capacitation.

Science.gov (United States)

Zhou, Ji; Chen, Li; Li, Jie; Li, Hongjun; Hong, Zhiwei; Xie, Min; Chen, Shengrong; Yao, Bing

2015-01-01

As the chemical environment of semen can have a profound effect on sperm quality, we examined the effect of pH on the motility, viability and capacitation of human sperm. The sperm in this study was collected from healthy males to avoid interference from other factors. The spermatozoa cultured in sperm nutrition solution at pH 5.2, 6.2, 7.2 and 8.2 were analyzed for sperm total motility, progressive motility (PR), hypo-osmotic swelling (HOS) rate, and sperm penetration. Our results showed that these parameters were similar in pH 7.2 and 8.2 sperm nutrition solutions, but decreased in pH 5.2 and 6.2 solutions. The HOS rate exhibited positive correlation with the sperm total motility and PR. In addition, the sperm Na(+)/K(+)-ATPase activity at different pHs was measured, and the enzyme activity was significantly lower in pH 5.2 and 6.2 media, comparing with that in pH 8.2 and pH 7.2 solutions. Using flow cytometry (FCM) and laser confocal scanning microscopy (LCSM) analysis, the intracellular Ca2(+ )concentrations of sperm cultured in sperm capacitation solution at pH 5.2, 6.2, 7.2 and 8.2 were determined. Compared with that at pH 7.2, the mean fluorescence intensity of sperm in pH 5.2 and 6.2 media decreased significantly, while that of pH 8.2 group showed no difference. Our results suggested that the declined Na(+)/K(+)-ATPase activity at acidic pHs result in decreased sperm movement and capacitation, which could be one of the mechanisms of male infertility.

On the relative effect of spawning asynchrony, sperm quantity and sperm quality on paternity under sperm competition in an external fertilizer

Directory of Open Access Journals (Sweden)

Torvald Blikra Egeland

2015-07-01

Full Text Available How much of a fitness benefit is obtained by dominant males of external fertilizers from releasing ejaculates in synchrony with female egg-release when engaging in sperm competition, and what is the most important sperm trait for paternity in these situations? The Arctic charr (Salvelinus alpinus is an external fertilizer experiencing intense male-male competition over reproductive opportunities including sperm competition. To compensate for their disadvantage the sneaker males, which often spawn out of synchrony with the female, produce more and faster sperm than the guarding males. We used controlled in vitro fertilization trials with experimentally produced dominant and subordinate, sneaker males to test what effect relative synchrony in gamete release, sperm quality (i.e., motility and velocity and sperm quantity have on a male’s fertilization success in pair-wise sperm competitions. When the sneaker males released ejaculates after the guarding male there was no overall difference in fertilization success. The quality (i.e., motility and velocity of a male’s sperm relative to that of the competing male was the best predictor of male fertilization success regardless of their mating tactic and spawning synchrony. The relative number of sperm cells also had an effect on fertilization success, but mainly when the dominant and sneaker male ejaculated synchronously. Our close imitation of natural sperm competition in charr shows that the sneaker males of external fertilizing species may fully compensate for their disadvantaged mating role by producing ejaculates of higher quality - an adjustment strangely not met by dominants.

Use of FTA® classic cards for epigenetic analysis of sperm DNA.

Science.gov (United States)

Serra, Olga; Frazzi, Raffaele; Perotti, Alessio; Barusi, Lorenzo; Buschini, Annamaria

2018-02-01

FTA® technologies provide the most reliable method for DNA extraction. Although FTA technologies have been widely used for genetic analysis, there is no literature on their use for epigenetic analysis yet. We present for the first time, a simple method for quantitative methylation assessment based on sperm cells stored on Whatman FTA classic cards. Specifically, elution of seminal DNA from FTA classic cards was successfully tested with an elution buffer and an incubation step in a thermocycler. The eluted DNA was bisulfite converted, amplified by PCR, and a region of interest was pyrosequenced.

Assesment of sperm quality using monoclonal antibodies and proteomic analysis

Czech Academy of Sciences Publication Activity Database

Čapková, Jana; Kubátová, Alena; Margaryan, Hasmik; Pěknicová, Jana

2012-01-01

Roč. 67, Issue Supplement s1 (2012), s. 28-28 ISSN 1046-7408. [13th International Symposium for Immunology of reproduction "From the roots to the tops of Reproductive Immunology". 22.06.2012-24.06.2012, Varna] R&D Projects: GA ČR(CZ) GA523/09/1793; GA ČR(CZ) GAP503/12/1834 Institutional research plan: CEZ:AV0Z50520701 Keywords : human sperm * immunofluorescence test * human seminal plasma proteins * flow cytometry * proteomic analysis Subject RIV: EC - Immunology

Gut microbiome may contribute to insulin resistance and systemic inflammation in obese rodents: a meta-analysis.

Science.gov (United States)

Jiao, Na; Baker, Susan S; Nugent, Colleen A; Tsompana, Maria; Cai, Liting; Wang, Yong; Buck, Michael J; Genco, Robert J; Baker, Robert D; Zhu, Ruixin; Zhu, Lixin

2018-04-01

A number of studies have associated obesity with altered gut microbiota, although results are discordant regarding compositional changes in the gut microbiota of obese animals. Herein we used a meta-analysis to obtain an unbiased evaluation of structural and functional changes of the gut microbiota in diet-induced obese rodents. The raw sequencing data of nine studies generated from high-fat diet (HFD)-induced obese rodent models were processed with QIIME to obtain gut microbiota compositions. Biological functions were predicted and annotated with KEGG pathways with PICRUSt. No significant difference was observed for alpha diversity and Bacteroidetes-to-Firmicutes ratio between obese and lean rodents. Bacteroidia, Clostridia, Bacilli, and Erysipelotrichi were dominant classes, but gut microbiota compositions varied among studies. Meta-analysis of the nine microbiome data sets identified 15 differential taxa and 57 differential pathways between obese and lean rodents. In obese rodents, increased abundance was observed for Dorea, Oscillospira, and Ruminococcus, known for fermenting polysaccharide into short chain fatty acids (SCFAs). Decreased Turicibacter and increased Lactococcus are consistent with elevated inflammation in the obese status. Differential functional pathways of the gut microbiome in obese rodents included enriched pyruvate metabolism, butanoate metabolism, propanoate metabolism, pentose phosphate pathway, fatty acid biosynthesis, and glycerolipid metabolism pathways. These pathways converge in the function of carbohydrate metabolism, SCFA metabolism, and biosynthesis of lipid. HFD-induced obesity results in structural and functional dysbiosis of gut microbiota. The altered gut microbiome may contribute to obesity development by promoting insulin resistance and systemic inflammation.

Fins improve the swimming performance of fish sperm: a hydrodynamic analysis of the Siberian sturgeon Acipenser baerii.

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Gillies, Eric A; Bondarenko, Volodymyr; Cosson, Jacky; Pacey, Allan A

2013-02-01

The flagella of sturgeon sperm have an ultrastructure comprising paddle-like fins extending along most of their length. These fins are seen in several other marine and freshwater fish. The sperm of these fish are fast swimmers and are relatively short lived: it is therefore tempting to think of these fins as having evolved for hydrodynamic advantage, but the actual advantage they impart, at such a small length scale and slow speed, is unclear. The phrase "the fins improve hydrodynamic efficiency" is commonly found in biological literature, yet little hydrodynamic analysis has previously been used to support such conjectures. In this paper, we examine various hydrodynamic models of sturgeon sperm and investigate both swimming velocity and energy expenditure. All of the models indicate a modest hydrodynamic advantage of finned sperm, in both straight line swimming speed and a hydrodynamic efficiency measure. We find a hydrodynamic advantage for a flagellum with fins, over one without fins, of the order of 15-20% in straight line propulsive velocity and 10-15% in a hydrodynamic efficiency measure. Copyright © 2012 Wiley Periodicals, Inc.

Identification Male Fertility Through Abnormalities Sperm Based Morphology (Teratospermia) using Invariant Moment Method

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Syahputra, M. F.; Chairani, R.; Seniman; Rahmat, R. F.; Abdullah, D.; Napitupulu, D.; Setiawan, M. I.; Albra, W.; Erliana, C. I.; Andayani, U.

2018-03-01

Sperm morphology is still a standard laboratory analysis in diagnosing infertility in men. Manually identification of sperm form is still not accurate, the difficulty in seeing the form of the invisible sperm from the digital microscope image is often a weakness in the process of identification and takes a long time. Therefore, male fertility identification application system is needed Through sperm abnormalities based on sperm morphology (teratospermia). The method used is invariant moment method. This study uses 15 data testing and 20 data training sperm image. That the process of male fertility identification through sperm abnormalities based on sperm morphology (teratospermia) has an accuracy rate of 80.77%. Use of time to process Identification of male fertility through sperm abnormalities Based on sperm morphology (teratospermia) during 0.4369 seconds.

A content analysis of posthumous sperm procurement protocols with considerations for developing an institutional policy.

Science.gov (United States)

Bahm, Sarah M; Karkazis, Katrina; Magnus, David

2013-09-01

To identify and analyze existing posthumous sperm procurement (PSP) protocols in order to outline central themes for institutions to consider when developing future policies. Qualitative content analysis. Large academic institutions across the United States. We performed a literature search and contacted 40 institutions to obtain nine full PSP protocols. We then performed a content analysis on these policies to identify major themes and factors to consider when developing a PSP protocol. Presence of a PSP policy. We identified six components of a thorough PSP protocol: Standard of Evidence, Terms of Eligibility, Sperm Designee, Restrictions on Use in Reproduction, Logistics, and Contraindications. We also identified two different approaches to policy structure. In the Limited Role approach, institutions have stricter consent requirements and limit their involvement to the time of procurement. In the Family-Centered approach, substituted judgment is permitted but a mandatory wait period is enforced before sperm use in reproduction. Institutions seeking to implement a PSP protocol will benefit from considering the six major building blocks of a thorough protocol and where they would like to fall on the spectrum from a Limited Role to a Family-Centered approach. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Leukocytospermia and sperm preparation - a flow cytometric study

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Perticarari Sandra

2009-01-01

Full Text Available Abstract Background Leukocytes represent the predominant source of reactive oxygen species both in seminal plasma and in sperm suspensions and have been demonstrated to negatively influence sperm function and fertilization rate in assisted reproduction procedures. Peroxidase test is the standard method recommended by WHO to detect semen leukocytes but it may be inaccurate. The aims of this study were (i to compare the efficiency of swim-up and density-gradient centrifugation techniques in removing seminal leukocytes, (ii to examine the effect of leukocytes on sperm preparation, and (iii to compare flow cytometry and peroxidase test in determining leukocyte concentration in semen using a multiparameter flow cytometric method. Methods Semen samples from 126 male partners of couples undergoing infertility investigations were analyzed for leukocytospermia using standard optical microscopy and flow cytometry. Sixty-nine out of 126 samples were also processed using simultaneously the swim-up and density-gradient centrifugation techniques. A multiparameter flow cytometric analysis to assess simultaneously sperm concentration, sperm viability, sperm apoptosis, and leukocyte concentration was carried out on neat and prepared sperm. Results Both sperm preparation methods removed most seminal leukocytes. However, the concentration of leukocytes was significantly lower after swim-up compared to that after density-gradient centrifugation preparation. Leukocytes concentration, either initial or in prepared fractions, was not correlated with sperm parameters (optical microscopy and flow cytometry parameters after semen processing. There was no correlation between leukocyte concentration in the ejaculate and sperm recovery rate, whereas a significant correlation was found between the concentration of the residual leukocytes in prepared fractions and viable sperm recovery rate. Although the overall concordance between the flow cytometry and the optical

Sperm motility in fish: technical applications and perspectives through CASA-Mot systems.

Science.gov (United States)

Gallego, V; Asturiano, J F

2018-03-09

Although a relatively high number of sperm quality biomarkers have been reported over the years in several fish species, sperm motility is nowadays considered the best biomarker for fish spermatozoa. The first scientific reports focusing on fish sperm motility date from a century ago, but the objective assessment allowed by computer-aided sperm analysis (CASA-Mot) systems was not applied to fish species until the mid-1980s. Since then, a high number of sperm kinetic parameters from more than 170 fish species have been reported in more than 700 scientific articles, covering a wide range of topics, such as sperm physiology, sperm storage, broodstock management, the phenomenon of sperm competition, ecotoxicology and understanding the life cycle of the species. The sperm kinetic parameters provided by CASA-Mot systems can serve as powerful and useful tools for aquaculture and ecological purposes, and this review provides an overview of the major research areas in which fish sperm motility assessment by a CASA-Mot system has been used successfully.

Sperm proteins in teleostean and chondrostean (sturgeon) fishes.

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Li, Ping; Hulak, Martin; Linhart, Otomar

2009-11-01

Sperm proteins in the seminal plasma and spermatozoa of teleostean and chondrostean have evolved adaptations due to the changes in the reproductive environment. Analysis of the composition and functions of these proteins provides new insights into sperm motility and fertilising abilities, thereby creating possibilities for improving artificial reproduction and germplasm resource conservation technologies (e.g. cryopreservation). Seminal plasma proteins are involved in the protection of spermatozoa during storage in the reproductive system, whereas all spermatozoa proteins contribute to the swimming and fertilising abilities of sperm. Compared to mammalian species, little data are available on fish sperm proteins and their functions. We review here the current state of the art in this field and focus on relevant subjects that require attention. Future research should concentrate on protein functions and their mode of action in fish species, especially on the role of spermatozoa surface proteins during fertilisation and on a description of sturgeon sperm proteins.

Ascidian Sperm Lysin System

OpenAIRE

Hitoshi, Sawada; Department of Biochemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University

2002-01-01

Fertilization is a precisely controlled process involving many gamete molecules in sperm binding to and penetration through the extracellular matrix of the egg. After sperm bind to the extracellular matrix (vitelline coat), they undergo the acrosome reaction which exposes and partially releases a lytic agent called "lysin" to digest the vitelline coat for the sperm penetration. The vitelline coat sperm lysin is generally a protease in deuterostomes. The molecular mechanism of the actual degra...

Seasonal changes in reproductive activity, sperm variables and sperm freezability in Blanca Andaluza bucks

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Lourdes Gallego-Calvo

2015-12-01

Full Text Available Interest in the preservation of endangered breeds such as the Blanca Andaluza goat, has increased and some steps should be therefore taken to ensure it. The study was designed to determine the seasonal reproductive pattern of Blanca Andaluza bucks, and whether this affects the quality of their semen and its freezability over the year. Seven bucks were used and their body weight, testicular weight, plasma testosterone concentration and fresh sperm quality determined every week. The collected sperm was cryopreserved and stored; it was then thawed and the same sperm quality variables measured every fortnight. High plasma testosterone concentrations were recorded during the summer and autumn, and low concentrations were recorded during winter and spring (p<0.001. No differences were seen between seasons in terms of the percentage of bucks ejaculating, the percentage of active bucks, or ejaculate volume. However, the sperm concentration, the total number of sperm per ejaculate, and the values for most fresh sperm variables were lower during the winter period (at least p<0.05. After freezing-thawing, the quality of winter-collected sperm was better, in some respects, than that of summer-collected sperm (at least p<0.05. These results reveal that Blanca Andaluza bucks show seasonal reproductive activity in terms of their plasma testosterone concentration, but no clear change in their sexual behaviour between seasons was observed. The values of fresh sperm variables also vary over the year, reaching their lowest during winter. However, after freezing-thawing, winter-collected sperm is of overall better quality than sperm collected during the summer.

Seasonal changes in reproductive activity, sperm variables and sperm freezability in Blanca Andaluza bucks

Energy Technology Data Exchange (ETDEWEB)

Gallego-Calvo, L.; Gatica, M.C.; Santiago-Moreno, J.; Guzmán, J.L.; Zarazaga, L.

2015-07-01

Interest in the preservation of endangered breeds such as the Blanca Andaluza goat, has increased and some steps should be therefore taken to ensure it. The study was designed to determine the seasonal reproductive pattern of Blanca Andaluza bucks, and whether this affects the quality of their semen and its freezability over the year. Seven bucks were used and their body weight, testicular weight, plasma testosterone concentration and fresh sperm quality determined every week. The collected sperm was cryopreserved and stored; it was then thawed and the same sperm quality variables measured every fortnight. High plasma testosterone concentrations were recorded during the summer and autumn, and low concentrations were recorded during winter and spring (p<0.001). No differences were seen between seasons in terms of the percentage of bucks ejaculating, the percentage of active bucks, or ejaculate volume. However, the sperm concentration, the total number of sperm per ejaculate, and the values for most fresh sperm variables were lower during the winter period (at least p<0.05). After freezing-thawing, the quality of winter-collected sperm was better, in some respects, than that of summer-collected sperm (at least p<0.05). These results reveal that Blanca Andaluza bucks show seasonal reproductive activity in terms of their plasma testosterone concentration, but no clear change in their sexual behaviour between seasons was observed. The values of fresh sperm variables also vary over the year, reaching their lowest during winter. However, after freezing-thawing, winter-collected sperm is of overall better quality than sperm collected during the summer. (Author)

Heritability of sperm length in the bumblebee Bombus terrestris

DEFF Research Database (Denmark)

Baer, Boris; de Jong, Gerdien; Schmid-Hempel, Regula

2006-01-01

estimates of narrow sense heritability of sperm length in a social insect, the bumblebee Bombus terrestris. In spite of a balanced and straightforward rearing design of colonies, and the possibility to replicate measurements of sperm within single males nested within colonies, the analysis proved...

Phenotypic plasticity in sperm traits in scorpionflies (Mecoptera : Panorpidae): Consequences of larval history and seasonality on sperm length and sperm transfer

NARCIS (Netherlands)

Vermeulen, Andreas; Engels, Sierk; Engqvist, Leif; Sauer, Klaus Peter

2009-01-01

We examined effects of seasonality, larval food availability and larval rearing density on sperm length, sperm transfer rates and body size in the bivoltine scorpionfly Panorpa vulgaris. Males of the first annual generation were larger and had larger sperm. Comparing individuals of two summer

Sperm preparation for fertilization

NARCIS (Netherlands)

Gadella, B.M.

2014-01-01

Description This book contains 19 chapters that discuss theoretical and applied andrology for domestic, zoo and wild animals. Topics include semen and its constituents; sperm production and harvest; determinants of sperm morphology; sperm preparation for fertilization; practical aspects of semen

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

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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.

A dictionary learning approach for human sperm heads classification.

Science.gov (United States)

Shaker, Fariba; Monadjemi, S Amirhassan; Alirezaie, Javad; Naghsh-Nilchi, Ahmad Reza

2017-12-01

To diagnose infertility in men, semen analysis is conducted in which sperm morphology is one of the factors that are evaluated. Since manual assessment of sperm morphology is time-consuming and subjective, automatic classification methods are being developed. Automatic classification of sperm heads is a complicated task due to the intra-class differences and inter-class similarities of class objects. In this research, a Dictionary Learning (DL) technique is utilized to construct a dictionary of sperm head shapes. This dictionary is used to classify the sperm heads into four different classes. Square patches are extracted from the sperm head images. Columnized patches from each class of sperm are used to learn class-specific dictionaries. The patches from a test image are reconstructed using each class-specific dictionary and the overall reconstruction error for each class is used to select the best matching class. Average accuracy, precision, recall, and F-score are used to evaluate the classification method. The method is evaluated using two publicly available datasets of human sperm head shapes. The proposed DL based method achieved an average accuracy of 92.2% on the HuSHeM dataset, and an average recall of 62% on the SCIAN-MorphoSpermGS dataset. The results show a significant improvement compared to a previously published shape-feature-based method. We have achieved high-performance results. In addition, our proposed approach offers a more balanced classifier in which all four classes are recognized with high precision and recall. In this paper, we use a Dictionary Learning approach in classifying human sperm heads. It is shown that the Dictionary Learning method is far more effective in classifying human sperm heads than classifiers using shape-based features. Also, a dataset of human sperm head shapes is introduced to facilitate future research. Copyright © 2017 Elsevier Ltd. All rights reserved.

Quantitative Assessment of Mammary Gland Density in Rodents Using Digital Image Analysis

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Thompson Henry J

2011-06-01

Full Text Available Abstract Background Rodent models have been used extensively to study mammary gland development and for studies of toxicology and carcinogenesis. Mammary gland gross morphology can visualized via the excision of intact mammary gland chains following fixation and staining with carmine using a tissue preparation referred to as a whole mount. Methods are described for the automated collection of digital images from an entire mammary gland whole mount and for the interrogation of digital data using a "masking" technique available with Image-Pro® plus image analysis software (Mediacybernetics. Silver Spring, MD. Results Parallel to mammographic analysis in humans, measurements of rodent mammary gland density were derived from area-based or volume-based algorithms and included: total circumscribed mammary fat pad mass, mammary epithelial mass, and epithelium-free fat pad mass. These values permitted estimation of absolute mass of mammary epithelium as well as breast density. The biological plausibility of these measurements was evaluated in mammary whole mounts from rats and mice. During mammary gland development, absolute epithelial mass increased linearly without significant changes in mammographic density. Treatment of rodents with tamoxifen, 9-cis-retinoic acid, or ovariectomy, and occurrence of diet induced obesity decreased both absolute epithelial mass and mammographic density. The area and volumetric methods gave similar results. Conclusions Digital image analysis can be used for screening agents for potential impact on reproductive toxicity or carcinogenesis as well as for mechanistic studies, particularly for cumulative effects on mammary epithelial mass as well as translational studies of mechanisms that explain the relationship between epithelial mass and cancer risk.

On sperm competition games: raffles and roles revisited.

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Mesterton-Gibbons, M

1999-08-01

In principle there are two approaches to modelling a trade-off between the positive and negative outcomes of a behavior: after suitably defining a value for the behavior in the absence of any trade-off, one can either multiply that value by an appropriate discount or subtract an appropriate cost. In a prospective analysis of sperm competition, Parker (Proc. Roy. Soc. Lond. B (1990) 242, 120-126) adopted the multiplicative approach to model the trade-off between the value of a mating and the cost of its acquisition. He obtained two paradoxical results. First, if two males 'know' whether they are first or second to mate, but these roles are assigned randomly, then sperm numbers should be the same for both males whether the 'raffle' for fertilization is fair or unfair. Second, if mating order is constant, then a favored male should expend less on sperm. His results are puzzling not only in terms of intuition about nature, but also in terms of his model's consistency. In other words, they present both an external and an internal paradox. Parker assumed the fairness of the raffle to a disfavored male to be independent of how much sperm a favored male deposits. This article both generalizes Parker's analysis by allowing fairness to decrease with sperm expenditure by the favored male and compares Parker's results to those obtained by the additive approach. In many respects, results are similar. Nevertheless, if the costs of mating are assumed to increase with sperm expenditure but not to depend on the role in which sperm is expended, as Parker assumed, then the additive approach is more fundamentally correct. In particular, Parker's constant-role paradox is an artifact of his approach. His random-role paradox is internally rationalized in terms of standard microeconomic theory. When fairness decreases, however slightly, with sperm expenditure by the favored male, both models demonstrate that the evolutionarily stable strategy is for more sperm to be deposited during a

More Pitfalls with Sperm Viability Staining and a Viability-Based Stress Test to Characterize Sperm Quality

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Barbara A. Eckel

2017-12-01

Full Text Available Sperm viability (SV, the proportion of live sperm in a sample, is a widely applied measure of sperm quality but few studies test its robustness. At least three reasons make SV problematic as a surrogate for sperm quality. First, reviewing the ecological literature revealed that previously identified methodological pitfalls have not been overcome, including low cross-study standardization of protocols, inadequate statistical treatment, and unaccounted for within-sample heterogeneity. Second, SV is affected by biological variation such as between species, reproductive organs, or sperm age cohorts. Third, the proportion of live sperm extracted from males appears more related to male than to sperm quality in the sense of the future performance of sperm. We propose an alternative method to assess sperm quality by characterizing the temporal decrease of SV in a stressor medium and illustrate in two species, the common bedbug (Cimex lectularius and the fruit fly (Drosophila melanogaster how some common methodological pitfalls may be circumvented. Our data empirically support the well-known but little-considered facts that (i non-blind measurements may alter SV and (ii that SV frequently have non-significant repeatability within one sample. (iii Cross-sectional sampling of ejaculates showed that this heterogeneity even masked a biological pattern—the sperm stratification within males. We show (iv that this shortcoming can be overcome by following the temporal decline of SV of a sperm subsample in a stress test. Finally, (v comparing the staining pattern of sperm between Cimex and Drosophila, we found that in the latter, the visibility of sperm is substantially delayed (30 min when sperm density is high. We show that this delay in stained sperm visibility was, however, not biased toward dead or live sperm. To measure sperm quality, we advocate analyzing the temporal decline in SV in a stressor medium over current protocols that use SV per se and

Rheotaxis guides mammalian sperm

Science.gov (United States)

Miki, Kiyoshi; Clapham, David E

2013-01-01

Background In sea urchins, spermatozoan motility is altered by chemotactic peptides, giving rise to the assumption that mammalian eggs also emit chemotactic agents that guide spermatozoa through the female reproductive tract to the mature oocyte. Mammalian spermatozoa indeed undergo complex adaptations within the female (the process of capacitation) that are initiated by agents ranging from pH to progesterone, but these factors are not necessarily taxic. Currently, chemotaxis, thermotaxis, and rheotaxis have not been definitively established in mammals. Results Here, we show that positive rheotaxis, the ability of organisms to orient and swim against the flow of surrounding fluid, is a major taxic factor for mouse and human sperm. This flow is generated within 4 hours of sexual stimulation and coitus in female mice; prolactin-triggered oviductal fluid secretion clears the oviduct of debris, lowers viscosity, and generates the stream that guides sperm migration in the oviduct. Rheotaxic movement is demonstrated in capacitated and uncapacitated spermatozoa in low and high viscosity medium. Finally, we show that a unique sperm motion we quantify using the sperm head's rolling rate reflects sperm rotation that generates essential force for positioning the sperm in the stream. Rotation requires CatSper channels, presumably by enabling Ca2+ influx. Conclusions We propose that rheotaxis is a major determinant of sperm guidance over long distances in the mammalian female reproductive tract. Coitus induces fluid flow to guide sperm in the oviduct. Sperm rheotaxis requires rotational motion during CatSper channel-dependent hyperactivated motility. PMID:23453951

A microhistological technique for analysis of food habits of mycophagous rodents.

Science.gov (United States)

Patrick W. McIntire; Andrew B. Carey

1989-01-01

We present a technique, based on microhistological analysis of fecal pellets, for quantifying the diets of forest rodents. This technique provides for the simultaneous recording of fungal spores and vascular plant material. Fecal samples should be freeze dried, weighed, and rehydrated with distilled water. We recommend a minimum sampling intensity of 50 fields of view...

Post Thawing Sperm Quality and Ca+2 Intensity Characters of Local Goat Sperm After Freezing by Simple Method Using Deep Freezing

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Gatot Ciptadi

2018-02-01

Full Text Available The objective of this research was to determine the effect of the simple modified freezing method, 1°C/minute freezing rate with different diluter ration on a post-thawing quality of local goat sperm namely Peranakan Etawah (PE. This work is aimed to study the quality of post-thawing sperm and to characterize the calcium intensity profile of both fresh and post thawing goat sperm. The method used is the experimental design of a laboratory. Freezing semen was performed in 2 main temperatures of -45°C then -196°C respectively using Mr. Frosty (® System. Early Sperm characters of Ca+2 intensity was performed by Confocal Laser Scanning Microscope (CLSM through Fluo-3 staining and Ca++ intensity was analysis descriptively. The result showed that post-thawing qualities are considered as good as standard qualities, at least, more than 40% based on Indonesian National Standard (SNI, 2014. The different level diluents commercial of Andromeda used were influenced highly significant (P<0.01. The best diluents ration is 1:4 (v/v for final sperms stocked at -196°C. However freezing sperm conserved in -196°C is better than in -45°C. Meanwhile, the sperm characters of two condition showed the important variation of Ca+2 intensity, with the length of region measurement of 39.06±4.595 and 32.696±9.011 µm each.  It was concluded that the calcium intensity pattern was varied more and higher in fresh sperm than in freezing sperms. This simple modified method of a freezing system was considered as a feasible alternative method for goat semen in a reason for both for sperm post-thawing quality and practical purposes.

Vitrification of neat semen alters sperm parameters and DNA integrity.

Science.gov (United States)

Khalili, Mohammad Ali; Adib, Maryam; Halvaei, Iman; Nabi, Ali

2014-05-06

Our aim was to evaluate the effect of neat semen vitrification on human sperm vital parameters and DNA integrity in men with normal and abnormal sperm parameters. Semen samples were 17 normozoospermic samples and 17 specimens with abnormal sperm parameters. Semen analysis was performed according to World Health Organization (WHO) criteria. Then, the smear was provided from each sample and fixed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Vitrification of neat semen was done by plunging cryoloops directly into liquid nitrogen and preserved for 7 days. The samples were warmed and re-evaluated for sperm parameters as well as DNA integrity. Besides, the correlation between sperm parameters and DNA fragmentation was assessed pre- and post vitrification. Cryopreserved spermatozoa showed significant decrease in sperm motility, viability and normal morphology after thawing in both normal and abnormal semen. Also, the rate of sperm DNA fragmentation was significantly higher after vitrification compared to fresh samples in normal (24.76 ± 5.03 and 16.41 ± 4.53, P = .002) and abnormal (34.29 ± 10.02 and 23.5 ± 8.31, P < .0001), respectively. There was negative correlation between sperm motility and sperm DNA integrity in both groups after vitrification. Vitrification of neat ejaculates has negative impact on sperm parameters as well as DNA integrity, particularly among abnormal semen subjects. It is, therefore, recommend to process semen samples and vitrify the sperm pellets.

Cryopreservation of microencapsulated canine sperm.

Science.gov (United States)

Shah, Shambhu; Otsuki, Tsubasa; Fujimura, Chika; Yamamoto, Naoki; Yamashita, Yasuhisa; Higaki, Shogo; Hishinuma, Mitsugu

2011-03-01

The objective was to develop a method for cryopreserving microencapsulated canine sperm. Pooled ejaculates from three beagle dogs were extended in egg yolk tris extender and encapsulated using alginate and poly-L-lysine at room temperature. The microcapsules were cooled at 4 °C, immersed in pre-cooled extender (equivalent in volume to the microcapsules) to reach final concentration of 7% (v/v) glycerol and 0.75% (v/v) Equex STM paste, and equilibrated for 5, 30 and 60 min at 4 °C. Thereafter, microcapsules were loaded into 0.5 mL plastic straws and frozen in liquid nitrogen. In Experiment 1, characteristics of microencapsulated canine sperm were evaluated after glycerol addition at 4 °C. Glycerol exposure for 5, 30 and 60 min did not significantly affect progressive motility, viability, or acrosomal integrity of microencapsulated sperm compared with pre-cooled unencapsulated sperm (control). In Experiment 2, characteristics of frozen-thawed canine microencapsulated sperm were evaluated at 0, 3, 6, and 9 h of culture at 38.5 °C. Pre-freeze glycerol exposure for 5, 30, and 60 min at 4 °C did not influence post-thaw quality in unencapsulated sperm. Post-thaw motility and acrosomal integrity of microencapsulated sperm decreased more than those of unencapsulated sperm (P < 0.05) following glycerol exposure for 5 min. However, motility, viability and acrosomal integrity of microencapsulated sperm after 30 and 60 min glycerol exposure were higher than unencapsulated sperm cultured for 6 or 9 h (P < 0.05). In conclusion, since microencapsulated canine sperm were successfully cryopreserved, this could be a viable alternative to convention sperm cryopreservation in this species. Copyright © 2011 Elsevier Inc. All rights reserved.

Thyroxin Is Useful to Improve Sperm Motility

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Mendeluk Gabriela Ruth

2016-07-01

Full Text Available Background The aim of this study was to evaluate the non-genomic action of thyroxin on sperm kinetic and its probable use to improve sperm recovery after applying an en- richment method like “swim-up” in comparison with the available one, pentoxifylline. Materials and Methods This is an experimental study. A total of 50 patients were re- cruited, followed by infertility consultation. Conventional sperm assays were performed according to World Health Organization criteria-2010 (WHO-2010. A Computer Aided Semen Analysis System was employed to assess kinetic parameters and concentrations. Number of the motile sperm recovered after preparation technique was calculated. Results Addition of T4 (0.002 µg/ml to semen samples increased hypermotility at 20 minutes (control: 14.18 ± 5.1% vs. 17.66 ± 8.88%, P<0.03, data expressed as mean ± SD and remained unchanged after 40 minutes. Significant differences were found in the motile sperm recovered after swim-up (control: 8.93×106 ± 9.52× 06vs. 17.20×106 ± 21.16×106, P<0.03, achieving all of the tested samples a desirable threshold value for artificial insemination outcome, while adding pentoxifylline increased the number of recovered sperm after swim-up in 60% of the studied cases. No synergism between two treatments could be determined. Conclusion We propose a new physiological tool to artificially improve insemination. The discussion opens windows to investigate unknown pathways involved in sperm ca- pacitation and gives innovative arguments to better understand infertility mechanisms.

Sperm Na+, K+-ATPase and Ca2+-ATPase activity: A preliminary study of comparison of swim up and density gradient centrifugation methods for sperm preparation

Science.gov (United States)

Lestari, Silvia W.; Larasati, Manggiasih D.; Asmarinah, Mansur, Indra G.

2018-02-01

As one of the treatment for infertility, the success rate of Intrauterine Insemination (IUI) is still relatively low. Several sperm preparation methods, swim-up (SU) and the density-gradient centrifugation (DGC) are frequently used to select for better sperm quality which also contribute to IUI failure. Sperm selection methods mainly separate the motile from the immotile sperm, eliminating the seminal plasma. The sperm motility involves the structure and function of sperm membrane in maintaining the balance of ion transport system which is regulated by the Na+, K+-ATPase, and Ca2+-ATPase enzymes. This study aims to re-evaluate the efficiency of these methods in selecting for sperm before being used for IUI and based the evaluation on sperm Na+,K+-ATPase and Ca2+-ATPase activities. Fourteen infertile men from couples who underwent IUI were involved in this study. The SU and DGC methods were used for the sperm preparation. Semen analysis was performed based on the reference value of World Health Organization (WHO) 2010. After isolating the membrane fraction of sperms, the Na+, K+-ATPase activity was defined as the difference in the released inorganic phosphate (Pi) with and without the existence of 10 mM ouabain in the reaction, while the Ca2+-ATPase was determined as the difference in Pi contents with and without the existence of 55 µm CaCl2. The prepared sperm demonstrated a higher percentage of motile sperm compared to sperm from the whole semen. Additionally, the percentage of motile sperm of post-DGC showed higher result than the sperm from post-SU. The velocity of sperm showed similar pattern with the percentage of motile sperm, in which the velocity of prepared sperm was higher than the sperm from whole semen. Furthermore, the sperm velocity of post-DGC was higher compared to the sperm from post-SU. The Na+, K+-ATPase activity of prepared sperm was higher compared to whole semen, whereas Na+, K+-ATPase activity in the post DGC was higher than post SU. The Ca2

Pulmonary exposure to carbonaceous nanomaterials and sperm quality.

Science.gov (United States)

Skovmand, Astrid; Jacobsen Lauvås, Anna; Christensen, Preben; Vogel, Ulla; Sørig Hougaard, Karin; Goericke-Pesch, Sandra

2018-01-31

Semen quality parameters are potentially affected by nanomaterials in several ways: Inhaled nanosized particles are potent inducers of pulmonary inflammation, leading to the release of inflammatory mediators. Small amounts of particles may translocate from the lungs into the lung capillaries, enter the systemic circulation and ultimately reach the testes. Both the inflammatory response and the particles may induce oxidative stress which can directly affect spermatogenesis. Furthermore, spermatogenesis may be indirectly affected by changes in the hormonal milieu as systemic inflammation is a potential modulator of endocrine function. The aim of this study was to investigate the effects of pulmonary exposure to carbonaceous nanomaterials on sperm quality parameters in an experimental mouse model. Effects on sperm quality after pulmonary inflammation induced by carbonaceous nanomaterials were investigated by intratracheally instilling sexually mature male NMRI mice with four different carbonaceous nanomaterials dispersed in nanopure water: graphene oxide (18 μg/mouse/i.t.), Flammruss 101, Printex 90 and SRM1650b (0.1 mg/mouse/i.t. each) weekly for seven consecutive weeks. Pulmonary inflammation was determined by differential cell count in bronchoalveolar lavage fluid. Epididymal sperm concentration and motility were measured by computer-assisted sperm analysis. Epididymal sperm viability and morphological abnormalities were assessed manually using Hoechst 33,342/PI flourescent and Spermac staining, respectively. Epididymal sperm were assessed with regard to sperm DNA integrity (damage). Daily sperm production was measured in the testis, and testosterone levels were measured in blood plasma by ELISA. Neutrophil numbers in the bronchoalveolar fluid showed sustained inflammatory response in the nanoparticle-exposed groups one week after the last instillation. No significant changes in epididymal sperm parameters, daily sperm production or plasma testosterone levels

Evidence for decreasing sperm count in African population from ...

African Journals Online (AJOL)

Purpose: This meta-analysis, following our previous reports those documented an overall 57% diminution in mean sperm concentration around the globe over past 35 years and 32.5% decline in past 50 years in European population, attempts to report the declining trend of sperm concentrations in African population ...

Quantitative evaluation of radiation-induced changes in sperm morphology and chromatin distribution

International Nuclear Information System (INIS)

Aubele, M.; Juetting, U.R.; Rodenacker, K.; Gais, P.; Burger, G.; Hacker-Klom, U.

1990-01-01

Sperm head cytometry provides a useful assay for the detection of radiation-induced damage in mouse germ cells. Exposure of the gonads to radiation is known to lead to an increase of diploid and higher polyploid sperm and of sperm with head shape abnormalities. In the pilot studies reported here quantitative analysis of the total DNA content, the morphology, and the chromatin distribution of mouse sperm was performed. The goal was to evaluate the discriminative power of features derived by high resolution image cytometry in distinguishing sperm of control and irradiated mice. Our results suggest that besides the induction of the above mentioned variations in DNA content and shape of sperm head, changes of the nonhomogeneous chromatin distribution within the sperm may also be used to quantify the radiation effect on sperm cells. Whereas the chromatin distribution features show larger variations for sperm 21 days after exposure (dpr), the shape parameters seem to be more important to discriminate sperm 35 dpr. This may be explained by differentiation processes, which take place in different stages during mouse spermatogenesis

EDC IMPACT: Reduced sperm counts in rats exposed to human relevant mixtures of endocrine disrupters

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M Axelstad

2018-01-01

Full Text Available Human semen quality is declining in many parts of the world, but the causes are ill defined. In rodents, impaired sperm production can be seen with early life exposure to certain endocrine-disrupting chemicals, but the effects of combined exposures are not properly investigated. In this study, we examined the effects of early exposure to the painkiller paracetamol and mixtures of human relevant endocrine-disrupting chemicals in rats. One mixture contained four estrogenic compounds; another contained eight anti-androgenic environmental chemicals and a third mixture contained estrogens, anti-androgens and paracetamol. All exposures were administered by oral gavage to time-mated Wistar dams rats (n = 16–20 throughout gestation and lactation. In the postnatal period, testicular histology was affected by the total mixture, and at the end of weaning, male testis weights were significantly increased by paracetamol and the high doses of the total and the anti-androgenic mixture, compared to controls. In all dose groups, epididymal sperm counts were reduced several months after end of exposure, i.e. at 10  months of age. Interestingly, the same pattern of effects was seen for paracetamol as for mixtures with diverse modes of action. Reduced sperm count was seen at a dose level reflecting human therapeutic exposure to paracetamol. Environmental chemical mixtures affected sperm count at the lowest mixture dose indicating an insufficient margin of safety for the most exposed humans. This causes concern for exposure of pregnant women to paracetamol as well as environmental endocrine disrupters.

Characterization and possible function of glyceraldehyde-3-phosphate dehydrogenase-spermatogenic protein GAPDHS in mammalian sperm.

Science.gov (United States)

Margaryan, Hasmik; Dorosh, Andriy; Capkova, Jana; Manaskova-Postlerova, Pavla; Philimonenko, Anatoly; Hozak, Pavel; Peknicova, Jana

2015-03-08

Sperm proteins are important for the sperm cell function in fertilization. Some of them are involved in the binding of sperm to the egg. We characterized the acrosomal sperm protein detected by a monoclonal antibody (MoAb) (Hs-8) that was prepared in our laboratory by immunization of BALB/c mice with human ejaculated sperms and we tested the possible role of this protein in the binding assay. Indirect immunofluorescence and immunogold labelling, gel electrophoresis, Western blotting and protein sequencing were used for Hs-8 antigen characterization. Functional analysis of GAPDHS from the sperm acrosome was performed in the boar model using sperm/zona pellucida binding assay. Monoclonal antibody Hs-8 is an anti-human sperm antibody that cross-reacts with the Hs-8-related protein in spermatozoa of other mammalian species (boar, mouse). In the immunofluorescence test, Hs-8 antibody recognized the protein localized in the acrosomal part of the sperm head and in the principal piece of the sperm flagellum. In immunoblotting test, MoAb Hs-8 labelled a protein of 45 kDa in the extract of human sperm. Sequence analysis identified protein Hs-8 as GAPDHS (glyceraldehyde 3-phosphate dehydrohenase-spermatogenic). For this reason, commercial mouse anti-GAPDHS MoAb was applied in control tests. Both antibodies showed similar staining patterns in immunofluorescence tests, in electron microscopy and in immunoblot analysis. Moreover, both Hs-8 and anti-GAPDHS antibodies blocked sperm/zona pellucida binding. GAPDHS is a sperm-specific glycolytic enzyme involved in energy production during spermatogenesis and sperm motility; its role in the sperm head is unknown. In this study, we identified the antigen with Hs8 antibody and confirmed its localization in the apical part of the sperm head in addition to the principal piece of the flagellum. In an indirect binding assay, we confirmed the potential role of GAPDHS as a binding protein that is involved in the secondary sperm

Ovarian fluid mediates the temporal decline in sperm viability in a fish with sperm storage.

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Clelia Gasparini

Full Text Available A loss of sperm viability and functionality during sperm transfer and storage within the female reproductive tract can have important fitness implications by disrupting fertilization and impairing offspring development and survival. Consequently, mechanisms that mitigate the temporal decline in sperm function are likely to be important targets of selection. In many species, ovarian fluid is known to regulate and maintain sperm quality. In this paper, we use the guppy Poecilia reticulata, a highly polyandrous freshwater fish exhibiting internal fertilization and sperm storage, to determine whether ovarian fluid (OF influences the decline in sperm viability (the proportion of live sperm in the ejaculate over time and whether any observed effects depend on male sexual ornamentation. To address these questions we used a paired experimental design in which ejaculates from individual males were tested in vitro both in presence and absence of OF. Our results revealed that the temporal decline in sperm viability was significantly reduced in the presence of OF compared to a saline control. This finding raises the intriguing possibility that OF may play a role in mediating the decline in sperm quality due to the deleterious effects of sperm ageing, although other possible explanations for this observation are discussed. Interestingly, we also show that the age-related decline in sperm viability was contingent on male sexual ornamentation; males with relatively high levels of iridescence (indicating higher sexual attractiveness exhibited a more pronounced decline in sperm viability over time than their less ornamented counterparts. This latter finding offers possible insights into the functional basis for the previously observed trade-off between these key components of pre- and postcopulatory sexual selection.

Effect of chamber characteristics, loading and analysis time on motility and kinetic variables analysed with the CASA-mot system in goat sperm.

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Del Gallego, R; Sadeghi, S; Blasco, E; Soler, C; Yániz, J L; Silvestre, M A

2017-02-01

Several factors unrelated to the semen samples could be influencing in the sperm motility analysis. The aim of the present research was to study the effect of four chambers with different characteristics, namely; slide-coverslip, Spermtrack, ISAS D4C10, and ISAS D4C20 on the sperm motility. The filling procedure (drop or capillarity) and analysis time (0, 120 and 240s), depth of chamber (10 or 20μm) and field on motility variables were analysed by use of the CASA-mot system in goat sperm. Use of the drop-filling chambers resulted in greater values than capillarity-filling chambers for all sperm motility and kinetic variables, except for LIN (64.5% compared with 56.3% of motility for drop- and capillarity-filling chambers respectively, PCASA-mot system with a drop-loaded chamber within 2min after filling the chamber. Copyright © 2016 Elsevier B.V. All rights reserved.

Is the quality of donated semen deteriorating? Findings from a 15 year longitudinal analysis of weekly sperm samples.

Science.gov (United States)

Haimov-Kochman, Ronit; Har-Nir, Ruth; Ein-Mor, Eliana; Ben-Shoshan, Vered; Greenfield, Caryn; Eldar, Ido; Bdolah, Yuval; Hurwitz, Arye

2012-06-01

Studies suggest that global semen quality is declining, but the debate remains open owing to geographic variation. To evaluate temporal trends of sperm parameters - namely concentration, motility and total motile sperm count - in sperm donated during the period 1995-2009. In a retrospective longitudinal cohort study we analyzed the sperm count and motility of 2182 semen samples provided on a weekly basis by 58 young, healthy, fertile, university-educated, paid donors. Despite the lowering of criteria for sperm parameters satisfactory for donation that were implemented in 2004, 38% of applicants for sperm donation are now rejected based on semen quality as compared to a third of applicants 10-15 years ago (P average sperm parameters dropped from a concentration of 106 +/- 25 million spermatozoa/ml with 79% +/- 4.3% motility to 68 +/- 14 million/ ml with 66% +/- 4.5% motile sperm (P sperm count per ejaculate also decreased, from 66.4 +/- 18.2 million to 48.7 +/- 12 million (P sperm donors, only 18% of donors had an acceptable sperm quality, with an average concentration of 87 +/- 12 million spermatozoa/ml, 73% +/- 2.6% motile sperm and total motile sperm count of 53.1 +/- 3.8 million per ejaculate - still significantly lower than 15 years ago (P= 0.01, P= 0.003, P= 0.058 respectively). The rapid deterioration of sperm quality among fertile semen donors is alarming and may lead to cessation of sperm donation programs.

Impaired Sperm Maturation in Rnase9 Knockout Mice1

Science.gov (United States)

Westmuckett, Andrew D.; Nguyen, Edward B.; Herlea-Pana, Oana M.; Alvau, Antonio; Salicioni, Ana M.; Moore, Kevin L.

2014-01-01

ABSTRACT Ribonuclease, RNase A family, 9 (RNASE9) is a ribonuclease A superfamily member that is expressed only in the epididymis. It is a small, secreted polypeptide, it lacks ribonuclease activity, and its function(s) is unknown. However, epididymis-specific expression suggests a role in sperm maturation. We generated Rnase9−/− mice to study RNASE9 function in vivo. We confirm that RNASE9 expression is restricted to the epididymis. Within the epididymis, RNASE9 is first detected in midcaput, persists through the distal caput and corpus, and wanes in the cauda. Rnase9−/− mice are born at the expected Mendelian ratio, have normal postnatal growth and development, and have no outwardly apparent phenotype. Spermatogenesis is normal, and Rnase9-null sperm are morphologically normal. Rnase9−/− males have normal fertility in unrestricted mating trials, and fertilization rates in in vitro fertilization assays are indistinguishable from wild-type mice. Visual observations coupled with analyses of sperm velocities shortly after swim out from the corpus shows that motility of Rnase9-null sperm is significantly impaired. However, no differences between wild-type and Rnase9-null sperm are detected by computer-assisted sperm analysis 10–90 min after sperm isolation from the corpus or cauda. Assessment of capacitation-dependent signaling pathways in Rnase9-null sperm showed that, while levels of tyrosine phosphorylation of sperm proteins were normal, there was decreased phosphorylation of protein kinase A substrates upon capacitation compared to wild-type mice. In conclusion, RNASE9 is dispensable for fertility, but the absence of RNASE9 during epididymal transit results in impaired sperm maturation. PMID:24719258

Identification of phosphoproteins coupled to initiation of motility in live epididymal mouse sperm

Science.gov (United States)

Tash, J. S.; Bracho, G. E.

1998-01-01

A method for collecting live immotile cauda epididymal mouse sperm that initiate motility by dilution into an activation buffer is described. Sperm in collection buffer showed low percent motility (MOT) and population progression (PRG) that increased 10-fold and 9-fold, respectively, during the first 2 min after dilution into activation buffer. Western phosphoserine (pS), phosphothreonine (pT), and phosphotyrosine (pY) analysis revealed a 120 kDa protein that markedly increased in pT content during initiation of motility and may be related to FP130, the motility-coupled axonemal protein of sea urchin sperm. A prominent 82 kDa protein that was pS and pT-phosphorylated in immotile and motile sperm is likely the fibrous sheath component AKAP82 that is phosphorylated during spermatogenesis. Analysis of live human sperm also identified a prominent 120 kDa pT protein. Thus it appears that phosphorylation of FP130 and related 120 kDa proteins in mouse, and perhaps human sperm, represent common targets during motility initiation in sperm. Copyright 1998 Academic Press.

Sperm freezing to address the risk of azoospermia on the day of ICSI.

Science.gov (United States)

Montagut, M; Gatimel, N; Bourdet-Loubère, S; Daudin, M; Bujan, L; Mieusset, R; Isus, F; Parinaud, J; Leandri, R

2015-11-01

In which cases is freezing of ejaculated sperm indicated before ICSI? Sperm freezing should be performed only when out of two analyses at least one total sperm count in the ejaculate is lower than 10(6). Due to variations in individual sperm parameters, in cases of severe oligozoospermia there is a risk of absence of spermatozoa on the day of ICSI, leading to cancellation of the attempt. Sperm freezing can avoid this problem but little is known of the parameters governing the decision to freeze sperm or not. This retrospective study included 247 men who underwent sperm cryopreservation to prevent the risk of azoospermia on the day of ICSI, from 2000 to 2012. Receiver operating characteristic curve analysis was used to define the threshold value. The lowest total sperm count per ejaculate was studied as a predictive factor for the use of frozen sperm in a total of 593 ICSI attempts. Moreover, 2003 patients who had at least 4 semen analyses for andrological diagnosis have been studied to evaluate the reproducibility of sperm count. To evaluate the psychological impact of sperm freezing, a questionnaire was administered to 84 men who attended for sperm cryopreservation between June and December 2014. The cost of sperm freezing was analysed according to the French prices. When at least one total sperm count was counts were ≥10(5) (P sperm freezing is recommended when one analysis from at least two, showed a sperm count sperm freezings. The psychological impact of sperm freezing was good since >70% of men had positive feelings about this technique. This was a fairly short-term study and preservation of future fertility was not assessed. It appeared impossible to find a threshold that would predict the risk of azoospermia with 100% accuracy. Therefore there is still a risk of absence of spermatozoa on the day of ICSI despite a good negative predictive value when no total sperm count was lower than 10(5). These data suggest that sperm freezing should be proposed when

Network Analysis of Rodent Transcriptomes in Spaceflight

Science.gov (United States)

Ramachandran, Maya; Fogle, Homer; Costes, Sylvain

2017-01-01

Network analysis methods leverage prior knowledge of cellular systems and the statistical and conceptual relationships between analyte measurements to determine gene connectivity. Correlation and conditional metrics are used to infer a network topology and provide a systems-level context for cellular responses. Integration across multiple experimental conditions and omics domains can reveal the regulatory mechanisms that underlie gene expression. GeneLab has assembled rich multi-omic (transcriptomics, proteomics, epigenomics, and epitranscriptomics) datasets for multiple murine tissues from the Rodent Research 1 (RR-1) experiment. RR-1 assesses the impact of 37 days of spaceflight on gene expression across a variety of tissue types, such as adrenal glands, quadriceps, gastrocnemius, tibalius anterior, extensor digitorum longus, soleus, eye, and kidney. Network analysis is particularly useful for RR-1 -omics datasets because it reinforces subtle relationships that may be overlooked in isolated analyses and subdues confounding factors. Our objective is to use network analysis to determine potential target nodes for therapeutic intervention and identify similarities with existing disease models. Multiple network algorithms are used for a higher confidence consensus.

Changes in Sperm Motility and Capacitation Induce Chromosomal Aberration of the Bovine Embryo following Intracytoplasmic Sperm Injection.

Directory of Open Access Journals (Sweden)

Yoku Kato

Full Text Available Intracytoplasmic sperm injection (ICSI has become the method of choice to treat human male infertility. One of the outstanding problems associated with this technique is our current lack of knowledge concerning the effect of sperm capacitation and motility upon the subsequent development of oocytes following ICSI. In the present study, we first examined the capacitation state of sperm exhibiting normal motility, along with sperm that had been activated, and examined the effect of reactive oxygen species (ROS produced by these sperm types upon embryogenesis following bovine in vitro fertilization (IVF and ICSI. Data showed that activated sperm reduced the chromosomal integrity of IVF/ICSI embryos at the blastocyst stage, while capacitated sperm produced ROS in capacitation media. Secondly, we treated sperm with carbonyl cyanide m-chlorophenyl hydrazine (CCCP, a chemical known to uncouple cell respiration within the mitochondria, and investigated the effect of this treatment upon blastocyst formation and chromosomal integrity at the blastocyst stage. Activated sperm in which the mitochondria had been treated with CCCP reduced levels of chromosomal aberration at the blastocyst stage following ICSI, by reducing mitochondrial activity in activated sperm. In conclusion, these findings suggest that capacitated sperm exhibiting activated motility induced chromosomal aberration during development to the blastocyst stage following ICSI. The injection of sperm exhibiting normal motility, or activated sperm in which mitochondrial activity had been reduced, improved the quality of ICSI-derived embryos. Therefore, the selection of sperm exhibiting progressive motility may not always be better for early embryo development and fetal growth following human ICSI, and that the use of a bovine model may contribute to a deeper understanding of sperm selection for human ICSI embryo development.

Low Sperm Count

Science.gov (United States)

... that the fluid (semen) you ejaculate during an orgasm contains fewer sperm than normal. A low sperm ... ejaculation occurs when semen enters the bladder during orgasm instead of emerging out of the tip of ...

Association between chronic pain and the sperm motion characteristics

DEFF Research Database (Denmark)

Dardmeh, Fereshteh; Alipour, Hiva; Nielsen, Hans Ingolf

2015-01-01

significantly higher in the processed sample of the CP group. This study demonstrated that chronic pain does not affect the sperm morphology, total concentration and motility based on conventional analysis but has significant influence at the level of sperm motion kinetics which could prove to be clinically...... stimulation, it can be speculated that the observed difference in sperm kinematic parameters could be related to the alterations in serum sex hormone levels emanating from the chronic pain. Further studies are required to explain the possible mechanism of action of chronic pain on male fertility....

Sperm quality assessment via separation and sedimentation in a microfluidic device.

Science.gov (United States)

Chen, Chang-Yu; Chiang, Tsun-Chao; Lin, Cheng-Ming; Lin, Shu-Sheng; Jong, De-Shien; Tsai, Vincent F-S; Hsieh, Ju-Ton; Wo, Andrew M

2013-09-07

A major reason for infertility is due to male factors, including the quality of spermatozoa, which is a primary factor and often difficult to assess, particularly the total sperm concentration and its motile percentage. This work presents a simple microfluidic device to assess sperm quality by quantifying both total and motile sperm counts. The key design feature of the microfluidic device is two channels separated by a permeative phase-guide structure, where one channel is filled with raw semen and the other with pure buffer. The semen sample was allowed to reach equilibrium in both chambers, whereas non-motile sperms remained in the original channel, and roughly half of the motile sperms would swim across the phase-guide barrier into the buffer channel. Sperms in each channel agglomerated into pellets after centrifugation, with the corresponding area representing total and motile sperm concentrations. Total sperm concentration up to 10(8) sperms per ml and motile percentage in the range of 10-70% were tested, encompassing the cutoff value of 40% stated by World Health Organization standards. Results from patient samples show compact and robust pellets after centrifugation. Comparison of total sperm concentration between the microfluidic device and the Makler chamber reveal they agree within 5% and show strong correlation, with a coefficient of determination of R(2) = 0.97. Motile sperm count between the microfluidic device and the Makler chamber agrees within 5%, with a coefficient of determination of R(2) = 0.84. Comparison of results from the Makler Chamber, sperm quality analyzer, and the microfluidic device revealed that results from the microfluidic device agree well with the Makler chamber. The sperm microfluidic chip analyzes both total and motile sperm concentrations in one spin, is accurate and easy to use, and should enable sperm quality analysis with ease.

Quantitative evaluation of radiation-induced changes in sperm morphology and chromatin distribution

International Nuclear Information System (INIS)

Aubele, M.; Burger, G.; Gais, P.; Juetting, V.; Rodenacker, K.; Hacker-Klom, V.

1993-01-01

Sperm head cytometry provides a useful assay for the detection of radiation induced damage in mouse germ cells. Exposure of the gonads to radiation is long known to lead to an increase of diploid and higher polyploid sperm and of sperm with head shape abnormalities. In the pilot studies reported here quantitative analysis of the total DNA content, the morphology, and the chromatin distribution of mouse sperm were performed. The goal was to evaluate the discriminative power of features derived by high resolution image cytometry in distinguishing sperm of control and irradiated mice. Our results suggest that besides the induction of the above mentioned variations in DNA content and shape of sperm head changes of the nonhomogeneous chromatin distribution within the sperm may also be used to quantify the radiation effect on sperm cells. Whereas the chromatin distribution features show bigger variations for sperm 21 days after exposure (dpr), the shape parameters seem to be more important to discriminate sperm 35 dpr. This may be explained by differentiation processes, which take place in different stages during mouse spermatogenesis. (authors). 25 refs., 4 tabs., 7 figs

Quantitative evaluation of radiation-induced changes in sperm morphology and chromatin distribution

Energy Technology Data Exchange (ETDEWEB)

Aubele, M; Burger, G; Gais, P; Juetting, V; Rodenacker, K [Gesellschaft fuer Strahlen- und Umweltforschung mbH Muenchen, Neuherberg (Germany); Hacker-Klom, V [Muenster Univ. (Germany). Inst. fuer Strahlenbiologie

1994-12-31

Sperm head cytometry provides a useful assay for the detection of radiation induced damage in mouse germ cells. Exposure of the gonads to radiation is long known to lead to an increase of diploid and higher polyploid sperm and of sperm with head shape abnormalities. In the pilot studies reported here quantitative analysis of the total DNA content, the morphology, and the chromatin distribution of mouse sperm were performed. The goal was to evaluate the discriminative power of features derived by high resolution image cytometry in distinguishing sperm of control and irradiated mice. Our results suggest that besides the induction of the above mentioned variations in DNA content and shape of sperm head changes of the nonhomogeneous chromatin distribution within the sperm may also be used to quantify the radiation effect on sperm cells. Whereas the chromatin distribution features show bigger variations for sperm 21 days after exposure (dpr), the shape parameters seem to be more important to discriminate sperm 35 dpr. This may be explained by differentiation processes, which take place in different stages during mouse spermatogenesis. (authors). 25 refs., 4 tabs., 7 figs.

Validation of image cytometry for sperm concentration measurement

DEFF Research Database (Denmark)

Egeberg Palme, Dorte L.; Johannsen, Trine Holm; Petersen, Jørgen Holm

2017-01-01

Sperm concentration is an essential parameter in the diagnostic evaluation of men from infertile couples. It is usually determined by manual counting using a hemocytometer, and is therefore both laborious and subjective. We have earlier shown that a newly developed image cytometry (IC) method may...... be used to determine sperm concentration. Here we present a validation of the IC method by analysis of 4010 semen samples. There was high agreement between IC and manual counting at sperm concentrations above 3 mill/ml and in samples with concentrations above 12 mill/ml the two methods can be used...... a lower coefficient of variation than the manual method (5% vs 10%), indicating a better precision of the IC method. In conclusion, measurement of sperm concentration by IC can be used at concentrations above 3 mill/ml and seems more accurate and precise than manual counting, making it an attractive...

Milk proteins interact with goat Binder of SPerm (BSP) proteins and decrease their binding to sperm.

Science.gov (United States)

de Menezes, Erika Bezerra; van Tilburg, Mauricio; Plante, Geneviève; de Oliveira, Rodrigo V; Moura, Arlindo A; Manjunath, Puttaswamy

2016-11-01

Seminal plasma Binder of SPerm (BSP) proteins bind to sperm at ejaculation and promote capacitation. When in excess, however, BSP proteins damage the sperm membrane. It has been suggested that milk components of semen extenders associate with BSP proteins, potentially protecting sperm. Thus, this study was conducted to investigate if milk proteins interact with BSP proteins and reduce BSP binding to goat sperm. Using gel filtration chromatography, milk was incubated with goat seminal plasma proteins and loaded onto columns with and without calcium. Milk was also fractionated into parts containing mostly whey proteins or mostly caseins, incubated with seminal plasma proteins and subjected to gel filtration. Eluted fractions were evaluated by immunoblot using anti-goat BSP antibodies, confirming milk protein-BSP protein interactions. As determined by ELISA, milk proteins coated on polystyrene wells bound to increasing of goat BSP proteins. Far-western dot blots confirmed that BSP proteins bound to caseins and β-lactoglobulin in a concentration-dependent manner. Then, cauda epididymal sperm from five goats was incubated with seminal plasma; seminal plasma followed by milk; and milk followed by seminal plasma. Sperm membrane proteins were extracted and evaluated by immunoblotting. The pattern of BSP binding to sperm membrane proteins was reduced by 59.3 % when epididymal sperm were incubated with seminal plasma and then with skimmed milk (p  0.05). In conclusion, goat BSP proteins have an affinity for caseins and whey proteins. Milk reduces BSP binding to goat sperm, depending whether or not sperm had been previously exposed to seminal plasma. Such events may explain the protective effect of milk during goat sperm preservation.

Monitoring sperm mitochondrial respiration response in a laser trap using ratiometric fluorescence

Science.gov (United States)

Mei, Adrian; Botvinick, Elliot; Berns, Michael

2005-08-01

Sperm motility is an important area in understanding male infertility. Various techniques, such as the Computer Assisted Sperm Analysis (CASA), have been used to understand sperm motility. Sperm motility is related to the energy (ATP) production of sperm. ATP is produced by the depolarization of the membrane potential of the inner membrane of the mitochondria. In this study, a mitochondrial dye, JC-1, has been used to monitor the energetics of the mitochondria. This fluorescent dye can emit at two different wavelengths, depending on the membrane potential of the mitochondria. It can fluoresce green at low membrane potential and red at high membrane potential. The ratio of the two colors (red/green) allows for an accurate measurement of the change of membrane potential. Various experiments were conducted to quantify the behavior of the dye within the sperm and the reaction of the sperm to trap. Sperm were trapped using laser tweezers. Results have shown that the ratio drops dramatically when sperm are trapped, indicating a depolarization of the membrane. The physiological response to this depolarization is yet to be determined, but the studies indicate that the sperm could have been slightly damaged by the laser. However, knowing that sperm depolarizes their membrane when trapped can help understand how sperm react to their environment and consequently help treat male infertility.

Sperm whale long-range echolocation sounds revealed by ANTARES, a deep-sea neutrino telescope

Science.gov (United States)

André, M.; Caballé, A.; van der Schaar, M.; Solsona, A.; Houégnigan, L.; Zaugg, S.; Sánchez, A. M.; Castell, J. V.; Solé, M.; Vila, F.; Djokic, D.; Adrián-Martínez, S.; Albert, A.; Anghinolfi, M.; Anton, G.; Ardid, M.; Aubert, J.-J.; Avgitas, T.; Baret, B.; Barrios-Martí, J.; Basa, S.; Bertin, V.; Biagi, S.; Bormuth, R.; Bouwhuis, M. C.; Bruijn, R.; Brunner, J.; Busto, J.; Capone, A.; Caramete, L.; Carr, J.; Celli, S.; Chiarusi, T.; Circella, M.; Coleiro, A.; Coniglione, R.; Costantini, H.; Coyle, P.; Creusot, A.; Deschamps, A.; de Bonis, G.; Distefano, C.; di Palma, I.; Donzaud, C.; Dornic, D.; Drouhin, D.; Eberl, T.; El Bojaddaini, I.; Elsässer, D.; Enzenhöfer, A.; Fehn, K.; Felis, I.; Fusco, L. A.; Galatà, S.; Gay, P.; Geißelsöder, S.; Geyer, K.; Giordano, V.; Gleixner, A.; Glotin, H.; Gracia-Ruiz, R.; Graf, K.; Hallmann, S.; van Haren, H.; Heijboer, A. J.; Hello, Y.; Hernandez-Rey, J. J.; Hößl, J.; Hofestädt, J.; Hugon, C.; Illuminati, G.; James, C. W.; de Jong, M.; Jongen, M.; Kadler, M.; Kalekin, O.; Katz, U.; Kießling, D.; Kouchner, A.; Kreter, M.; Kreykenbohm, I.; Kulikovskiy, V.; Lachaud, C.; Lahmann, R.; Lefèvre, D.; Leonora, E.; Loucatos, S.; Marcelin, M.; Margiotta, A.; Marinelli, A.; Martínez-Mora, J. A.; Mathieu, A.; Melis, K.; Michael, T.; Migliozzi, P.; Moussa, A.; Mueller, C.; Nezri, E.; Păvălaş, G. E.; Pellegrino, C.; Perrina, C.; Piattelli, P.; Popa, V.; Pradier, T.; Racca, C.; Riccobene, G.; Roensch, K.; Saldaña, M.; Samtleben, D. F. E.; Sanguineti, M.; Sapienza, P.; Schnabel, J.; Schüssler, F.; Seitz, T.; Sieger, C.; Spurio, M.; Stolarczyk, Th.; Sánchez-Losa, A.; Taiuti, M.; Trovato, A.; Tselengidou, M.; Turpin, D.; Tönnis, C.; Vallage, B.; Vallée, C.; van Elewyck, V.; Vivolo, D.; Wagner, S.; Wilms, J.; Zornoza, J. D.; Zuñiga, J.

2017-04-01

Despite dedicated research has been carried out to adequately map the distribution of the sperm whale in the Mediterranean Sea, unlike other regions of the world, the species population status is still presently uncertain. The analysis of two years of continuous acoustic data provided by the ANTARES neutrino telescope revealed the year-round presence of sperm whales in the Ligurian Sea, probably associated with the availability of cephalopods in the region. The presence of the Ligurian Sea sperm whales was demonstrated through the real-time analysis of audio data streamed from a cabled-to-shore deep-sea observatory that allowed the hourly tracking of their long-range echolocation behaviour on the Internet. Interestingly, the same acoustic analysis indicated that the occurrence of surface shipping noise would apparently not condition the foraging behaviour of the sperm whale in the area, since shipping noise was almost always present when sperm whales were acoustically detected. The continuous presence of the sperm whale in the region confirms the ecological value of the Ligurian sea and the importance of ANTARES to help monitoring its ecosystems.

Sperm whale long-range echolocation sounds revealed by ANTARES, a deep-sea neutrino telescope

Science.gov (United States)

André, M.; Caballé, A.; van der Schaar, M.; Solsona, A.; Houégnigan, L.; Zaugg, S.; Sánchez, A. M.; Castell, J. V.; Solé, M.; Vila, F.; Djokic, D.; Adrián-Martínez, S.; Albert, A.; Anghinolfi, M.; Anton, G.; Ardid, M.; Aubert, J.-J.; Avgitas, T.; Baret, B.; Barrios-Martí, J.; Basa, S.; Bertin, V.; Biagi, S.; Bormuth, R.; Bouwhuis, M. C.; Bruijn, R.; Brunner, J.; Busto, J.; Capone, A.; Caramete, L.; Carr, J.; Celli, S.; Chiarusi, T.; Circella, M.; Coleiro, A.; Coniglione, R.; Costantini, H.; Coyle, P.; Creusot, A.; Deschamps, A.; De Bonis, G.; Distefano, C.; Di Palma, I.; Donzaud, C.; Dornic, D.; Drouhin, D.; Eberl, T.; El Bojaddaini, I.; Elsässer, D.; Enzenhöfer, A.; Fehn, K.; Felis, I.; Fusco, L. A.; Galatà, S.; Gay, P.; Geißelsöder, S.; Geyer, K.; Giordano, V.; Gleixner, A.; Glotin, H.; Gracia-Ruiz, R.; Graf, K.; Hallmann, S.; van Haren, H.; Heijboer, A. J.; Hello, Y.; Hernandez-Rey, J. J.; Hößl, J.; Hofestädt, J.; Hugon, C.; Illuminati, G.; James, C. W.; de Jong, M.; Jongen, M.; Kadler, M.; Kalekin, O.; Katz, U.; Kießling, D.; Kouchner, A.; Kreter, M.; Kreykenbohm, I.; Kulikovskiy, V.; Lachaud, C.; Lahmann, R.; Lefèvre, D.; Leonora, E.; Loucatos, S.; Marcelin, M.; Margiotta, A.; Marinelli, A.; Martínez-Mora, J. A.; Mathieu, A.; Melis, K.; Michael, T.; Migliozzi, P.; Moussa, A.; Mueller, C.; Nezri, E.; Păvălaş, G. E.; Pellegrino, C.; Perrina, C.; Piattelli, P.; Popa, V.; Pradier, T.; Racca, C.; Riccobene, G.; Roensch, K.; Saldaña, M.; Samtleben, D. F. E.; Sanguineti, M.; Sapienza, P.; Schnabel, J.; Schüssler, F.; Seitz, T.; Sieger, C.; Spurio, M.; Stolarczyk, Th.; Sánchez-Losa, A.; Taiuti, M.; Trovato, A.; Tselengidou, M.; Turpin, D.; Tönnis, C.; Vallage, B.; Vallée, C.; Van Elewyck, V.; Vivolo, D.; Wagner, S.; Wilms, J.; Zornoza, J. D.; Zuñiga, J.

2017-01-01

Despite dedicated research has been carried out to adequately map the distribution of the sperm whale in the Mediterranean Sea, unlike other regions of the world, the species population status is still presently uncertain. The analysis of two years of continuous acoustic data provided by the ANTARES neutrino telescope revealed the year-round presence of sperm whales in the Ligurian Sea, probably associated with the availability of cephalopods in the region. The presence of the Ligurian Sea sperm whales was demonstrated through the real-time analysis of audio data streamed from a cabled-to-shore deep-sea observatory that allowed the hourly tracking of their long-range echolocation behaviour on the Internet. Interestingly, the same acoustic analysis indicated that the occurrence of surface shipping noise would apparently not condition the foraging behaviour of the sperm whale in the area, since shipping noise was almost always present when sperm whales were acoustically detected. The continuous presence of the sperm whale in the region confirms the ecological value of the Ligurian sea and the importance of ANTARES to help monitoring its ecosystems. PMID:28401960

Sperm morphological and morphometric evaluation in captive collared peccaries (Pecari tajacu

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Patrícia C. Sousa

2013-07-01

Full Text Available The aim of this study was to compare different staining methods for the evaluation of sperm morphology by light microscopy and also to describe the morphometry of the entire sperm in collared peccaries (Pecari tajacu. Semen from 10 males was obtained by electroejaculation and evaluated for sperm motility, vigor, and concentration. Semen smears were prepared through three different staining methods: Bengal rose, brome-phenol blue, and eosin-nigrosin. Smears were evaluated under light microscopy and sperm morphologic alterations were determined in percentage. In addition, sperm morphometric analysis was conducted by light microscopy coupled to image analyzer software. The smears stained with Bengal Rose provide the best results for the visualization of the sperm tail, midpiece, and head. The use of eosin-nigrosin stain did not allow an adequate impregnation, and some sperm presented a few contrasts with the background. A higher incidence of bent coiled tails was verified in the use of brome-phenol blue staining (P<0.05. Through morphometric evaluation, it was observed that the tail occupies the greatest proportion (89% of the sperm which presents a discretely elongated head. According to the results, the use of the Bengal Rose stain is recommended for the morphologic evaluation of the collared peccary sperm.

Chlamydiae in the ejaculate: their influence on the quality and morphology of sperm.

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Veznik, Zdenek; Pospisil, Leopold; Svecova, Drahomira; Zajicova, Atanaska; Unzeitig, Vit

2004-07-01

Given the lack of information concerning the role of Chlamydia trachomatis in male fertility, the aim of this study was to ascertain and analyze the quality of Chlamydiae-positive and -negative semen. Sperm count was performed according to the 1999 World Health Organization (WHO) laboratory manual for examination of human semen and sperm-cervical mucus interaction, and sperm survival was assessed by a 120-min test. The evaluation of the morphological examination of ejaculates was carried out using the sasmo (strict morphological analysis of ejaculates) computer program. Chlamydiae were detected by immunofluorescent reaction using the Progen Biotechnik GmbH diagnostic set. Fisher's exact test and the chi-quadrate test were used for statistical analysis. Of the total of 627 sperm samples examined, Chlamydiae were detected in 136 cases (21.7%). Sperm analysis showed significant differences between Chlamydiae-positive and -negative samples. The Chlamydiae-contaminated group showed normal sperm morphology 14.4% lower, volume 6.4% lower, concentration 8.3% lower, motility 7.8% and velocity 9.3% lower than in Chlamydiae-negative samples. The average values for normal spermatozoa and motility in the Chlamydiae-positive group were also significantly reduced. Chlamydia trachomatis was found to be a possible factor in sperm pathology. These results could help to elucidate the role of Chlamydia trachomatis in male infertility.

Variance in total levels of phospholipase C zeta (PLC-ζ) in human sperm may limit the applicability of quantitative immunofluorescent analysis as a diagnostic indicator of oocyte activation capability.

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Kashir, Junaid; Jones, Celine; Mounce, Ginny; Ramadan, Walaa M; Lemmon, Bernadette; Heindryckx, Bjorn; de Sutter, Petra; Parrington, John; Turner, Karen; Child, Tim; McVeigh, Enda; Coward, Kevin

2013-01-01

To examine whether similar levels of phospholipase C zeta (PLC-ζ) protein are present in sperm from men whose ejaculates resulted in normal oocyte activation, and to examine whether a predominant pattern of PLC-ζ localization is linked to normal oocyte activation ability. Laboratory study. University laboratory. Control subjects (men with proven oocyte activation capacity; n = 16) and men whose sperm resulted in recurrent intracytoplasmic sperm injection failure (oocyte activation deficient [OAD]; n = 5). Quantitative immunofluorescent analysis of PLC-ζ protein in human sperm. Total levels of PLC-ζ fluorescence, proportions of sperm exhibiting PLC-ζ immunoreactivity, and proportions of PLC-ζ localization patterns in sperm from control and OAD men. Sperm from control subjects presented a significantly higher proportion of sperm exhibiting PLC-ζ immunofluorescence compared with infertile men diagnosed with OAD (82.6% and 27.4%, respectively). Total levels of PLC-ζ in sperm from individual control and OAD patients exhibited significant variance, with sperm from 10 out of 16 (62.5%) exhibiting levels similar to OAD samples. Predominant PLC-ζ localization patterns varied between control and OAD samples with no predictable or consistent pattern. The results indicate that sperm from control men exhibited significant variance in total levels of PLC-ζ protein, as well as significant variance in the predominant localization pattern. Such variance may hinder the diagnostic application of quantitative PLC-ζ immunofluorescent analysis. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Sperm chemorepulsion, a supplementary mechanism to regulate fertilization.

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Guidobaldi, H A; Cubilla, M; Moreno, A; Molino, M V; Bahamondes, L; Giojalas, L C

2017-08-01

Are human spermatozoa able of chemorepulsive behaviour? Capacitated human spermatozoa are able to be chemorepelled by synthetic Progesterone Receptor Ligands (sPRL, known as contraceptives) and zinc (a cation released by the oocyte upon fertilization). Moving cells can be oriented towards or against a molecular gradient, processes called chemoattraction and chemorepulsion, respectively, which have been described in unicellular organisms such as amoebas and bacteria, to organismic cells such macrophages and developmental cells. In the case of spermatozoa, chemoattraction may help the finding of an oocyte and has been widely studied in various invertebrate and mammalian species; however, chemorepulsion has not yet been verified in spermatozoa. This is an in vitro study involving human, rabbit and mouse spermatozoa which were used to perform 3-30 experiments per treatment. Human sperm samples were obtained by masturbation from healthy donors who gave written consent. Only those samples exhibiting normal semen parameters according to current WHO criteria were included in the study. Rabbit spermatozoa were obtained by artificial vagina whereas mice spermatozoa were obtained from epididymis. The sperm selection assay (SSA), originally designed to evaluate sperm chemoattraction towards progesterone (P), and a video-microscopy and computer motion analysis system were used to test sperm chemorepulsion. Additional kinetic parameters were also determined by video-microscopy and computer motion analysis. In some experiments, the level of induced acrosome-reacted spermatozoa was determined. Rabbit mating manipulation was achieved to perform the sperm-oocyte co-incubation assay. Sperm accumulation in the well containing 100 pg/ml of sPRL was lower than the culture medium negative control (P financial interests. N/A. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions

Presence and function of dopamine transporter (DAT in stallion sperm: dopamine modulates sperm motility and acrosomal integrity.

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Javier A Urra

Full Text Available Dopamine is a catecholamine with multiple physiological functions, playing a key role in nervous system; however its participation in reproductive processes and sperm physiology is controversial. High dopamine concentrations have been reported in different portions of the feminine and masculine reproductive tract, although the role fulfilled by this catecholamine in reproductive physiology is as yet unknown. We have previously shown that dopamine type 2 receptor is functional in boar sperm, suggesting that dopamine acts as a physiological modulator of sperm viability, capacitation and motility. In the present study, using immunodetection methods, we revealed the presence of several proteins important for the dopamine uptake and signalling in mammalian sperm, specifically monoamine transporters as dopamine (DAT, serotonin (SERT and norepinephrine (NET transporters in equine sperm. We also demonstrated for the first time in equine sperm a functional dopamine transporter using 4-[4-(Dimethylaminostyryl]-N-methylpyridinium iodide (ASP(+, as substrate. In addition, we also showed that dopamine (1 mM treatment in vitro, does not affect sperm viability but decreases total and progressive sperm motility. This effect is reversed by blocking the dopamine transporter with the selective inhibitor vanoxerine (GBR12909 and non-selective inhibitors of dopamine reuptake such as nomifensine and bupropion. The effect of dopamine in sperm physiology was evaluated and we demonstrated that acrosome integrity and thyrosine phosphorylation in equine sperm is significantly reduced at high concentrations of this catecholamine. In summary, our results revealed the presence of monoamine transporter DAT, NET and SERT in equine sperm, and that the dopamine uptake by DAT can regulate sperm function, specifically acrosomal integrity and sperm motility.

Human sperm degradation of zona pellucida proteins contributes to fertilization.

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Saldívar-Hernández, Analilia; González-González, María E; Sánchez-Tusié, Ana; Maldonado-Rosas, Israel; López, Pablo; Treviño, Claudia L; Larrea, Fernando; Chirinos, Mayel

2015-09-02

The mammalian oocyte extracellular matrix known as the zona pellucida (ZP) acts as a barrier to accomplish sperm fusion with the female gamete. Although penetration of the ZP is a limiting event to achieve fertilization, this is one of the least comprehended stages of gamete interaction. Even though previous studies suggest that proteases of sperm origin contribute to facilitate the passage of sperm through the ZP, in human this process is not yet fully understood. The aim of this study was to determine the ability of human sperm to degrade recombinant human ZP (rhZPs) proteins and to characterize the proteases involved in this process. Purified rhZP2, rhZP3 and rhZP4 proteins were incubated with capacitated sperm and the proteolytic activity was determined by Western blot analysis. To further characterize the proteases involved, parallel incubations were performed in the presence of the protease inhibitors o-phenanthroline, benzamidine and MG-132 meant to block the activity of metalloproteases, serine proteases and the proteasome, respectively. Additionally, protease inhibitors effect on sperm-ZP binding was evaluated by hemizona assay. The results showed that rhZPs were hydrolyzed in the presence of capacitated sperm. O-phenanthroline inhibited the degradation of rhZP3, MG-132 inhibited the degradation of rhZP4 and benzamidine inhibited the degradation of the three proteins under investigation. Moreover, hemizona assays demonstrated that sperm proteasome inhibition impairs sperm interaction with human native ZP. This study suggests that sperm proteasomes could participate in the degradation of ZP, particularly of the ZP4 protein. Besides, metalloproteases may be involved in specific degradation of ZP3 while serine proteases may contribute to unspecific degradation of the ZP. These findings suggest that localized degradation of ZP proteins by sperm is probably involved in ZP penetration and may be of help in understanding the mechanisms of fertilization in humans.

An immunological approach of sperm sexing and different methods for identification of X- and Y-chromosome bearing sperm

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Shiv Kumar Yadav

2017-05-01

Full Text Available Separation of X- and Y-chromosome bearing sperm has been practiced for selection of desired sex of offspring to increase the profit in livestock industries. At present, fluorescence-activated cell sorter is the only successful method for separation of X- and Y-chromosome bearing sperm. This technology is based on the differences in DNA content between these two types of sperm and has been commercialized for bovine sperm. However, this technology still has problems in terms of high economic cost, sperm damage, and lower pregnancy rates compared to unsorted semen. Therefore, an inexpensive, convenient, and non-invasive approach for sperm sexing would be of benefit to agricultural sector. Within this perspective, immunological sperm sexing method is one of the attractive choices to separate X- and Y-chromosome bearing sperm. This article reviews the current knowledge about immunological approaches, viz., H-Y antigen, sex-specific antigens, and differentially expressed proteins for sperm sexing. Moreover, this review also highlighted the different methods for identification of X- and Y-sperm.

Confocal fluorescence microscopy in a murine model of microdissection testicular sperm extraction to improve sperm retrieval.

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Smith, Ryan P; Lowe, Greg J; Kavoussi, Parviz K; Steers, William D; Costabile, Raymond A; Herr, John C; Shetty, Jagathpala; Lysiak, Jeffrey J

2012-05-01

Microdissection testicular sperm extraction markedly improves the sperm retrieval rates in men with nonobstructive azoospermia. However, localizing sperm foci can be time-consuming and it is not always successful. Fiberoptic confocal fluorescent microscopy offers the advantage of rapid in vivo detection of fluorescently labeled sperm in the seminiferous tubules. After establishing the feasibility of fiberoptic confocal fluorescent microscopy to identify antibody labeled sperm in vivo C57/B6 mice underwent intraperitoneal injection of busulfan to induce azoospermia. During spermatogenesis reestablishment at approximately 16 weeks the mice were anesthetized and the testes were delivered through a low midline incision. Fluorescein isothiocyanate labeled antibody to intra-acrosomal protein Hs-14 was injected retrograde into a single murine rete testis. The testes were imaged in vivo with fiberoptic confocal fluorescent microscopy and sperm foci were detected. The respective seminiferous tubules were excised and squash prepared for immunofluorescence microscopy. Sperm foci were identified in the testis injected with fluorescently tagged antibody by in vivo fiberoptic confocal fluorescence microscopy. The contralateral control testis of each mouse showed no specific signal. Immunofluorescence microscopy of the excised tubules provided morphological confirmation of the presence of labeled sperm with an absence in controls. Findings were consistent in the feasibility portion of the study and in the busulfan model of nonobstructive azoospermia. Fiberoptic confocal fluorescent microscopy was feasible during microdissection testicular sperm extraction in an azoospermic mouse model to identify fluorescently labeled sperm in vivo. Translation to the clinical setting could decrease operative time and improve the sperm harvest rate. Copyright © 2012 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

Effect of different monosaccharides and disaccharides on boar sperm quality after cryopreservation.

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Gómez-Fernández, José; Gómez-Izquierdo, Emilio; Tomás, Cristina; Mocé, Eva; de Mercado, Eduardo

2012-07-01

The aim of the present study was to evaluate the cryoprotectant effect of different non-permeating sugars for boar sperm. Pooled semen from three boars was used for the experiments. In the first experiment, the sperm quality of boar sperm cryopreserved with an egg-yolk based extender supplemented with different monosaccharides (glucose, galactose or fructose) was compared to a control cryopreserved in lactose-egg yolk extender. In the second experiment, the effect of five disaccharides (lactose, sucrose, lactulose, trehalose or melibiose) on boar sperm cryosurvival was studied. Several sperm quality parameters were assessed by flow cytometry in samples incubated for 30 and 150 min at 37°C after thawing: percentages of sperm with intact plasma membrane (SIPM), sperm presenting high plasma membrane fluidity (HPMF), sperm with intracellular reactive oxygen substances production (IROSP) and apoptotic sperm (AS). In addition, the percentages of total motile (TMS) and progressively motile sperm (PMS) were assessed at the same incubation times with a computer-assisted sperm analysis system. Freezing extenders supplemented with each of the monosaccharide presented smaller cryoprotective effect than the control extender supplemented with lactose (Pextender supplemented with lactulose exhibited in general the lowest sperm quality, except for the percentage of capacitated sperm, which was highest (Pextender. Our results suggest that disaccharides have higher cryoprotective effect than monosaccharides, although the monosaccharide composition of the disaccharides is also important, since the best results were obtained with those disaccharides presenting glucose in their composition. Copyright © 2012 Elsevier B.V. All rights reserved.

Sperm DNA fragmentation in boars is delayed or abolished by using sperm extenders.

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Pérez-Llano, Begoña; Enciso, María; García-Casado, Pedro; Sala, Rubén; Gosálvez, Jaime

2006-12-01

The semen quality of seven young adult boars was assessed for percentages of sperm motility, normal acrosomes, abnormal sperm, cells positive to sHOST (short Hipoosmotic Swelling Test), HPNA cells (sHOST Positive with Normal Acrosome cells) and the percentage of sperm heads, which exhibited DNA fragmentation using the Sperm Chromatin Dispersion test (SCD). These parameters were analysed in sperm samples both undiluted and diluted using a commercial extender and stored at 15 degrees C for 21 days. Results showed that semen quality decreases faster in the undiluted semen samples from day 0 to day 7 compared to diluted semen samples that remained with a high quality up to day 11. The undiluted semen exhibited a low DNA fragmentation index (DFI) during the first days and then a significant increase from day 7 up to day 21. This increase in the DFI coincided with the lowest levels of the other semen quality parameters. On the contrary, the samples diluted in the commercial extender showed very low levels of DNA fragmentation in all boars during the preservation period. When the evolution of DNA fragmentation was analysed in the undiluted samples, differences were found among boars. These differences were not shown in the samples diluted in the extender where the basal DFI remained stable during the 21 days. The main conclusion of this study was that some sperm extenders delay or partially prevent sperm DNA fragmentation.

Manual vs. computer-assisted sperm analysis: can CASA replace manual assessment of human semen in clinical practice?

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Talarczyk-Desole, Joanna; Berger, Anna; Taszarek-Hauke, Grażyna; Hauke, Jan; Pawelczyk, Leszek; Jedrzejczak, Piotr

2017-01-01

The aim of the study was to check the quality of computer-assisted sperm analysis (CASA) system in comparison to the reference manual method as well as standardization of the computer-assisted semen assessment. The study was conducted between January and June 2015 at the Andrology Laboratory of the Division of Infertility and Reproductive Endocrinology, Poznań University of Medical Sciences, Poland. The study group consisted of 230 men who gave sperm samples for the first time in our center as part of an infertility investigation. The samples underwent manual and computer-assisted assessment of concentration, motility and morphology. A total of 184 samples were examined twice: manually, according to the 2010 WHO recommendations, and with CASA, using the program set-tings provided by the manufacturer. Additionally, 46 samples underwent two manual analyses and two computer-assisted analyses. The p-value of p CASA and manually. In the group of patients where all analyses with each method were performed twice on the same sample we found no significant differences between both assessments of the same probe, neither in the samples analyzed manually nor with CASA, although standard deviation was higher in the CASA group. Our results suggest that computer-assisted sperm analysis requires further improvement for a wider application in clinical practice.

Evaluation of methods to determine sperm density for the european eel, anguilla anguilla

DEFF Research Database (Denmark)

Sørensen, Sune Riis; Gallego, V.; Pérez, L.

2013-01-01

, computer-assisted sperm analysis (CASA) and flow cytometry (FCM), using Neubauer Improved haemocytometer as benchmark. Initially, relationships between spermatocrit, haemocytometer counts and sperm motility were analysed, as well as the effect of sperm dilution on haemocytometer counts. Furthermore......, accuracy and precision of spermatocrit, applying a range of G-forces, were tested and the best G-force used in method comparisons. We found no effect of dilution on haemocytometer sperm density estimates, whereas motility associated positively with haemocytometer counts, but not with spermatocrit. Results......European eel, Anguilla anguilla, is a target species for future captive breeding, yet best methodology to estimate sperm density for application in in vitro fertilization is not established. Thus, our objectives were to evaluate methods to estimate European eel sperm density including spermatocrit...

Tamoxifen is a potent antioxidant modulator for sperm quality in patients with idiopathic oligoasthenospermia.

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Guo, Li; Jing, Jun; Feng, Yu-Ming; Yao, Bing

2015-09-01

To explore the new mechanisms of tamoxifen (TAM) in the treatment for patients with idiopathic oligoasthenospermia-antioxidation. In a prospective, randomized, controlled clinical trial, 120 cases of idiopathic oligoasthenospermia were enrolled and randomly assigned to the indomethacin group (n = 60) treated with indomethacin (25 mg, bid) and TAM group (n = 60) treated with TAM (10 mg, bid) for 3 months. Before and after treatment, we evaluated semen parameters, serum malondialdehyde (MDA) and total antioxidant capacity (TAC), seminal plasma MDA and TAC, spermatozoa intracellular reactive oxygen species (ROS), sperm succinate dehydrogenase (SDH) activity, sperm mitochondrial membrane potential (MMP), and sperm adenosine triphosphate (ATP) content. The independent t test and one-way repeated measures analysis of variance were used to compare the variables between and within two groups. In the indomethacin group, the percentage of progressive motile sperms, total motility, sperm MMP, and ATP content were increased significantly after 3-month treatment (P sperm count, sperm concentration, the percentage of progressive motile sperms, total motility, serum and seminal plasma TAC, sperm MMP, and ATP content were significantly improved or increased (P sperm count, sperm concentration, serum TAC, seminal plasma TAC, spermatozoa intracellular ROS, and sperm SDH activity. TAM treatment can significantly improve sperm quality, which is achieved through alleviating oxidative stress, improving sperm mitochondrial functionality, and subsequently increasing sperm motility.

Severe Fertility Effects of sheepish Sperm Caused by Failure To Enter Female Sperm Storage Organs in Drosophila melanogaster

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Masatoshi Tomaru

2018-01-01

Full Text Available In Drosophila, mature sperm are transferred from males to females during copulation, stored in the sperm storage organs of females, and then utilized for fertilization. Here, we report a gene named sheepish (shps of Drosophila melanogaster that is essential for sperm storage in females. shps mutant males, although producing morphologically normal and motile sperm that are effectively transferred to females, produce very few offspring. Direct counts of sperm indicated that the primary defect was correlated to failure of shps sperm to migrate into the female sperm storage organs. Increased sperm motion parameters were seen in the control after transfer to females, whereas sperm from shps males have characteristics of the motion parameters different from the control. The few sperm that occasionally entered the female sperm storage organs showed no obvious defects in fertilization and early embryo development. The female postmating responses after copulation with shps males appeared normal, at least with respect to conformational changes of uterus, mating plug formation, and female remating rates. The shps gene encodes a protein with homology to amine oxidases, including as observed in mammals, with a transmembrane region at the C-terminal end. The shps mutation was characterized by a nonsense replacement in the third exon of CG13611, and shps was rescued by transformants of the wild-type copy of CG13611. Thus, shps may define a new class of gene responsible for sperm storage.

ANALYSIS OF INDIVIDUAL BIOLOGICAL CHARACTERISTICS OF AMUR CARP (CYPRINUS CARPIO HAEMATOPTERUS REPRODUCED USING CRYOPRESERVED SPERM

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N. Kolisnyk

2014-12-01

Full Text Available Purpose. To reproduce Amur carp population using cryopreserved sperm and analyze some biological and fish culture peculiarities of the reproduced fish stock. Methodology. Generally accepted methods for fish culture [1]. Experimental reproduction was carried out in pond conditions of «Carpathian vodogray» LTD (Lisnevychi village, Pustomytivsky district, Lviv region. Hydrochemical analysis was carried out classically by O. Alуokin (1970 [2], hydrobiological studies in the fatting ponds according to V. Zhadin (1956, 1960 [3, 4]. Haemoglobin concentration was determined by hemocyanin method of G. Dervis, A. Vorobiov [5]. Blood for this method was collected from fish heart with the use of Pasteur pipettes in Eppendorf tubes with heparin. Following exterior morphometric parameters were analysed: body weight (m, g, standard fish body length (l, cm, largest body height (H, cm and body circumference (O cm. Following exterior indices were calculated based on these parameters: body depth index (l/H, body circumference index (l/O and Fulton’s condition factor (Kv. The study was carried out using two groups of carp: control and experimental. The first group was reproduced from the native sperm, the second from the cryopreserved sperm. Findings. Carp reproduction and growing was carried out using native and cryopreserved sperm. This work contains the results of growing 1+ Amur carp of experimental and control groups. Hydrochemical and hydrobiological parameters of the fattening ponds were studied. Peculiarities of the exterior and some hematological parameters of the carp of different origin were characterized. Originality. For the first time we performed a comparison of some biological parameters of Amur carp reproduced using native and cryopreserved sperm. Practical Value. Considering the economic importance of Amur carp due to its use in hybridization, reproduction of its population plays an important role in the development of the stocks of the pure

Birefringence characteristics in sperm heads allow for the selection of reacted spermatozoa for intracytoplasmic sperm injection.

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Gianaroli, Luca; Magli, M Cristina; Ferraretti, Anna P; Crippa, Andor; Lappi, Michela; Capitani, Serena; Baccetti, Baccio

2010-02-01

To verify clinical outcome after injection of spermatozoa that have undergone the acrosome reaction (reacted spermatozoa) vs. those still having an intact acrosome (nonreacted spermatozoa). Prospective, randomized study. Reproductive Medicine Unit, Italian Society for the Study of Reproductive Medicine, Bologna, Italy. According to a prospective randomization including 71 couples with severe male factor infertility, intracytoplasmic sperm injection (ICSI) was performed under polarized light that permitted analysis of the pattern of birefringence in the sperm head. Twenty-three patients had their oocytes injected with reacted spermatozoa, 26 patient's oocytes were injected with nonreacted spermatozoa, and in 22 patients both reacted and nonreacted spermatozoa were injected. Intracytoplasmic sperm injection was performed under polarized light to selectively inject acrosome-reacted and acrosome-nonreacted spermatozoa. Rates of fertilization, cleavage, pregnancy, implantation, and ongoing implantation. There was no effect on the fertilizing capacity and embryo development of either type of sperm, whereas the implantation rate was higher in oocytes injected with reacted spermatozoa (39.0%) vs. those injected with nonreacted spermatozoa (8.6%). The implantation rate was 24.4% in the group injected with both reacted and nonreacted spermatozoa. The delivery rate per cycle followed the same trend. Spermatozoa that have undergone the acrosome reaction seem to be more prone to supporting the development of viable ICSI embryos. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Sperm origins and concentration do not impact the clinical outcomes in intracytoplasmic sperm injection cycles.

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Yang, Cen; Zhou, Ze-Hong; Zheng, Dan-Ni; Xu, Xiao-Fei; Huang, Jin; Lian, Ying; Qiao, Jie

2018-05-25

In the present study, we evaluated the impact of sperm origins and concentration on the clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles. A total of 1201 ICSI cycles were retrospectively analyzed for male azoospermia or oligozoospermia between January 2015 and December 2015 in the Peking University Third Hospital. Patients were divided into three groups (Group 1 vs Group 2/3; surgically extracted sperm vs ejaculated sperms): Group 1 included 343 ICSI cycles and Group 2 analyzed 388 cycles on semen with sperm concentration cycles with sperm concentration between 5 × 10 6 ml -1 and 15 × 10 6 ml -1 (mild oligozoospermia group). Fertilization rates, clinical pregnancy rates, and live birth rates were analyzed and compared among groups of different semen origins and concentrations on the oocyte retrieval day. Group 2 showed a lower fertilization rate than Group 3 (62.9% ± 21.6% vs 66.8% ± 22.1%,Pcycles.

Deduction of a calcium ion circuit affecting rooster sperm in vitro.

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Froman, D P

2016-08-01

Four premises for rooster sperm preservation were outlined previously. Understanding mitochondrial Ca cycling in terms of whole-cell Ca flux was one premise. The present work tested the hypothesis that sperm mitochondria can be damaged by intracellular as well as extracellular Ca. Sperm were washed by centrifugation through 12% (wt/vol) Sperm were washed by centrifugation through 12%(at/vol) Accudenz to procure sperm at a physiological concentration within a chemically-defined suspension. Five solutions were tested. Each solution contained 30 m glucose, and had an osmolality of 320 mmol/kg and a pH of 7.4. Washed sperm were diluted to 2.0 × 10 sperm/mL. Each replicate sperm suspension was cooled to 10°C. Sperm mobility was measured after 1, 2, 4, 8, 12, and 24 h. Data were plotted as a function of time in each experiment. Function type was confirmed by lack of fit analysis. A parabola with a maximum at 3.7 h was observed when sperm were suspended in 205 m taurine buffered with 50 m-tris[hydroxyl-methyl]methyl-2-amino-ethanesulfonic acid (TES). This effect was attributed to a Ca flux from the nuclear envelope into mitochondria. An exponential decay was observed when TES-buffered taurine contained 2 m Ca. This effect was attributed to mitochondrial Ca overload induced by uptake of extracellular Ca. Exponential decay also was observed when TES-buffered taurine contained a Ca chelator. This effect was attributed to a Ca flux from the nuclear envelope through mitochondria and then into an extracellular Ca sink. This possibility was supported by the response of sperm to thapsigargin. Specifically, inhibition of sarcoendoplasmic reticulum Ca-ATPase compromised sperm mobility relative to a buffer control. Finally, a 60 m phosphate buffer containing 2 m citrate yielded a linear relationship in contrast to the TES-buffered solutions tested. Sperm mobility after 24 h of storage in the phosphate buffer was 92% of that observed for prewashed sperm. The linear response was

Correlation between adjusted late spermatid score and sperm count in normozoospermic and oligozoospermic men

OpenAIRE

Ibrahim Fahmy; Yasser Elkhiat; Mohamed Saad; Hussein Ghanem

2013-01-01

Objective: To correlate late spermatid score and sperm count in men with normal semen analysis and oligozoospermia undergoing intracytoplasmic sperm injection (ICSI) to set the average threshold of spermatids/tubule of testicular tissue needed for normal sperm count/ml of semen. Material and methods: This study was conducted on 24 normozoospermic subjects and 18 oligozoospermic patients who underwent wide bore needle biopsy because of failed sperm collection at the day of ICSI. Clinical da...

Impacts of ocean acidification on sperm develop with exposure time for a polychaete with long lived sperm.

Science.gov (United States)

Campbell, Anna L; Ellis, Robert P; Urbina, Mauricio A; Mourabit, Sulayman; Galloway, Tamara S; Lewis, Ceri

2017-08-01

The majority of marine invertebrate species release eggs and sperm into seawater for external fertilisation. Seawater conditions are currently changing at an unprecedented rate as a consequence of ocean acidification (OA). Sperm are thought to be particularly vulnerable to these changes and may be exposed to external environmental conditions for variable periods of time between spawning and fertilisation. Here, we undertook a mechanistic investigation of sperm swimming performance in the coastal polychaete Arenicola marina during an extended exposure to OA conditions (pH NBS 7.77, 1000 μatm pCO 2 ). We found that key fitness-related aspects of sperm functioning declined faster under OA conditions i.e. impacts became apparent with exposure time. Sperm swimming speed (VCL), the number of motile sperm and sperm path linearity all dropped significantly after 4 h under OA conditions whilst remaining constant under ambient conditions at this time point. Our results highlight the importance of sperm exposure duration in ocean acidification experiments and may help towards explaining species specific differences in response. Copyright © 2017 Elsevier Ltd. All rights reserved.

Sperm competition in bats.

OpenAIRE

Hosken, D J

1997-01-01

Sperm competition is a widespread phenomenon influencing the evolution of male anatomy, physiology and behaviour. Bats are an ideal group for studying sperm competition. Females store fertile sperm for up to 200 days and the size of social groups varies from single animals to groups of hundreds of thousands. This study examines the relationship between social group size and investment in spermatogenesis across 31 species of microchiropteran bat using new and published data on testis mass and ...

Effect of Vitrification on Sperm Parameters and Apoptosis in Fertile Men

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M Adib

2011-01-01

Full Text Available Introduction & Objective: Today, cryopreservation of the human sperm is a common technique for treating infertility. It has been indicated that cryopreservation by different methods decrease the sperm motility and viability in fertile men, but still effect of freezing of the sperm by vitrification method have not been evaluated on sperm parameters and apoptosis. The aim of this study was to evaluate the effect of vitrification of sperm of fertile men on different sperm parameters (motility, morphology, viability and count and apoptosis after thawing. Materials & Methods: In this experimental study which was conducted at Yazd Infertility Research and Clinical Center in 2009, seventeen semen samples were collected by masturbation from people who came to this centre. Semen analysis was performed according to WHO standards. Smear was provided from these samples and fixed for TUNEL staining. Some samples were directly cryopreserved by cryoloope in liquid nitrogen and stored at least for Seven days. After thawing, samples were evaluated for sperm parameters. The collected data was analyzed by the SPSS software using paired T-test and Willcoxon statistical test. Results: The progressive movement of sperm was significantly decreased by vitrification. Also significant decrease in viability and morphology of the sperm and increase in the rate of apoptosis was observed after vitrification. The amount of apoptosis had negatively correlated with normal parameters of spermatozoa (especially progressive motility and viability. Conclusion: These results indicated that vitrification is harmful for sperm parameters and of apoptosis rate in fertile men. However, the apoptosis rate was lower compared to other freezing methods.

Urban resident attitudes toward rodents, rodent control products, and environmental effects

Science.gov (United States)

Rodent control in urban areas can result in the inadvertent mortality of non-target species (e.g., bobcats). However, there is little detailed information about rodent control practices of urban residents. Our objective was to evaluate urban rodent control behaviors in two area...

Sperm DNA damage has a negative effect on early embryonic development following in vitro fertilization

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Wei-Wei Zheng

2018-01-01

Full Text Available Sperm DNA damage is recognized as an important biomarker of male infertility. To investigate this, sperm DNA damage was assessed by the sperm chromatin dispersion (SCD test in semen and motile spermatozoa harvested by combined density gradient centrifugation (DGC and swim-up in 161 couples undergoing in vitro fertilization (IVF. Semen analysis and sperm DNA damage results were compared between couples who did or did not achieve pregnancy. The sperm DNA damage level was significantly different between the two groups (P < 0.05 and was negatively correlated with IVF outcomes. Logistic regression analysis confirmed that it was an independent predictor for achieving clinical pregnancy. The effects of different levels of sperm DNA damage on IVF outcomes were also compared. There were significant differences in day 3 embryo quality, blastocyst formation rate, and implantation and pregnancy rates (P < 0.05, but not in the basic fertilization rate between the two groups. Thus, sperm DNA damage as measured by the SCD appears useful for predicting the clinical pregnancy rate following IVF.

Sperm motility of externally fertilizing fish and amphibians.

Science.gov (United States)

Browne, R K; Kaurova, S A; Uteshev, V K; Shishova, N V; McGinnity, D; Figiel, C R; Mansour, N; Agney, D; Wu, M; Gakhova, E N; Dzyuba, B; Cosson, J

2015-01-01

We review the phylogeny, sperm competition, morphology, physiology, and fertilization environments of the sperm of externally fertilizing fish and amphibians. Increased sperm competition in both fish and anurans generally increases sperm numbers, sperm length, and energy reserves. The difference between the internal osmolarity and iconicity of sperm cells and those of the aquatic medium control the activation, longevity, and velocity of sperm motility. Hypo-osmolarity of the aquatic medium activates the motility of freshwater fish and amphibian sperm and hyperosmolarity activates the motility of marine fish sperm. The average longevity of the motility of marine fish sperm (~550 seconds) was significantly (P amphibian sperm in general and anurans reversion from internal to external fertilization. Our findings provide a greater understanding of the reproductive biology of externally fertilizing fish and amphibians, and a biological foundation for the further development of reproduction technologies for their sustainable management.

Sperm Impairment by Sperm Agglutinating Factor Isolated from Escherichia coli: Receptor Specific Interactions

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Kiranjeet Kaur

2013-01-01

Full Text Available In an earlier work done in our laboratory, we have been able to isolate a sperm agglutinating strain of Escherichia coli from the semen sample of a male attending infertility clinic. Further, factor responsible for sperm agglutination (SAF was isolated and purified, and, using SAF as a tool, corresponding SAF binding receptor from human spermatozoa has been purified. Characterization of SAF and SAF binding receptor using MALDI-TOF showed homology to glutamate decarboxylase and MHC class I molecule, respectively. Coincubation of SAF with spermatozoa not only resulted in spermagglutination but could also compromise other sperm parameters, namely, Mg2+ dependent ATPase activity and apoptosis. Intravaginal administration of SAF could lead to infertility in Balb/c mice. SAF induced impairment of sperm parameters, and infertility was observed to be due to interaction of SAF with sperm surface receptor component as, when purified receptor was introduced, receptor completely inhibited all the detrimental effects induced by SAF. From these results, it could be concluded that interaction of SAF with spermatozoa is receptor mediated.

Sperm subpopulational dinamycs during the cryopreservation procedure in caprine (Capra aegagrus hircus ejaculates

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Barbas JP

2017-08-01

Full Text Available The objective of the present research was to determine specific sperm subpopulational dynamics in different processing steps during cryopreservation process by using objective functional sperm kinematic descriptors in goat ejaculates. Fresh ejaculates (n=40 collected from eight bucks were analised for volume, concentration, sperm viability, acrosome integrity, and sperm motility using computer-assisted sperm analysis (CASA system. Eight sperm kinematic descriptors (VCL, VSL, VAP, LIN, STR, BCF, ALH, and WOB were assessed using CASA system after five different handling step (1st: fresh semen collection (F; 2nd: 1st washing/centrifugation step (1WC; 3rd: 2nd washing / centrifugation step (2WC; 4th: cooling step at 4ºC (CL; and 5th: post-thawing step at 37ºC (PT during a standard cryopreservation protocol for goat semen. The results obtained from the kinematic parameters were analysed by using Principal Component Analysis (PCA and multivariate clustering procedures to identify specific kinematic subpopulations and establish the relationship between the distribution of the subpopulations found and the functional sperm motility in each step. Except for the 1st (SbpF1-SbpF3 and 4th (SbpCL1-SbpCL3 intervals, four sperm kinematic subpopulations (Sbp1LC1-Sbp1LC4, Sbp2LC1-Sbp2LC4 and SbpPD1-SbpPD4 were observed. Based on kinematic velocity parameters and the subpopulation disclosed, rapid, slow, vigorous, passive, non-progressive and progressive sperm were discerned. Moreover, based on kinematic linearity parameters and depending on the subpopulation uncovered, curvilinear, regular-linear, parabolic and erratic-non-linear trajectories were detected. Subpopulations remained varible throughout handling steps and multiple significant differences among the sperm kinematic parameters were observed (p<0.001 as well as in the frequency of distribution of kinematic subpopulations among steps (p<0.05. In conclusion, this study confirms the variability and

Abundance and Distribution of Sperm Whales in the Canary Islands: Can Sperm Whales in the Archipelago Sustain the Current Level of Ship-Strike Mortalities?

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Fais, Andrea; Lewis, Tim P.; Zitterbart, Daniel P.; Álvarez, Omar; Tejedor, Ana; Aguilar Soto, Natacha

2016-01-01

Sperm whales are present in the Canary Islands year-round, suggesting that the archipelago is an important area for this species in the North Atlantic. However, the area experiences one of the highest reported rates of sperm whale ship-strike in the world. Here we investigate if the number of sperm whales found in the archipelago can sustain the current rate of ship-strike mortality. The results of this study may also have implications for offshore areas where concentrations of sperm whales may coincide with high densities of ship traffic, but where ship-strikes may be undocumented. The absolute abundance of sperm whales in an area of 52933 km2, covering the territorial waters of the Canary Islands, was estimated from 2668 km of acoustic line-transect survey using Distance sampling analysis. Data on sperm whale diving and acoustic behaviour, obtained from bio-logging, were used to calculate g(0) = 0.92, this is less than one because of occasional extended periods when whales do not echolocate. This resulted in an absolute abundance estimate of 224 sperm whales (95% log-normal CI 120–418) within the survey area. The recruitment capability of this number of whales, some 2.5 whales per year, is likely to be exceeded by the current ship-strike mortality rate. Furthermore, we found areas of higher whale density within the archipelago, many coincident with those previously described, suggesting that these are important habitats for females and immature animals inhabiting the archipelago. Some of these areas are crossed by active shipping lanes increasing the risk of ship-strikes. Given the philopatry in female sperm whales, replacement of impacted whales might be limited. Therefore, the application of mitigation measures to reduce the ship-strike mortality rate seems essential for the conservation of sperm whales in the Canary Islands. PMID:26999791

Abundance and Distribution of Sperm Whales in the Canary Islands: Can Sperm Whales in the Archipelago Sustain the Current Level of Ship-Strike Mortalities?

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Andrea Fais

Full Text Available Sperm whales are present in the Canary Islands year-round, suggesting that the archipelago is an important area for this species in the North Atlantic. However, the area experiences one of the highest reported rates of sperm whale ship-strike in the world. Here we investigate if the number of sperm whales found in the archipelago can sustain the current rate of ship-strike mortality. The results of this study may also have implications for offshore areas where concentrations of sperm whales may coincide with high densities of ship traffic, but where ship-strikes may be undocumented. The absolute abundance of sperm whales in an area of 52933 km2, covering the territorial waters of the Canary Islands, was estimated from 2668 km of acoustic line-transect survey using Distance sampling analysis. Data on sperm whale diving and acoustic behaviour, obtained from bio-logging, were used to calculate g(0 = 0.92, this is less than one because of occasional extended periods when whales do not echolocate. This resulted in an absolute abundance estimate of 224 sperm whales (95% log-normal CI 120-418 within the survey area. The recruitment capability of this number of whales, some 2.5 whales per year, is likely to be exceeded by the current ship-strike mortality rate. Furthermore, we found areas of higher whale density within the archipelago, many coincident with those previously described, suggesting that these are important habitats for females and immature animals inhabiting the archipelago. Some of these areas are crossed by active shipping lanes increasing the risk of ship-strikes. Given the philopatry in female sperm whales, replacement of impacted whales might be limited. Therefore, the application of mitigation measures to reduce the ship-strike mortality rate seems essential for the conservation of sperm whales in the Canary Islands.

Abundance and Distribution of Sperm Whales in the Canary Islands: Can Sperm Whales in the Archipelago Sustain the Current Level of Ship-Strike Mortalities?

Science.gov (United States)

Fais, Andrea; Lewis, Tim P; Zitterbart, Daniel P; Álvarez, Omar; Tejedor, Ana; Aguilar Soto, Natacha

2016-01-01

Sperm whales are present in the Canary Islands year-round, suggesting that the archipelago is an important area for this species in the North Atlantic. However, the area experiences one of the highest reported rates of sperm whale ship-strike in the world. Here we investigate if the number of sperm whales found in the archipelago can sustain the current rate of ship-strike mortality. The results of this study may also have implications for offshore areas where concentrations of sperm whales may coincide with high densities of ship traffic, but where ship-strikes may be undocumented. The absolute abundance of sperm whales in an area of 52933 km2, covering the territorial waters of the Canary Islands, was estimated from 2668 km of acoustic line-transect survey using Distance sampling analysis. Data on sperm whale diving and acoustic behaviour, obtained from bio-logging, were used to calculate g(0) = 0.92, this is less than one because of occasional extended periods when whales do not echolocate. This resulted in an absolute abundance estimate of 224 sperm whales (95% log-normal CI 120-418) within the survey area. The recruitment capability of this number of whales, some 2.5 whales per year, is likely to be exceeded by the current ship-strike mortality rate. Furthermore, we found areas of higher whale density within the archipelago, many coincident with those previously described, suggesting that these are important habitats for females and immature animals inhabiting the archipelago. Some of these areas are crossed by active shipping lanes increasing the risk of ship-strikes. Given the philopatry in female sperm whales, replacement of impacted whales might be limited. Therefore, the application of mitigation measures to reduce the ship-strike mortality rate seems essential for the conservation of sperm whales in the Canary Islands.

Comparison of semen variables, sperm DNA damage and sperm membrane proteins in two male layer breeder lines.

Science.gov (United States)

M, Shanmugam; T R, Kannaki; A, Vinoth

2016-09-01

Semen variables are affected by the breed and strain of chicken. The present study was undertaken to compare the semen quality in two lines of adult chickens with particular reference to sperm chromatin condensation, sperm DNA damage and sperm membrane proteins. Semen from a PD3 and White Leghorn control line was collected at 46 and 47 weeks and 55 weeks of age. The semen was evaluated for gross variables and sperm chromatin condensation by aniline blue staining. Sperm DNA damage was assessed by using the comet assay at 47 weeks of age and sperm membrane proteins were assessed at 55 weeks of age. The duration of fertility was studied by inseminating 100 million sperm once into the hens of the same line as well as another line. The eggs were collected after insemination for 15days and incubated. The eggs were candled on 18th day of incubation for observing embryonic development. The White Leghorn control line had a greater sperm concentration and lesser percentage of morphologically abnormal sperm at the different ages where assessments occurred. There was no difference in sperm chromatin condensation, DNA damage and membrane proteins between the lines. Only low molecular weight protein bands of less than 95kDa were observed in samples of both lines. The line from which semen was used had no effect on the duration over which fertility was sustained after insemination either when used in the same line or another line. Thus, from the results of the present study it may be concluded that there was a difference in gross semen variables between the lines that were studied, however, the sperm chromatin condensation, DNA damage, membrane proteins and duration over which fertility was sustained after insemination did not differ between the lines. Copyright © 2016 Elsevier B.V. All rights reserved.

Ocean acidification impacts on sperm mitochondrial membrane potential bring sperm swimming behaviour near its tipping point.

Science.gov (United States)

Schlegel, Peter; Binet, Monique T; Havenhand, Jonathan N; Doyle, Christopher J; Williamson, Jane E

2015-04-01

Broadcast spawning marine invertebrates are susceptible to environmental stressors such as climate change, as their reproduction depends on the successful meeting and fertilization of gametes in the water column. Under near-future scenarios of ocean acidification, the swimming behaviour of marine invertebrate sperm is altered. We tested whether this was due to changes in sperm mitochondrial activity by investigating the effects of ocean acidification on sperm metabolism and swimming behaviour in the sea urchin Centrostephanus rodgersii. We used a fluorescent molecular probe (JC-1) and flow cytometry to visualize mitochondrial activity (measured as change in mitochondrial membrane potential, MMP). Sperm MMP was significantly reduced in ΔpH -0.3 (35% reduction) and ΔpH -0.5 (48% reduction) treatments, whereas sperm swimming behaviour was less sensitive with only slight changes (up to 11% decrease) observed overall. There was significant inter-individual variability in responses of sperm swimming behaviour and MMP to acidified seawater. We suggest it is likely that sperm exposed to these changes in pH are close to their tipping point in terms of physiological tolerance to acidity. Importantly, substantial inter-individual variation in responses of sperm swimming to ocean acidification may increase the scope for selection of resilient phenotypes, which, if heritable, could provide a basis for adaptation to future ocean acidification. © 2015. Published by The Company of Biologists Ltd.

Sperm quality but not relatedness predicts sperm competition success in threespine sticklebacks (Gasterosteus aculeatus).

Science.gov (United States)

Mehlis, Marion; Rahn, Anna K; Bakker, Theo C M

2015-04-26

Mating between close relatives often leads to a reduction of an individual's fitness, due to an increased expression of deleterious alleles. Thus, in many animal taxa pre- as well as postcopulatory inbreeding avoidance mechanisms have evolved. An increased risk of inbreeding and hence a loss of genetic variation may occur during founder events as in most cases only few individuals establish a new population. The threespine stickleback (Gasterosteus aculeatus) is a small externally fertilizing fish species subject to strong sperm competition. Sticklebacks inhabit both marine and freshwater environments and anadromous populations have repeatedly established new genetically less diverse freshwater populations. Previous studies showed that anadromous sticklebacks strongly suffer from inbreeding depression and when given the choice females prefer to mate with unrelated males. The present study aimed to address whether there exists a postcopulatory inbreeding avoidance mechanism solely based on sperm-egg interactions in sperm competition experiments. We used F1 individuals that originated either from a large, genetically heterogeneous anadromous population or from a small, genetically less diverse freshwater population. For each population, eggs of two different females were in vitro fertilized by the same two males' sperm in a paired study design. In the main experiment one male was the female's full-sib brother and in the control experiment all individuals were unrelated. The results revealed that fertilization success was independent of relatedness in both populations suggesting a general lack of a postcopulatory inbreeding avoidance mechanism. Instead, male quality (i.e. sperm morphology) predicted paternity success during competitive fertilization trials. In sticklebacks, there is no evidence for postcopulatory inbreeding avoidance. Sperm morphology predicted paternity instead, thus sperm quality traits are under strong sexual selection, presumably driven by the

BMI in relation to sperm count: an updated systematic review and collaborative meta-analysis

NARCIS (Netherlands)

Sermondade, N.; Faure, C.; Fezeu, L.; Shayeb, A. G.; Bonde, J. P.; Jensen, T. K.; van Wely, M.; Cao, J.; Martini, A. C.; Eskandar, M.; Chavarro, J. E.; Koloszar, S.; Twigt, J. M.; Ramlau-Hansen, C. H.; Borges, E.; Lotti, F.; Steegers-Theunissen, R. P. M.; Zorn, B.; Polotsky, A. J.; La Vignera, S.; Eskenazi, B.; Tremellen, K.; Magnusdottir, E. V.; Fejes, I.; Hercberg, S.; Lévy, R.; Czernichow, S.

2013-01-01

BACKGROUND The global obesity epidemic has paralleled a decrease in semen quality. Yet, the association between obesity and sperm parameters remains controversial. The purpose of this report was to update the evidence on the association between BMI and sperm count through a systematic review with

Ketotifen, a mast cell blocker improves sperm motility in asthenospermic infertile men

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Nasrin Saharkhiz

2013-01-01

Full Text Available Aim: This study aimed to evaluate the efficacy of ketotifen on sperm motility of asthenospermic infertile men. Setting and Design: It is a prospective study designed in vivo. Materials and Methods: In this interventional experimental study, a total of 40 infertile couples with asthenospermic infertility factor undergoing assisted reproductive technology (ART cycles were enrolled. The couples were randomly assigned to one of two groups at the starting of the cycle. In control group (n = 20, the men did not receive Ketotifen, while in experiment group (n = 20, the men received oraly ketotifen (1 mg Bid for 2 months. Semen analysis, under optimal circumferences, was obtained prior to initiation of treatment. The second semen analysis was done 2-3 weeks after stopped ketotifen treatment and sperm motility was defined. Clinical pregnancy was identified as the presence of a fetal sac by vaginal ultrasound examination. Statistical Analysis Used: All data are expressed as the mean ± standard error of mean (SEM. t test was used for comparing the data of the control and treated groups. Results: The mean sperm motility increased significantly (from 16.7% to 21.4% after ketotifen treatment (P < 0.001. This sperm motility improvement was more pronounced in the primary infertility cases (P < 0.003. The rate of pregnancy was 12.5% in infertile couples that their men receiving 1 mg/twice a day ketotifen. In 52% of infertile men′s semen, the percentage of sperm motility was increased from 5% to 35% and this sperm motility improvement was also observed in 33% of necrospermia (0% motility cases. Conclusion: These results suggest that ketotifen may represent as a novel therapeutic approach to improve sperm motility in the infertile men with cause of asthenospermia or necrospermia.

Varicocele Negatively Affects Sperm Mitochondrial Respiration.

Science.gov (United States)

Ferramosca, Alessandra; Albani, Denise; Coppola, Lamberto; Zara, Vincenzo

2015-10-01

To evaluate the effect of varicocele on oxidative stress, sperm mitochondrial respiratory efficiency, sperm morphology, and semen parameters. A total of 20 patients with varicocele and 20 normozoospermic subjects without varicocele (control group) were recruited from a medical center for reproductive biology. The levels of serum reactive oxygen metabolites and seminal lipid peroxides were assessed for both control and varicocele subjects. Sperm deoxyribonucleic acid fragmentation was measured by sperm chromatin dispersion test. Mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically treated sperm cells. In this study, varicocele patients were compared with men without varicoceles. Oxidative stress was observed in the serum and seminal fluid of varicocele patients. These patients showed an increase of 59% (P <.05) in serum reactive oxygen metabolites and a 3-fold increase in the level of sperm lipid peroxides. A parallel and significant increase (a 2-fold increase; P <.05) in the degree of sperm deoxyribonucleic acid fragmentation was also observed. Varicocele patients showed a 27% decrease (P <.05) in mitochondrial respiratory activity in comparison to the control group. A 32% increase (P <.05) in sperm midpiece defects and a 41% decrease (P <.05) in sperm concentration and motility were also observed. Men with varicocele have increased markers of oxidative stress and decreased mitochondrial respiratory activity. These results correlated with abnormalities in semen parameters. For morphology, these correlated with midpiece defects. Copyright © 2015 Elsevier Inc. All rights reserved.

Hypercholesterolemia Impaired Sperm Functionality in Rabbits

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Monclus, Maria A.; Cabrillana, Maria E.; Clementi, Marisa A.; Espínola, Leandro S.; Cid Barría, Jose L.; Vincenti, Amanda E.; Santi, Analia G.; Fornés, Miguel W.

2010-01-01

Hypercholesterolemia represents a high risk factor for frequent diseases and it has also been associated with poor semen quality that may lead to male infertility. The aim of this study was to analyze semen and sperm function in diet-induced hypercholesterolemic rabbits. Twelve adult White New Zealand male rabbits were fed ad libitum a control diet or a diet supplemented with 0.05% cholesterol. Rabbits under cholesterol-enriched diet significantly increased total cholesterol level in the serum. Semen examination revealed a significant reduction in semen volume and sperm motility in hypercholesterolemic rabbits (HCR). Sperm cell morphology was seriously affected, displaying primarily a “folded head”-head fold along the major axe-, and the presence of cytoplasmic droplet on sperm flagellum. Cholesterol was particularly increased in acrosomal region when detected by filipin probe. The rise in cholesterol concentration in sperm cells was determined quantitatively by Gas chromatographic-mass spectrometric analyses. We also found a reduction of protein tyrosine phosphorylation in sperm incubated under capacitating conditions from HCR. Interestingly, the addition of Protein Kinase A pathway activators -dibutyryl-cyclic AMP and iso-butylmethylxanthine- to the medium restored sperm capacitation. Finally, it was also reported a significant decrease in the percentage of reacted sperm in the presence of progesterone. In conclusion, our data showed that diet-induced hypercholesterolemia adversely affects semen quality and sperm motility, capacitation and acrosomal reaction in rabbits; probably due to an increase in cellular cholesterol content that alters membrane related events. PMID:20976152

Quantitative evaluation of sperm in men, based on an interval scale

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V. V. Danilov

2014-11-01

Full Text Available Modern methods of estimation spermogram are quite difficult and highly subjective. Were analyzed spermogram 215 men aged 20 to 46 years (mean age 29,7 ± 2,4 years. The analysis spermogram were selected 5 out of 21 key indicators of ejaculate that affect fertility: the total number, motility, viability, morphology of sperm, ejaculate volume. Based on these data the interval scale, which allows to quantify the sperm. Obtained as a result of an integral index is a reflection of pathological semen analysis based on deviations from the “ideal norm”, that allows to evaluate different sperm, and to evaluate the effectiveness of treatment, as with the use of conservative methods, and various options for surgical intervention.

Sperm DNA fragmentation affects epigenetic feature in human male pronucleus.

Science.gov (United States)

Rajabi, H; Mohseni-Kouchesfehani, H; Eslami-Arshaghi, T; Salehi, M

2018-02-01

To evaluate whether the sperm DNA fragmentation affects male pronucleus epigenetic factors, semen analysis was performed and DNA fragmentation was assessed by the method of sperm chromatin structure assay (SCSA). Human-mouse interspecies fertilisation was used to create human male pronucleus. Male pronucleus DNA methylation and H4K12 acetylation were evaluated by immunostaining. Results showed a significant positive correlation between the level of sperm DNA fragmentation and DNA methylation in male pronuclei. In other words, an increase in DNA damage caused an upsurge in DNA methylation. In the case of H4K12 acetylation, no correlation was detected between DNA damage and the level of histone acetylation in the normal group, but results for the group in which male pronuclei were derived from sperm cells with DNA fragmentation, increased DNA damage led to a decreased acetylation level. Sperm DNA fragmentation interferes with the active demethylation process and disrupts the insertion of histones into the male chromatin in the male pronucleus, following fertilisation. © 2017 Blackwell Verlag GmbH.

Comparison of two different extenders for cryopreservation of epididymal dog sperm.

Science.gov (United States)

Martins, Mim; Justino, R C; Sant'anna, M C; Trautwein, L G C; Souza, F F

2012-12-01

The collection of epididymal sperm is an option for preservation of germplasm of genetically superior animals that need to be orchiectomized or have died. The extender type used to freeze sperm is important to avoid spermatozoal membrane damage and to preserve semen quality after cryopreservation. The objective of this study was to verify the effects of a commercial bovine extender (Bovimix(®); Nutricell, Campinas) and a traditional TRIS-citric acid-glucose-egg yolk-7% glycerol extender on cryopreservation of canine epididymal sperm. The testes of 13 adult dogs were kept at 5 °C for 24 h in saline solution, and epididymal sperm was recovered in Ringers solution without lactate and were evaluated for motility. Samples with ≥ 80% motility were pooled and then divided before dilution and packaging in 0.5 ml plastic straws, equilibration at 4 °C for 1 h, freezing in nitrogen vapour for 20 min and storing at -196 °C. The straws were thawed at 56 °C for 10 s and were evaluated for motility by computer assisted analysis (CASA). The semen parameters, sperm movement index, linearity, total motility and rapid progressive motility were statistically higher in Bovimix(®) than TRIS. In contrast, amplitude of lateral head displacement, slow sperm and static sperm were lower in Bovimix(®). Despite the high percentage of sperm defects in epididymal cells, regardless of the extender, we concluded that Bovimix(®) is a viable alternative for the freezing of canine epididymal sperm. © 2012 Blackwell Verlag GmbH.

Evaluation of Lasting Effects of Heat Stress on Sperm Profile and Oxidative Status of Ram Semen and Epididymal Sperm

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Thais Rose dos Santos Hamilton

2016-01-01

Full Text Available Higher temperatures lead to an increase of testicular metabolism that results in spermatic damage. Oxidative stress is the main factor responsible for testicular damage caused by heat stress. The aim of this study was to evaluate lasting effects of heat stress on ejaculated sperm and immediate or long-term effects of heat stress on epididymal sperm. We observed decrease in motility and mass motility of ejaculated sperm, as well as an increase in the percentages of sperm showing major and minor defects, damaged plasma and acrosome membranes, and a decrease in the percentage of sperm with high mitochondrial membrane potential in the treated group until one spermatic cycle. An increased enzymatic activity of glutathione peroxidase and an increase of stressed cells were observed in ejaculated sperm of the treated group. A decrease in the percentage of epididymal sperm with high mitochondrial membrane potential was observed in the treated group. However, when comparing immediate and long-term effects, we observed an increase in the percentage of sperm with low mitochondrial membrane potential. In conclusion, testicular heat stress induced oxidative stress that led to rescuable alterations after one spermatic cycle in ejaculated sperm and also after 30 days in epididymal sperm.

Experimental evolution of sperm competitiveness in a mammal

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Simmons Leigh W

2011-01-01

Full Text Available Abstract Background When females mate with multiple partners, sperm from rival males compete to fertilise the ova. Studies of experimental evolution have proven the selective action of sperm competition on male reproductive traits. However, while reproductive traits may evolve in response to sperm competition, this does not necessarily provide evidence that sperm competitive ability responds to selection. Indeed, a study of Drosophila failed to observe divergence in sperm competitive ability of males in lines selected for enhanced sperm offence and defence. Results Adopting the naturally polygamous house mouse (Mus domesticus as our vertebrate model, we performed an experimental evolution study and observed genetic divergence in sperm quality; males from the polygamous selection lines produced ejaculates with increased sperm numbers and greater sperm motility compared to males from the monogamous lines. Here, after 12 generations of experimental evolution, we conducted competitive matings between males from lineages evolving under sperm competition and males from lineages subject to relaxed selection. We reduced variation in paternity arising from embryo mortality by genotyping embryos in utero at 14 days gestation. Our microsatellite data revealed a significant paternity bias toward males that evolved under the selective regime of sperm competition. Conclusion We provide evidence that the sperm competitiveness phenotype can respond to selection, and show that improved sperm quality translates to greater competitive fertilisation success in house mice.

Sperm-Hybrid Micromotor for Targeted Drug Delivery.

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Xu, Haifeng; Medina-Sánchez, Mariana; Magdanz, Veronika; Schwarz, Lukas; Hebenstreit, Franziska; Schmidt, Oliver G

2018-01-23

A sperm-driven micromotor is presented as a targeted drug delivery system, which is appealing to potentially treat diseases in the female reproductive tract. This system is demonstrated to be an efficient drug delivery vehicle by first loading a motile sperm cell with an anticancer drug (doxorubicin hydrochloride), guiding it magnetically, to an in vitro cultured tumor spheroid, and finally freeing the sperm cell to deliver the drug locally. The sperm release mechanism is designed to liberate the sperm when the biohybrid micromotor hits the tumor walls, allowing it to swim into the tumor and deliver the drug through the sperm-cancer cell membrane fusion. In our experiments, the sperm cells exhibited a high drug encapsulation capability and drug carrying stability, conveniently minimizing  toxic side effects and unwanted drug accumulation in healthy tissues. Overall, sperm cells are excellent candidates to operate in physiological environments, as they neither express pathogenic proteins nor proliferate to form undesirable colonies, unlike other cells or microorganisms. This sperm-hybrid micromotor is a biocompatible platform with potential application in gynecological healthcare, treating or detecting cancer or other diseases in the female reproductive system.

Predictive value of sperm morphology and progressively motile sperm count for pregnancy outcomes in intrauterine insemination.

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Lemmens, Louise; Kos, Snjezana; Beijer, Cornelis; Brinkman, Jacoline W; van der Horst, Frans A L; van den Hoven, Leonie; Kieslinger, Dorit C; van Trooyen-van Vrouwerff, Netty J; Wolthuis, Albert; Hendriks, Jan C M; Wetzels, Alex M M

2016-06-01

To investigate the value of sperm parameters to predict an ongoing pregnancy outcome in couples treated with intrauterine insemination (IUI), during a methodologically stable period of time. Retrospective, observational study with logistic regression analyses. University hospital. A total of 1,166 couples visiting the fertility laboratory for their first IUI episode, including 4,251 IUI cycles. None. Sperm morphology, total progressively motile sperm count (TPMSC), and number of inseminated progressively motile spermatozoa (NIPMS); odds ratios (ORs) of the sperm parameters after the first IUI cycle and the first finished IUI episode; discriminatory accuracy of the multivariable model. None of the sperm parameters was of predictive value for pregnancy after the first IUI cycle. In the first finished IUI episode, a positive relationship was found for ≤4% of morphologically normal spermatozoa (OR 1.39) and a moderate NIPMS (5-10 million; OR 1.73). Low NIPMS showed a negative relation (≤1 million; OR 0.42). The TPMSC had no predictive value. The multivariable model (i.e., sperm morphology, NIPMS, female age, male age, and the number of cycles in the episode) had a moderate discriminatory accuracy (area under the curve 0.73). Intrauterine insemination is especially relevant for couples with moderate male factor infertility (sperm morphology ≤4%, NIPMS 5-10 million). In the multivariable model, however, the predictive power of these sperm parameters is rather low. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

SPERM COUNT DISTRIBUTIONS IN FERTILE MEN

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Sperm concentration and count are often used as indicators of environmental impacts on male reproductive health. Existing clinical databases may be biased towards subfertile men with low sperm counts and less is known about expected sperm count distributions in cohorts of fertil...

Panax ginseng induces the expression of CatSper genes and sperm hyperactivation

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Eun Hwa Park

2014-12-01

Full Text Available The cation channel of sperm (CatSper protein family plays important roles in male reproduction and infertility. The four members of this family are expressed exclusively in the testis and are localized differently in sperm. To investigate the effects of Panax ginseng treatment on the expression of CatSper genes and sperm hyperactivation in male mice, sperm motility and CatSper gene expression were assessed using a computer-assisted semen analysis system, a Fluoroskan Ascent microplate fluorometer to assess Ca 2+ influx, real-time polymerase chain reaction, Western blotting and immunofluorescence. The results suggested that the Ca 2+ levels of sperm cells treated with P. ginseng were increased significantly compared with the normal group. The P. ginseng-treated groups showed increased sperm motility parameters, such as the curvilinear velocity and amplitude of lateral head displacement. Taken together, the data suggest that CatSper messenger ribonucleic acid levels were increased significantly in mouse testes in the P. ginseng-treated group, as was the protein level, with the exception of CatSper2. In conclusion, P. ginseng plays an important role in improving sperm hyperactivation via CatSper gene expression.

New permeable cryoprotectant-free vitrification method for native human sperm.

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Aizpurua, J; Medrano, L; Enciso, M; Sarasa, J; Romero, A; Fernández, M A; Gómez-Torres, M J

2017-10-01

Is permeable cryoprotectant-free vitrification of native sperm samples a good alternative to conventional slow freezing? The permeable cryoprotectant-free sperm vitrification protocol tested in this study renders considerably better recovery rates of good quality sperm compared to slow freezing. Slow freezing is currently the most commonly used technique for sperm cryopreservation, though this method has been repeatedly shown to have negative effects on both structural and functional sperm features. New alternative methods such as vitrification have been established as a successful alternative in other reproductive cell types, but vitrification of spermatozoa is still a rather unexplored methodology, with limited studies showing its efficacy in male gametes. This study included 18 normozoospermic sperm samples from patients seeking ART treatment between 2014 and 2015. The effects of a new vitrification protocol on functional and structural sperm quality parameters in comparison to fresh and slow-frozen samples were assessed. All samples were divided into three aliquots: fresh (F), slow freezing-thawing (S) and vitrification-warming (V). Sperm concentration, motility, morphology, vitality, DNA fragmentation, cytoskeleton integrity and spontaneous acrosome reaction were assessed and compared between the groups. Results showed improved preservation of sperm features after vitrification compared to conventional freezing. Permeable cryoprotectant-free vitrification presented a significantly higher percentage of live spermatozoa, than slow freezing, better preservation of acrosomes was achieved in vitrified samples and DNA fragmentation was reduced approximately one-third on average compared to slow freezing. Regarding tubulin assay, three different labelling patterns were observed. The frequency of these labelling patterns was similar in F and V groups but this was not the case of the S group. The multivariate analysis of all sperm quality parameters studied revealed

Relationship of Total Motile Sperm Count and Percentage Motile Sperm to Successful Pregnancy Rates Following Intrauterine Insemination

OpenAIRE

Pasqualotto, Eleonora B.; Daitch, James A.; Hendin, Benjamin N.; Falcone, Tommaso; Thomas, Anthony J.; Nelson, David R.; Agarwal, Ashok

1999-01-01

Purpose:This study sought (i) to investigate the relationship between postwash total motile sperm count and postwash percentage motile sperm in predicting successful intrauterine insemination and (ii) to determine the minimal postwash total motile sperm count required to achieve pregnancy with intrauterine insemination.

Mass spectrometry profiling of oxysterols in human sperm identifies 25-hydroxycholesterol as a marker of sperm function

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Chiara Zerbinati

2017-04-01

Full Text Available Cholesterol is a main lipid component of sperm cell that is essential for sperm membrane fluidity, capacitation, and acrosomal reaction. Recent data obtained in bovine sperm showed that sperm capacitation is associated to the formation of oxysterols, oxidized products of cholesterol. The aim of this study was to profile oxysterol content in human semen, and to investigate their potential role in sperm pathophysiology. Among the 12 oxysterols analyzed, 25-hydroxycholesterol (25-HC resulted the most represented in normozoospermic samples, and its concentration positively correlated with spermatozoa number. We detected Cholesterol 25-hydroxylase, the enzyme responsible for 25-HC production, in human spermatozoa at the level of the neck and the post acrosomal area. Upon incubation with spermatozoa, 25-HC induced calcium and cholesterol transients in connection with the acrosomal reaction. Our results support a role for 25-HC in sperm function.

Sperm competition, immunity, selfish genes and cancer.

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Lewis, Z; Price, T A R; Wedell, N

2008-10-01

Sperm competition is widespread and has played an important role in shaping male reproductive characters such as testis size and numbers of sperm produced, and this is reflected in the rapid evolution of many reproductive genes. Additionally, sperm competition has been implicated in the rapid evolution of seminal fluids. However, our understanding of the molecular basis of many traits thought to be important in sperm competition is rudimentary. Furthermore, links between sperm competition and a range of issues not directly related to reproduction are only just beginning to be explored. These include associations between sperm competition and selfish genes, immunity and diseases such as cancer.We briefly review these topics and suggest areas we consider worthy of additional research.

Proteins associated with critical sperm functions and sperm head shape are differentially expressed in morphologically abnormal bovine sperm induced by scrotal insulation

NARCIS (Netherlands)

Saadi, H.A.S.; Riemsdijk, van E.L.C.; Dance, A.L.; Rajamanickam, G.D.; Kastelic, J.P.; Thundathil, J.C.

2013-01-01

The objective was to investigate expression patterns of proteins in pyriform sperm, a common morphological abnormality in bull sperm. Ejaculates were collected from sexually mature Holstein bulls (n = 3) twice weekly for 10 weeks (pre-thermal insult samples). Testicular temperature was elevated in

Flow cytometry application in the assessment of sperm DNA integrity of men with asthenozoospermia.

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A Brodowska

2008-04-01

Full Text Available Sperm genomic integrity and ultrastructural features of ejaculated spermatozoa contributing to the assessment of gamete fertility potential in patients with asthenozoospermia are discussed. The proportion of TUNEL-positive cells was significantly higher in the semen of patients with low sperm motility (n=40; p<0.01 as compared to men with normal sperm motility (n=54. Sperm DNA fragmentation negatively correlated (n=94 with sperm motility, sperm concentration, and integrity of the sperm cellular membrane (HOS-test. Two categories of patients were distinguished: (1 patients (23 out of 94 subjects with < or = 4% of TUNEL-positive cells and (2 patients (71 subjects with 4% of TUNEL-positive cells. A significant difference was noted in the sperm motility and HOS-test results between patients from both groups. Large numbers of immature spermatozoa with extensive cytoplasmic retention, ultrastructural chromatin and midpiece abnormalities, and conglomerates containing sperm fragments were present more frequently in the semen of asthenozoospermic subjects with >4% of TUNEL-positive sperm cells. Low sperm motility seems to be accompanied by serious defects of gamete chromatin expressed as diminished sperm genomic integrity and abnormal DNA condensation and by defects of sperm midpiece. These abnormalities may reflect developmental failure during the spermatogenic remodeling process. The DNA fragmentation test may be considered as an additional assay for the evaluation of spermatozoa beside standard analysis and taken together with electron microscopy may help to determine the actual number of "healthy" spermatozoa thereby playing an important role during diagnosis and treatment of male infertility.

Oxidative stress negatively affects human sperm mitochondrial respiration.

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Ferramosca, Alessandra; Pinto Provenzano, Sara; Montagna, Daniela Domenica; Coppola, Lamberto; Zara, Vincenzo

2013-07-01

To correlate the level of oxidative stress in serum and seminal fluid and the level of sperm deoxyribonucleic acid (DNA) fragmentation with sperm mitochondrial respiratory efficiency. Sperm mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically treated sperm cells. A possible relationship between sperm mitochondrial respiratory efficiency, the level of oxidative stress, and the level of sperm DNA fragmentation was investigated. Sperm motility was positively correlated with mitochondrial respiration but negatively correlated with oxidative stress and DNA fragmentation. Interestingly, sperm mitochondrial respiratory activity was negatively affected by oxidative stress and DNA fragmentation. Our data indicate that sperm mitochondrial respiration is decreased in patients with high levels of reactive oxygen species by an uncoupling between electron transport and adenosine triphosphate synthesis. This reduction in mitochondrial functionality might be 1 of the reasons responsible for the decrease in spermatozoa motility. Copyright © 2013 Elsevier Inc. All rights reserved.

Formation of primary sperm conjugates in a haplogyne spider (Caponiidae, Araneae) with remarks on the evolution of sperm conjugation in spiders.

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Lipke, Elisabeth; Michalik, Peter

2012-11-01

Sperm conjugation, where two or more sperm are physically united, is a rare but widespread pheno-menon across the animal kingdom. One group well known for its different types of sperm conjugation are spiders. Particularly, haplogyne spiders show a high diversity of sperm traits. Besides individual cleistospermia, primary (synspermia) and secondary (coenospermia, "spermatophore") sperm conjugation occurs. However, the evolution of sperm conjugates and sperm is not understood in this group. Here, we look at how sperm are transferred in Caponiidae (Haplogynae) in pursuit of additional information about the evolution of sperm transfer forms in spiders. Additionally, we investigated the male reproductive system and spermatozoa using light- and transmission electron-microscopy and provide a 3D reconstruction of individual as of well as conjugated spermatozoa. Mature spermatozoa are characterized by an extremely elongated, helical nucleus resulting in the longest spider sperm known to date. At the end of spermiogenesis, synspermia are formed by complete fusion of four spermatids. Thus, synspermia might have evolved early within ecribellate Haplogynae. The fused sperm cells are surrounded by a prominent vesicular area. The function of the vesicular area remains still unknown but might be correlated with the capacitation process inside the female. Further phylogenetic and functional implications of the spermatozoa and sperm conjugation are discussed. Copyright © 2012 Elsevier Ltd. All rights reserved.

Microdissection Testicular Sperm Extraction (micro-TESE as a Sperm Acquisition Method for Men with Nonobstructive Azoospermia Seeking Fertility: Operative and Laboratory Aspects

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Sandro C. Esteves

2013-06-01

Full Text Available Introduction Rare foci of sperm production may be found in up to 60% of men with nonobstructive azoospermia (NOA. Sperm production, if present, is minimal for sperm appearance in the ejaculate. Given that there are no treatment options to restore fertility, sperm retrieval is the only alternative to find testicular sperm than then can be used for in vitro fertilization (IVF. Among sperm acquisition methods, micro-TESE has higher success rates at obtaining sperm compared with testicular sperm extraction and testicular sperm aspiration. Materials and Methods This video describes the operative aspects of micro-TESE, performed on an outpatient basis, in a man with NOA and history of cryptorchidism in whom orchidopexy was performed at age 6. The concept of micro-TESE is to identify areas of sperm production within the testes with the aid of optical magnification (15-25X and based on the size and appearance of the seminiferous tubules (ST. Conclusion Micro-TESE allowed the identification and extraction of sperm-containing STs with minimum tissue excision and marked reduction in time processing of testicular specimens for sperm injection.

Grape juice concentrate alleviates epididymis and sperm damage in cadmium-intoxicated rats.

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Lamas, Celina de A; Cuquetto-Leite, Livia; do Nascimento da Silva, Emanueli; Thomazini, Bruna F; Cordeiro, Gabriel da S; Predes, Fabrícia de S; Gollücke, Andrea P B; Dolder, Heidi

2017-04-01

The possibility of long-term grape juice concentrate (GJC) consumption conferring a protective effect against cadmium (Cd)-induced damage to the epididymis, completely preserving sperm profile, was evaluated here for the first time in the scientific literature. Male Wistar rats (n = 6/per group) received an intraperitoneal Cd injection (1.2 mg/Kg) at age 80 days and GJC (2 g/Kg) by gavage from 50 days until 136 days old. Groups receiving either Cd or GJC were added. An intraperitoneal injection of saline (0.9%) and water by gavage was administered in the absence of treatment with Cd or GJC. Animals were anaesthetized and exsanguinated at 136 days; the vas deferens, left testis and epididymis were removed; and perfusion continued with fixative. The right epididymis was collected for morphological analysis. Cd had a devastating effect demonstrated by reduced sperm count in testes and epididymis, sperm production and normal sperm count, besides increased epididymis sperm transit time and completely disorganized morphology. These alterations were attributed to higher Cd levels in the testes and a lipid peroxidation (LP) process. Consumption of GJC plus Cd intoxication was effective, reducing metal accumulation and LP. Consequently, we could identify a preserved sperm profile, with improvement in testis and epididymis sperm count, normal sperm structure and sperm transit time. Moreover, GJC extends its protective effect to the epididymis, allowing complete re-establishment of its morphology, ensuring successful sperm maturation process. In conclusion, our study indicates long-term GJC as a promising therapy against reproductive chemical intoxication injury damage, preserving sperm prior to ejaculation. © 2017 The Authors. International Journal of Experimental Pathology © 2017 International Journal of Experimental Pathology.

Seminal plasma and sperm proteome of ring-tailed coatis (Nasua nasua, Linnaeus, 1766).

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Silva, Herlon Victor Rodrigues; Rodriguez-Villamil, Paula; Magalhães, Francisco Felipe de; Nunes, Thalles Gothardo Pereira; Freitas, Luana Azevedo de; Ribeiro, Leandro Rodrigues; Silva, Alexandre Rodrigues; Moura, Arlindo A; Silva, Lúcia Daniel Machado da

2018-04-15

Ring-tailed coati is listed as a species of least concern in the International Union for Conservation of Nature (IUCN) Red List, however, there has been a sharp decline in their population. The present study was conducted to evaluate the major proteins of both seminal plasma and sperm in ring-tailed coatis. Semen sample was collected from three adult coatis and evaluated for their morphological characteristics. Further, the sample was centrifuged to separate spermatozoa from seminal plasma, and then stored in liquid nitrogen. The seminal plasma and sperm proteins were subjected to one-dimensional (1-D) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and identified by mass spectrometry. Gene ontology and protein networks were analyzed using bioinformatics tools. Based on sperm concentration and average protein content of the semen, the concentration of protein/spermatozoon was found to be 104.69 ± 44.43 μg. The analysis of SDS-PAGE gels showed 20.3 ± 3.1 and 17 ± 2 protein bands/lane for seminal plasma and sperm, respectively. In-gel protein digestion and peptide analysis by mass spectrometry revealed 238 and 246 proteins in the seminal plasma and sperm, respectively. The gene ontology analysis revealed that the proteins of seminal plasma mainly participated in cellular (35%) and regulatory (21%) processes. According to their cellular localization, seminal plasma proteins were categorized as structural (18%), extracellular (17%), and nuclear (14%) proteins with molecular functions, such as catalytic activity (43%) and binding (43%). The sperm proteins were also involved in cellular (38%) and regulatory (23%) processes, and mainly categorized as extracellular (17%), nuclear (13%), and cytoplasmic (10%) proteins. The major molecular functions of the sperm proteins were catalytic activity (44%) and binding (42%). These results indicated that the seminal plasma of ring-tailed coati has an array of proteins that can potentially

Activation of free sperm and dissociation of sperm bundles (spermatozeugmata) of an endangered viviparous fish, Xenotoca eiseni.

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Liu, Yue; Yang, Huiping; Torres, Leticia; Tiersch, Terrence R

2018-04-01

Knowledge of sperm motility activation for viviparous fishes has been limited to study of several species in Poeciliidae, and the dissociation of sperm bundles is even less understood. The goal of this study was to use the endangered Redtail Splitfin (Xenotoca eiseni) as a model to investigate the activation of sperm from viviparous fishes by study of free sperm and spermatozeugmata (unencapsulated sperm bundles). The specific objectives were to evaluate the effects of: (1) osmotic pressure and refrigerated storage (4 °C) on activation of free sperm, (2) osmotic pressure, ions, and pH on dissociation of spermatozeugmata, and (3) CaCl 2 concentration and pH on sperm membrane integrity. Free sperm were activated in Ca 2+ -free Hanks' balanced salt solution at 81-516 mOsmol/kg. The highest motility (19 ± 6%) was at 305 mOsmol/kg and swim remained for 84 h. Glucose (300-700 mOsmol/kg), NaCl (50-600 mOsmol/kg), and KCl, MgCl 2 , and MnCl 2 at 5-160 mM activated sperm within spermatozeugmata, but did not dissociate spermatozeugmata. CaCl 2 at 5-160 mM dissociated spermatozeugmata within 10 min. Solutions of NaCl-NaOH at pH 11.6 to 12.4 dissociated spermatozeugmata within 1 min. The percentage of viable cells had no significant differences (P = 0.2033) among different concentrations of CaCl 2 , but it was lower (P fishes, and for development of germplasm repositories for imperiled goodeids. Copyright © 2018 Elsevier Inc. All rights reserved.

Impairment of sperm DNA methylation in male infertility: a meta-analytic study.

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Santi, D; De Vincentis, S; Magnani, E; Spaggiari, G

2017-07-01

Considering the widespread use of assisted reproductive techniques (ART), DNA methylation of specific genes involved in spermatogenesis achieves increasingly clinical relevance, representing a possible explanation of increased incidence of syndromes related to genomic imprinting in medically assisted pregnancies. Several trials suggested a relationship between male sub-fertility and sperm DNA methylation, although its weight on seminal parameters alteration is still a matter of debate. To evaluate whether aberrant sperm DNA methylation of imprinted genes is associated with impaired sperm parameters. Meta-analysis of controlled clinical trials evaluating imprinted genes sperm DNA methylation comparing men with idiopathic infertility to fertile controls. Twenty-four studies were included, allowing a meta-analytic evaluation for H19, MEST, SNRPN, and LINE-1. When a high heterogeneity of the results was demonstrated, the random effect model was used. H19 methylation levels resulted significantly lower in 879 infertile compared with 562 fertile men (7.53%, 95% CI: 5.14-9.93%, p male infertility is associated with altered sperm methylation at H19, MEST, and SNRPN. Although its role in infertility remains unclear, sperm DNA methylation could be associated with the epigenetic risk in ART. In this setting, before proposing this analysis in clinical practice, an accurate identification of the most representative genes and a cost-effectiveness evaluation should be assessed in ad hoc prospective studies. © 2017 American Society of Andrology and European Academy of Andrology.

Effects of hydrostatic pressure on mouse sperm.

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Karimi, N; Kamangar, P Bahrami; Azadbakht, M; Amini, A; Amiri, I

2014-01-01

The objective of this study was to investigate the abnormalities in sperm after exposure to hydrostatic pressure. Hydrostatic pressure acting on the cells is one of the fundamental environmental mechanical forces. Disorders of relationship between the cells and this mechanical force, such as when pressure varies beyond physiological limits, can lead to disease or pathological states. Sperm exposed to different range of hydrostatic pressure within male reproductive system and after entering the female reproductive system. Sexually mature male NMRI mice, 8-12 weeks-old were sperm donors. Sperms were separated from the caudal epididymis and maintained in Ham's F-10 culture medium supplemented with 10 % FBS and divided into control and treatments. Sperm suspensions in the treatments were placed within pressure chamber and were subjected to increased hydrostatic pressure of 25, 50 and 100 mmHg (treatment I, II and III) above atmospheric pressure for 2 and 4 h. Sperm viability, motility, morphology, DNA integrity and fertilizing ability were assessed and compared with control. Results showed that hydrostatic pressure dependent on ranges and time manner reduced sperm quality due to adverse effect on viability, motility , morphology, DNA integrity and fertilizing ability in all of treatments, especially after 4h (phydrostatic pressure reduces sperm quality as a consequence of adverse effects on sperm parameters and may cause male infertility or subfertility (Tab. 5, Ref. 5).

Mitochondrial outer membrane permeabilization increases reactive oxygen species production and decreases mean sperm velocity but is not associated with DNA fragmentation in human sperm.

Science.gov (United States)

Treulen, F; Uribe, P; Boguen, R; Villegas, J V

2016-02-01

Does induction of mitochondrial outer membrane permeabilization (MOMP) in vitro affect specific functional parameters of human spermatozoa? Our findings show that MOMP induction increases intracellular reactive oxygen species (ROS) and decreases mean sperm velocity but does not alter DNA integrity. MOMP in somatic cells is related to a variety of apoptotic traits, such as alteration of mitochondrial membrane potential (ΔΨm), and increase in ROS production and DNA fragmentation. Although the presence of these apoptotic features has been reported in spermatozoa, to date the effects of MOMP on sperm function and DNA integrity have not been analysed. The study included spermatozoa from fertile donors. Motile sperm were obtained using the swim-up method. The highly motile sperm were collected and diluted with human tubal fluid to a final cell concentration of 5 × 10(6) ml(-1). To induce MOMP, selected sperm were treated at 37°C for 4 h with a mimetic of a Bcl-2 pro-apoptotic protein, ABT-737. MOMP was evaluated by relocating of cytochrome c. In addition, the effect of ABT-737 on mitochondrial inner membrane permeabilization was assessed using the calcein-AM/cobalt chloride method. In turn, ΔΨm was evaluated with JC-1 staining, intracellular ROS production with dihydroethidium, sperm motility was analysed by computer-assisted sperm analysis and DNA fragmentation by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) assay. Measurements were performed by flow cytometry. MOMP was associated with ΔΨm dissipation (P < 0.05), increased ROS production (P < 0.05) and decreased mean sperm velocity (P < 0.05), but it was not associated with DNA fragmentation. MOMP did not induce a large increase in ROS, which could explain the negligible effect of MOMP on sperm DNA fragmentation under our experimental conditions. The study was carried out in vitro using highly motile sperm, selected by swim-up, from healthy donors. The results obtained in this

Tactic-specific differences in seminal fluid influence sperm performance.

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Locatello, Lisa; Poli, Federica; Rasotto, Maria B

2013-03-22

Seminal fluid often makes up a large part of an ejaculate, yet most empirical and theoretical studies on sperm competition have focused on how sperm characteristics (number and quality) affect fertilization success. However, seminal fluid influences own sperm performance and may potentially influence the outcome of sperm competition, by also affecting that of rivals. As a consequence males may be expected to allocate their investment in both sperm and seminal fluid in relation to the potential level of competition. Grass goby (Zosterisessor ophiocephalus) is an external fertilizer with guard-sneaker mating tactics, where sperm competition risk varies according to the tactic adopted. Here, we experimentally manipulated grass goby ejaculates by separately combining sperm and seminal fluid from territorial and sneaker males. While sperm of sneaker and territorial males did not differ in their performance when they interacted with their own seminal fluid only, sperm of sneakers increased their velocity and fertilization rate in the presence of territorial males' seminal fluid. By contrast, sneaker males' seminal fluid had a detrimental effect on the performance of territorial males' sperm. Sperm velocity was unaffected by the seminal fluid of males employing the same tactic, suggesting that seminal fluid's effect on rival-tactic sperm is not based on a self/non-self recognition mechanism. Our findings show that cross interactions of sperm and seminal fluid may influence the fertilization success of competing ejaculates with males investing in both sperm and seminal fluid in response to sperm competition risk.

Study of Sperm Parameters and Sperm Fertility in Mice were Exposed to Tamoxifen during Embryonic Development

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J Soleimanirad

2017-05-01

Full Text Available Introduction: Tamoxifen is steroidal drug, which mainly treats breast cancer and also used to stimulate ovulation. The purpose of the present study was the evaluation of sperm parameters and fertility of mice whose mothers had received tamoxifen during pregnancy. Methods: In this study, 30 female and 15 male mice of NMRI were selected for mating. After mating female mice were randomly divided into two groups, the first group (control and second group (experimental. All of which contained 15 mice. From the day 13th day of pregnancy, experimental group has received tamoxifen with the dosage of 5 mg/kg for 7 days. After childbirth of the mated mice, male infants were selected. After reaching the age of puberty (6-8Weeks, adult mice were sacrificed by the cervical dislocation. After take sperm, sperm parameters (count, normality and motility, and sperm fertility was performed. In this study SPSS software and statistical t-test was used (p <0.001. Results: Studies showed that sperm parameters and sperm fertilization were significantly different. The number of sperm in the control group was 83.50±28.20 million, and in the experimental group was 60±14.14 million. There was a decrease in average sperm count in the experimental group compared with the control group (p <0.001. Our findings from in vitro fertilization culture media showed that embryos formation and oocyte disruption between control and experimental groups significantly different (p <0.001. Conclusion: The results showed that tamoxifen exposure during development can cause histological changes in the seminiferous tubules, which can lead to infertility.

Reproductive function in male endurance athletes: sperm analysis and hormonal profile.

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Lucía, A; Chicharro, J L; Pérez, M; Serratosa, L; Bandrés, F; Legido, J C

1996-12-01

The purpose of this investigation was to study the effects of endurance exercise on male reproductive function (sex hormones and seminograms). Professional cyclists [n = 12; mean age 24 +/- 2 (SD) yr], elite triathletes (n = 9; 26 +/- 3 yr), recreational marathon runners (n = 10; 32 +/- 6 yr), and sedentary subjects (control group; n = 9; 30 +/- 4 yr) were selected as subjects. for each group, the following parameters were measured three times during the sports season (training period: winter; competition period: spring; resting period: fall): percentage of body fat, hormonal profile (resting levels of follicle-stimulating hormone, luteinizing hormone, total and free testosterone, and cortisol), and seminograms (quantitative parameters sperm volume and sperm count; qualitative parameters: sperm motality and morphology). The following comparisons were made in the measured parameters: 1) within groups (longitudinal design) and 2) between groups in each of the three periods (cross-sectional design) and over time (mixed design). In addition, both the volume and the intensity of training of each subject during the season (except for the control group) were quantified. Despite significant differences in training characteristics and in body fat percent, in general no significant differences (P > 0.05) were found in hormonal profiles or in semen characteristics between or within groups. A lower sperm motility (46.2 +/- 19.5%), however, was observed in the cyclists during the competition period when compared either with the other groups during this same period (P < 0.05) or with themselves during the other two periods of study (P < 0.01). In any case, the later phenomenon was attributed to physical factors associated with cycling, such as mechanical trauma to the testis and/or increased gonadal temperature. In conclusion, our findings suggest that endurance exercise does not adversely affect the hypothalamic-pituitary-testis axis.

Sperm counts may have declined in young university students in Southern Spain

DEFF Research Database (Denmark)

Mendiola, Jaime; Jørgensen, Niels; Mínguez-Alarcón, Lidia

2013-01-01

Several studies have investigated temporal trends in semen quality in Northern Europe, but none has examined this question in Southern Europe. A prior study conducted in Almeria Province (Southern Spain) reported higher sperm count and concentration among Spanish young men recruited from 2001......, Body mass index (BMI), presence of varicocoele and prenatal exposure to tobacco) to look for a birth-cohort effect over the combined study period (2001-2011). Sperm concentration and total sperm count declined significantly with year of birth in the pooled analysis (β = -0.04 and β = -0...

Variation in sperm morphometry and sperm competition among barn swallow (Hirundo rustica) populations

Czech Academy of Sciences Publication Activity Database

Laskemoen, T.; Albrecht, Tomáš; Bonisoli-Alquati, A.; Cepák, J.; De Lope, F.; Hermosell, I. G.; Johannessen, L. E.; Kleven, O.; Marzal, A.; Mousseau, T. A.; Moller, A. P.; Robertson, R. J.; Rudolfsen, G.; Saino, N.; Vortman, Y.; Lifjeld, J. T.

2013-01-01

Roč. 67, č. 2 (2013), s. 301-309 ISSN 0340-5443 R&D Projects: GA ČR(CZ) GAP506/12/2472 Institutional support: RVO:68081766 Keywords : Barn swallow * Extrapair paternity * Hirundo rustica * Sperm competition * Sperm size Subject RIV: EG - Zoology Impact factor: 3.049, year: 2013

Delta opioid receptor on equine sperm cells: subcellular localization and involvement in sperm motility analyzed by computer assisted sperm analyzer (CASA

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Lacalandra Giovanni M

2010-06-01

Full Text Available Abstract Background Opioid receptors and endogenous opioid peptides act not only in the control of nociceptive pathways, indeed several reports demonstrate the effects of opiates on sperm cell motility and morphology suggesting the importance of these receptors in the modulation of reproduction in mammals. In this study we investigated the expression of delta opioid receptors on equine spermatozoa by western blot/indirect immunofluorescence and its relationship with sperm cell physiology. Methods We analyzed viability, motility, capacitation, acrosome reaction and mitochondrial activity in the presence of naltrindole and DPDPE by means of a computer assisted sperm analyzer and a fluorescent confocal microscope. The evaluation of viability, capacitation and acrosome reaction was carried out by the double CTC/Hoechst staining, whereas mitochondrial activity was assessed by means of MitoTracker Orange dye. Results We showed that in equine sperm cells, delta opioid receptor is expressed as a doublet of 65 and 50 kDa molecular mass and is localized in the mid piece of tail; we also demonstrated that naltrindole, a delta opioid receptor antagonist, could be utilized in modulating several physiological parameters of the equine spermatozoon in a dose-dependent way. We also found that low concentrations of the antagonist increase sperm motility whereas high concentrations show the opposite effect. Moreover low concentrations hamper capacitation, acrosome reaction and viability even if the percentage of cells with active mitochondria seems to be increased; the opposite effect is exerted at high concentrations. We have also observed that the delta opioid receptor agonist DPDPE is scarcely involved in affecting the same parameters at the employed concentrations. Conclusions The results described in this paper add new important details in the comprehension of the mammalian sperm physiology and suggest new insights for improving reproduction and for

Sex-sorting sperm using flow cytometry/cell sorting.

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Garner, Duane L; Evans, K Michael; Seidel, George E

2013-01-01

The sex of mammalian offspring can be predetermined by flow sorting relatively pure living populations of X- and Y-chromosome-bearing sperm. This method is based on precise staining of the DNA of sperm with the nucleic acid-specific fluorophore, Hoechst 33342, to differentiate between the subpopulations of X- and Y-sperm. The fluorescently stained sperm are then sex-sorted using a specialized high speed sorter, MoFlo(®) SX XDP, and collected into biologically supportive media prior to reconcentration and cryopreservation in numbers adequate for use with artificial insemination for some species or for in vitro fertilization. Sperm sorting can provide subpopulations of X- or Y-bearing bovine sperm at rates in the 8,000 sperm/s range while maintaining; a purity of 90% such that it has been applied to cattle on a commercial basis. The sex of offspring has been predetermined in a wide variety of mammalian species including cattle, swine, horses, sheep, goats, dogs, cats, deer, elk, dolphins, water buffalo as well as in humans using flow cytometric sorting of X- and Y-sperm.

Studies on varicocele III: ultrastructural sperm evaluation and 18, X and Y aneuploidies.

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Baccetti, Baccio M; Bruni, Emanuele; Capitani, Serena; Collodel, Giulia; Mancini, Stefano; Piomboni, Paola; Moretti, Elena

2006-01-01

The idea that varicocele plays a detrimental role in fertility is supported by the presence of a higher frequency of affected men among the infertile population than among men with normal semen parameters. In this research we examined ejaculates from a large group of selected men affected by varicocele by light and electron microscopy. The effect of varicocele on chromosome meiotic segregation was investigated by fluorescence in situ hybridization (FISH). The potential benefits of varicocelectomy on sperm quality were evaluated by analyzing sperm characteristics before and after surgical correction of varicocele. Transmission electron microscopy (TEM) analysis, elaborated previously, showed that the incidence of immaturity, apoptosis, and necrosis was higher in the varicocele group than in controls. FISH analysis performed on sperm nuclei from selected patients with varicocele showed that the mean frequencies of disomies and diploidies were generally out of the normal range, indicating a severe disturbance in meiotic segregation. Sperm characteristics evaluated before and after varicocele repair showed a general improvement. As a consequence, the varicocele seem to affect sperm morphology and function concomitantly with meiotic segregation derangement. In consideration of these data, we suggest that TEM and FISH analyses should be performed for all varicocele patients.

Relationship between conventional semen characteristics, sperm motility patterns and fertility of Andalusian donkeys (Equus asinus).

Science.gov (United States)

Dorado, J; Acha, D; Ortiz, I; Gálvez, M J; Carrasco, J J; Díaz, B; Gómez-Arrones, V; Calero-Carretero, R; Hidalgo, M

2013-12-01

Sperm quality has an important role in determining fertility. The aims of this study were to compare the conventional sperm parameters, plus the characteristics of the motility patterns of the different sperm subpopulations, of donkey donors with different fertility level, and to determine their relationships to fertility. Thirty ejaculates from 6 Andalusian donkeys were assessed for gel-free volume, pH, sperm concentration, motility and morphology. The fertility of donkeys was classified on the basis of pregnancy rates per cycle, where donkeys with a per cycle pregnancy rate ≥60% were considered to be "fertile" (n=3) and those with a per cycle pregnancy rate donkeys with in vitro measures of sperm motility using computer-assisted sperm analysis techniques. Copyright © 2013 Elsevier B.V. All rights reserved.

Sperm DNA quality evaluated by comet assay and sperm chromatin structure assay in stallions after unilateral orchiectomy.

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Serafini, R; Varner, D D; Bissett, W; Blanchard, T L; Teague, S R; Love, C C

2015-09-15

Unilateral orchiectomy (UO) may interfere with thermoregulation of the remaining testis caused by inflammation surrounding the incision site, thus altering normal spermatogenesis and consequently sperm quality. Two measures of sperm DNA quality (neutral comet assay and the sperm chromatin structure assay [SCSA]) were compared before UO (0 days) and at 14, 30, and 60 days after UO to determine whether sperm DNA changed after a mild testis stress (i.e., UO). The percent DNA in the comet tail was higher at 14 and 60 days compared to 0 days (P comet tail measures (i.e., length, moment, migration) were higher at all time periods after UO compared to 0 days (P comet assay and the SCSA, which was not identified using traditional measures of sperm quality. Copyright © 2015 Elsevier Inc. All rights reserved.

Effect of method of euthanasia on sperm motility of mature Sprague-Dawley rats.

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Stutler, Shannon A; Johnson, Eric W; Still, Kenneth R; Schaeffer, David J; Hess, Rex A; Arfsten, Darryl P

2007-03-01

Euthanasia is one of the most commonly performed procedures in laboratory animal settings. The method of euthanasia may affect experimental results in studies using animals and must be compatible with research objectives including subsequent tissue analyses. Our present study was performed to evaluate the effects of 7 euthanasia methods on sperm motility in mature rats. Rats were euthanized using CO2, 2 commercially available euthanasia solutions (Beuthanasia-D and Sleepaway), and 4 volatile anesthetics (enflurane, halothane, isoflurane, and sevoflurane). Rats euthanized by rapid decapitation alone served as negative controls, and a-chlorohydrin-treated rats euthanized by rapid decapitation were positive controls for sperm impairment. For 5 of these methods, we also measured time to ataxia, recumbency, respiratory arrest, and no auscultable heartbeat. Immediately after euthanasia of each rat, distal caudal epididymides were removed; 1 was processed for automated sperm motility analysis, and the other was frozen for subsequent concentration analysis. Time to all measured parameters was less for volatile anesthetics than for Beuthanasia-D. Times to last respiration and no heartbeat were less for halothane and isoflurane than for enflurane and sevoflurane. Percentage motile sperm did not differ significantly between methods. Percentage progressively motile sperm did not vary significantly between methods except for Beuthanasia-D, for which it was significantly less than the negative control value. Specific sperm motion parameters for each euthanasia method except CO2 and Sleepaway varied significantly from the negative control. Our results indicate that the method of euthanasia is an important consideration when rat sperm motility parameters must be evaluated.

Validation of the FACSCount AF system for determination of sperm concentration in boar semen

DEFF Research Database (Denmark)

Hansen, C.; Christensen, P.; Stryhn, H.

2002-01-01

microspheres ( beads). Satisfactory staining can be achieved within 2-3 min and the following flow cytometric analysis on the FACSCount AF System rapidly provides the user with a precise and accurate assessment of the sperm concentration. In this study, the FACSCount AF System and Sperm Counting Reagent ( BD...... Biosciences) was compared with microscopic counting using a Burker-Turk haemocytometer. In addition, sperm concentration was determined using the Corning 254 spectrophotometer which is used routinely by Danish artificial insemination stations for boars. The results show that the agreement between flow...... cytometry and microscopic counting is very high. The slope for the regression line was 1.12 (SE = 0.03) with an estimated intercept with the Y-axis of 22 x 10(6) sperm/ml (SE = 10 x 10(6) sperm/ml) and an estimated error of the model of 10 x 10(6) sperm/ml. For the spectrophotometer, the slope...

Prudent sperm use by leaf-cutter ant queens

DEFF Research Database (Denmark)

Den Boer, Susanne Petronella A; Baer, Boris; Dreier, Stéphanie Agnès Jeanine

2009-01-01

are able to fertilize close to 100 per cent of the eggs and that the average sperm use per egg is very low, but increases with queen age. The robustness of stored sperm was found to decrease with years of storage, signifying that senescence affects sperm either directly or indirectly via the declining...... glandular secretions or deteriorating sperm-storage organs. We evaluate our findings with a heuristic model, which suggests that the average queen has sperm for almost 9 years of normal colony development. We discuss the extent to which leaf-cutter ant queens have been able to optimize their sperm...

Testicular Damage following Testicular Sperm Retrieval

DEFF Research Database (Denmark)

Fedder, Jens; Marcussen, Niels; Fedder, Maja D.K.

2017-01-01

The aim of this study was to evaluate the possible development of histological abnormalities such as fibrosis and microcalcifications after sperm retrieval in a ram model. Fourteen testicles in nine rams were exposed to open biopsy, multiple TESAs, or TESE, and the remaining four testicles were...... left unoperated on as controls. Three months after sperm retrieval, the testicles were removed, fixed, and cut into 1/2 cm thick slices and systematically put onto a glass plate exposing macroscopic abnormalities. Tissue from abnormal areas was cut into 3 μm sections and stained for histological...... evaluation. Pathological abnormalities were observed in testicles exposed to sperm retrieval (≥11 of 14) compared to 0 of 4 control testicles. Testicular damage was found independently of the kind of intervention used. Therefore, cryopreservation of excess sperm should be considered while retrieving sperm....

The predictive value of parameters of clinical presentations for sperm yield in patients with nonobstructive azoospermia receiving microdissection testicular sperm extraction

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Ming-Hsuan Ku

2017-12-01

Conclusion: Clinical presentations or phenotypes can be used as predictive factors for successful sperm retrieval during mTESE in patients with NOA. Hypogonadotropic hypogonadism and cases with UDT history have a higher chance of sperm retrieval. Initial testicular needle biopsy, if available, can provide valuable information about chances of sperm retrieval. Hypospermatogenesis predicts high sperm yield rate, and LMA can have best upgrade results of sperm yield after mTESE.

Sperm DNA damage in relation to lipid peroxidation following freezing-thawing of boar semen

OpenAIRE

Fraser, L.; Strzeżek, J.; Wasilewska, K.; Pareek, C.S.

2017-01-01

This study investigated the relationships between lipid peroxidation (LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based extenders after single and repeated freezing and thawing. As well as an analysis of sperm motion characteristics, mitochondrial function, membrane integrity, and lipid peroxidation (LPO) were assessed simulta...

Role of Trace Elements for Oxidative Status and Quality of Human Sperm.

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Nenkova, Galina; Petrov, Lubomir; Alexandrova, Albena

2017-08-04

Oxidative stress affects sperm quality negatively. To maintain the pro/antioxidant balance, some metal ions (e.g. copper, zink, iron, selenium), which are co-factors of the antioxidant enzymes, are essential. However, iron and copper could act as prooxidants inducing oxidative damage of spermatozoa. To reveal a possible correlation between the concentrations of some metal ions (iron, copper, zinc, and selenium) in human seminal plasma, oxidative stress, assessed by malondialdehyde and total glutathione levels, and semen quality, assessed by the parameters count, motility, and morphology. Descriptive study. The semen analysis for volume, count, and motility was performed according to World Health Organization (2010) guidelines, using computer-assisted semen analysis. For the determination of spermatozoa morphology, a SpermBlue staining method was applied. Depending on their parameters, the sperm samples were categorized into normozoospermic, teratozoospermic, asthenoteratozoospermic, and oligoteratozoospermic. The seminal plasma content of iron, copper, zinc, and selenium was estimated by atomic absorption spectroscopy. The malondialdehyde and total glutathione levels were quantified spectrophotometrically. In the groups with poor sperm quality, the levels of Fe were higher, whereas those of Zn and Se were significantly lower than in the normozoospermic group. In all groups with poor sperm quality, increased levels of malondialdehyde and decreased glutathione levels were detected as evidence of oxidative stress occurrence. All these differences are most pronounced in the asthenoteratozoospermic group where values differ nearly twice as much compared to the normozoospermic group. The Fe concentration correlated positively with the malondialdehyde (r=0.666, p=0.018), whereas it showed a negative correlation with the level of total glutathione (r=-0.689, p=0.013). The total glutathione level correlated positively with the sperm motility (r=0.589, p=0.044). The elevated

Role of Trace Elements for Oxidative Status and Quality of Human Sperm

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Galina Nenkova

2017-08-01

Full Text Available Background: Oxidative stress affects sperm quality negatively. To maintain the pro/antioxidant balance, some metal ions (e.g. copper, zink, iron, selenium, which are co-factors of the antioxidant enzymes, are essential. However, iron and copper could act as prooxidants inducing oxidative damage of spermatozoa. Aims: To reveal a possible correlation between the concentrations of some metal ions (iron, copper, zinc, and selenium in human seminal plasma, oxidative stress, assessed by malondialdehyde and total glutathione levels, and semen quality, assessed by the parameters count, motility, and morphology. Study Design: Descriptive study. Methods: The semen analysis for volume, count, and motility was performed according to World Health Organization (2010 guidelines, using computer-assisted semen analysis. For the determination of spermatozoa morphology, a SpermBlue staining method was applied. Depending on their parameters, the sperm samples were categorized into normozoospermic, teratozoospermic, asthenoteratozoospermic, and oligoteratozoospermic. The seminal plasma content of iron, copper, zinc, and selenium was estimated by atomic absorption spectroscopy. The malondialdehyde and total glutathione levels were quantified spectrophotometrically. Results: In the groups with poor sperm quality, the levels of Fe were higher, whereas those of Zn and Se were significantly lower than in the normozoospermic group. In all groups with poor sperm quality, increased levels of malondialdehyde and decreased glutathione levels were detected as evidence of oxidative stress occurrence. All these differences are most pronounced in the asthenoteratozoospermic group where values differ nearly twice as much compared to the normozoospermic group. The Fe concentration correlated positively with the malondialdehyde (r=0.666, p=0.018, whereas it showed a negative correlation with the level of total glutathione (r=-0.689, p=0.013. The total glutathione level correlated

Sperm DNA fragmentation, recurrent implantation failure and recurrent miscarriage

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Carol Coughlan

2015-01-01

Full Text Available Evidence is increasing that the integrity of sperm DNA may also be related to implantation failure and recurrent miscarriage (RM. To investigate this, the sperm DNA fragmentation in partners of 35 women with recurrent implantation failure (RIF following in vitro fertilization, 16 women diagnosed with RM and seven recent fathers (control were examined. Sperm were examined pre- and post-density centrifugation by the sperm chromatin dispersion (SCD test and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL assay. There were no significant differences in the age of either partner or sperm concentration, motility or morphology between three groups. Moreover, there were no obvious differences in sperm DNA fragmentation measured by either test. However, whilst on average sperm DNA fragmentation in all groups was statistically lower in prepared sperm when measured by the SCD test, this was not seen with the results from the TUNEL assay. These results do not support the hypothesis that sperm DNA fragmentation is an important cause of RIF or RM, or that sperm DNA integrity testing has value in such patients. It also highlights significant differences between test methodologies and sperm preparation methods in interpreting the data from sperm DNA fragmentation tests.

Total motile sperm count has a superior predictive value over the WHO 2010 cut-off values for the outcomes of intracytoplasmic sperm injection cycles.

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Borges, E; Setti, A S; Braga, D P A F; Figueira, R C S; Iaconelli, A

2016-09-01

The objective of this study was to compare (i) the intracytoplasmic sperm injection outcomes among groups with different total motile sperm count ranges, (ii) the intracytoplasmic sperm injection outcomes between groups with normal and abnormal total motile sperm count, and (iii) the predictive values of WHO 2010 cut-off values and pre-wash total motile sperm count for the intracytoplasmic sperm injection outcomes, in couples with male infertility. This study included data from 518 patients undergoing their first intracytoplasmic sperm injection cycle as a result of male infertility. Couples were divided into five groups according to their total motile sperm count: Group I, total motile sperm count sperm count 1-5 × 10(6) ; group III, total motile sperm count 5-10 × 10(6) ; group IV, total motile sperm count 10-20 × 10(6) ; and group V, total motile sperm count >20 × 10(6) (which was considered a normal total motile sperm count value). Then, couples were grouped into an abnormal and normal total motile sperm count group. The groups were compared regarding intracytoplasmic sperm injection outcomes. The predictive values of WHO 2010 cut-off values and total motile sperm count for the intracytoplasmic sperm injection outcomes were also investigated. The fertilization rate was lower in total motile sperm count group I compared to total motile sperm count group V (72.5 ± 17.6 vs. 84.9 ± 14.4, p = 0.011). The normal total motile sperm count group had a higher fertilization rate (84.9 ± 14.4 vs. 81.1 ± 15.8, p = 0.016) and lower miscarriage rate (17.9% vs. 29.5%, p = 0.041) compared to the abnormal total motile sperm count group. The total motile sperm count was the only parameter that demonstrated a predictive value for the formation of high-quality embryos on D2 (OR: 1.18, p = 0.013), formation of high-quality embryos on D3 (OR: 1.12, p = 0.037), formation of blastocysts on D5 (OR: 1.16, p = 0.011), blastocyst expansion grade on D5

Sperm motility and morphology as changing parameters linked to sperm count variations.

OpenAIRE

Dua A; Vaidya S

1996-01-01

Variations in semen analyses of 177 males over a 1 year period were assessed. The average means of total counts, motility, morphology, total motile count and non-motile % were determined for 5 classes of patients ranging from azoospermic to normospermic. Positive relationships between a falling sperm count, a decrease in motility and total motile counts were seen. Also, increasingly, abnormal forms were found with lower sperm counts.

Polyclonal VDAC3 antibody decreases human sperm motility: a novel approach to male contraception

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Asmarinah Asmarinah

2011-02-01

Full Text Available Background: Voltage dependent anion channels (VDAC mediate transport of anions, cations and ATP which play an important role in sperm motility. This study was aimed to examine the effect of polyclonal VDAC3 antiserum to human sperm motility.Methods: Polyclonal VDAC3 antiserum used in this study was produced in rabbits by immunization of VDAC3-specific synthetic peptides.  Preimmunserum was collected before immunization and used for control experiment. Recognition of VDAC3 antiserum to antigen in human sperm was performed by western blot. Thirty sperm samples obtained from fertile men which had high quality of sperm motility were washed and collected by Percoll gradient. Sperm motility was assessed by means of evaluation of sperm velocity (seconds per 0.1 mm distance and the number of unmoved sperm (million per ml which were observed 0 minute, 30 minutes and 60 minutes after addition of VDAC3 antiserum and preimmunserum as a control. Both data were analyzed by SPSS 13.0 software.Results: VDAC3 antiserum recognized VDAC3 protein in human sperm. Statistical analysis demonstrated that there were increasing numbers of unmoved spermatozoa after addition of anti-VDAC3 antiserum in vitro for 60 minutes observation compared with preimmunserum (control. We found also that sperm velocity decreased signifi cantly after giving anti-VDAC3 antiserum in vitro for 0 minute, 30 minutes, and 60 minutes compared with pre-immunee serum (control.Conclusion: VDAC3 antiserum can decrease motility of human sperm. and may provide a novel principle of male contraception in the future. (Med J Indones 2011; 20:5-10Keywords: VDAC3 antiserum, sperm, motility, contraception

Automated Sperm Head Detection Using Intersecting Cortical Model Optimised by Particle Swarm Optimization.

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Tan, Weng Chun; Mat Isa, Nor Ashidi

2016-01-01

In human sperm motility analysis, sperm segmentation plays an important role to determine the location of multiple sperms. To ensure an improved segmentation result, the Laplacian of Gaussian filter is implemented as a kernel in a pre-processing step before applying the image segmentation process to automatically segment and detect human spermatozoa. This study proposes an intersecting cortical model (ICM), which was derived from several visual cortex models, to segment the sperm head region. However, the proposed method suffered from parameter selection; thus, the ICM network is optimised using particle swarm optimization where feature mutual information is introduced as the new fitness function. The final results showed that the proposed method is more accurate and robust than four state-of-the-art segmentation methods. The proposed method resulted in rates of 98.14%, 98.82%, 86.46% and 99.81% in accuracy, sensitivity, specificity and precision, respectively, after testing with 1200 sperms. The proposed algorithm is expected to be implemented in analysing sperm motility because of the robustness and capability of this algorithm.

A comparison of sperm agglutination and immobilization assays with a quantitative ELISA for anti-sperm antibody in serum.

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Lynch, D M; Leali, B A; Howe, S E

1986-08-01

An enzyme-linked immunosorbent assay (ELISA) that quantitates antisperm antibody in serum was compared with standard sperm agglutination and immobilization assays with the use of sera from 40 normal and 292 subfertile individuals. Quantitation of the assay was accomplished by standardizing assay parameters, including the incorporation of a standard reference curve, the number of whole target sperm, the optimal dilution of serum, the selection of microtiter plate, and the time and temperatures involved in the adsorption and incubation phases. With this method, the level of antisperm antibody binding to target sperm in 40 normal fertile individuals was found to be 2.3 (+/- 1.1 standard deviation [SD]) fg immunoglobulin (Ig)/sperm. An increased mean level of 7.4 +/- 3.7 fg Ig/sperm was determined in 84 infertile patients with positive agglutination and/or immobilization tests. In 208 individuals with negative agglutination and immobilization tests the mean concentration of antisperm antibody was 2.5 +/- 1.3 fg Ig/sperm. Postvasectomy patients assayed by this method had a mean Ig binding value of 7.1 +/- 2.4 fg Ig/sperm. The infertile group with positive agglutination and/or immobilization tests had a significantly higher mean antisperm antibody level than the normal fertile group, according to the Student's t-test for independent samples (P less than 0.001). This indirect serum-based assay reproducibly quantitates antisperm antibody binding to whole target sperm, suggests the normal and abnormal levels of antisperm antibody, and correlates with standard functional assays.

Mitochondrial respiratory efficiency is positively correlated with human sperm motility.

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Ferramosca, Alessandra; Provenzano, Sara Pinto; Coppola, Lamberto; Zara, Vincenzo

2012-04-01

To correlate sperm mitochondrial respiratory efficiency with variations in sperm motility and with sperm morphologic anomalies. Sperm mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically-treated sperm cells. A possible relationship among sperm mitochondrial respiratory efficiency, sperm motility, and morphologic anomalies was investigated. Mitochondrial respiratory efficiency was positively correlated with sperm motility and negatively correlated with the percentage of immotile spermatozoa. Moreover, midpiece defects impaired mitochondrial functionality. Our data indicate that an increase in sperm motility requires a parallel increase in mitochondrial respiratory capacity, thereby supporting the fundamental role played by mitochondrial oxidative phosphorylation in sperm motility of normozoospermic subjects. These results are of physiopathological relevance because they suggest that disturbances of sperm mitochondrial function and of energy production could be responsible for asthenozoospermia. Copyright © 2012 Elsevier Inc. All rights reserved.

Office-based sperm concentration: A simplified method for ...

African Journals Online (AJOL)

Methods: Semen samples from 51 sperm donors were used. Following swim-up separation, the sperm concentration of the retrieved motile fraction was counted, as well as progressive motile sperm using a standardised wet preparation. The number of sperm in a 10 μL droplet covered with a 22 × 22 mm coverslip was ...

Sperm flagellum volume determines freezability in red deer spermatozoa.

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José Luis Ros-Santaella

Full Text Available The factors affecting the inter-individual differences in sperm freezability is a major line of research in spermatology. Poor sperm freezability is mainly characterised by a low sperm velocity, which in turn is associated with low fertility rates in most animal species. Studies concerning the implications of sperm morphometry on freezability are quite limited, and most of them are based on sperm head size regardless of the structural parts of the flagellum, which 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. Sperm samples from thirty-three stags were frozen and classified as good freezers (GF or bad freezers (BF at two hours post-thawing using three sperm kinetic parameters which are strongly correlated with fertility in this species. Fourteen stags were clearly identified as GF, whereas nineteen were BF. No significant difference in sperm head size between the two groups was found. On the contrary, the GF exhibited a lower principal piece volume than the BF (6.13 µm3 vs 6.61 µm3, respectively, p = 0.006. The volume of the flagellum structures showed a strong negative relationship with post-thawing sperm velocity. For instance, the volume of the sperm principal piece was negatively correlated with sperm velocity at two hours post-thawing (r = -0.60; p<0.001. Our results clearly show that a higher volume of the sperm principal piece results in poor freezability, and highlights the key role of flagellum size in sperm cryopreservation success.

Branchial Cilia and Sperm Flagella Recruit Distinct Axonemal Components

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Konno, Alu; Shiba, Kogiku; Cai, Chunhua; Inaba, Kazuo

2015-01-01

Eukaryotic cilia and flagella have highly conserved 9 + 2 structures. They are functionally diverged to play cell-type-specific roles even in a multicellular organism. Although their structural components are therefore believed to be common, few studies have investigated the molecular diversity of the protein components of the cilia and flagella in a single organism. Here we carried out a proteomic analysis and compared protein components between branchial cilia and sperm flagella in a marine invertebrate chordate, Ciona intestinalis. Distinct feature of protein recruitment in branchial cilia and sperm flagella has been clarified; (1) Isoforms of α- and β-tubulins as well as those of actins are distinctly used in branchial cilia or sperm flagella. (2) Structural components, such as dynein docking complex, tektins and an outer dense fiber protein, are used differently by the cilia and flagella. (3) Sperm flagella are specialized for the cAMP- and Ca2+-dependent regulation of outer arm dynein and for energy metabolism by glycolytic enzymes. Our present study clearly demonstrates that flagellar or ciliary proteins are properly recruited according to their function and stability, despite their apparent structural resemblance and conservation. PMID:25962172

Sperm motility and morphology as changing parameters linked to sperm count variations.

Directory of Open Access Journals (Sweden)

Dua A

1996-10-01

Full Text Available Variations in semen analyses of 177 males over a 1 year period were assessed. The average means of total counts, motility, morphology, total motile count and non-motile % were determined for 5 classes of patients ranging from azoospermic to normospermic. Positive relationships between a falling sperm count, a decrease in motility and total motile counts were seen. Also, increasingly, abnormal forms were found with lower sperm counts.

The different types of sperm morphology and behavior within a single species: Why do sperm of squid sneaker males form a cluster?

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Hirohashi, Noritaka; Iwata, Yoko

2013-11-01

Some coastal squids exhibit male dimorphism (large and small body size) that is linked to mating behaviors. Large "consort" males compete with other, rival males to copulate with a female, and thereby transfer their spermatophores to her internal site around the oviduct. Small "sneaker" males rush to a single female or copulating pair and transfer spermatophores to her external body surface around the seminal receptacle near the mouth. We previously found that in Loligo bleekeri, sneaker sperm are ~50% longer than consort sperm, and only the sneaker sperm, once ejaculated from the spermatophore (sperm mass), form a cluster because of chemoattraction toward their own respiratory CO2. Here, we report that sperm clusters are able to move en masse. Because a fraction of ejaculated sperm from a sneaker's spermatophore are eventually located in the female's seminal receptacle, we hypothesize that sperm clustering facilitates collective migration to the seminal receptacle or an egg micropyle. Sperm clustering is regarded as a cooperative behavior that may have evolved by sperm competition and/or physical and physiological constraints imposed by male mating tactics.

Glycolysis and mitochondrial respiration in mouse LDHC-null sperm.

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Odet, Fanny; Gabel, Scott; London, Robert E; Goldberg, Erwin; Eddy, Edward M

2013-04-01

We demonstrated previously that a knockout (KO) of the lactate dehydrogenase type C (Ldhc) gene disrupted male fertility and caused a considerable reduction in sperm glucose consumption, ATP production, and motility. While that study used mice with a mixed genetic background, the present study used C57BL/6 (B6) and 129S6 (129) Ldhc KO mice. We found that B6 KO males were subfertile and 129 KO males were infertile. Sperm from 129 wild-type (WT) mice have a lower glycolytic rate than sperm from B6 WT mice, resulting in a greater reduction in ATP production in 129 KO sperm than in B6 KO sperm. The lower glycolytic rate in 129 sperm offered a novel opportunity to examine the role of mitochondrial respiration in sperm ATP production and motility. We observed that in media containing a mitochondrial substrate (pyruvate or lactate) as the sole energy source, ATP levels and progressive motility in 129 KO sperm were similar to those in 129 WT sperm. However, when glucose was added, lactate was unable to maintain ATP levels or progressive motility in 129 KO sperm. The rate of respiration (ZO2) was high when 129 KO or WT sperm were incubated with lactate alone, but addition of glucose caused a reduction in ZO2. These results indicate that in the absence of glucose, 129 sperm can produce ATP via oxidative phosphorylation, but in the presence of glucose, oxidative phosphorylation is suppressed and the sperm utilize aerobic glycolysis, a phenomenon known as the Crabtree effect.

Sperm with large nuclear vacuoles and semen quality in the evaluation of male infertility.

Science.gov (United States)

Komiya, Akira; Watanabe, Akihiko; Kawauchi, Yoko; Fuse, Hideki

2013-02-01

This study compared the sperm nuclear vacuoles and semen quality in the evaluation of male infertility. One hundred and forty-two semen samples were obtained from patients who visited the Male Infertility Clinic at Toyama University Hospital. Semen samples were evaluated by conventional semen analyses and the Sperm Motility Analysis System (SMAS). In addition, spermatozoa were analyzed at 3,700-6,150x magnification on an inverted microscope equipped with DIC/Nomarski differential interference contrast optics. A large nuclear vacuole (LNV) was defined as one or more vacuoles with the maximum diameter showing > 50% width of the sperm head. The percentage of spermatozoa with LNV (% LNV) was calculated for each sample. Correlations between the % LNV and parameters in SMAS and conventional semen analyses were analyzed. Processed motile spermatozoa from each sample were evaluated. The mean age of patients was 35 years old. Semen volume was 2.9 ± 1.6mL (0.1-11.0; mean ± standard deviation, minimum-maximum), sperm count was 39.3 ± 54.9 (x10(6)/mL, 0.01-262.0), sperm motility was 25.1 ± 17.8% (0-76.0), and normal sperm morphology was 10.3 ± 10.1% (0-49.0). After motile spermatozoa selection, we could evaluate % LNV in 125 ejaculates (88.0%) and at least one spermatozoon with LNV was observed in 118 ejaculates (94.4%). The percentage of spermatozoa with LNV was 28.0 ± 22.4% (0-100) and % LNV increased significantly when semen quality decreased. The correlation between the % LNV and the semen parameters was weak to moderate; correlation coefficients were -0.3577 in sperm count (p sperm motility (p = 0.0084), -0.2769 in motile sperm count (p = 0.019), -0.2419 in total motile sperm count (p = 0.0070), and -0.1676 in normal sperm morphology (p = 0.0639). The % LNV did not show a significant correlation with the SMAS parameters except for weak correlation to beat/cross frequency (r = -0.2414, p = 0.0071). The percentage of

Relationship of leptin administration with production of reactive oxygen species, sperm DNA fragmentation, sperm parameters and hormone profile in the adult rat.

Science.gov (United States)

Abbasihormozi, Shima; Shahverdi, Abdolhossein; Kouhkan, Azam; Cheraghi, Javad; Akhlaghi, Ali Asghar; Kheimeh, Abolfazl

2013-06-01

Leptin, an adipose tissue-derived hormone, plays an important role in energy homeostasis and metabolism, and in the neuroendocrine and reproductive systems. The function of leptin in male reproduction is unclear; however, it is known to affect sex hormones, sperm motility and its parameters. Leptin induces mitochondrial superoxide production in aortic endothelia and may increase oxidative stress and abnormal sperm production in leptin-treated rats. This study aims to evaluate whether exogenous leptin affects sperm parameters, hormone profiles, and the production of reactive oxygen species (ROS) in adult rats. A total of 65 Sprague-Dawley rats were divided into three treated groups and a control group. Treated rats received daily intraperitoneal injections of 5, 10 and 30 μg/kg of leptin administered for a duration of 7, 15, and 42 days. Control rats were given 0.1 mL of 0.9 % normal saline for the same period. One day after final drug administration, we evaluated serum specimens for follicle-stimulating hormone (FSH), leutinizing hormone (LH), free testosterone (FT), and total testosterone (TT) levels. Samples from the rat epididymis were also evaluated for sperm parameters and motility characteristics by a Computer-Aided Semen Analysis (CASA) system. Samples were treated with 2',7'-dichlorofluorescein-diacetate (DCFH-DA) and analyzed using flow cytometry and TUNEL to determine the impact of leptin administration on sperm DNA fragmentation. According to CASA, significant differences in all sperm parameters in leptin-treated rats and their age-matched controls were detected, except for TM, ALH and BCF. Serum FSH and LH levels were significantly higher in rats that received 10 and 30 μg/kg of leptin compared to those treated with 5 μg/kg of leptin in the same group and control rats (P control group (P hormone profile modulation.

Intracellular pH in sperm physiology.

Science.gov (United States)

Nishigaki, Takuya; José, Omar; González-Cota, Ana Laura; Romero, Francisco; Treviño, Claudia L; Darszon, Alberto

2014-08-01

Intracellular pH (pHi) regulation is essential for cell function. Notably, several unique sperm ion transporters and enzymes whose elimination causes infertility are either pHi dependent or somehow related to pHi regulation. Amongst them are: CatSper, a Ca(2+) channel; Slo3, a K(+) channel; the sperm-specific Na(+)/H(+) exchanger and the soluble adenylyl cyclase. It is thus clear that pHi regulation is of the utmost importance for sperm physiology. This review briefly summarizes the key components involved in pHi regulation, their characteristics and participation in fundamental sperm functions such as motility, maturation and the acrosome reaction. Copyright © 2014 Elsevier Inc. All rights reserved.

Sperm subpopulations in avian species: a comparative study between the rooster (Gallus domesticus) and Guinea fowl (Numida meleagris).

Science.gov (United States)

García-Herreros, Manuel

2016-01-01

The main aims of this research were to study possible differences in objective morphometric sperm characteristics, establish normative sperm morphometry standards, and evaluate the presumed different subpopulation distribution of avian spermatozoa from the rooster (Gallus domesticus ) and Guinea fowl (Numida meleagris ) as model avian species. Seventy-two ejaculates (36 per species studied) were obtained manually, following a training period involving gently combined dorso-abdominal and lumbo-sacral massage of the birds. Ejaculates were processed for volume, sperm concentration, viability, motility, and morphology. Moreover, samples were submitted for sperm morphometric assessment using objective Computer-Assisted Semen Analysis for Morphometry (CASA-Morph) methods, with sperm morphometric descriptors evaluated by Principal Component Analysis (PCA) and multivariate clustering analyses. There were several differences observed between the avian species in values obtained for ejaculate volume and sperm concentration (P rooster and Guinea fowl, respectively). Moreover, the distribution of the sperm subpopulations was found to be structurally different between species. In conclusion, our findings from using CASA-Morph methods indicate pronounced sperm morphometric variation between these two avian species. Because of the strong differences observed in morphometric parameter values and their subpopulation distribution, these results suggest that application of objective analytical methods such as CASA-Morph could substantially improve the reliability of comparative studies and help establish valid normative sperm morphological values for avian species.

Single-cell-based evaluation of sperm progressive motility via fluorescent assessment of mitochondria membrane potential.

Science.gov (United States)

Moscatelli, Natalina; Spagnolo, Barbara; Pisanello, Marco; Lemma, Enrico Domenico; De Vittorio, Massimo; Zara, Vincenzo; Pisanello, Ferruccio; Ferramosca, Alessandra

2017-12-20

Sperm cells progressive motility is the most important parameter involved in the fertilization process. Sperm middle piece contains mitochondria, which play a critical role in energy production and whose proper operation ensures the reproductive success. Notably, sperm progressive motility is strictly related to mitochondrial membrane potential (MMP) and consequently to mitochondrial functionality. Although previous studies presented an evaluation of mitochondrial function through MMP assessment in entire sperm cells samples, a quantitative approach at single-cell level could provide more insights in the analysis of semen quality. Here we combine laser scanning confocal microscopy and functional fluorescent staining of mitochondrial membrane to assess MMP distribution among isolated spermatozoa. We found that the sperm fluorescence value increases as a function of growing progressive motility and that such fluorescence is influenced by MMP disruptors, potentially allowing for the discrimination of different quality classes of sperm cells in heterogeneous populations.

Reduction in sperm count and increase in abnormal sperm in the mouse following x-irradiation or injection of 22Na

International Nuclear Information System (INIS)

Harrison, A.; Moore, P.C.

1980-01-01

The accumulated reduction in sperm count and increase in the number of aberrant sperm in the mouse were used to compare acute X-irradiation with protracted radiation from an injection of 22 Na. The effects per unit of absorbed dose of acute and protracted radiation were similar on sperm count, but in respect of numbers of abnormal sperm the 22 Na induced a significantly greater maximum than X-rays. (author)

Semen quality parameters, their inter-relationship and post-washing sperm attributes of Rhode Island Red roosters

Directory of Open Access Journals (Sweden)

R. Richard Churchil

2014-12-01

Full Text Available Aim: The present experiments were conducted (a to evaluate the semen attributes of older Rhode Island Red (RIR roosters and the inter-trait relationships, (b to test sperm washing and storage duration suitable for gene transfer experiments. Materials and Methods: The semen characteristics of older RIR roosters were studied, and Pearson correlation analysis was done to demonstrate the inter-trait relationships. Progressive motility and percent live sperms were tested at different post-washing intervals to identify suitable sperm processing conditions for gene transfer experiments. Results: The volume, appearance score, initial motility, sperm count and percent live and abnormal spermatozoa were 0.38 ml, 3.58, 80.34%, 4.03 × 109 sperms/ml, 83.18% and 4.52% respectively. Positive correlation was observed among appearance score, motility, live sperm and sperm count. Semen volume is negatively correlated with all the other characters except live sperms, whereas, percent abnormal sperms negatively associated with all the other traits. Significant (p<0.05 decrease in terms of motility and live sperm was recorded at 60 min post-washing. Conclusion: The semen attributes of RIR roosters compares well with the other breeds of chicken. The appearance score can be used to assess fertility where microscopic evaluation facilities are limited. The sperm washing protocol tested in the experiment is suitable for gene transfer experiments.

Sperm cryopreservation in fish and shellfish.

Science.gov (United States)

Tiersch, Terrence R; Yang, Huiping; Jenkins, Jill A; Dong, Qiaoxiang

2007-01-01

Initial success in sperm cryopreservation came at about the same time for aquatic species and livestock. However, in the 50-plus years since then cryopreserved sperm of livestock has grown into a billion-dollar global industry, while despite work in some 200 species with well over 200 published reports, cryopreservation of aquatic species sperm remains essentially a research activity with little commercial application. Most research has focused on large-bodied culture and sport fishes, such as salmonids, carps, and catfishes, and mollusks such as commercially important oyster and abalone species. However, only a handful of studies have addressed sperm cryopreservation in small fishes, such as zebrafish, and in endangered species. Overall, this work has yielded techniques that are being applied with varying levels of success around the world. Barriers to expanded application include a diverse and widely distributed literature base, technical problems, small sperm volumes, variable results, a general lack of access to the technology, and most importantly, the lack of standardization in practices and reporting. The benefits of cryopreservation include at least five levels of improvements for existing industries and for creation of new industries. First, cryopreservation can be used to improve existing hatchery operations by providing sperm on demand and simplifying the timing of induced spawning. Second, frozen sperm can enhance efficient use of facilities and create new opportunities in the hatchery by eliminating the need to maintain live males, potentially freeing resources for use with females and larvae. Third, valuable genetic lineages such as endangered species, research models, or improved farmed strains can be protected by storage of frozen sperm. Fourth, cryopreservation opens the door for rapid genetic improvement. Frozen sperm can be used in breeding programs to create improved lines and shape the genetic resources available for aquaculture. Finally

21 CFR 173.275 - Hydrogenated sperm oil.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Hydrogenated sperm oil. 173.275 Section 173.275... CONSUMPTION Solvents, Lubricants, Release Agents and Related Substances § 173.275 Hydrogenated sperm oil. The food additive hydrogenated sperm oil may be safely used in accordance with the following prescribed...

COMP-1 promotes competitive advantage of nematode sperm.

Science.gov (United States)

Hansen, Jody M; Chavez, Daniela R; Stanfield, Gillian M

2015-03-19

Competition among sperm to fertilize oocytes is a ubiquitous feature of sexual reproduction as well as a profoundly important aspect of sexual selection. However, little is known about the cellular mechanisms sperm use to gain competitive advantage or how these mechanisms are regulated genetically. In this study, we utilize a forward genetic screen in Caenorhabditis elegans to identify a gene, comp-1, whose function is specifically required in competitive contexts. We show that comp-1 functions in sperm to modulate their migration through and localization within the reproductive tract, thereby promoting their access to oocytes. Contrary to previously described models, comp-1 mutant sperm show no defects in size or velocity, thereby defining a novel pathway for preferential usage. Our results indicate not only that sperm functional traits can influence the outcome of sperm competition, but also that these traits can be modulated in a context-dependent manner depending on the presence of competing sperm.

Immature germ cells in semen ? correlation with total sperm count and sperm motility

OpenAIRE

Patil, Priya S.; Humbarwadi, Rajendra S.; Patil, Ashalata D.; Gune, Anita R.

2013-01-01

Background: Current data regarding infertility suggests that male factor contributes up to 30% of the total cases of infertility. Semen analysis reveals the presence of spermatozoa as well as a number of non-sperm cells, presently being mentioned in routine semen report as "round cells" without further differentiating them into leucocytes or immature germ cells. Aim: The aim of this work was to study a simple, cost-effective, and convenient method for differentiating the round cells in se...

An automatic system to study sperm motility and energetics.

Science.gov (United States)

Shi, Linda Z; Nascimento, Jaclyn M; Chandsawangbhuwana, Charlie; Botvinick, Elliot L; Berns, Michael W

2008-08-01

An integrated robotic laser and microscope system has been developed to automatically analyze individual sperm motility and energetics. The custom-designed optical system directs near-infrared laser light into an inverted microscope to create a single-point 3-D gradient laser trap at the focal spot of the microscope objective. A two-level computer structure is described that quantifies the sperm motility (in terms of swimming speed and swimming force) and energetics (measuring mid-piece membrane potential) using real-time tracking (done by the upper-level system) and fluorescent ratio imaging (done by the lower-level system). The communication between these two systems is achieved by a gigabit network. The custom-built image processing algorithm identifies the sperm swimming trajectory in real-time using phase contrast images, and then subsequently traps the sperm by automatically moving the microscope stage to relocate the sperm to the laser trap focal plane. Once the sperm is stably trapped (determined by the algorithm), the algorithm can also gradually reduce the laser power by rotating the polarizer in the laser path to measure the trapping power at which the sperm is capable of escaping the trap. To monitor the membrane potential of the mitochondria located in a sperm's mid-piece, the sperm is treated with a ratiometrically-encoded fluorescent probe. The proposed algorithm can relocate the sperm to the center of the ratio imaging camera and the average ratio value can be measured in real-time. The three parameters, sperm escape power, sperm swimming speed and ratio values of the mid-piece membrane potential of individual sperm can be compared with respect to time. This two-level automatic system to study individual sperm motility and energetics has not only increased experimental throughput by an order of magnitude but also has allowed us to monitor sperm energetics prior to and after exposure to the laser trap. This system should have application in both the

Localized accumulation of cytosolic calcium near the fused sperm is associated with the calcium- and voltage-dependent block of sperm entry in the sea urchin egg.

Science.gov (United States)

Ivonnet, Pedro I; Mohri, Tatsuma; McCulloh, David H

2017-10-01

Interaction of the sperm and egg depolarizes the egg membrane, allowing the sperm to enter; however, if the egg membrane is not allowed to depolarize from its resting potential (e.g., by voltage-clamp), the sperm will not enter. Previous studies demonstrated that sperm entry into sea urchin eggs that are voltage-clamped at negative membrane potentials is regulated both by the egg's membrane potential and a voltage-dependent influx of calcium into the egg. In these cases, electrical or cytoplasmic continuity (sperm-egg membrane fusion) occurs at negative membrane potentials, but subsequent loss of cytoplasmic continuity results in failure of sperm entry (unfusion). The work presented herein examined where, in relation to the sperm, and when, in relation to the sperm-induced electrophysiological events, the egg's calcium influx occurs, and how these events relate to successful or failed sperm entry. When sperm entered the egg, elevation of intracellular calcium concentration ([Ca 2+ ] i ) began near the fused sperm on average 5.9 s after sperm-egg membrane fusion. Conversely, when sperm failed to enter the egg, [Ca 2+ ] i elevated near the site of sperm-egg fusion on average 0.7 s after sperm-egg membrane fusion, which is significantly earlier than in eggs for which sperm entered. Therefore, the accumulation of calcium near the site of sperm-egg fusion is spatially and temporally consistent with the mechanism that may be responsible for loss of cytoplasmic continuity and failure of sperm entry. © 2017 Wiley Periodicals, Inc.

[Impact of sperm DNA and acrosome integrity and acrosome reaction rate on outcomes of rescue intracytoplasmic sperm injection].

Science.gov (United States)

He, Yongzhi; Li, Dawen; Cheng, Junping; Huo, Zhongchao; Huang, Hongyi; Xiao, Xin

2016-01-01

Objective To explore the effects of sperm DNA integrity rate, acrosome integrity rate and acrosome reaction rate on the outcomes of rescue intracytoplasmic sperm injection (ICSI). This retrospective analysis was conducted among 97 infertile couples receiving rescue ICSI due to failure of in vitro fertilization procedures in our Reproductive Medicine Center. Of these 97 women, 41 had clinical pregnancy and 56 did not, and the effects of sperm DNA integrity rate (estimated by DNA fragmentation index, DFI), acrosome integrity rate and acrosome reaction rate on rescue ICSI outcomes were analyzed. No significant difference was found in paternal age, testosterone value, testicular volume, FSH, female patient' age or the number of eggs retrieved between the two groups (P>0.05), but the infertility years was significantly shorter in the pregnancy group than in the non-pregnancy group (Prate and cleavage rate were similar between the two groups (P>0.05), but the good embryo rate was significantly higher in the pregnancy group (Preaction rate did not differ significantly between the two groups (P>0.05), but the acrosome integrity rate was significantly higher in the pregnancy group (Prate, acrosome integrity or acrosome reaction rate were not correlated with the fertilization rate, cleavage rate or good embryo rate (P>0.05). The pregnancy rate, twin and single fetus rates were 42.3%, 10.3% and 32.0% in this cohort after recue ICSI, respectively. Rescue ICSI is an effective treatment after failed in vitro fertilization procedure, and sperm acrosome integrity rate is associated with the outcome of rescue ICSI.

Time bound changes (in 24 h) in human sperm motility and level of ...

African Journals Online (AJOL)

A detailed sperm motility study for 24 h after collection was done. The level of calcium and magnesium in seminal plasma during this period was also seen to understand the role of these electrolytes on sperm motility. Good care was taken in selection of subjects (young and healthy), collection and pre-physical analysis of ...

Tolerance to environmental desiccation in moss sperm.

Science.gov (United States)

Shortlidge, Erin E; Rosenstiel, Todd N; Eppley, Sarah M

2012-05-01

• Sexual reproduction in mosses requires that sperm be released freely into the environment before finding and fertilizing a receptive female. After release from the male plant, moss sperm may experience a range of abiotic stresses; however, few data are available examining stress tolerance of moss sperm and whether there is genetic variation for stress tolerance in this important life stage. • Here, we investigated the effects of environmental desiccation and recovery on the sperm cells of three moss species (Bryum argenteum, Campylopus introflexus, and Ceratodon purpureus). • We found that a fraction of sperm cells were tolerant to environmental desiccation for extended periods (d) and that tolerance did not vary among species. We found that this tolerance occurs irrespective of ambient dehydration conditions, and that the addition of sucrose during dry-down improved cell recovery. Although we observed no interspecific variation, significant variation among individuals within species in sperm cell tolerance to environmental desiccation was observed, suggesting selection could potentially act on this basic reproductive trait. • The observation of desiccation-tolerant sperm in multiple moss species has important implications for understanding bryophyte reproduction, suggesting the presence of a significant, uncharacterized complexity in the ecology of moss mating systems. © 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.

Are there intracellular Ca2+ oscillations correlated with flagellar beating in human sperm? A three vs. two-dimensional analysis.

Science.gov (United States)

Corkidi, G; Montoya, F; Hernández-Herrera, P; Ríos-Herrera, W A; Müller, M F; Treviño, C L; Darszon, A

2017-09-01

Are there intracellular Ca2+ ([Ca2+]i) oscillations correlated with flagellar beating in human sperm? The results reveal statistically significant [Ca2+]i oscillations that are correlated with the human sperm flagellar beating frequency, when measured in three-dimensions (3D). Fast [Ca2+]i oscillations that are correlated to the beating flagellar frequency of cells swimming in a restricted volume have been detected in hamster sperm. To date, such findings have not been confirmed in any other mammalian sperm species. An important question that has remained regarding these observations is whether the fast [Ca2+]i oscillations are real or might they be due to remaining defocusing effects of the Z component arising from the 3D beating of the flagella. Healthy donors whose semen samples fulfill the WHO criteria between the age of 18-28 were selected. Cells from at least six different donors were utilized for analysis. Approximately the same number of experimental and control cells were analyzed. Motile cells were obtained by the swim-up technique and were loaded with Fluo-4 (Ca2+ sensitive dye) or with Calcein (Ca2+ insensitive dye). Ni2+ was used as a non-specific plasma membrane Ca2+ channel blocker. Fluorescence data and flagella position were acquired in 3D. Each cell was recorded for up to 5.6 s within a depth of 16 microns with a high speed camera (coupled to an image intensifier) acquiring at a rate of 3000 frames per second, while an oscillating objective vibrated at 90 Hz via a piezoelectric device. From these samples, eight experimental and nine control sperm cells were analyzed in both 2D and 3D. We have implemented a new system that allows [Ca2+]i measurements of the human sperm flagellum beating in 3D. These measurements reveal statistically significant [Ca2+]i oscillations that correlate with the flagellar beating frequency. These oscillations may arise from intracellular sources and/or Ca2+ transporters, as they were insensitive to external Ni2+, a non

Sperm storage induces an immunity cost in ants

DEFF Research Database (Denmark)

Baer, Boris; Armitage, Sophie A O; Boomsma, Jacobus J

2006-01-01

Ant queens are among the most long-lived insects known. They mate early in adult life and maintain millions of viable sperm in their sperm storage organ until they die many years later. Because they never re-mate, the reproductive success of queens is ultimately sperm-limited, but it is not known...... what selective forces determine the upper limit to sperm storage. Here we show that sperm storage carries a significant cost of reduced immunity during colony founding. Newly mated queens of the leaf-cutting ant Atta colombica upregulate their immune response shortly after completing their nest burrow...

CASAnova: a multiclass support vector machine model for the classification of human sperm motility patterns.

Science.gov (United States)

Goodson, Summer G; White, Sarah; Stevans, Alicia M; Bhat, Sanjana; Kao, Chia-Yu; Jaworski, Scott; Marlowe, Tamara R; Kohlmeier, Martin; McMillan, Leonard; Zeisel, Steven H; O'Brien, Deborah A

2017-11-01

The ability to accurately monitor alterations in sperm motility is paramount to understanding multiple genetic and biochemical perturbations impacting normal fertilization. Computer-aided sperm analysis (CASA) of human sperm typically reports motile percentage and kinematic parameters at the population level, and uses kinematic gating methods to identify subpopulations such as progressive or hyperactivated sperm. The goal of this study was to develop an automated method that classifies all patterns of human sperm motility during in vitro capacitation following the removal of seminal plasma. We visually classified CASA tracks of 2817 sperm from 18 individuals and used a support vector machine-based decision tree to compute four hyperplanes that separate five classes based on their kinematic parameters. We then developed a web-based program, CASAnova, which applies these equations sequentially to assign a single classification to each motile sperm. Vigorous sperm are classified as progressive, intermediate, or hyperactivated, and nonvigorous sperm as slow or weakly motile. This program correctly classifies sperm motility into one of five classes with an overall accuracy of 89.9%. Application of CASAnova to capacitating sperm populations showed a shift from predominantly linear patterns of motility at initial time points to more vigorous patterns, including hyperactivated motility, as capacitation proceeds. Both intermediate and hyperactivated motility patterns were largely eliminated when sperm were incubated in noncapacitating medium, demonstrating the sensitivity of this method. The five CASAnova classifications are distinctive and reflect kinetic parameters of washed human sperm, providing an accurate, quantitative, and high-throughput method for monitoring alterations in motility. © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Cryopreservation of sperm bundles (spermatozeugmata) from endangered livebearing goodeids.

Science.gov (United States)

Liu, Yue; Torres, Leticia; Tiersch, Terrence R

2018-04-14

More than half of fishes in the family Goodeidae are considered to be endangered, threatened, or vulnerable. Sperm cryopreservation is an effective tool for conserving genetic resources of imperiled populations, but development of protocols with livebearing fishes faces numerous challenges including the natural packaging of sperm into bundles. In this study the cryopreservation of sperm bundles (spermatozeugmata) of three goodeids species was evaluated. Sperm quality was evaluated by activation with NaCl-NaOH solution (at 300 mOsmol/kg and pH 11.8), and analysis of dissociable bundles and dissociation duration. Using Redtail Splitfin (Xenotoca eiseni) as a model, the effects of cryoprotectants (dimethyl sulfoxide, methanol, and glycerol) with different concentrations (5-15% v/v %), equilibration exposure times (1-60 min), cooling rates (5-40 °C/min), concentrations (4 × 10 4 -4 × 10 6 bundles/ml), buffers (HBSS, PBS and NaCl), and buffer osmolalities (200-400 mOsmol/kg) were investigated. After cooling and thawing, sperm bundles maintained their packed form. A specific protocol was developed (10% dimethyl sulfoxide, 20-min equilibration, 10 °C/min cooling rate, 4 × 10 6 bundles/ml, and 300 mOsmol/kg HBSS). This protocol yielded 89 ± 5% of post-thaw dissociable bundles with 209 ± 10 s of dissociation duration for X. eiseni, 96 ± 9% with 814 ± 14 s for Blackfin Goodea (Goodea atripinni), and 66 ± 2% with 726 ± 25 s for Striped Goodeid (Ataeniobius toweri). This is the first study of cryopreservation of sperm within bundles for livebearing fishes and provides a basis for establishment of germplasm repositories for goodeids and other livebearers. Copyright © 2018 Elsevier Inc. All rights reserved.

[The effects of exposure to ozone on sperm quality in Wuhan].

Science.gov (United States)

Tian, X J; Wang, X C; Ye, B; Li, C L; Zhang, Y; Ma, L

2017-03-06

Objective: To evaluate the effects of exposure to ozone (O(3)) on sperm quality during different stages of spermatogenesis. Methods: All 1 780 subjects attending to the Reproductive Medicine Center in Renmin Hospital of Wuhan University were recruited from April, 4, 2013 to June, 30, 2015. The subjects were living in Wuhan more than 3 months before attending to the program, aged 20 to 40 years. Semen quality (sperm concentration and sperm count) were measured according to standardized protocols. Corresponding daily 8 hours average concentration of O(3), other polluted concentration, average temperature and relative humidity were collected in different time, including lag 0, 10, 70 and 90 d, and lag 0-9 d, lag 10-14 d, lag 70-90 d and lag 0-90 d. After controlling the age, BMI, education level and other confounders, generalized linear Model was used to investigate the association between O(3) and sperm quality during different stages of spermatogenesis. Results: Average daily concentration of O(3) during the study period was (114.20±74.88) μg/m(3) and the mean values of sperm concentration and count were (76.32±50.17) millions/ml and (164.77 ± 133.05) millions/sample, respectively. Exposure to O(3) was associated with decreasing sperm concentration and count. For every 1 μg/m(3) increase of O(3), the decrease of sperm concentration during lag 10, lag 0-9 and lag 10-14 days exposure windows were 0.040 (95% CI: 0.004-0.077) millions/ml, 0.081 (95% CI: 0.003-0.158) millions/ml and 0.059 (95% CI: 0.001-0.116) millions/ml, respectively. And the decrease of sperm count during lag 10, lag 0-9 days exposure windows were 0.105 (95% CI: 0.008-0.202) millions/sample and 0.221 (95% CI: 0.016-0.426) millions/sample. After stratification, in the ozone concentration sperm concentration was 0.249 (95% CI: 0.028-0.470) millions/ml. After sensitivity analysis, the effects of exposure to ozone on sperm concentration and sperm count remained relatively unchanged. Conclusion: Our

Factor V Leiden is associated with increased sperm count.

Science.gov (United States)

van Mens, T E; Joensen, U N; Bochdanovits, Z; Takizawa, A; Peter, J; Jørgensen, N; Szecsi, P B; Meijers, J C M; Weiler, H; Rajpert-De Meyts, E; Repping, S; Middeldorp, S

2017-11-01

Is the thrombophilia mutation factor V Leiden (FVL) associated with an increased total sperm count? Carriers of FVL have a higher total sperm count than non-FVL-carriers, which could not be explained by genetic linkage or by observations in a FVL-mouse model. FVL has a high prevalence in Caucasians despite detrimental health effects. Carriers have been shown to have higher fecundity, which might partly explain this evolutionary paradox. We determined FVL status in two cohorts (Dutch, n = 627; Danish, n = 854) of consecutively included men without known causes for spermatogenic failure, and performed an individual patient data meta-analysis of these two cohorts together with one previously published (Dutch, n = 908) cohort. We explored possible biological underpinnings for the relation between sperm count and FVL, by use of a FVL-mouse model and investigations of genetic linkage. Participants were male partners of subfertile couples (two Dutch cohorts) and young men from the general population (Danish cohort): FVL carrier rate was 4.0%, 4.6% and 7.3%, respectively. There were differences in smoking, abstinence time and age between the cohorts. We corrected for these in the primary analysis, which consisted of a mixed linear effects model, also incorporating unobjectified population differences. In public haplotype data from subjects of European descent, we explored linkage disequilibrium of FVL with all known single nucleotide polymorphisms in a 1.5 MB region around the F5 gene with an R2 cutoff of 0.8. We sequenced exons of four candidate genes hypothesized to be linked to FVL in a subgroup of FVL carriers with extreme sperm count values. The animal studies consisted of never mated 15-18-week-old C57BL/J6 mice heterozygous and homozygous for FVL and wild-type mice. We compared spermatogenesis parameters (normalized internal genitalia weights, epididymis sperm content and sperm motility) between FVL and wild-type mice. Human FVL carriers have a higher total sperm

A Method for Preparation, Storage and Activation of Large Populations of Immotile Sea Urchin Sperm

Science.gov (United States)

Bracho, Geracimo E.; Fritch, Jennifer J.; Tash, Joseph S.

1997-01-01

Reversible protein phosphorylation is associated with initiation and modulation of sperm flagellar motility. Many studies aimed at examining the signal transduction mechanisms underlying the expression of motility have relied on detergent-permeabilized sperm reactivated with exogenous 32 P-ATP. However, the reactivation conditions allow variable levels of motility to be expressed and phosphorylation of many proteins that appear to be unrelated to sperm motility. Thus, identification of the few relevant proteins is difficult. We have developed a method to collect and keep sperm immotile until reactivated for analysis to normal motility levels. Artificial sea water (ASW) buffered with 5 mM 2-[N-morpholino]ethanesulfonic acid at pH 6.0 and containing 50 mM KCI, allows collection and storage of immotile sea urchin sperm for up to 96 h at 4-5 C. Motility under these conditions is essentially zero, but sperm is rapidly reactivated to normal motility by diluting with ASW to standard pH (8.0) and KCI concentration (10 mM).

Radiolabelling of sperm cells with 99mTc-HMPAO. In vivo visualization of sperm cell migration in rabbits

International Nuclear Information System (INIS)

Bockisch, A.; Tennessee Univ., Knoxville, TN; Tennessee Univ., Knoxville, TN; Al-Hasani, S.; Ven, H.V.D.; Diedrich, K.; Krebs, D.; Posch, C.; Hotze, A.; Biersack, H.J.

1989-01-01

The present paper is the first descriptive radiolabeling of sperm cells in order to visualize their in vivo migration and imaging by scintigraphic technique, 99m Tc-HMPAO was used which combines favourable characteristics of both imaging modalities and radiation exposure. The radiolabeling yield was optimised for human sperm cells, and was increasing with the number of sperm cells, the amount of HMPAO, the 99m Tc-HMPAO concentration and the duration of the incubation. Incubation periods greater than 20 min, however, resulted only in a minor increase of labeling yield. A delay of more than 5 min between the labeling of the HMPAO with 99m Tc and initiation of the incubation of the sperm cells with the 99m Tc-HMPAO also decreased the maximum labeling yield. The radiolabeled cells were found to be stable and after 18 h > 93% of the activity was still bound to the sperm cells. After insemination of labeled sperm cells in ovulating rabbits the accumulation of the cells in the Fallopian tubes and their subsequent migration could be clearly visualized by scintigraphic techniques in vivo. (orig.) [de

A new approach to sperm preservation based on bioenergetic theory.

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Froman, D P; Feltmann, A J

2010-04-01

To date, attempts to preserve chicken sperm have been based on a trial-and-error experimental approach. The present work outlines the development of an alternative approach based on empiricism and bioenergetic theory. In previous work, we found fowl sperm motility to be dependent on mitochondrial calcium cycling, phospholipase A(2), and long-chain fatty acids as an endogenous energy source. It is noteworthy that fowl sperm reside within the sperm storage tubules (SST) of the oviduct over an interval of days to weeks after insemination. In this regard, a model for in vivo sperm storage was developed and tested in additional previous research. Sperm penetration of the SST, sperm residence within the SST, and sperm egress from the SST can be explained in terms mitochondrial function. Understanding sperm function and longevity in terms of bioenergetics presented the possibility that sperm could be inactivated by disrupting mitochondrial calcium cycling and could thereby be preserved. However, this possibility also posed a problem: maintenance of the inner membrane potential of the mitochondrion within inactivated sperm. This report describes a series of experiments in which fowl sperm were inactivated by treatment with the calcium chelator tetrasodium 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, and then reactivated by treatment with calcium ions. The effect of tetrasodium 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid on mitochondrial calcium cycling was confirmed by flow cytometry and confocal microscopy. When treated sperm were cooled to 10 degrees C, inactivated sperm could be reactivated throughout a 5-h storage interval. When stored sperm were held for 3 h before reactivation and insemination, fertility was 88% of the control. Storage did not affect hatchability. In summary, short-term storage was realized by manipulating mitochondrial function. We propose that 1) complex V consumes ATP within inactivated sperm and, by doing so, maintains

Effect of sperm concentration on characteristics and fertilization capacity of rooster sperm frozen in the presence of the antioxidants catalase and vitamin E.

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Moghbeli, Morteza; Kohram, Hamid; Zare-Shahaneh, Ahmad; Zhandi, Mahdi; Sharideh, Hossein; Sharafi, Mohsen

2016-10-01

The objective of this study conducted was to determine the influence of different levels of sperm concentration, including catalase (CAT) and vitamin E (VitE) in rooster semen extender on postthawed quality and fertility of rooster semen. Semen was collected twice a week from six roosters (Arian) and diluted according to experimental treatments consisting of sperm suspensions containing different sperm concentrations (200, 400, and 600 × 106 sperm/mL) without antioxidant supplementation as control (Con) groups (Con200, Con400, and Con600, respectively), sperm suspensions containing different sperm concentrations (200, 400, and 600 × 106 sperm/mL) supplemented with 5-μg/mL VitE (VitE200, VitE400, and VitE600, respectively) and different sperm concentrations (200, 400, and 600 × 106 sperm/mL) supplementation with 100 IU/mL CAT (CAT200, CAT400, and CAT600, respectively). After thawing; sperm motility, membrane integrity, and mitochondrial function were assessed. Fertility and hatchability rates were determined by using 100 artificially inseminated hens. The percentage of total motility (TM) and activity of mitochondria decreased (P rooster extender with different level sperm concentrations had no effect (P > 0.05) on fertility and hatchability rates. In conclusion, although adding VitE and CAT in extender with different levels of sperm concentration improved postthawed quality of rooster semen, but adding VitE and CAT in the extender have no effect on fertility rate. Copyright © 2016 Elsevier Inc. All rights reserved.

Dietary supplementation of antioxidants improves semen quality of IVF patients in terms of motility, sperm count, and nuclear vacuolization.

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Wirleitner, Barbara; Vanderzwalmen, Pierre; Stecher, Astrid; Spitzer, Dietmar; Schuff, Maximilian; Schwerda, Delf; Bach, Magnus; Schechinger, Birgit; Herbert Zech, Nicolas

2012-12-01

This study aimed to investigate the influence of an oral antioxidative supplementation on sperm quality of in vitro fertilization (IVF) patients, as analyzed by sperm motility according to the WHO criteria and motile sperm organelle morphology examination (MSOME). Semen samples were collected from 147 patients before undergoing an IVF/intracytoplasmic morphologically-selected sperm injection (IMSI) cycle and 2 - 12 months after an antioxidative supplementation. Semen analysis was evaluated according to WHO and MSOME criteria. Spermatozoa were grouped according to the size of nuclear vacuoles within the sperm's heads. Patients were divided into oligoasthenoteratozoospermic (OAT) and non-OAT men. Between first and second semen analysis, patients were supplemented orally with an antioxidative preparation. After the antioxidative therapy we observed a significant reduction in the percentage of immotile sperm cells in the patients. Additionally, the percentage of class I spermatozoa according to MSOME criteria was significantly higher after antioxidative supplementation. In OAT patients the percentage of class I sperm was found to be increased, although not significantly. However, we observed a drastic improvement in sperm motility as well as in total sperm count in this group. The results demonstrated a considerable improvement in semen quality, notably in OAT patients. Considering the putative relationship between semen quality on the one hand and reactive oxygen species on the other, the observed changes in the sperm parameters indicate that a decline in semen quality, and even subtle morphological changes, might be associated with oxidative stress. Our findings suggest that an antioxidative and micronutrient supplementation has a remarkable benefit for IVF patients having restricted sperm parameters, in particular.

Sperm cryopreservation in live-bearing Xiphophorus fishes: offspring production from Xiphophorus variatus and strategies for establishment of sperm repositories.

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Yang, Huiping; Cuevas-Uribe, Rafael; Savage, Markita G; Walter, Ronald B; Tiersch, Terrence R

2012-09-01

Cryopreservation of sperm from Xiphophorus fishes has produced live young in three species: X. hellerii, X. couchianus, and X. maculatus. In this study, the goal was to establish protocols for sperm cryopreservation and artificial insemination to produce live young in X. variatus, and to identify needs for repository development. The objectives were to: 1) collect basic biological characteristics of males; 2) cryopreserve sperm from X. variatus, 3) harvest live young from cryopreserved sperm, and 4) discuss the requirements for establishment of sperm repositories. The 35 males used in this study had a body weight of 0.298±0.096 g (mean±SD), body length of 2.5±0.2 cm, and testis weight of 6.4±3.4 mg. The sperm production per gram of testis was 2.33±1.32×10(9) cells. After freezing, the post-thaw motility decreased significantly to 37%±17% (ranging from 5% to 70%) (p=0.000) from 57%±14% (40%-80%) of fresh sperm (N=20). Artificial insemination of post-thaw sperm produced confirmed offspring from females of X. hellerii and X. variatus. This research, taken together with previous studies, provides a foundation for development of strategies for sperm repositories of Xiphophorus fishes. This includes: 1) the need for breeding strategies for regeneration of target populations, 2) identification of minimum fertilization capacity of frozen samples, 3) identification of fish numbers necessary for sampling and their genetic relationships, 4) selection of packaging containers for labeling and biosecurity, 5) assurance of quality control and standardization of procedures, 6) information systems that can manage the data associated with cryopreserved samples, including the genetic data, 7) biological data of sampled fish, 8) inventory data associated with frozen samples, and 9) data linking germplasm samples with other related materials such as body tissues or cells saved for DNA and RNA analyses.

Hunting, Food Preparation, and Consumption of Rodents in Lao PDR.

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Kanokwan Suwannarong

Full Text Available A cross-sectional study was conducted in 29 villages of Khamkeuth District in Bolikhamxay Province in the Lao PDR during March to May 2013. The study aimed to determine the characteristics associated with rodent consumption and related behaviors among different ethnic groups, ages, and genders. Five-hundred-eighty-four (584 males and females from 18-50 years of age participated in this study. Half of them were Hmong (292, 50% while 152 respondents were Lao-Tai (26% or other ethnic groups (140, 24%. Most of the respondents (79.5% had farming as their main occupation. Prevalences of the studied outcomes were high: 39.9 for hunting or capturing rodents in the previous year, 77.7% for preparing rodents as food, and 86.3% for rodent consumption. Multivariable logistic regression analysis showed that likelihood of these types of rodent contact was more consistently associated with behavioral factors (gathering things from the forest and elsewhere, cultivation-related activities, and taking measures to prevent rodent-borne disease than with socio-demographic, environmental, or cultural factors. The strongest associations were observed for gathering things; these associations were consistently positive and statistically significant. Although this study did not directly assess rodent-borne zoonosis risk, we believe that study findings raise concern that such risk may be substantial in the study area and other similar areas. Further epidemiological studies on the association between rodent-borne disease infection and rodent hunting, preparation for food, and consumption are recommended. Moreover, further studies are needed on the association between these potential exposure factors (i.e., rodent hunting, preparation for food, and consumption and rodent-borne infections, especially among ethnic groups like the Hmong in Lao PDR and those in neighboring countries with similar socio-demographic, environmental, behavioral and cultural contexts.

COMP-1 promotes competitive advantage of nematode sperm

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Hansen, Jody M; Chavez, Daniela R; Stanfield, Gillian M

2015-01-01

Competition among sperm to fertilize oocytes is a ubiquitous feature of sexual reproduction as well as a profoundly important aspect of sexual selection. However, little is known about the cellular mechanisms sperm use to gain competitive advantage or how these mechanisms are regulated genetically. In this study, we utilize a forward genetic screen in Caenorhabditis elegans to identify a gene, comp-1, whose function is specifically required in competitive contexts. We show that comp-1 functions in sperm to modulate their migration through and localization within the reproductive tract, thereby promoting their access to oocytes. Contrary to previously described models, comp-1 mutant sperm show no defects in size or velocity, thereby defining a novel pathway for preferential usage. Our results indicate not only that sperm functional traits can influence the outcome of sperm competition, but also that these traits can be modulated in a context-dependent manner depending on the presence of competing sperm. DOI: http://dx.doi.org/10.7554/eLife.05423.001 PMID:25789512

Parallel Evolution of Sperm Hyper-Activation Ca2+ Channels.

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Cooper, Jacob C; Phadnis, Nitin

2017-07-01

Sperm hyper-activation is a dramatic change in sperm behavior where mature sperm burst into a final sprint in the race to the egg. The mechanism of sperm hyper-activation in many metazoans, including humans, consists of a jolt of Ca2+ into the sperm flagellum via CatSper ion channels. Surprisingly, all nine CatSper genes have been independently lost in several animal lineages. In Drosophila, sperm hyper-activation is performed through the cooption of the polycystic kidney disease 2 (pkd2) Ca2+ channel. The parallels between CatSpers in primates and pkd2 in Drosophila provide a unique opportunity to examine the molecular evolution of the sperm hyper-activation machinery in two independent, nonhomologous calcium channels separated by > 500 million years of divergence. Here, we use a comprehensive phylogenomic approach to investigate the selective pressures on these sperm hyper-activation channels. First, we find that the entire CatSper complex evolves rapidly under recurrent positive selection in primates. Second, we find that pkd2 has parallel patterns of adaptive evolution in Drosophila. Third, we show that this adaptive evolution of pkd2 is driven by its role in sperm hyper-activation. These patterns of selection suggest that the evolution of the sperm hyper-activation machinery is driven by sexual conflict with antagonistic ligands that modulate channel activity. Together, our results add sperm hyper-activation channels to the class of fast evolving reproductive proteins and provide insights into the mechanisms used by the sexes to manipulate sperm behavior. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Initiation of male sperm-transfer behavior in Caenorhabditis elegans requires input from the ventral nerve cord

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Gharib Shahla

2006-08-01

Full Text Available Abstract Background The Caenorhabditis elegans male exhibits a stereotypic behavioral pattern when attempting to mate. This behavior has been divided into the following steps: response, backing, turning, vulva location, spicule insertion, and sperm transfer. We and others have begun in-depth analyses of all these steps in order to understand how complex behaviors are generated. Here we extend our understanding of the sperm-transfer step of male mating behavior. Results Based on observation of wild-type males and on genetic analysis, we have divided the sperm-transfer step of mating behavior into four sub-steps: initiation, release, continued transfer, and cessation. To begin to understand how these sub-steps of sperm transfer are regulated, we screened for ethylmethanesulfonate (EMS-induced mutations that cause males to transfer sperm aberrantly. We isolated an allele of unc-18, a previously reported member of the Sec1/Munc-18 (SM family of proteins that is necessary for regulated exocytosis in C. elegans motor neurons. Our allele, sy671, is defective in two distinct sub-steps of sperm transfer: initiation and continued transfer. By a series of transgenic site-of-action experiments, we found that motor neurons in the ventral nerve cord require UNC-18 for the initiation of sperm transfer, and that UNC-18 acts downstream or in parallel to the SPV sensory neurons in this process. In addition to this neuronal requirement, we found that non-neuronal expression of UNC-18, in the male gonad, is necessary for the continuation of sperm transfer. Conclusion Our division of sperm-transfer behavior into sub-steps has provided a framework for the further detailed analysis of sperm transfer and its integration with other aspects of mating behavior. By determining the site of action of UNC-18 in sperm-transfer behavior, and its relation to the SPV sensory neurons, we have further defined the cells and tissues involved in the generation of this behavior. We

Initiation of male sperm-transfer behavior in Caenorhabditis elegans requires input from the ventral nerve cord.

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Schindelman, Gary; Whittaker, Allyson J; Thum, Jian Yuan; Gharib, Shahla; Sternberg, Paul W

2006-08-15

The Caenorhabditis elegans male exhibits a stereotypic behavioral pattern when attempting to mate. This behavior has been divided into the following steps: response, backing, turning, vulva location, spicule insertion, and sperm transfer. We and others have begun in-depth analyses of all these steps in order to understand how complex behaviors are generated. Here we extend our understanding of the sperm-transfer step of male mating behavior. Based on observation of wild-type males and on genetic analysis, we have divided the sperm-transfer step of mating behavior into four sub-steps: initiation, release, continued transfer, and cessation. To begin to understand how these sub-steps of sperm transfer are regulated, we screened for ethylmethanesulfonate (EMS)-induced mutations that cause males to transfer sperm aberrantly. We isolated an allele of unc-18, a previously reported member of the Sec1/Munc-18 (SM) family of proteins that is necessary for regulated exocytosis in C. elegans motor neurons. Our allele, sy671, is defective in two distinct sub-steps of sperm transfer: initiation and continued transfer. By a series of transgenic site-of-action experiments, we found that motor neurons in the ventral nerve cord require UNC-18 for the initiation of sperm transfer, and that UNC-18 acts downstream or in parallel to the SPV sensory neurons in this process. In addition to this neuronal requirement, we found that non-neuronal expression of UNC-18, in the male gonad, is necessary for the continuation of sperm transfer. Our division of sperm-transfer behavior into sub-steps has provided a framework for the further detailed analysis of sperm transfer and its integration with other aspects of mating behavior. By determining the site of action of UNC-18 in sperm-transfer behavior, and its relation to the SPV sensory neurons, we have further defined the cells and tissues involved in the generation of this behavior. We have shown both a neuronal and non-neuronal requirement for

Is sperm banking of interest to patients with nongerm cell urological cancer before potentially fertility damaging treatments?

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Salonia, Andrea; Gallina, Andrea; Matloob, Rayan; Rocchini, Lorenzo; Saccà, Antonino; Abdollah, Firas; Colombo, Renzo; Suardi, Nazareno; Briganti, Alberto; Guazzoni, Giorgio; Rigatti, Patrizio; Montorsi, Francesco

2009-09-01

We assessed the opinions of patients with nongerm cell urological cancer on sperm banking before undergoing surgical or nonsurgical therapy that could potentially endanger subsequent fertility. Between April 2007 and July 2008, 753 patients visited a urological office and were invited to complete a brief self-administered questionnaire to assess opinions on sperm banking before undergoing any eventual therapy potentially dangerous for male fertility. Logistic regression models tested the association between predictors (age, educational level, relationship status, previous fatherhood and benign disorder vs nongerm cell urological cancer) and patient wishes for sperm banking. Median patient age was 65 years (mean 61.6, range 18 to 76). Overall 522 patients (69.3%) had nongerm cell urological cancer and only 242 (32.1%) were in favor of pretreatment sperm banking. On univariate analysis age (OR 0.961, p banking, whereas having cancer and educational status were not significantly correlated. Multivariate analysis indicated that aging (OR 0.966, p = 0.001) and previous fatherhood (OR 0.587, p = 0.029) maintained inverse associations. Having urological cancer was positively (OR 1.494, p = 0.045) associated with the wish for sperm banking. In urological patients there is a low rate of willingness to bank sperm before any potential fertility damaging therapeutic approach. Having nongerm cell urological cancer is an independent predictor that is positively associated with the wish to bank sperm. It is vitally important to provide comprehensive information about pretreatment sperm banking to young adults with nongerm cell urological cancer.

Percoll gradient-centrifuged capacitated mouse sperm have increased fertilizing ability and higher contents of sulfogalactosylglycerolipid and docosahexaenoic acid-containing phosphatidylcholine compared to washed capacitated mouse sperm.

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Furimsky, Anna; Vuong, Ngoc; Xu, Hongbin; Kumarathasan, Premkumari; Xu, Min; Weerachatyanukul, Wattana; Bou Khalil, Maroun; Kates, Morris; Tanphaichitr, Nongnuj

2005-03-01

Although Percoll gradient centrifugation has been used routinely to prepare motile human sperm, its use in preparing motile mouse sperm has been limited. Here, we showed that Percoll gradient-centrifuged (PGC) capacitated mouse sperm had markedly higher fertilizing ability (sperm-zona pellucida [ZP] binding and in vitro fertilization) than washed capacitated mouse sperm. We also showed that the lipid profiles of PGC capacitated sperm and washed capacitated sperm differed significantly. The PGC sperm had much lower contents of cholesterol and phospholipids. This resulted in relative enrichment of male germ cell-specific sulfogalactosylglycerolipid (SGG), a ZP-binding ligand, in PGC capacitated sperm, and this would explain, in part, their increased ZP-binding ability compared with that of washed capacitated sperm. Analyses of phospholipid fatty acyl chains revealed that PGC capacitated sperm were enriched in phosphatidylcholine (PC) molecular species containing highly unsaturated fatty acids (HUFAs), with docosahexaenoic acid (DHA; C22: 6n-3) being the predominant HUFA (42% of total hydrocarbon chains of PC). In contrast, the level of PC-HUFAs comprising arachidonic acid (20:4n-6), docosapentaenoic acid (C22:5n-6), and DHA in washed capacitated sperm was only 27%. Having the highest unsaturation degree among all HUFAs in PC, DHA would enhance membrane fluidity to the uppermost. Therefore, membranes of PGC capacitated sperm would undergo fertilization-related fusion events at higher rates than washed capacitated sperm. These results suggested that PGC mouse sperm should be used in fertilization experiments and that SGG and DHA should be considered to be important biomarkers for sperm fertilizing ability.

Secular variations in sperm quality: fact or science fiction?

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Luc Multigner

Full Text Available The debate concerning the possible degradation in human sperm quality began in the 1970s, was revived at the beginning of the 1990s and has continued to mobilize the scientific community ever since. After the meta-analysis by Carlsen et al. (1992 showing a decline in human semen quality over the last 50 years, several groups investigated the sperm characteristics of more or less homogeneous groups of men who had provided semen at the same center for 10 to 20 years. A significant decrease in sperm concentration was reported in some studies, but not in others. Meanwhile, there is an increasing number of reports suggesting that physical and chemical factors introduced and spread by human activity in the environment may have contributed to sperm decline. At the end of the 20th century the debate on declining semen quality is not closed. The lack of certainty and the serious consequences that such a decline would have on the fertility of human populations make this an important public health issue at the start of the 21st century. For this reason, intensive research should be developed in both fundamental and epidemiological domains, particularly in South America, where industrial and agricultural pollution pose a serious threat to the population.

Evolutionary genetics and ecology of sperm-dependent parthenogenesis

NARCIS (Netherlands)

Beukeboom, L.W.; Vrijenhoek, R.C.

1998-01-01

Sperm-dependent (or pseudogamous) forms of parthenogenetic reproduction occur in a wide variety of animals. Inheritance is typically clonal and matroclinous (of female descent), but sperm are needed to initiate normal development. As opposed to true parthenogenesis (i.e., sperm-independent

Exposure of rainbow trout milt to mercury and cadmium alters sperm motility parameters and reproductive success

International Nuclear Information System (INIS)

Dietrich, Grzegorz J.; Dietrich, Mariola; Kowalski, R.K.; Dobosz, Stefan; Karol, Halina; Demianowicz, Wieslaw; Glogowski, Jan

2010-01-01

In the current work, seminal plasma was used for the first time as an incubation medium for monitoring short-time exposure effects of sublethal concentrations of mercury and cadmium ions on rainbow trout sperm. Sperm motility parameters (CASA) and hatching rates were used as gamete quality markers. Additionally live/dead sperm viability test and comet assay of DNA fragmentation were performed. We demonstrated that computer-assisted sperm motility analysis (CASA) may serve as a predictor of reproductive success, when milt contaminated with heavy metals is used. Results presented in this study demonstrate that mercury ions altered sperm motility characteristics at 1-10 mg Hg 2+ /l and 10 mg Cd 2+ /l and hatching rates at 10 mg Hg 2+ /l and 10 mg Cd 2+ /l after 4 h of exposure. Although mercury ions affected sperm motility parameters immediately after dilution with milt as well as at 4 h of exposure, no differences in sperm motility parameters were found between intact and mercury-treated milt after 24 h of exposure. Our results suggest that rainbow trout seminal plasma has a protective role against the toxic effects of mercury ions of rainbow trout sperm motility.

The effects of increased testicular temperature on testis-specific isoform of Na+/K+ -ATPase in sperm and its role in spermatogenesis and sperm function.

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Thundathil, J C; Rajamanickam, G D; Kastelic, J P; Newton, L D

2012-08-01

Impaired testicular thermoregulation is commonly implicated in abnormal spermatogenesis and impaired sperm function in animals and humans, with outcomes ranging from subclinical infertility to sterility. Bovine testes must be maintained 4-5 °C below body-core temperature for normal spermatogenesis. The effects of elevated testicular temperature have been extensively studied in cattle using a scrotal insulation model, which results in abnormal spermatogenesis and impaired sperm morphology and function. Using this model and proteomic approaches, we compared normal and abnormal sperm (from the same bulls) to elucidate the molecular basis of impaired function. We identified a cohort of sperm functional proteins differentially expressed between normal vs abnormal sperm, including a testis-specific isoform of Na(+) /K(+) -ATPase. In addition to its role as a sodium pump regulating sperm motility, Na(+) /K(+) -ATPase is also involved as a signalling molecule during sperm capacitation. In conclusion, because of its involvement in regulation of sperm function, this protein has potential as a fertility marker. Furthermore, comparing normal vs abnormal sperm (induced by scrotal insulation) is a useful model for identifying proteins regulating sperm function. © 2012 Blackwell Verlag GmbH.

Association between obesity and sperm quality.

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Ramaraju, G A; Teppala, S; Prathigudupu, K; Kalagara, M; Thota, S; Kota, M; Cheemakurthi, R

2018-04-01

There is awareness of likelihood of abnormal spermatozoa in obese men; however, results from previous studies are inconclusive. Advances in computer-aided sperm analysis (CASA) enable precise evaluation of sperm quality and include assessment of several parameters. We studied a retrospective cohort of 1285 men with CASA data from our infertility clinic during 2016. Obesity (BMI ≥30) was associated with lower (mean ± SE) volume (-0.28 ± 0.12, p-value = .04), sperm count (48.36 ± 16.51, p-value = .002), concentration (-15.83 ± 5.40, p-value = .01), progressive motility (-4.45 ± 1.92, p-value = .001), total motility (-5.50 ± 2.12, p-value = .002), average curve velocity (μm/s) (-2.09 ± 0.85, p-value = .001), average path velocity (μm/s) (-1.59 ± 0.75, p-value = .006), and higher per cent head defects (0.92 ± 0.81, p-value = .02), thin heads (1.12 ± 0.39, p-value = .007) and pyriform heads (1.36 ± 0.65, p-value = .02). Obese men were also more likely to have (odds ratio, 95% CI) oligospermia (1.67, 1.15-2.41, p-value = .007) and asthenospermia (1.82, 1.20-2.77, p-value = .005). This is the first report of abnormal sperm parameters in obese men based on CASA. Clinicians may need to factor in paternal obesity prior to assisted reproduction. © 2017 Blackwell Verlag GmbH.

In vivo test of bitter (andrographis paniculata nees.) extract to ejaculated sperm quality

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Sumarmin, R.; Huda, NK; Yuniarti, E.; Violita

2018-03-01

Sambiloto or Bitter (Andrographis paniculata Nees.), are often used to treat various diseases, such as influenza, cancer, anti-inflammation, anti-HIV, anti-mitotic and anti-fertility. This study aimed to determine the effects of the bitter (Andrographis paniculata Nees.) extract to ejaculated sperm mice quality (Mus musculus L. Swiss Webster). This research was conducted using Completely Randomized Design with 4 treatments, which are 0.0 g/b.w., (P0), 0.2 g/b.w., (P1), 0,4 g/b.w., (P3), or 0.6 g/b.w., (P4) bitter extract orally for 36 days. After treatment, the mice decapitated, dissected and collected the sperm from vas deferens. Then, the number of sperm counted by used the improved Neubauer and then stained by Eosin to count the abnormal sperm. Data analyzed by ANOVA (Analysis of Variance) then DNMRT. The results showed that the average numbers of sperm are 28.80 x 105 (P0), 19.50 x 105 (P1), 12.50 x105 (P2) and 9.50 x 105 (P3). The average abnormal sperm numbers are 18.33 x 105 (P0), 22.50 x 105 (P1), 31.50 x105 (P2) and 39.33 x 105 (P3). It showed that the effective treatment to decrease sperm number was 0.2 g/b.w., of bitter extract. It can conclude that the bitter (Andrographis paniculata Nees.) extract decreases the quality of the ejaculated sperm of mice (Mus musculus L.)

Effect of superoxide dismutase supplementation on sperm DNA fragmentation

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Luciano Negri

2017-10-01

Full Text Available Background: antioxidants supplementation improves sperm quality, but few trials have analyzed the effects on sperm DNA fragmentation (SDF. This study compares the effectiveness of SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol in reducing SDF with other antioxidants without SOD, hydroxytyrosol, and carnosol. Materials and methods: men with high SDF at baseline were selected in our clinical database. The patients taken into account had a 2-month control. SDF was measured by Sperm Chromatin Dispersion test (SCD. Untreated men were used as a control group. The remaining subjects received some oral antioxidant supplements (12 different combinations of both hydrophilic and lipophilic antioxidants, with some of them receiving nutritional support with a SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol. Results: 118 men were selected for a retrospective study. Mean age 39.3 ± 5.4 years. Fifteen had no treatment, 55 were treated with a SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol, and 48 took some antioxidant supplements for 2 months. Clinically, variations of at least 10% in baseline values of classic semen parameters and sperm DNA fragmentation were taken into consideration. Classic seminal parameters did not vary significantly in the three groups, with the exception of viability (p = 0.001. We assessed which of the active substances (no. 19 in different formulations were associated with variations in SDF. In the multivariable analysis of the 7 active substances that passed the univariable analysis, only the SOD molecule appeared to be linked to an improvement in SDF (< 0.0001. In detail, only one patient in the control group showed a spontaneous improvement in SDF (6%, compared to 16/48 (33% of those taking various oral antioxidant supplements, and 31/55 (56% of those taking a SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol. Conclusions: SOD

Short communication Relationship between sperm plasma ...

African Journals Online (AJOL)

Matshidiso MB. Masenya

2017-01-04

Jan 4, 2017 ... observed between sperm plasma membrane integrity and fertility. There was a weak positive correlation between normal sperm morphology and conception rate (r = 0.11). Additionally ..... been approved by all of us. Authors' ...

Sperm count as a surrogate endpoint for male fertility control.

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Benda, Norbert; Gerlinger, Christoph

2007-11-30

When assessing the effectiveness of a hormonal method of fertility control in men, the classical approach used for the assessment of hormonal contraceptives in women, by estimating the pregnancy rate or using a life-table analysis for the time to pregnancy, is difficult to apply in a clinical development program. The main reasons are the dissociation of the treated unit, i.e. the man, and the observed unit, i.e. his female partner, the high variability in the frequency of male intercourse, the logistical cost and ethical concerns related to the monitoring of the trial. A reasonable surrogate endpoint of the definite endpoint time to pregnancy is sperm count. In addition to the avoidance of the mentioned problems, trials that compare different treatments are possible with reasonable sample sizes, and study duration can be shorter. However, current products do not suppress sperm production to 100 per cent in all men and the sperm count is only observed with measurement error. Complete azoospermia might not be necessary in order to achieve an acceptable failure rate compared with other forms of male fertility control. Therefore, the use of sperm count as a surrogate endpoint must rely on the results of a previous trial in which both the definitive- and surrogate-endpoint results were assessed. The paper discusses different estimation functions of the mean pregnancy rate (corresponding to the cumulative hazard) that are based on the results of sperm count trial and a previous trial in which both sperm count and time to pregnancy were assessed, as well as the underlying assumptions. Sample size estimations are given for pregnancy rate estimation with a given precision.

Some Reflections on Intracytoplasmic Morphologically Selected Sperm Injection

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Thomas Ebner

2014-07-01

Full Text Available Although intracytoplasmic sperm injection (ICSI allows proper fertilization in most cases of male sub fertility, it is one of the most unphysiological techniques in assisted reproductive technologies (ART. Thus, over the last decade, researchers have tried to improve sperm observation with higher-resolution microscopy techniques such as the intracytoplasmic morphologically selected sperm injection (IMSI technique. In order to identify literatures for this review, the PubMed database was searched from 2000 onwards using the terms IMSI, motile sperm organelle morphology examination (MSOME and sperm vacuole. Approximately 10 years after the introduction of the MSOME and IMSI procedures, several questions related to the prevalence, origin, location, and clinical consequences of sperm vacuoles have not yet been clarified. It seems that IMSI as a routine application is not state of the art and the only confirmed indications for IMSI are recurrent implantation failure following ICSI and severe male factor.

Attitudes of infertile couples to sperm donation: the role of religious beliefs

Directory of Open Access Journals (Sweden)

نرگس باقری لنکرانی

2016-09-01

Full Text Available Half of infertile couples referring to health centers are infertile males. The aim of present study was considering the attitudes of infertile couples to sperm donation. This study is a qualitative content analysis research which concentrates on the opinions of the people who discuss about sperm donation in a cyberspace social network. In this field the opinions of jurists and scientific issues based on priorities of each available option for the treatment of infertility have been compared and prioritized. In order to analyze these attitudes, and according to the religion and its impact on the approach and decision making, particularly in the treatment of infertility as the completion of married life, the shadow of religion in decision-making is considered. Finally, three main themes and four sub-themes were extracted. Main themes are: the most frequent methods i.e. sperm donation, embryo donation and adoption. Due to the more negative view of sperm donation comparing to embryo donation it seems that the existence of sperm donation just increases the number of treatment options, but adds many restrictions in the whole Islamic community.

Methods of sperm vitality assessment.

Science.gov (United States)

Moskovtsev, Sergey I; Librach, Clifford L

2013-01-01

Sperm vitality is a reflection of the proportion of live, membrane-intact spermatozoa determined by either dye exclusion or osmoregulatory capacity under hypo-osmotic conditions. In this chapter we address the two most common methods of sperm vitality assessment: eosin-nigrosin staining and the hypo-osmotic swelling test, both utilized in clinical Andrology laboratories.

Characterization of Na+K+-ATPase in bovine sperm.

Science.gov (United States)

Hickey, Katie D; Buhr, Mary M

2012-04-15

Existing as a ubiquitous transmembrane protein, Na(+)K(+)-ATPase affects sperm fertility and capacitation through ion transport and a recently identified signaling function. Functional Na(+)K(+)-ATPase is a dimer of α and β subunits, each with isoforms (four and three, respectively). Since specific isoform pairings and locations may influence or indicate function, the objective of this study was to identify and localize subunits of Na(+)K(+)-ATPase in fresh bull sperm by immunoblotting and immunocytochemistry using antibodies against α1 and 3, and all β isoforms. Relative quantity of Na(+)K(+)-ATPase in head plasma membranes (HPM's) from sperm of different bulls was determined by densitometry of immunoblot bands, and compared to bovine kidney. Sperm and kidney specifically bound all antibodies at kDa equivalent to commercial controls, and to additional lower kDa bands in HPM. Immunofluorescence of intact sperm confirmed that all isoforms were present in the head region of sperm and that α3 was also uniformly distributed post-equatorially. Permeabilization exposing internal membranes typically resulted in an increase in fluorescence, indicating that some antibody binding sites were present on the inner surface of the HPM or the acrosomal membrane. Deglycosylation of β1 reduced the kDa of bands in sperm, rat brain and kidney, with the kDa of the deglycosylated bands differing among tissues. Two-dimensional blots of β1 revealed three distinct spots. Based on the unique quantity, location and structure Na(+)K(+)-ATPase subunits in sperm, we inferred that this protein has unique functions in sperm. Copyright © 2012 Elsevier Inc. All rights reserved.

Semen Analysis Test

Science.gov (United States)

... Sources Ask Us Also Known As Sperm Analysis Sperm Count Seminal Fluid Analysis Formal Name Semen Analysis This ... semen Viscosity—consistency or thickness of the semen Sperm count—total number of sperm Sperm concentration (density)—number ...

Sperm immobilization by dental focus microorganisms.

Science.gov (United States)

Linossier, A; Thumann, A; Bustos-Obregon, E

1982-01-01

Focal infections and their ability to produce alterations in different tissues have been in dispute for long time. The purpose of this work was to observe "in vitro" the effect of an Escherichia coli filtrate obtained from open pulpar necrosis on human sperm motility. It was observed that the E. coli filtrate produced a loss in sperm motility. The immobilizating factor was studied and characterized as a heat-stable, resistant to lyophilization and non-dializable substance, which could via blood stream reach the male reproductive system and affect sperm motility.

Sperm subpopulations in avian species: a comparative study between the rooster (Gallus domesticus and Guinea fowl (Numida meleagris

Directory of Open Access Journals (Sweden)

Manuel García-Herreros

2016-01-01

Full Text Available The main aims of this research were to study possible differences in objective morphometric sperm characteristics, establish normative sperm morphometry standards, and evaluate the presumed different subpopulation distribution of avian spermatozoa from the rooster (Gallus domesticus and Guinea fowl (Numida meleagris as model avian species. Seventy-two ejaculates (36 per species studied were obtained manually, following a training period involving gently combined dorso-abdominal and lumbo-sacral massage of the birds. Ejaculates were processed for volume, sperm concentration, viability, motility, and morphology. Moreover, samples were submitted for sperm morphometric assessment using objective Computer-Assisted Semen Analysis for Morphometry (CASA-Morph methods, with sperm morphometric descriptors evaluated by Principal Component Analysis (PCA and multivariate clustering analyses. There were several differences observed between the avian species in values obtained for ejaculate volume and sperm concentration (P < 0.001. Irrespective of species, PCA revealed two Principal Components (PCs explaining more than 80% of the variance. In addition, the number of subpopulations differed with species (three and five subpopulations for rooster and Guinea fowl, respectively. Moreover, the distribution of the sperm subpopulations was found to be structurally different between species. In conclusion, our findings from using CASA-Morph methods indicate pronounced sperm morphometric variation between these two avian species. Because of the strong differences observed in morphometric parameter values and their subpopulation distribution, these results suggest that application of objective analytical methods such as CASA-Morph could substantially improve the reliability of comparative studies and help establish valid normative sperm morphological values for avian species.

Sperm depletion as a consequence of increased sperm competition risk in the European bitterling, Rhodeus amarus

Czech Academy of Sciences Publication Activity Database

Smith, C.; Pateman-Jones, C.; Zieba, G.; Przybylski, M.; Reichard, Martin

2009-01-01

Roč. 77, č. 5 (2009), s. 1227-1233 ISSN 0003-3472 Institutional research plan: CEZ:AV0Z60930519 Keywords : alternative mating tactics * European bitterling * mating system * sexual conflict * sexual selection * sneaking * sperm competition * sperm economy Subject RIV: EG - Zoology Impact factor: 2.890, year: 2009

Correlation analysis of the progesterone-induced sperm acrosome reaction rate and the fertilisation rate in vitro.

Science.gov (United States)

Jiang, T; Qin, Y; Ye, T; Wang, Y; Pan, J; Zhu, Y; Duan, L; Li, K; Teng, X

2015-10-01

In this study, we aimed to investigate whether progesterone-induced acrosome reaction (AR) rate could be an indicator for fertilisation rate in vitro. Twenty-six couples with unexplained infertility and undergoing in vitro fertilisation (IVF) treatment were involved. On the oocytes retrieval day after routine IVF, residual sperm samples were collected to receive progesterone induction (progesterone group) or not (control group). AR rate was calculated and fertilisation rate was recorded. The correlation between progesterone-induced AR and fertilisation rate and between sperm normal morphology and 3PN (tripronuclear) were analysed using the Spearman correlation analysis. The AR rate of progesterone group was statistically higher than that of the control group (15.6 ± 5.88% versus 9.66 ± 5.771%, P rate (r = -0.053, P > 0.01) or rate of high-quality embryo development (r = -0.055, P > 0.01). Normal sperm morphology also showed no significant correlation with the amount of 3PN zygotes (r = 0.029, P > 0.01), rate of 3PN zygotes production (r = 0.20, P > 0.01), rate of 3PN embryo development (r = -0.406, P > 0.01), fertilisation rate (r = -0.148, P > 0.01) or progesterone-induced AR rate (r = 0.214, P > 0.01). Progesterone can induce AR in vitro significantly; however, the progesterone-induced AR may not be used to indicate fertilisation rate. © 2014 Blackwell Verlag GmbH.

Reliable collection of Caspian brown trout (Salmo trutta caspius) sperm using a catheter.

Science.gov (United States)

Aramli, M S; Golshahi, K; Banan, A; Sotoudeh, E

2016-10-01

The traditional stripping procedure for collecting fish semen is associated with the risk of urine contamination, which may significantly affect semen quality and quantity. The use of a catheter as an alternative method for semen collection may overcome this problem. Therefore, this study compared Caspian brown trout (Salmo trutta caspius) semen parameters (i.e. sperm density, seminal plasma osmolality, motility parameters of spermatozoa analysed using computer-assisted sperm analysis and fertility) between the traditional stripping method and the use of a catheter. All parameter values of the semen collected with a catheter were significantly higher (p brown trout sperm. © 2016 Blackwell Verlag GmbH.

Challenges in Development of Sperm Repositories for Biomedical Fishes: Quality Control in Small-Bodied Species.

Science.gov (United States)

Torres, Leticia; Liu, Yue; Guitreau, Amy; Yang, Huiping; Tiersch, Terrence R

2017-12-01

Quality control (QC) is essential for reproducible and efficient functioning of germplasm repositories. However, many biomedical fish models present significant QC challenges due to small body sizes (<5 cm) and miniscule sperm volumes (<5 μL). Using minimal volumes of sperm, we used Zebrafish to evaluate common QC endpoints as surrogates for fertilization success along sequential steps of cryopreservation. First, concentrations of calibration bead suspensions were evaluated with a Makler ® counting chamber by using different sample volumes and mixing methods. For sperm analysis, samples were initially diluted at a 1:30 ratio with Hanks' balanced salt solution (HBSS). Motility was evaluated by using different ratios of sperm and activation medium, and membrane integrity was analyzed with flow cytometry at different concentrations. Concentration and sperm motility could be confidently estimated by using volumes as small as 1 μL, whereas membrane integrity required a minimum of 2 μL (at 1 × 10 6 cells/mL). Thus, <5 μL of sperm suspension (after dilution to 30-150 μL with HBSS) was required to evaluate sperm quality by using three endpoints. Sperm quality assessment using a combination of complementary endpoints enhances QC efforts during cryopreservation, increasing reliability and reproducibility, and reducing waste of time and resources.

Aggregation of human sperm at higher temperature is due to hyperactivation.

Science.gov (United States)

Keppler, E L; Chan, P J; Patton, W C; King, A

1999-01-01

Chemotaxis of sperm cells to chemicals and hormones, such as progesterone, helps us to understand the concept of sperm transport. Here, the hypothesis was that heat increased sperm hyperactive motility, which caused the sperm to aggregate at the higher temperature. The objectives were (1) to determine the concentration of sperm at both halves of an artificial female reproductive tract made from a hermetically sealed cryopreservation straw filled with culture medium and placed with each end at different temperatures, and (2) to analyze the motility or kinematic parameters and hyperactivation of sperm found at the different temperatures. Cryopreserved-thawed human donor sperm (N = 6) were pooled and processed through 2-layer colloid solution. Analyses of the motile sperm were carried out and the washed sperm were homogeneously mixed and pipetted into several 0.5-mL French cryopreservation straws and heat-sealed. The control substance, consisting of acid-treated sperm, was also placed in several straws. The plastic straws of sperm were placed half at 23 degrees C and half was at either 37 or 40 degrees C. After 4 h, sperm at different sections of the straws were analyzed using the Hamilton Thorn motility analyzer (HTM-C). After 4 h of incubation, the concentration of sperm was doubled at the 40 degrees C heated half of the straw when compared with the other half of the straw at 23 degrees C. There were no differences in sperm concentration in the straw kept half at 37 degrees C and half at 23 degrees C. There were significantly higher percent motility, mean average path velocity, straight line velocity, lateral head displacement, and percent hyperactivation in sperm at the 40 degrees C temperature. The aggregation of sperm at the higher temperature of 40 degrees C may be due to enhanced motility, increased sperm velocities, and a 10-fold increase in hyperactivation at that temperature. The 37 degrees C temperature was not sufficient to attract sperm. Sperm cells

An extract of pomegranate fruit and galangal rhizome increases the numbers of motile sperm: a prospective, randomised, controlled, double-blinded trial.

Science.gov (United States)

Fedder, Maja D K; Jakobsen, Henrik B; Giversen, Ina; Christensen, Lars P; Parner, Erik T; Fedder, Jens

2014-01-01

Pomegranate fruit (Punica granatum) and galangal (Alpinia galanga) have separately been shown to stimulate spermatogenesis and to increase sperm counts and motility in rodents. Within traditional medicine, pomegranate fruit has long been used to increase fertility, however studies on the effect on spermatogenesis in humans have never been published. With this study we investigated whether oral intake of tablets containing standardised amounts of extract of pomegranate fruit and powder of greater galangal rhizome (Punalpin) would increase the total number of motile spermatozoa. The study was designed as a prospective, randomized, controlled, double-blinded trial. Enrolment was based on the mean total number of motile spermatozoa of two ejaculates. The participants delivered an ejaculate after 4-8 days of tablet intake and two ejaculates just before they stopped taking the tablets. Seventy adult men with a semen quality not meeting the standards for commercial application at Nordic Cryobank, but without azoospermia, were included in the study. Participants were randomized to take tablets containing extract of pomegranate fruit (standardised with respect to punicalagin A+B, punicalin and ellagic acid) and freeze-dried rhizome of greater galangal (standardised with respect to 1'S-1'-acetoxychavicol acetate) or placebo on a daily basis for three months. Sixty-six participants completed the intervention (active treatment: n = 34; placebo: n = 32). After the intervention the total number of motile spermatozoa was increased in participants treated with plant extracts compared with the placebo group (p = 0.026). After three months of active treatment, the average total number of motile sperm increased by 62% (from 23.4 to 37.8 millions), while for the placebo group, the number of motile sperm increased by 20%. Sperm morphology was not affected by the treatment. Our findings may help subfertile men to gain an improved amount of motile ejaculated sperm by taking tablets

Different Levels of DNA Methylation Detected in Human Sperms after Morphological Selection Using High Magnification Microscopy

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Nino Guy Cassuto

2016-01-01

Full Text Available Objective. To analyze DNA methylation levels between two groups of spermatozoa taken from the same sample, following morphological selection by high magnification (HM at 6100x microscopy. A prospective study was conducted and studied 876 spermatozoa from 10 randomly selected men. Sperm morphology was characterized at HM according to criteria previously established. High-scoring Score 6 and low-scoring Score 0 sperm were selected. Sperm DNA methylation level was assessed using an immunoassay method targeting 5-methylcytosine residues by fluorescence microscopy with imaging analysis system to detect DNA methylation in single spermatozoon. Results. In total, 448 S6 spermatozoa and 428 S0 spermatozoa were analyzed. A strong relationship was found between sperm DNA methylation levels and sperm morphology observed at HM. Sperm DNA methylation level in the S6 group was significantly lower compared with that in the S0 group (p<10-6, OR = 2.4; and p<0.001, as determined using the Wilcoxon test. Conclusion. Differences in DNA methylation levels are associated with sperm morphology variations as observed at HM, which allows spermatozoa with abnormal levels to be discarded and ultimately decrease birth defects, malformations, and epigenetic diseases that may be transmitted from sperm to offspring in ICSI.

The beneficial role of Naringin- a citrus bioflavonoid, against oxidative stress-induced neurobehavioral disorders and cognitive dysfunction in rodents: A systematic review and meta-analysis.

Science.gov (United States)

Viswanatha, Gollapalle Lakshminarayanashastry; Shylaja, H; Moolemath, Yogananda

2017-10-01

Naringin is a bioflavonoid, very abundantly found in citrus species. In literature, naringin has been scientifically well documented for its beneficial effects in various neurological disorders. In this systematic review and meta-analysis, we have made an attempt to correlate the protective role of naringin against oxidative stress-induced neurological disorders in rodents. The systematic search was performed using electronic databases; the search was mainly focused on the role of naringin in oxidative stress-induced neuropathological conditions in rodents. While, the meta-analysis was performed on the effect of naringin on oxidative stress markers [superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), reduced glutathione (GSH), lipid peroxidation (LPO)], nitrite, mitochondrial complexes (I to IV) and enzymes (acetylcholinesterase, Na + -K + -ATPase, Ca 2+ -ATPase, and Mg 2+ -ATPase) in the rodent brain. The data was analyzed using Review Manager Software. Based on the inclusion and exclusion criteria, twenty studies were selected. The meta-analysis revealed that, naringin could significantly inhibit various physical and chemical stimuli- induced neurological perturbances in the rodent brain, mediated through oxidative stress. Further, naringin also significantly restored the levels of all the oxidative stress markers (oxidative, nitrosative, enzymes, and mitochondrial complexes) in different parts of the rodent brain. This systematic review and meta-analysis supports the available scientific evidence on the beneficial role of naringin in the management of various neurological ailments. However, further studies involving human subjects is recommended to establish the safety and therapeutic efficacy in humans. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Accountings of Selecting Sperm on the (ethical) border

DEFF Research Database (Denmark)

Willum Adrian, Stine

During the past years, Denmark has become a destination for fertility travellers, in need for donated sperm. Today, treatment is possible no matter of marital status or sexuality. Furthermore, users of sperm donation can choose between anonymous and non-anonymous sperm, with either basic...... will analyze how women and their partners crossing the Danish border accounts for their donor choice, and their practices of choosing. I will be exploring the question: How are women and their partners accounting for choosing a sperm donor, crossing the (ethical) border to Denmark? The use of the term...

Patterns of sperm damage in Chernobyl passerine birds suggest a trade-off between sperm length and integrity

Czech Academy of Sciences Publication Activity Database

Hermosell, I. G.; Laskemoen, T.; Rowe, M.; Moller, A. P.; Mousseau, T. A.; Albrecht, Tomáš; Lifjeld, J. T.

2013-01-01

Roč. 9, č. 5 (2013), s. 20130530 ISSN 1744-9561 R&D Projects: GA ČR(CZ) GAP506/12/2472 Institutional support: RVO:68081766 Keywords : acrosome * radiation * sperm evolution * sperm size Subject RIV: EG - Zoology Impact factor: 3.425, year: 2013

Modelling a tethered mammalian sperm cell undergoing hyperactivation

KAUST Repository

Curtis, M.P.

2012-09-01

The beat patterns of mammalian sperm flagella can be categorised into two different types. The first involves symmetric waves propagating down the flagellum with a net linear propulsion of the sperm cell. The second, hyperactive, waveform is classified by vigorous asymmetric waves of higher amplitude, lower wavenumber and frequency propagating down the flagellum resulting in highly curved trajectories. The latter beat pattern is part of the capacitation process whereby sperm prepare for the prospective penetration of the zona pellucida and fusion with the egg. Hyperactivation is often observed to initiate as sperm escape from epithelial and ciliary bindings formed within the isthmic regions of the female oviducts, leading to a conjecture in the literature that this waveform is mechanically important for sperm escape. Hence, we explore the mechanical effects of hyperactivation on a tethered sperm, focussing on a Newtonian fluid. Using a resistive force theory model we demonstrate that hyperactivation can indeed generate forces that pull the sperm away from a tethering point and consequently a hyperactivated sperm cell bound to an epithelial surface need not always be pushed by its flagellum. More generally, directions of the forces generated by tethered flagella are insensitive to reductions in beat frequency and the detailed flagellar responses depend on the nature of the binding at the tethering point. Furthermore, waveform asymmetry and amplitude increases enhance the tendency for a tethered flagellum to start tugging on its binding. The same is generally predicted to be true for reductions in the wavenumber of the flagellum beat, but not universally so, emphasising the dynamical complexity of flagellar force generation. Finally, qualitative observations drawn from experimental data of human sperm bound to excised female reproductive tract are also presented and are found to be consistent with the theoretical predictions. © 2012 Elsevier Ltd.

Modelling a tethered mammalian sperm cell undergoing hyperactivation

KAUST Repository

Curtis, M.P.; Kirkman-Brown, J.C.; Connolly, T.J.; Gaffney, E.A.

2012-01-01

The beat patterns of mammalian sperm flagella can be categorised into two different types. The first involves symmetric waves propagating down the flagellum with a net linear propulsion of the sperm cell. The second, hyperactive, waveform is classified by vigorous asymmetric waves of higher amplitude, lower wavenumber and frequency propagating down the flagellum resulting in highly curved trajectories. The latter beat pattern is part of the capacitation process whereby sperm prepare for the prospective penetration of the zona pellucida and fusion with the egg. Hyperactivation is often observed to initiate as sperm escape from epithelial and ciliary bindings formed within the isthmic regions of the female oviducts, leading to a conjecture in the literature that this waveform is mechanically important for sperm escape. Hence, we explore the mechanical effects of hyperactivation on a tethered sperm, focussing on a Newtonian fluid. Using a resistive force theory model we demonstrate that hyperactivation can indeed generate forces that pull the sperm away from a tethering point and consequently a hyperactivated sperm cell bound to an epithelial surface need not always be pushed by its flagellum. More generally, directions of the forces generated by tethered flagella are insensitive to reductions in beat frequency and the detailed flagellar responses depend on the nature of the binding at the tethering point. Furthermore, waveform asymmetry and amplitude increases enhance the tendency for a tethered flagellum to start tugging on its binding. The same is generally predicted to be true for reductions in the wavenumber of the flagellum beat, but not universally so, emphasising the dynamical complexity of flagellar force generation. Finally, qualitative observations drawn from experimental data of human sperm bound to excised female reproductive tract are also presented and are found to be consistent with the theoretical predictions. © 2012 Elsevier Ltd.

Axicon-based annular laser trap for studies on sperm activity

Science.gov (United States)

Shao, Bing; Vinson, Jaclyn M.; Botvinick, Elliot L.; Esener, Sadik C.; Berns, Michael W.

2005-08-01

As a powerful and noninvasive tool, laser trapping has been widely applied for the confinement and physiological study of biological cells and organelles. Researchers have used the single spot laser trap to hold individual sperm and quantitatively evaluated the motile force generated by a sperm. Early studies revealed the relationship between sperm motility and swimming behavior and helped the investigations in medical aspects of sperm activity. As sperm chemotaxis draws more and more interest in fertilization research, the studies on sperm-egg communication may help to explain male or female infertility and provide exciting new approaches to contraception. However, single spot laser trapping can only be used to investigate an individual target, which has limits in efficiency and throughput. To study the chemotactic response of sperm to eggs and to characterize sperm motility, an annular laser trap with a diameter of several hundred microns is designed, simulated with ray tracing tool, and implemented. An axicon transforms the wavefront such that the laser beam is incident on the microscope objective from all directions while filling the back aperture completely for high efficiency trapping. A trapping experiment with microspheres is carried out to evaluate the system performance. The power requirement for annular sperm trapping is determined experimentally and compared with theoretical calculations. With a chemo-attractant located in the center and sperm approaching from all directions, the annular laser trapping could serve as a speed bump for sperm so that motility characterization and fertility sorting can be performed efficiently.

Quantification of leptin in seminal plasma of buffalo bulls and its correlation with antioxidant status, conventional and computer-assisted sperm analysis (CASA) semen variables.

Science.gov (United States)

Kumar, Pradeep; Saini, Monika; Kumar, Dharmendra; Jan, M H; Swami, Dheer Singh; Sharma, R K

2016-03-01

The present study is the first to quantify leptin in seminal plasma of buffalo and investigate its relationship with seminal attributes. Ten ejaculates each from 10 Murrah buffalo bulls were collected. Semen quality variables such as semen volume, sperm concentration, sperm abnormalities, membrane integrity, antioxidant enzyme activities (superoxide dismutase, catalase and total antioxidant capacity), malondialdehyde (MDA) concentration, as well as sperm kinetics and motility variables were evaluated. The leptin concentration in serum and seminal plasma were estimated by the ELISA method. Bulls were classified in two groups on the basis of sperm concentration with Group I having >800 million sperm/mL and Group II <500 million sperm/mL. Greater (P<0.05) mean sperm abnormalities, seminal leptin concentrations and MDA concentrations were recorded in Group II than Group I. The seminal leptin was positively correlated with sperm abnormalities and MDA concentration while being negatively correlated with sperm concentration, but there was no correlation with sperm kinetic and motility variables, sperm membrane integrity and seminal plasma antioxidant enzyme activity. Thus, the data suggest that seminal leptin has a role in spermatogenesis and can be used as a marker for spermatogenesis to predict the capacity of buffalo bulls for semen production. Copyright © 2016 Elsevier B.V. All rights reserved.

Aspermy, Sperm Quality and Radiation in Chernobyl Birds

Science.gov (United States)

Møller, Anders Pape; Bonisoli-Alquati, Andrea; Mousseau, Timothy A.; Rudolfsen, Geir

2014-01-01

Background Following the Chernobyl nuclear power plant accident, large amounts of radionuclides were emitted and spread in the environment. Animals living in such contaminated areas are predicted to suffer fitness costs including reductions in the quality and quantity of gametes. Methodology/Principal Findings We studied whether aspermy and sperm quality were affected by radioactive contamination by examining ejaculates from wild caught birds breeding in areas varying in background radiation level by more than three orders of magnitude around Chernobyl, Ukraine. The frequency of males with aspermy increased logarithmically with radiation level. While 18.4% of males from contaminated areas had no sperm that was only the case for 3.0% of males from uncontaminated control areas. Furthermore, there were negative relationships between sperm quality as reflected by reduced sperm velocity and motility, respectively, and radiation. Conclusions/Significance Our results suggest that radioactive contamination around Chernobyl affects sperm production and quality. We are the first to report an interspecific difference in sperm quality in relation to radioactive contamination. PMID:24963711

Effect of ionizing radiations on sperm of mice

International Nuclear Information System (INIS)

Khattab, F.I.; Hassan, N.H.A.; Eldawt, H.A.; Roushdy, H.M.

1995-01-01

Effects of both gamma rays and reactor beams of mixed neutrons and gamma rays on sperms were investigated. Caudal sperms of adult male mice were examined 3 and 6 weeks post-exposure to 0.5 , 1.5, 3.0 and 6.0 Gy gamma rays and 1, 2, 3, and 4 weeks in animals exposed to 42.5 and 58.5 R of mixed neutrons and gamma-rays. Sperms were seriously affected due to exposure to both types of irradiation. There were dose dependent changes in the mean frequency of abnormal sperms. The peak values were at dose level of 3.0 Gy recording 26.43% and 29.82% in both groups sacrificed 3 and 6 weeks respectively compared with 1.57% for the control group. The increase in the abnormal sperms in the groups of the animals exposed to the reactor beam was always related to the time interval between treatment and observation, the longer the time the more sperms with abnormal head were encountered. 14 figs., 2 tabs

Modified expression of several sperm proteins after chronic exposure to the antiandrogenic compound vinclozolin.

Science.gov (United States)

Auger, Jacques; Eustache, Florence; Maceiras, Paula; Broussard, Cédric; Chafey, Philippe; Lesaffre, Corinne; Vaiman, Daniel; Camoin, Luc; Auer, Jana

2010-10-01

Little is known about the molecular impact of in vivo exposure to endocrine disruptors (EDs) on sperm structures and functions. We recently reported that the lifelong exposure of rats to the antiandrogenic compound vinclozolin results in low epididymal weight, changes in sperm kinematic parameters, and immature sperm chromatin condensation, together with the impairment of several fertility end points. These results led us to focus specifically on possible molecular abnormalities in sperm. Sperm samples were recovered from the frozen epididymides of rats exposed during the previous study. The proteins present in the samples from six exposed and six control rats were analyzed in pairs, by two-dimensional fluorescence difference gel electrophoresis, to investigate possible exposure-induced changes to sperm protein profiles. Twelve proteins, from the 380 matched spots observed in at least five gels, were present in larger or smaller amounts after vinclozolin exposure. These proteins were identified by mass spectrometry, and several are known to play a crucial role in the sperm fertilizing ability, among which, two mitochondrial enzymes, malate dehydrogenase 2 and aldehyde dehydrogenase (both of which were present in smaller amounts after treatment) and A-kinase anchor protein 4 (larger amounts of precursor after treatment). Finally, Ingenuity Pathway Analysis revealed highly significant interactions between proteins over- and underexpressed after treatment. This is the first study to show an association between in vivo exposure to an ED and changes to the sperm protein profile. These modifications may be at least partly responsible for the reproductive abnormalities and impaired fertility recently reported in this rat model of vinclozolin exposure.

Sperm variables as predictors of fertility in Black Castellana roosters; use in the selection of sperm donors for genome resource banking purposes

Energy Technology Data Exchange (ETDEWEB)

Santiago Moreno, J.; Lopez Sebastian, A.; Castano, C.; Coloma, M. A.; Gomez Brunet, A.; Toledano Diaz, A.; Prieto, M. T.; Campo, J. L.

2009-07-01

Semen was collected from 10 Black Castellana roosters and the classic sperm variables (ejaculate volume, sperm concentration and sperm motility) examined. In addition, the hypo-osmotic swelling test was used to investigate sperm cell membrane integrity, and acidic aniline blue staining used to screen for morphological abnormalities (including acrosome integrity) and to examine the condensation status of the chromatin. The latter was also examined by Gram staining. Large and small semen volumes were associated high and low sperm concentrations respectively (R2=0.04, P<0.05). The percentage of motile spermatozoa correlated strongly with the percentage of sperm cells showing an intact acrosome (R2=0.13, P<0.001) and with the percentage of morphologically normal spermatozoa (R2=0.04, P<0.05). The percentage of Gram positive spermatozoa was positively correlated with semen appearance (R2=0.12, P<0.05), sperm cell concentration (R2=0.13, P<0.05), and with the sperm motility variables studied (R2=0.14, P<0.05 for percentage mobility, and R2=0.12, P<0.05 for quality of movement). Only three of the 10 roosters, all with fertilisation potentials of 80-90%, were considered potential sperm donors for genome resource banking purposes. The remaining birds were all of low fertility (. 50%); in fact, some produced semen volumes too small to perform fertility tests. Semen volume and membrane integrity were found to be the best variables for predicting the fertilisation potential of rooster ejaculates. (Author) 37 refs.

FEMALE CONTROL OF SPERM TRANSFER AND INTRASPECIFIC VARIATION IN SPERM PRECEDENCE: ANTECEDENTS TO THE EVOLUTION OF A COURTSHIP FOOD GIFT.

Science.gov (United States)

Sakaluk, Scott K; Eggert, Anne-Katrin

1996-04-01

Manipulation of ejaculates is believed to be an important avenue of female choice throughout the animal kingdom, but evidence of its importance to sexual selection remains scarce. In crickets, such manipulation is manifest in the premature removal of the externally attached spermatophore, which may afford females an important means of postcopulatory mate choice. We tested the hypothesis that premature spermatophore removal contributes significantly to intraspecific variation in sperm precedence by (1) experimentally manipulating spermatophore attachment durations of competing male Gryllodes sigillatus and (2) employing protein electrophoresis to determine the paternity of doubly mated females. The relative spermatophore attachment durations of competing males had a significant influence on male paternity, but the pattern of sperm precedence deviated significantly from the predictions of an ideal lottery. Instead, paternity data and morphological evidence accorded best with a model of partial sperm displacement derived here. Our model is similar to a displacement model of Parker et al. in that sperm of the second male mixes instantaneously with that of the first throughout the displacement process, but the novel feature of our model is that the number of sperm displaced is only a fraction of the number of sperm transferred by the second male. Regardless of the underlying mechanism, female G. sigillatus can clearly alter the paternity of their offspring through their spermatophore-removal behavior, and employ such cryptic choice in favoring larger males and those providing larger courtship food gifts. We discuss how female control of sperm transfer and intraspecific variation in sperm precedence may be important precursors to the evolution of gift giving in insects. © 1996 The Society for the Study of Evolution.

Mouse Y-Encoded Transcription Factor Zfy2 Is Essential for Sperm Head Remodelling and Sperm Tail Development

NARCIS (Netherlands)

Vernet, Nadege; Mahadevaiah, Shantha K.; Decarpentrie, Fanny; Longepied, Guy; de Rooij, Dirk G.; Burgoyne, Paul S.; Mitchell, Michael J.

2016-01-01

A previous study indicated that genetic information encoded on the mouse Y chromosome short arm (Yp) is required for efficient completion of the second meiotic division (that generates haploid round spermatids), restructuring of the sperm head, and development of the sperm tail. Using mouse models

A systematic review on sperm DNA fragmentation in male factor infertility: Laboratory assessment

Directory of Open Access Journals (Sweden)

Manesh Kumar Panner Selvam

2018-03-01

Full Text Available Objective: To review sperm DNA fragmentation (SDF testing as an important sperm function test in addition to conventional semen analysis. High SDF is negatively associated with semen quality, the fertilisation process, embryo quality, and pregnancy outcome. Over recent decades, different SDF assays have been developed and reviewed extensively to assess their applicability and accuracy as advanced sperm function tests. Amongst them, the standardisation of the terminal deoxynucleotidyl transferased UTP nick-end labelling (TUNEL assay with a bench top flow cytometer in clinical practice deserves special mention with a threshold value of 16.8% to differentiate infertile men with DNA damage from fertile men. Materials and methods: A systematic literature search was performed through the PubMed, Medline, and ScienceDirect databases using the keywords ‘sperm DNA fragmentation’ and ‘laboratory assessment’. Non-English articles were excluded and studies related to humans were only included. Results: Of the 618 identified, 87 studies (original research and reviews and in addition eight book chapters meeting the selection criteria were included in this review. In all, 366 articles were rejected in the preliminary screening and a further 165 articles related to non-human subjects were excluded. Conclusion: There are pros and cons to all the available SDF assays. TUNEL is a reliable technique with greater accuracy and as an additional diagnostic test in Andrology laboratories along with basic semen analysis can predict fertility outcome, and thus direct the choice of an assisted reproductive technology procedure for infertile couples. Also, the TUNEL assay can be used as a prognostic test and results are beneficial in deciding personalised treatment for infertile men. Keywords: Sperm DNA fragmentation (SDF, Terminal deoxynucleotidyl transferased UTP nick-end labelling (TUNEL, DNA damage, Sperm DNA fragmentation (SDF assay

Mammalian Sperm Motility: Observation and Theory

KAUST Repository

Gaffney, E.A.

2011-01-21

Mammalian spermatozoa motility is a subject of growing importance because of rising human infertility and the possibility of improving animal breeding. We highlight opportunities for fluid and continuum dynamics to provide novel insights concerning the mechanics of these specialized cells, especially during their remarkable journey to the egg. The biological structure of the motile sperm appendage, the flagellum, is described and placed in the context of the mechanics underlying the migration of mammalian sperm through the numerous environments of the female reproductive tract. This process demands certain specific changes to flagellar movement and motility for which further mechanical insight would be valuable, although this requires improved modeling capabilities, particularly to increase our understanding of sperm progression in vivo. We summarize current theoretical studies, highlighting the synergistic combination of imaging and theory in exploring sperm motility, and discuss the challenges for future observational and theoretical studies in understanding the underlying mechanics. © 2011 by Annual Reviews. All rights reserved.

Sperm quality in New Zealand: Is the downward trend continuing?

Science.gov (United States)

Birdsall, Mary A; Peek, John; Valiapan, Sumithra

2015-10-16

To investigate whether the decline in sperm concentration in New Zealand sperm donors observed from 1987 to 2007 continued in the period 2008-2014. A retrospective study from 2008 to 2014. The first semen sample of 285 men presenting as sperm donors in Auckland and Wellington was analysed for sperm concentration, seminal fluid volume and the percentage of motile sperm. These results were compared to results from 1987 to 2007 from the same clinics. The decline in semen volume and sperm concentration observed between 1987 and 2007 did not continue in 2008-2014. Sperm concentration decreased from 1987 until some time between 1997 and 2001, and has remained stable at an average of 62x106/ml between 2001 and 2014. Sperm motility declined significantly (8%) in the period 2008-2014, but there was no significant change over the total period studied, between 1987 and 2014. After a decline between 1987 and sometime during 1997-2001, the sperm concentration in men presenting as donors remained unchanged between 2002 and 2014, suggesting semen quality has not changed in New Zealand men over the last decade.

Pronounced within-individual plasticity in sperm morphometry across social environments.

Science.gov (United States)

Immler, Simone; Pryke, Sarah R; Birkhead, Tim R; Griffith, Simon C

2010-06-01

Sperm morphometry (i.e., size and shape) and function are important determinants of male reproductive success and are thought to be under stabilizing selection. However, recent studies suggest that sperm morphometry can be a phenotypically plastic trait, which can be adjusted to varying conditions. We tested whether different behavioral strategies in aggression between aggressive red and nonaggressive black males of the color polymorphic Gouldian finch (Erythrura gouldiae) can influence sperm morphometry. We show pronounced within-individual phenotypic plasticity in sperm morphometry of male Gouldian finches in three different social environments. Both red and black males placed in intermediate to high competitive environments (high frequency of red males) increased the relative length of their sperm midpiece. By contrast, red males placed in low to intermediate competitive environments (higher frequency of black males) increased the length of the sperm flagellum. Significant changes in stress and sex steroid hormone levels (in response to the competitive environment) appear to influence sperm traits in red but not in black males, suggesting that changes in hormonal levels are not solely responsible for the observed changes in sperm morphometry. These findings imply that males can adjust sperm morphometry across social environments.

Acute toxicity effects of perfluorooctane sulfonate on sperm vitality, kinematics and fertilization success in zebrafish

Science.gov (United States)

Xia, Jigang; Niu, Cuijuan

2017-07-01

Perfluorooctane sulfonate (PFOS) has emerged as one of the most concerning contaminants in recent years. This study aimed to investigate the acute toxicity effect of PFOS on sperm viability, kinematics and fertilization success in zebrafish ( Danio rerio). Sperm were activated in aqueous media containing a range of PFOS concentrations (0, 0.09, 0.9 and 9 mg/L). Viabilities and kinematics of the sperm exposed to different PFOS treatments were assessed via computer-assisted sperm analysis (CASA) at 20, 40, 60, and 80 s after activation. PFOS exposure decreased the percentage of motile sperm, the curvilinear velocity (VCL), and the mean angular displacement (MAD) of spermatozoa, but showed no influence on the straight-line velocity (VSL) or the angular path velocity (VAP). Furthermore, a significant decrease in fertilization success was observed in spermatozoa that were exposed to 0.9 mg/L PFOS or more. These findings indicate that PFOS pollution in natural aquatic environment may be a potential threaten to successful reproduction of fish.

21 CFR 1250.96 - Rodent control.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Rodent control. 1250.96 Section 1250.96 Food and... SANITATION Sanitation Facilities and Conditions on Vessels § 1250.96 Rodent control. Vessels shall be... of rodent control. ...

Effect of Vitrification on Sperm Parameters and Apoptosis in Fertile Men

OpenAIRE

M Adib; M Ramezani; MA Khalili

2011-01-01

Introduction & Objective: Today, cryopreservation of the human sperm is a common technique for treating infertility. It has been indicated that cryopreservation by different methods decrease the sperm motility and viability in fertile men, but still effect of freezing of the sperm by vitrification method have not been evaluated on sperm parameters and apoptosis. The aim of this study was to evaluate the effect of vitrification of sperm of fertile men on different sperm parameters (motility, m...

Sperm competition risk drives rapid ejaculate adjustments mediated by seminal fluid.

Science.gov (United States)

Bartlett, Michael J; Steeves, Tammy E; Gemmell, Neil J; Rosengrave, Patrice C

2017-10-31

In many species, males can make rapid adjustments to ejaculate performance in response to sperm competition risk; however, the mechanisms behind these changes are not understood. Here, we manipulate male social status in an externally fertilising fish, chinook salmon ( Oncorhynchus tshawytscha ), and find that in less than 48 hr, males can upregulate sperm velocity when faced with an increased risk of sperm competition. Using a series of in vitro sperm manipulation and competition experiments, we show that rapid changes in sperm velocity are mediated by seminal fluid and the effect of seminal fluid on sperm velocity directly impacts paternity share and therefore reproductive success. These combined findings, completely consistent with sperm competition theory, provide unequivocal evidence that sperm competition risk drives plastic adjustment of ejaculate quality, that seminal fluid harbours the mechanism for the rapid adjustment of sperm velocity and that fitness benefits accrue to males from such adjustment.

A novel cross-species inhibitor to study the function of CatSper Ca2+ channels in sperm.

Science.gov (United States)

Rennhack, Andreas; Schiffer, Christian; Brenker, Christoph; Fridman, Dmitry; Nitao, Elis T; Cheng, Yi-Min; Tamburrino, Lara; Balbach, Melanie; Stölting, Gabriel; Berger, Thomas K; Kierzek, Michelina; Alvarez, Luis; Wachten, Dagmar; Zeng, Xu-Hui; Baldi, Elisabetta; Publicover, Stephen; Kaupp, U Benjamin; Strünker, Timo

2018-05-03

Sperm from many species share the sperm-specific Ca 2+ channel CatSper (cation channel of sperm) that controls the intracellular Ca 2+ concentration and, thereby, the swimming behaviour. A growing body of evidence suggests that the mechanisms controlling CatSper activity and the role of the channel during fertilization differ among species. However, a lack of suitable pharmacological tools has hampered the elucidation of the function of CatSper. Known CatSper inhibitors exhibit considerable side effects and inhibit also Slo3, the K + channel in mammalian sperm. The drug RU1968 was reported to suppress Ca 2+ signaling in human sperm by an unknown mechanism. We resynthesized the drug and revisited its mechanism of action in sperm form humans, mice, and sea urchins. We show by Ca 2+ fluorimetry, single-cell Ca 2+ imaging, electrophysiology, opto-chemistry, and motility analysis that RU1968 inhibits CatSper in sperm from invertebrates and mammals. The drug lacks toxic side effects in human sperm, does not affect mouse Slo3, and inhibits human Slo3 with about 15-fold lower potency than CatSper. Moreover, in human sperm, the inhibitor mimics CatSper dysfunction and suppresses motility responses evoked by progesterone, an oviductal steroid that activates CatSper. Finally, we show that the drug abolishes CatSper-mediated chemotactic navigation in sea urchin sperm. We propose RU1968 as a novel tool to elucidate the function of CatSper in sperm across species. This article is protected by copyright. All rights reserved.

Sperm-egg penetration assay assessment of the contraceptive effects of glycerol and egg yolk in rooster sperm diluents.

Science.gov (United States)

Abouelezz, F M K; Castaño, C; Toledano-Díaz, A; Esteso, M C; López-Sebastián, A; Campo, J L; Santiago-Moreno, J

2015-06-01

Glycerol (GLY) and egg yolk (EY) are good cryoprotectants of avian and mammalian sperm, but in birds, they strongly inhibit the eventual fertilization of ova. Using the sperm penetration (SP-holes) assay and fertility trials, the present study investigates (1) the possible mechanism by which this contraceptive effect occurs in chickens and (2) the maximum concentrations of GLY and EY tolerated by fresh rooster sperm. Seventy Black-Barred Andaluza hens (five per treatment) were inseminated four times (twice per week) with 0.1 mL of fresh semen from roosters of the same breed diluted 1:1 (v:v) with Lake and Ravie medium containing different concentrations of GLY or EY. No adverse effects on acrosome integrity, sperm motility, or viability were seen with any concentration of GLY or EY. The number of SP-holes on perivitelline layer samples taken from above the germinal disc became progressively lower at GLY concentrations of 1.5% or greater (P > 0.05). No holes caused by sperms were seen in unfertilized eggs. The corresponding fertility results showed similar reductions when the GLY concentration was 1.5% or greater. No changes in the number of SP-holes were seen with increasing EY concentrations (0%-7.5%), nor were any differences in fertility observed, except for a reduction when 15% EY was used. The results therefore reveal that GLY affects the transit of sperms through the oviduct in their attempt to reach the infundibulum area, limiting their access to the ovum perivitelline layer. Egg yolk had no such effect, nor did it influence acrosome reaction capacity; its mechanism of contraceptive action therefore remains unknown. The maximum GLY and EY concentrations tolerated by the rooster sperm were 0.75% and 7.5%, respectively. Copyright © 2015 Elsevier Inc. All rights reserved.

Label-Free Imaging and Biochemical Characterization of Bovine Sperm Cells

Science.gov (United States)

Ferrara, Maria Antonietta; Di Caprio, Giuseppe; Managò, Stefano; De Angelis, Annalisa; Sirleto, Luigi; Coppola, Giuseppe; De Luca, Anna Chiara

2015-01-01

A full label-free morphological and biochemical characterization is desirable to select spermatozoa during preparation for artificial insemination. In order to study these fundamental parameters, we take advantage of two attractive techniques: digital holography (DH) and Raman spectroscopy (RS). DH presents new opportunities for studying morphological aspect of cells and tissues non-invasively, quantitatively and without the need for staining or tagging, while RS is a very specific technique allowing the biochemical analysis of cellular components with a spatial resolution in the sub-micrometer range. In this paper, morphological and biochemical bovine sperm cell alterations were studied using these techniques. In addition, a complementary DH and RS study was performed to identify X- and Y-chromosome-bearing sperm cells. We demonstrate that the two techniques together are a powerful and highly efficient tool elucidating some important criterions for sperm morphological selection and sex-identification, overcoming many of the limitations associated with existing protocols. PMID:25836358

The Persistence of Sperm and the Development of Time Since Intercourse (TSI) Guidelines in Sexual Assault Cases at Forensic Science Ireland, Dublin, Ireland.

Science.gov (United States)

Casey, David G; Domijan, Katarina; MacNeill, Sarah; Rizet, Damien; O'Connell, Declan; Ryan, Jennifer

2017-05-01

The persistence of sperm using confirmatory microscopic analysis, the persistence of sperm with tails, time since intercourse (TSI) analysis, and results from the acid phosphatase (AP) reaction from approximately 5581 swabs taken from circa 1450 sexual assault cases are presented. The observed proportions of sperm in the vagina and anus declines significantly after 48 h TSI, and sperm on oral swabs were observed up to 15 h TSI. The AP reaction as a predictor of sperm on intimate swabs is questioned. All AP reaction times gave a low true positive rate; 23% of sperm-positive swabs gave a negative AP reaction time. We show the AP reaction is an unsafe and an unreliable predictor of sperm on intimate swabs. We propose that TSI not AP informs precase assessment and the evaluative approach for sexual assault cases. To help inform an evaluative approach, TSI guidelines are presented. © 2016 American Academy of Forensic Sciences.

Sperm banking is of key importance in patients with prostate cancer.

Science.gov (United States)

Salonia, Andrea; Capogrosso, Paolo; Castiglione, Fabio; Russo, Andrea; Gallina, Andrea; Ferrari, Matteo; Clementi, Maria Chiara; Castagna, Giulia; Briganti, Alberto; Cantiello, Francesco; Damiano, Rocco; Montorsi, Francesco

2013-08-01

To assess the need for sperm banking among patients with prostate cancer (PCa) who are candidates for radical prostatectomy (RP). Cross-sectional study. Urologic department. Cohort of 510 Caucasian-European candidates for RP. A 10-item self-administered questionnaire to assess opinions on sperm banking before RP, to which descriptive statistics and logistic regression models were applied. PCa patients' wishes for preoperative sperm banking. Data collection was completed for 495 patients (97.1%). Ninety-nine (20%) expressed a wish for preoperative sperm banking. Men who wanted to bank sperm were younger (mean 62.2 vs. 65.1 years), were more frequently childless (21.2% vs. 8.8%), and more frequently had a more intense desire for fatherhood (64.7% vs. 9.3%) than the patients not interested in banking sperm. Willingness to bank sperm was not affected by the patient's educational or relationship status. Moreover, the interest for sperm banking was maintained regardless of cost issues. Overall, 84% of the patients considered it necessary to have a dedicated service of preoperative sperm cryopreservation. One out of five PCa patients would bank sperm before RP. Most patients considered it necessary to establish a dedicated service for preoperative sperm cryopreservation, regardless of their own motivation to bank sperm. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

[Analysis of correlative factors of sterility in males undergoing routine sperm inspection by masturbation].

Science.gov (United States)

Zhou, Liyuan; Shi, Xiaobo; Wang, Xin; Liu, Dan

2012-07-01

To investigate the factors influencing sterility in males undergoing routine sperm inspection by masturbation. Scales for demographic data, self-compiled infertility questionnaire, Symptom Checklist-90 (SCL-90) , and sexual life subscale of Olson Marital Quality Questionnaire (ENRICH) were assessed in 220 cases of sterility in males who had undergone sperm examination after ejaculation. The total SCL-90 scores and the factor scores of anxiety, phobia, somatization, obsessive compulsive behavior, interpersonal-sensitivity, hostility, and depression were significantly higher than the norm (Pmasturbation have negative emotions such as anxiety, phobia, somatization, and interpersonal sensitivities. The defective ejaculation may be the influential factor at the stage.

Sperm cryopreservation affects postthaw motility, but not embryogenesis or larval growth in the Brazilian fish Brycon insignis (Characiformes).

Science.gov (United States)

Viveiros, A T M; Isaú, Z A; Caneppele, D; Leal, M C

2012-09-01

Sperm cryopreservation is an important method for preserving genetic information and facilitating artificial reproduction. The objective was to investigate whether the cryopreservation process affects postthaw sperm motility, embryogenesis, and larval growth in the fish Brycon insignis. Sperm was diluted in methyl glycol and Beltsville Thawing solution, frozen in a nitrogen vapor vessel (dry shipper) and stored in liquid nitrogen. Half of the samples were evaluated both subjectively (% of motile sperm and motility quality score-arbitrary grading system from 0 [no movement] to 5 [rapidly swimming sperm]) and in a computer-assisted sperm analyzer (CASA; percentage of motile sperm and velocity). The other half was used for fertilization and the evaluation of embryogenesis (cleavage and gastrula stages), hatching rate, percentage of larvae with normal development and larval growth up to 112 days posthatching (dph). Fresh sperm was analyzed subjectively (percentage of motile sperm and motility quality score) and used as the control. In the subjective analysis, sperm motility significantly decreased from 100% motile sperm and quality score of 5 in fresh sperm to 54% motile sperm and quality score of 3 after thawing. Under computer-assisted sperm analyzer evaluation, postthaw sperm had 67% motile sperm, 122 μm/sec of curvilinear velocity, 87 μm/sec of straight-line velocity and 103 μm/sec of average path velocity. There were no significant differences between progenies (pooled data) for the percentage of viable embryos in cleavage (62%) or gastrula stages (24%) or in the hatching rate (24%), percentage of normal hatched larvae (93%), larval body weight (39.8 g), or standard length (12.7 cm) at 112 days posthatching. Based on these findings, cryopreserved sperm can be used as a tool to restore the population of endangered species, such as B. insignis, as well as for aquaculture purposes, without any concern regarding quality of the offspring. Copyright © 2012 Elsevier

Postmortem sperm procurement: a legal perspective.

Science.gov (United States)

Kahan, S E; Seftel, A D; Resnick, M I

1999-06-01

Postmortem sperm procurement with subsequent artificial insemination has become a technically feasible method for posthumous conception. A variety of legal questions exist involving the rights and relationships of the deceased, his family and his issue. We addressed these questions and designed a workable protocol for postmortem sperm procurement. MEDLINE, WESTLAW and LEXIS medical literature, and case law searches were conducted. United States and international case law, United States (federal and state) statutes, Uniform Law Commissions Acts, and law review commentaries and articles were reviewed. While postmortem sperm procurement is being requested throughout the United States, no standard protocol or procedural guidelines have been established by federal or state statute. Furthermore, the courts have not yet addressed this specific scenario in reported case law. Statutes and case law do address related factual scenarios and issues, including property rights in human bodies, rules governing transplantation of human organs/body parts, rights of parties in in vivo sperm bank donations and responsibilities of parents to the conceptus of artificial insemination. A workable protocol can be established by analyzing case law and statutes addressing factually similar scenarios. Urologists must focus on the express intent of the decedent and limit any postmortem sperm retrieval to the specific requests made by the decedent. Decedent requests should be documented in writing. The decedent must be competent and of majority age. In the absence of decedent expressed affirmative directive calling for sperm retrieval, no other relative or guardian may authorize this retrieval. Issues regarding the legitimacy and inheritance rights of the conceptus will most consistently be addressed when explicitly provided for in the will of the decedent.

Semi-automated scoring of triple-probe FISH in human sperm using confocal microscopy.

Science.gov (United States)

Branch, Francesca; Nguyen, GiaLinh; Porter, Nicholas; Young, Heather A; Martenies, Sheena E; McCray, Nathan; Deloid, Glen; Popratiloff, Anastas; Perry, Melissa J

2017-09-01

Structural and numerical sperm chromosomal aberrations result from abnormal meiosis and are directly linked to infertility. Any live births that arise from aneuploid conceptuses can result in syndromes such as Kleinfelter, Turners, XYY and Edwards. Multi-probe fluorescence in situ hybridization (FISH) is commonly used to study sperm aneuploidy, however manual FISH scoring in sperm samples is labor-intensive and introduces errors. Automated scoring methods are continuously evolving. One challenging aspect for optimizing automated sperm FISH scoring has been the overlap in excitation and emission of the fluorescent probes used to enumerate the chromosomes of interest. Our objective was to demonstrate the feasibility of combining confocal microscopy and spectral imaging with high-throughput methods for accurately measuring sperm aneuploidy. Our approach used confocal microscopy to analyze numerical chromosomal abnormalities in human sperm using enhanced slide preparation and rigorous semi-automated scoring methods. FISH for chromosomes X, Y, and 18 was conducted to determine sex chromosome disomy in sperm nuclei. Application of online spectral linear unmixing was used for effective separation of four fluorochromes while decreasing data acquisition time. Semi-automated image processing, segmentation, classification, and scoring were performed on 10 slides using custom image processing and analysis software and results were compared with manual methods. No significant differences in disomy frequencies were seen between the semi automated and manual methods. Samples treated with pepsin were observed to have reduced background autofluorescence and more uniform distribution of cells. These results demonstrate that semi-automated methods using spectral imaging on a confocal platform are a feasible approach for analyzing numerical chromosomal aberrations in sperm, and are comparable to manual methods. © 2017 International Society for Advancement of Cytometry. © 2017

Comparison of sperm motility subpopulation structure among wild anadromous and farmed male Atlantic salmon (Salmo salar) parr using a CASA system.

Science.gov (United States)

Caldeira, Carina; García-Molina, Almudena; Valverde, Anthony; Bompart, Daznia; Hassane, Megan; Martin, Patrick; Soler, Carles

2018-04-13

Atlantic salmon (Salmo salar) is an endangered freshwater species that needs help to recover its wild stocks. However, the priority in aquaculture is to obtain successful fertilisation and genetic variability to secure the revival of the species. The aims of the present work were to study sperm subpopulation structure and motility patterns in wild anadromous males and farmed male Atlantic salmon parr. Salmon sperm samples were collected from wild anadromous salmon (WS) and two generations of farmed parr males. Sperm samples were collected from sexually mature males and sperm motility was analysed at different times after activation (5 and 35s). Differences among the three groups were analysed using statistical techniques based on Cluster analysis the Bayesian method. Atlantic salmon were found to have three sperm subpopulations, and the spermatozoa in ejaculates of mature farmed parr males had a higher velocity and larger size than those of WS males. This could be an adaptation to high sperm competition because salmonid species are naturally adapted to this process. Motility analysis enables us to identify sperm subpopulations, and it may be useful to correlate these sperm subpopulations with fertilisation ability to test whether faster-swimming spermatozoa have a higher probability of success.

Performance of the postwash total motile sperm count as a predictor of pregnancy at the time of intrauterine insemination: a meta-analysis

NARCIS (Netherlands)

van Weert, Janne-Meije; Repping, Sjoerd; van Voorhis, Bradley J.; van der Veen, Fulco; Bossuyt, Patrick M. M.; Mol, Ben W. J.

2004-01-01

Objective: To assess the performance and clinical value of the postwash total motile sperm count (postwash TMC) as a test to predict intrauterine insemination (IUI) outcome. Design: Meta-analysis of diagnostic tests. Setting: Tertiary fertility center. Patient(s): Patients undergoing IUI.

Efficacy of two sperm preparation techniques in reducing non-specific bacterial species from human semen

Directory of Open Access Journals (Sweden)

Prabath K Abeysundara

2013-01-01

Full Text Available Context: Artificial reproductive techniques using seminal preparations with bacteria may cause pelvic inflammatory disease and its sequalae. Aims: To assess efficacy of two sperm preparation techniques to clear bacteria and the effect of bacteriospermia on sperm recovery rates. Settings and Design: A descriptive cross-sectional study was carried out among males of subfertile couples. Subjects and Methods: Semen samples were randomly allocated into swim-up method (group S, n = 68 and density gradient method (group D, n = 50 for sperm preparation. Seminal fluid analysis and bacterial cultures were performed in each sample before and after sperm preparation. Statistical Analysis: McNemar′s chi-squared test and independent samples t-test in SPSS version 16.0 were used. Results: Organisms were found in 86 (72.88% out of 118 samples, before sperm preparation; Streptococcus species (n = 40, 46.51% of which 14 were Group D Streptococcus species, Coagulase negative Staphylococcus species (n = 17, 19.76%, Staphylococcus aureus (n = 13, 15.11%, Coliform species (n = 11, 12.79% of which 09 were Escherichia coli and Corynebacterium species (n = 5, 5.81%. There was a statistically significant reduction of culture positive samples in raw vs. processed samples; in group S, 49 (72.05% vs. 16 (23.52% and in group D, 37 (74% vs. 18 (36%. In group S and D, mean (SD recovery rates of culture positive vs. culture negative samples were 39.44% (SD-14.02 vs. 44.22% (SD-22.38, P = 0.39 and 52.50% (SD-37.16 vs. 49.58% (SD-40.32, P = 0.82 respectively. Conclusions: Both sperm preparation methods significantly reduced bacteria in semen, but total clearance was not achieved. Sperm recovery rate was not affected by bacteriospermia.

Analysis of an Hertwig effect in the rainbow trout (Salmo gairdneri Richardson) after fertilization with γ-irradiated sperm

International Nuclear Information System (INIS)

Chourrout, D.; Chevassus, B.; Herioux, F.

1980-01-01

In this work, we tried to optimize γ-treatment of sperm to obtain large numbers of haploid gynogenetic embryos after fertilization. The analysis of increasing doses showed a typical 'Hertwig effect' in all cases, but we found wide between-male variations in the most efficient dose. This variability was reduced by mixing several irradiated samples collected from different males or given different efficient doses

The chromosomal risk in sperm from heterozygous Robertsonian translocation carriers is related to the sperm count and the translocation type.

Science.gov (United States)

Ferfouri, Fatma; Selva, Jacqueline; Boitrelle, Florence; Gomes, Denise Molina; Torre, Antoine; Albert, Martine; Bailly, Marc; Clement, Patrice; Vialard, François

2011-12-01

To study the chromosomal risk in sperm from Robertsonian translocation (RobT) carriers as a function of the sperm count and translocation type. Prospective study. Departments of reproductive biology, cytogenetics, gynecology, and obstetrics. A total of 29 RobT patients (8 normozoospermic and 21 oligozoospermic) and 20 46,XY patients (10 normozoospermic and 10 oligozoospermic). Sperm fluorescence in situ hybridization with probes for translocation malsegregation and chromosome 13, 18, 21, X, and Y probes for studying the interchromosomal effect (ICE). Translocation malsegregation and ICE aneuploidy rates. In RobT carriers, the sperm translocation malsegregation rate was significantly lower in normozoospermic patients (9.7%) than in oligozoospermic patients (18.0%). Considering only oligozoospermic patients, sperm malsegregation rates were significantly lower for rob(14;21) than for rob(13;14) (11.4% vs. 18.9%). In turn, the rates were significantly lower for rob(13;14) than for rare RobTs (18.9% vs. 25.3%). In sperm from normozoospermic RobT, an ICE was suggested by higher chromosome 13 and 21 aneuploidy rates than in control sperm. Conversely, chromosome 13 and 21 sperm aneuploidy rates were lower in oligozoospermic RobT patients than in oligozoospermic 46,XY patients, but higher than in control subjects. Both translocation type and sperm count influence the RobT malsegregation risk. Of the chromosomes analyzed (13, 18, 21, X, and Y), only chromosomes 13 and 21 were found to be associated with an ICE. Relative to the RobT effect, idiopathic alterations in spermatogenesis in 46,XY patients appear to be more harmful for meiosis. Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Performance Analysis of Exam Gloves Used for Aseptic Rodent Surgery

Science.gov (United States)

LeMoine, Dana M; Bergdall, Valerie K; Freed, Carrie

2015-01-01

Aseptic technique includes the use of sterile surgical gloves for survival surgeries in rodents to minimize the incidence of infections. Exam gloves are much less expensive than are surgical gloves and may represent a cost-effective, readily available option for use in rodent surgery. This study examined the effectiveness of surface disinfection of exam gloves with 70% isopropyl alcohol or a solution of hydrogen peroxide and peracetic acid (HP–PA) in reducing bacterial contamination. Performance levels for asepsis were met when gloves were negative for bacterial contamination after surface disinfection and sham ‘exertion’ activity. According to these criteria, 94% of HP–PA-disinfected gloves passed, compared with 47% of alcohol-disinfected gloves. In addition, the effect of autoclaving on the integrity of exam gloves was examined, given that autoclaving is another readily available option for aseptic preparation. Performance criteria for glove integrity after autoclaving consisted of: the ability to don the gloves followed by successful simulation of wound closure and completion of stretch tests without tearing or observable defects. Using this criteria, 98% of autoclaved nitrile exam gloves and 76% of autoclaved latex exam gloves met performance expectations compared with the performance of standard surgical gloves (88% nitrile, 100% latex). The results of this study support the use of HP–PA-disinfected latex and nitrile exam gloves or autoclaved nitrile exam gloves as viable cost-effective alternatives to sterile surgical gloves for rodent surgeries. PMID:26045458

Performance analysis of exam gloves used for aseptic rodent surgery.

Science.gov (United States)

LeMoine, Dana M; Bergdall, Valerie K; Freed, Carrie

2015-05-01

Aseptic technique includes the use of sterile surgical gloves for survival surgeries in rodents to minimize the incidence of infections. Exam gloves are much less expensive than are surgical gloves and may represent a cost-effective, readily available option for use in rodent surgery. This study examined the effectiveness of surface disinfection of exam gloves with 70% isopropyl alcohol or a solution of hydrogen peroxide and peracetic acid (HP-PA) in reducing bacterial contamination. Performance levels for asepsis were met when gloves were negative for bacterial contamination after surface disinfection and sham 'exertion' activity. According to these criteria, 94% of HP-PA-disinfected gloves passed, compared with 47% of alcohol-disinfected gloves. In addition, the effect of autoclaving on the integrity of exam gloves was examined, given that autoclaving is another readily available option for aseptic preparation. Performance criteria for glove integrity after autoclaving consisted of: the ability to don the gloves followed by successful simulation of wound closure and completion of stretch tests without tearing or observable defects. Using this criteria, 98% of autoclaved nitrile exam gloves and 76% of autoclaved latex exam gloves met performance expectations compared with the performance of standard surgical gloves (88% nitrile, 100% latex). The results of this study support the use of HP-PA-disinfected latex and nitrile exam gloves or autoclaved nitrile exam gloves as viable cost-effective alternatives to sterile surgical gloves for rodent surgeries.

Does breastfeeding influence future sperm quality and reproductive hormones?

DEFF Research Database (Denmark)

Laustsen, J M; Jensen, M S; Thulstrup, Ane Marie

2011-01-01

was not statistically significantly associated with sperm concentration, total sperm count, sperm motility or morphology, oligozoospermia, follicle-stimulating hormone, inhibin B, luteinizing hormone, sex hormone-binding globulin (SHBG), the calculated level of free testosterone, free oestradiol, the free testosterone...... testosterone nor free oestradiol was different between the two groups. This study shows no association between breastfeeding and sperm quality or reproductive hormones and a strong association is unlikely. A larger study would be needed to detect more subtle effects....

Upward swimming of a sperm cell in shear flow.

Science.gov (United States)

Omori, Toshihiro; Ishikawa, Takuji

2016-03-01

Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation.