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Sample records for xenopus egg fertilization

  1. Cortical Isolation from Xenopus laevis Oocytes and Eggs.

    Science.gov (United States)

    Sive, Hazel L; Grainger, Robert M; Harland, Richard M

    2007-06-01

    INTRODUCTIONIn Xenopus laevis, the cortex is the layer of gelatinous cytoplasm that lies just below the plasma membrane of the egg. Rotation of the cortex relative to the deeper cytoplasm soon after fertilization is intimately linked to normal dorsal axis specification. The cortex can be dissected from the egg to analyze its composition and activity or to clone associated RNAs. This protocol describes a procedure for isolating the vegetal cortex of the fertilized egg.

  2. Fertilization and development of eggs of the South African clawed toad, Xenopus laevis, on sounding rockets in space.

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    Ubbels, G A; Berendsen, W; Kerkvliet, S; Narraway, J

    1992-01-01

    Egg rotation and centrifugation experiments strongly suggest a role for gravity in the determination of the spatial structure of amphibian embryos. Decisive experiments can only be made in Space. Eggs of Xenopus laevis, the South African clawed toad, were the first vertebrate eggs which were successfully fertilized on Sounding Rockets in Space. Unfixed, newly fertilized eggs survived reentry, and a reasonable number showed a seemingly normal gastrulation but died between gastrulation and neurulation. Only a few reached the larval stage, but these developed abnormally. In the future, we intend to test whether this abnormal morphogenesis is due to reentry perturbations, or due to a real microgravity effect, through perturbation of the reinitiation of meiosis and other processes, or started by later sperm penetration.

  3. The Sperm-surface glycoprotein, SGP, is necessary for fertilization in the frog, Xenopus laevis.

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    Nagai, Keita; Ishida, Takuya; Hashimoto, Takafumi; Harada, Yuichirou; Ueno, Shuichi; Ueda, Yasushi; Kubo, Hideo; Iwao, Yasuhiro

    2009-06-01

    To identify a molecule involved in sperm-egg plasma membrane binding at fertilization, a monoclonal antibody against a sperm-surface glycoprotein (SGP) was obtained by immunizing mice with a sperm membrane fraction of the frog, Xenopus laevis, followed by screening of the culture supernatants based on their inhibitory activity against fertilization. The fertilization of both jellied and denuded eggs was effectively inhibited by pretreatment of sperm with intact anti-SGP antibody as well as its Fab fragment, indicating that the antibody recognizes a molecule on the sperm's surface that is necessary for fertilization. On Western blots, the anti-SGP antibody recognized large molecules, with molecular masses of 65-150 kDa and minor smaller molecules with masses of 20-28 kDa in the sperm membrane vesicles. SGP was distributed over nearly the entire surface of the sperm, probably as an integral membrane protein in close association with microfilaments. More membrane vesicles containing SGP bound to the surface were found in the animal hemisphere compared with the vegetal hemisphere in unfertilized eggs, but the vesicle-binding was not observed in fertilized eggs. These results indicate that SGP mediates sperm-egg membrane binding and is responsible for the establishment of fertilization in Xenopus.

  4. Xenopus egg cytoplasm with intact actin.

    Science.gov (United States)

    Field, Christine M; Nguyen, Phuong A; Ishihara, Keisuke; Groen, Aaron C; Mitchison, Timothy J

    2014-01-01

    We report optimized methods for preparing Xenopus egg extracts without cytochalasin D, that we term "actin-intact egg extract." These are undiluted egg cytoplasm that contains abundant organelles, and glycogen which supplies energy, and represents the least perturbed cell-free cytoplasm preparation we know of. We used this system to probe cell cycle regulation of actin and myosin-II dynamics (Field et al., 2011), and to reconstitute the large, interphase asters that organize early Xenopus embryos (Mitchison et al., 2012; Wühr, Tan, Parker, Detrich, & Mitchison, 2010). Actin-intact Xenopus egg extracts are useful for analysis of actin dynamics, and interaction of actin with other cytoplasmic systems, in a cell-free system that closely mimics egg physiology, and more generally for probing the biochemistry and biophysics of the egg, zygote, and early embryo. Detailed protocols are provided along with assays used to check cell cycle state and tips for handling and storing undiluted egg extracts. © 2014 Elsevier Inc. All rights reserved.

  5. Fertilization stimulates an increase in inositol trisphosphate and inositol lipid levels in Xenopus eggs.

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    Snow, P; Yim, D L; Leibow, J D; Saini, S; Nuccitelli, R

    1996-11-25

    Previous experiments from our lab have suggested that the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) is required for sperm-induced egg activation in Xenopus laevis. Here we measure the endogenous production of both Ins(1,4,5)P3 and PIP2 during the sperm-induced and ionomycin-induced calcium wave in the egg and find that both increase following fertilization. Ins(1,4,5)P3 increases 3.2-fold from an unfertilized egg level of 0.13 pmole per egg (0.29 microM) to a peak of 0.42 pmole per egg (0.93 microM) as the calcium wave reaches the antipode in the fertilized egg. This continuous production of Ins(1,4,5)P3 during the time that the Ca2+ wave is propagating across the egg suggests the involvement of Ins(1,4,5)P3 in wave propagation. This increase in Ins(1,4,5)P3 is smaller in ionomycin-activated eggs than in sperm-activated eggs, suggesting that the sperm-induced production of Ins(1,4,5)P3 involves a PIP2 hydrolysis pathway that is not simply raising intracellular Ca2+. While one might expect PIP2 levels to fall as a result of hydrolysis, we find that PIP2 actually increases 2-fold. The total lipid fraction in unfertilized egg exhibits 0.8 pmole PIP2 per egg and this increases to 1.5 pmole as the calcium wave reaches the antipode. The PIP2 concentration peaks 2 min after the completion of the calcium wave at 1.8 pmole per egg. The amount of PIP2 in the animal and vegetal hemispheres of the egg was also measured by cutting frozen eggs in half. The vegetal hemisphere contained twice the amount of PIP2 as the animal hemisphere but it also contained twice the amount of lipid. Thus, there was an equivalent amount of PIP2 normalized to lipid in each hemisphere. Isolated animal and vegetal hemisphere cortices exhibit similar PIP2 concentrations, suggesting that the 2-fold higher total PIP2 in the vegetal half is not due to a gradient of PIP2 in the plasma membrane, but rather implies that cytoplasmic organelle membranes also contain PIP2.

  6. Lateral mobility of plasma membrane lipids in Xenopus eggs: regional differences related to animal/vegetal polarity become extreme upon fertilization.

    Science.gov (United States)

    Dictus, W J; van Zoelen, E J; Tetteroo, P A; Tertoolen, L G; de Laat, S W; Bluemink, J G

    1984-01-01

    Regional differences in the lateral mobility properties of plasma membrane lipids have been studied in unfertilized and fertilized Xenopus eggs by fluorescence photobleaching recovery (FPR) measurements. Out of a variety of commonly used lipid probes only the aminofluorescein-labeled fatty acids HEDAF (5-(N-hexadecanoyl)-aminofluorescein) and TEDAF (5-(N-tetradecanoyl)-aminofluorescein) appear to partition into the plasma membrane. Under all experimental conditions used these molecules show partial recovery upon photobleaching indicating the existence of lipidic microdomains. In the unfertilized egg the mobile fraction of plasma membrane lipids (approximately 50%) has a fivefold smaller lateral diffusion coefficient (D = 1.5 X 10(-8) cm2/sec) in the animal than in the vegetal plasma membrane (D = 7.6 X 10(-8) cm2/sec). This demonstrates the presence of an animal/vegetal polarity within the Xenopus egg plasma membrane. Upon fertilization this polarity is strongly (greater than 100X) enhanced leading to the formation of two distinct macrodomains within the plasma membrane. At the animal side of the egg lipids are completely immobilized on the time scale of FPR measurements (D less than 10(-10) cm2/sec), whereas at the vegetal side D is only slightly reduced (D = 4.4 X 10(-8) cm2/sec). The immobilization of animal plasma membrane lipids, which could play a role in the polyspermy block, probably arises by the fusion of cortical granules which are more numerous here. The transition between the animal and the vegetal domain is sharp and coincides with the boundary between the presumptive ecto- and endoderm. The role of regional differences in the plasma membrane is discussed in relation to cell diversification in early development.

  7. Properties of the chromatin assembled on DNA injected into Xenopus oocytes and eggs

    International Nuclear Information System (INIS)

    Gargiulo, G.; Wasserman, W.; Worcel, A.

    1983-01-01

    The onset of DNA synthesis occurs between 10 and 30 minutes after activation of the egg and thus the transition from nuclease-sensitive to nuclease-resistant supercoils may take place on the newly replicated DNA. To test this possibility, the nonradioactive circular 5-kb DNA carrying the Drosophila histone gene repeat and [α -32 P]dCTP were coinjected into fertilized eggs. Such protocol labels both the injected, replicated heterologous DNA and the replicated endogenous, maternal Xenopus DNA. The labeled, presumably replicated, supercoiled DNA is resistant to micrococcal nuclease as expected. The endogenous, high-molecular-weight Xenopus DNA is degraded to 180-bp nucleosomal DNA. Thus, the nuclease resistance is not a general property of chromatin during the cleavage stage of the Xenopus embryo but is a peculiar feature of the injected DNA. 42 references, 5 figures

  8. Analysis of signal transduction in cell-free extracts and rafts of Xenopus eggs.

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    Tokmakov, Alexander A; Iwasaki, Tetsushi; Sato, Ken-Ichi; Fukami, Yasuo

    2010-05-01

    Intracellular signaling during egg activation/fertilization has been extensively studied using intact eggs, which can be manipulated by microinjection of different mRNAs, proteins, or chemical drugs. Furthermore, egg extracts, which retain high CSF activity (CSF-arrested extracts), were developed for studying fertilization/activation signal transduction, which have significant advantages as a model system. The addition of calcium to CSF-arrested extracts initiates a plethora of signaling events that take place during egg activation. Hence, the signaling downstream of calcium mobilization has been successfully studied in the egg extracts. Moreover, despite disruption of membrane-associated signaling compartments and ordered compartmentalization during extract preparation, CSF-arrested extracts can be successfully used to study early signaling events, which occur upstream of calcium release during egg activation/fertilization. In combination with the CSF-arrested extracts, activated egg rafts can reproduce some events of egg activation, including PLCgamma activation, IP3 production, transient calcium release, MAPK inactivation, and meiotic exit. This becomes possible due to complementation of the sperm-induced egg activation signaling machinery present in the rafts with the components of signal transduction system localized in the extracts. Herein, we describe protocols for studying molecular mechanisms of egg fertilization/activation using cell-free extracts and membrane rafts prepared from metaphase-arrested Xenopus eggs.

  9. Membrane junctions in Xenopus eggs: their distribution suggests a role in calcium regulation.

    Science.gov (United States)

    Gardiner, D M; Grey, R D

    1983-04-01

    We have observed the presence of membrane junctions formed between the plasma membrane and cortical endoplasmic reticulum of mature, unactivated eggs of xenopus laevis. The parallel, paired membranes of the junction are separated by a 10-mn gap within which electron-dense material is present. This material occurs in patches with an average center-to-center distance of approximately 30 nm. These junctions are rare in immature (but fully grown) oocytes (approximately 2 percent of the plasma membrane is associated with junctions) and increase dramatically during progesterone-induced maturation. Junctions in the mature, unactivated egg are two to three times more abundant in the animal hemisphere (25-30 percent of the plasma membrane associated with junction) as compared with the vegetal hemisphere (10-15 percent). Junction density decreases rapidly to values characteristic of immature oocytes in response to egg activation. The plasma membrane-ER junctions of xenopus eggs are strikingly similar in structure to membrane junctions in muscle cells thought to be essential in the triggering of intracellular calcium release from the sarcoplasmic reticulum. In addition, the junctions' distinctive, animal-vegetal polarity of distribution, their dramatic appearance during maturation, and their disapperance during activation are correlated with previously documented patterns of calcium-mediated events in anuran eggs. We discuss several lines of evidence supporting the hypothesis that these junctions in xenopus eggs are sites that transduce extracellular events into intracellular calcium release during fertilization and activation of development.

  10. Lateral mobility of plasma membrane lipids in Xenopus eggs: Regional differences related to animal/vegetal polarity

    NARCIS (Netherlands)

    Laat, S.W. de; Bluemink, J.G.; Dictus, W.J.A.G.; Zoelen, E.J.J. van; Tetteroo, P.A.T.; Tertoolen, L.G.J.

    1984-01-01

    Regional differences in the lateral mobility properties of plasma membrane lipids were studied in unfertilized and fertilized Xenopus eggs by fluorescence photobleaching recovery (FPR) measurements. Out of a variety of commonly used lipid probes only the aminofluorescein- -1abelled fatty

  11. Subcellular metabolite and lipid analysis of Xenopus laevis eggs by LAESI mass spectrometry.

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    Shrestha, Bindesh; Sripadi, Prabhakar; Reschke, Brent R; Henderson, Holly D; Powell, Matthew J; Moody, Sally A; Vertes, Akos

    2014-01-01

    Xenopus laevis eggs are used as a biological model system for studying fertilization and early embryonic development in vertebrates. Most methods used for their molecular analysis require elaborate sample preparation including separate protocols for the water soluble and lipid components. In this study, laser ablation electrospray ionization (LAESI), an ambient ionization technique, was used for direct mass spectrometric analysis of X. laevis eggs and early stage embryos up to five cleavage cycles. Single unfertilized and fertilized eggs, their animal and vegetal poles, and embryos through the 32-cell stage were analyzed. Fifty two small metabolite ions, including glutathione, GABA and amino acids, as well as numerous lipids including 14 fatty acids, 13 lysophosphatidylcholines, 36 phosphatidylcholines and 29 triacylglycerols were putatively identified. Additionally, some proteins, for example thymosin β4 (Xen), were also detected. On the subcellular level, the lipid profiles were found to differ between the animal and vegetal poles of the eggs. Radial profiling revealed profound compositional differences between the jelly coat vitelline/plasma membrane and egg cytoplasm. Changes in the metabolic profile of the egg following fertilization, e.g., the decline of polyamine content with the development of the embryo were observed using LAESI-MS. This approach enables the exploration of metabolic and lipid changes during the early stages of embryogenesis.

  12. Transmembrane Signal Transduction in Oocyte Maturation and Fertilization: Focusing on Xenopus laevis as a Model Animal

    Directory of Open Access Journals (Sweden)

    Ken-ichi Sato

    2014-12-01

    Full Text Available Fertilization is a cell biological phenomenon of crucial importance for the birth of new life in a variety of multicellular and sexual reproduction species such as algae, animal and plants. Fertilization involves a sequence of events, in which the female gamete “egg” and the male gamete “spermatozoon (sperm” develop, acquire their functions, meet and fuse with each other, to initiate embryonic and zygotic development. Here, it will be briefly reviewed how oocyte cytoplasmic components are orchestrated to undergo hormone-induced oocyte maturation and sperm-induced activation of development. I then review how sperm-egg membrane interaction/fusion and activation of development in the fertilized egg are accomplished and regulated through egg coat- or egg plasma membrane-associated components, highlighting recent findings and future directions in the studies using Xenopus laevis as a model experimental animal.

  13. Cytoskeleton and gravity at work in the establishment of dorso-ventral polarity in the egg of Xenopus laevis

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    Ubbels, Geertje A.; Brom, Tim G.

    The establishment of polarities during early embryogenesis is essential for normal development. Amphibian eggs are appropriate models for studies on embryonic pattern formation. The animal-vegetal axis of the axially symmetrical amphibian egg originates during oogenesis and foreshadows the main body axis of the embryo. The dorso-ventral polarity is epigenetically established before first cleavage. Recent experiments strongly suggest that in the monospermic eggs of the anuran Xenopus laevis both the cytoskeleton and gravity act in the determination of the dorso-ventral polarity. In order to test the role of gravity in this process, eggs will be fertilized under microgravity conditions during the SL-D1 flight in 1985. In a fully automatic experiment container eggs will be kept under well-defined conditions and artificially fertilized as soon as microgravity is reached; eggs and embryos at different stages will then be fixed for later examination. Back on earth the material will be analysed and we will know whether fertilization under microgravity conditions is possible. If so, the relation of the dorso-ventral axis to the former sperm entry point will be determined on the whole embryos; in addition eggs and embryos will be analysed cytologically.

  14. Lateral mobility of plasma membrane lipids in Xenopus eggs: Regional differences related to animal/vegetal polarity

    OpenAIRE

    Laat, S.W. de; Bluemink, J.G.; Dictus, W.J.A.G.; Zoelen, E.J.J. van; Tetteroo, P.A.T.; Tertoolen, L.G.J.

    1984-01-01

    Regional differences in the lateral mobility properties of plasma membrane lipids were studied in unfertilized and fertilized Xenopus eggs by fluorescence photobleaching recovery (FPR) measurements. Out of a variety of commonly used lipid probes only the aminofluorescein- -1abelled fatty acids HEDAF (5-(N-hexadecanoyl)- aminofluorescein) and TEDAF (5-(N-tetradecanoyl)-aminofluorescein) appear to distribute itself in the plasma membrane. Under all experimental conditions used these molecules s...

  15. Hyperdorsoanterior embryos from Xenopus eggs treated with D2O

    International Nuclear Information System (INIS)

    Scharf, S.R.; Rowning, B.; Wu, M.; Gerhart, J.C.

    1989-01-01

    Excessively dorsalized embryos of Xenopus laevis develop from eggs treated with 30-70% D 2 O for a few minutes within the first third of the cell cycle following fertilization. As the concentration of D 2 O and the duration of exposure are increased, the anatomy of these embryos shifts in the direction of enlarged dorsal and anterior structures and reduced ventral and posterior ones. Twinning of dorsoanterior structures is frequent. Intermediate forms include embryos with large heads but no trunks or tails. The limit form of the series has cylindrical symmetry, with circumferential bands of eye pigment and cement gland, a core of notochord-like tissue, and a centrally located beating heart. D 2 O treatment seems to increase the egg's sensitivity to the dorsalizing effects of cortical rotation and to stimulate the egg to initiate two or more directions of rotation. Such eggs probably establish thereafter a widened and/or duplicated Nieuwkoop center in the vegetal hemisphere, with the subsequent induction of a widened and/or duplicated Spemann organizer region in the marginal zone, which leads to excessive dorsal development. The existence of these anatomical forms indicates the potential of the egg to undertake dorsal development at all positions of its circumference and suggests that normal patterning depends on the limited and localized activation or disinhibition of this widespread potential

  16. Remodeling of ribosomal genes in somatic cells by Xenopus egg extract

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    Ostrup, Olga, E-mail: osvarcova@gmail.com [Institute of Basic Animal and Veterinary Sciences, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C (Denmark); Stem Cell Epigenetics Laboratory, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, Oslo (Norway); Norwegian Center for Stem Cell Research, Oslo (Norway); Hyttel, Poul; Klaerke, Dan A. [Institute of Basic Animal and Veterinary Sciences, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C (Denmark); Collas, Philippe, E-mail: philc@medisin.uio.no [Stem Cell Epigenetics Laboratory, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, Oslo (Norway); Norwegian Center for Stem Cell Research, Oslo (Norway)

    2011-09-02

    Highlights: {yields} Xenopus egg extract remodels nuclei and alter cell growth characteristics. {yields} Ribosomal genes are reprogrammed within 6 h after extract exposure. {yields} rDNA reprogramming involves promoter targeting of SNF2H remodeling complex. {yields} Xenopus egg extract does not initiate stress-related response in somatic cells. {yields} Aza-cytidine elicits a stress-induced response in reprogrammed cells. -- Abstract: Extracts from Xenopus eggs can reprogram gene expression in somatic nuclei, however little is known about the earliest processes associated with the switch in the transcriptional program. We show here that an early reprogramming event is the remodeling of ribosomal chromatin and gene expression. This occurs within hours of extract treatment and is distinct from a stress response. Egg extract elicits remodeling of the nuclear envelope, chromatin and nucleolus. Nucleolar remodeling involves a rapid and stable decrease in ribosomal gene transcription, and promoter targeting of the nucleolar remodeling complex component SNF2H without affecting occupancy of the transcription factor UBF and the stress silencers SUV39H1 and SIRT1. During this process, nucleolar localization of UBF and SIRT1 is not altered. On contrary, azacytidine pre-treatment has an adverse effect on rDNA remodeling induced by extract and elicits a stress-type nuclear response. Thus, an early event of Xenopus egg extract-mediated nuclear reprogramming is the remodeling of ribosomal genes involving nucleolar remodeling complex. Condition-specific and rapid silencing of ribosomal genes may serve as a sensitive marker for evaluation of various reprogramming methods.

  17. Xenopus egg extract: A powerful tool to study genome maintenance mechanisms.

    Science.gov (United States)

    Hoogenboom, Wouter S; Klein Douwel, Daisy; Knipscheer, Puck

    2017-08-15

    DNA repair pathways are crucial to maintain the integrity of our genome and prevent genetic diseases such as cancer. There are many different types of DNA damage and specific DNA repair mechanisms have evolved to deal with these lesions. In addition to these repair pathways there is an extensive signaling network that regulates processes important for repair, such as cell cycle control and transcription. Despite extensive research, DNA damage repair and signaling are not fully understood. In vitro systems such as the Xenopus egg extract system, have played, and still play, an important role in deciphering the molecular details of these processes. Xenopus laevis egg extracts contain all factors required to efficiently perform DNA repair outside a cell, using mechanisms conserved in humans. These extracts have been used to study several genome maintenance pathways, including mismatch repair, non-homologous end joining, ICL repair, DNA damage checkpoint activation, and replication fork stability. Here we describe how the Xenopus egg extract system, in combination with specifically designed DNA templates, contributed to our detailed understanding of these pathways. Copyright © 2017. Published by Elsevier Inc.

  18. Remodeling of ribosomal genes in somatic cells by Xenopus egg extract

    DEFF Research Database (Denmark)

    Østrup, Olga; Hyttel, Poul; Klærke, Dan Arne

    2011-01-01

    Extracts from Xenopus eggs can reprogram gene expression in somatic nuclei, however little is known about the earliest processes associated with the switch in the transcriptional program. We show here that an early reprogramming event is the remodeling of ribosomal chromatin and gene expression....... This occurs within hours of extract treatment and is distinct from a stress response. Egg extract elicits remodeling of the nuclear envelope, chromatin and nucleolus. Nucleolar remodeling involves a rapid and stable decrease in ribosomal gene transcription, and promoter targeting of the nucleolar remodeling...... and elicits a stress-type nuclear response. Thus, an early event of Xenopus egg extract-mediated nuclear reprogramming is the remodeling of ribosomal genes involving nucleolar remodeling complex. Condition-specific and rapid silencing of ribosomal genes may serve as a sensitive marker for evaluation...

  19. Lateral mobility of plasma membrane lipids in dividing Xenopus eggs.

    Science.gov (United States)

    Tetteroo, P A; Bluemink, J G; Dictus, W J; van Zoelen, E J; de Laat, S W

    1984-07-01

    The lateral mobility of plasma membrane lipids was analyzed during first cleavage of Xenopus laevis eggs by fluorescence photobleaching recovery (FPR) measurements, using the lipid analogs 5-(N-hexadecanoyl)aminofluorescein ("HEDAF") and 5-(N-tetradecanoyl)aminofluorescein ("TEDAF") as probes. The preexisting plasma membrane of the animal side showed an inhomogeneous, dotted fluorescence pattern after labeling and the lateral mobility of both probes used was below the detection limits of the FPR method (D much less than 10(-10) cm2/sec). In contrast, the preexisting plasma membrane of the vegetal side exhibited homogeneous fluorescence and the lateral diffusion coefficient of both probes used was relatively high (HEDAF, D = 2.8 X 10(-8) cm2/sec; TEDAF, D = 2.4 X 10(-8) cm2/sec). In the cleaving egg visible transfer of HEDAF or TEDAF from prelabeled plasma membrane to the new membrane in the furrow did not occur, even on the vegetal side. Upon labeling during cleavage, however, the new membrane was uniformly labeled and both probes were mobile, as in the vegetal preexisting plasma membrane. These data show that the membrane of the dividing Xenopus egg comprises three macrodomains: (i) the animal preexisting plasma membrane; (ii) the vegetal preexisting plasma membrane; (iii) the new furrow membrane.

  20. Differential nuclear remodeling of mammalian somatic cells by Xenopus laevis oocyte and egg cytoplasm

    International Nuclear Information System (INIS)

    Alberio, Ramiro; Johnson, Andrew D.; Stick, Reimer; Campbell, Keith H.S.

    2005-01-01

    The mechanisms governing nuclear reprogramming have not been fully elucidated yet; however, recent studies show a universally conserved ability of both oocyte and egg components to reprogram gene expression in somatic cells. The activation of genes associated with pluripotency by oocyte/egg components may require the remodeling of nuclear structures, such that they can acquire the features of early embryos and pluripotent cells. Here, we report on the remodeling of the nuclear lamina of mammalian cells by Xenopus oocyte and egg extracts. Lamin A/C is removed from somatic cells incubated in oocyte and egg extracts in an active process that requires permeable nuclear pores. Removal of lamin A/C is specific, since B-type lamins are not changed, and it is not dependent on the incorporation Xenopus egg specific lamin III. Moreover, transcriptional activity is differentially regulated in somatic cells incubated in the extracts. Pol I and II transcriptions are maintained in cells in oocyte extracts; however, both activities are abolished in egg extracts. Our study shows that components of oocyte and egg extracts can modify the nuclear lamina of somatic cells and that this nuclear remodeling induces a structural change in the nucleus which may have implications for transcriptional activity. These experiments suggest that modifications in the nuclear lamina structure by the removal of somatic proteins and the incorporation of oocyte/egg components may contribute to the reprogramming of somatic cell nuclei and may define a characteristic configuration of pluripotent cells

  1. DNA is a co-factor for its own replication in Xenopus egg extracts

    NARCIS (Netherlands)

    Lebofsky, Ronald; van Oijen, Antoine M.; Walter, Johannes C.

    Soluble Xenopus egg extracts efficiently replicate added plasmids using a physiological mechanism, and thus represent a powerful system to understand vertebrate DNA replication. Surprisingly, DNA replication in this system is highly sensitive to plasmid concentration, being undetectable below

  2. Calcium-responsive contractility during fertilization in sea urchin eggs.

    Science.gov (United States)

    Stack, Christianna; Lucero, Amy J; Shuster, Charles B

    2006-04-01

    Fertilization triggers a reorganization of oocyte cytoskeleton, and in sea urchins, there is a dramatic increase in cortical F-actin. However, the role that myosin II plays during fertilization remains largely unexplored. Myosin II is localized to the cortical cytoskeleton both before and after fertilization and to examine myosin II contractility in living cells, Lytechinus pictus eggs were observed by time-lapse microscopy. Upon sperm binding, a cell surface deflection traversed the egg that was followed by and dependent on the calcium wave. The calcium-dependence of surface contractility could be reproduced in unfertilized eggs, where mobilization of intracellular calcium in unfertilized eggs under compression resulted in a marked contractile response. Lastly, inhibition of myosin II delayed absorption of the fertilization cone, suggesting that myosin II not only responds to the same signals that activate eggs but also participates in the remodeling of the cortical actomyosin cytoskeleton during the first zygotic cell cycle. (c) 2006 Wiley-Liss, Inc.

  3. Evidence for an RNA polymerization activity in axolotl and Xenopus egg extracts.

    Directory of Open Access Journals (Sweden)

    Hélène Pelczar

    2010-12-01

    Full Text Available We have previously reported a post-transcriptional RNA amplification observed in vivo following injection of in vitro synthesized transcripts into axolotl oocytes, unfertilized (UFE or fertilized eggs. To further characterize this phenomenon, low speed extracts (LSE from axolotl and Xenopus UFE were prepared and tested in an RNA polymerization assay. The major conclusions are: i the amphibian extracts catalyze the incorporation of radioactive ribonucleotide in RNase but not DNase sensitive products showing that these products correspond to RNA; ii the phenomenon is resistant to α-amanitin, an inhibitor of RNA polymerases II and III and to cordycepin (3'dAMP, but sensitive to cordycepin 5'-triphosphate, an RNA elongation inhibitor, which supports the existence of an RNA polymerase activity different from polymerases II and III; the detection of radiolabelled RNA comigrating at the same length as the exogenous transcript added to the extracts allowed us to show that iii the RNA polymerization is not a 3' end labelling and that iv the radiolabelled RNA is single rather than double stranded. In vitro cell-free systems derived from amphibian UFE therefore validate our previous in vivo results hypothesizing the existence of an evolutionary conserved enzymatic activity with the properties of an RNA dependent RNA polymerase (RdRp.

  4. Identification of metalloprotease/disintegrins in Xenopus laevis testis with a potential role in fertilization.

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    Shilling, F M; Krätzschmar, J; Cai, H; Weskamp, G; Gayko, U; Leibow, J; Myles, D G; Nuccitelli, R; Blobel, C P

    1997-06-15

    Proteins containing a membrane-anchored metalloprotease domain, a disintegrin domain, and a cysteine-rich region (MDC proteins) are thought to play an important role in mammalian fertilization, as well as in somatic cell-cell interactions. We have identified PCR sequence tags encoding the disintegrin domain of five distinct MDC proteins from Xenopus laevis testis cDNA. Four of these sequence tags (xMDC9, xMDC11.1, xMDC11.2, and xMDC13) showed strong similarity to known mammalian MDC proteins, whereas the fifth (xMDC16) apparently represents a novel family member. Northern blot analysis revealed that the mRNA for xMDC16 was only expressed in testis, and not in heart, muscle, liver, ovaries, or eggs, whereas the mRNAs corresponding to the four other PCR products were expressed in testis and in some or all somatic tissues tested. The xMDC16 protein sequence, as predicted from the full-length cDNA, contains a metalloprotease domain with the active-site sequence HEXXH, a disintegrin domain, a cysteine-rich region, an EGF repeat, a transmembrane domain, and a short cytoplasmic tail. To study a potential role for these xMDC proteins in fertilization, peptides corresponding to the predicted integrin-binding domain of each protein were tested for their ability to inhibit X. laevis fertilization. Cyclic and linear xMDC16 peptides inhibited fertilization in a concentration-dependent manner, whereas xMDC16 peptides that were scrambled or had certain amino acid replacements in the predicted integrin-binding domain did not affect fertilization. Cyclic and linear xMDC9 peptides and linear xMDC13 peptides also inhibited fertilization similarly to xMDC16 peptides, whereas peptides corresponding to the predicted integrin-binding site of xMDC11.1 and xMDC11.2 did not. These results are discussed in the context of a model in which multiple MDC protein-receptor interactions are necessary for fertilization to occur.

  5. Quantification of egg proteome changes during fertilization in sterlet Acipenser ruthenus.

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    Niksirat, Hamid; Andersson, Liselotte; Golpour, Amin; Chupani, Latifeh; James, Peter

    2017-08-19

    Eggs of sterlet are discharged outside into ambient aquatic environment where egg activation and fertilization occur. Effects of different activation media including freshwater and clay suspension on protein abundances of egg were quantified in sterlet Acipenser ruthenus. In-gel digestion and high resolution mass spectrometry were used for label-free protein quantification in the eggs of five females. No significant (p > 0.05) difference was found between protein abundances in eggs activated with different media. However, results showed significant (p eggs as control. The fact that abundance of proteasome subunit alpha significantly reduced only in eggs which were activated by clay suspension suggests that activation medium can somehow intervene with protein regulation during fertilization. In conclusion, external fertilization in sturgeon egg is accompanied by huge release of proteins into the external environment that may participate in the construction of a transient microenvironment around egg for attraction and protection of spermatozoa to ensure ensuing fertilization. Data are available via ProteomeXchange with identifier PXD006232. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Membrane junctions in xenopus eggs: their distribution suggests a role in calcium regulation

    OpenAIRE

    Gardiner, DM; Grey, RD

    1983-01-01

    We have observed the presence of membrane junctions formed between the plasma membrane and cortical endoplasmic reticulum of mature, unactivated eggs of xenopus laevis. The parallel, paired membranes of the junction are separated by a 10-mn gap within which electron-dense material is present. This material occurs in patches with an average center-to-center distance of approximately 30 nm. These junctions are rare in immature (but fully grown) oocytes (approximately 2 percent of the plasma mem...

  7. Slow fertilization of stickleback eggs: the result of sexual conflict?

    Directory of Open Access Journals (Sweden)

    Frommen Joachim G

    2006-05-01

    Full Text Available Abstract Background The fertilization success in sperm competition in externally fertilizing fish depends on number and quality of sperm. The time delay between sequential ejaculations may further influence the outcome of sperm competition. Such a time interval can load the raffle over fertilization if fertilization takes place very fast. Short fertilization times are generally assumed for externally fertilizing fish such as the three-spined stickleback (Gasterosteus aculeatus. In this pair-spawning fish, territorial males often try to steal fertilizations in nests of neighbouring males. This sneaking behaviour causes sperm competition. Sneakers will only get a share of paternity when eggs are not fertilized immediately after sperm release. Contrary to males, females may be interested in multiple paternity of their clutch of eggs. There thus may be a sexual conflict over the speed of fertilization. Results In this study we used two different in vitro fertilization experiments to assess how fast eggs are fertilized in sticklebacks. We show that complete fertilization takes more than 5 min which is atypically long for externally fertilizing fishes. Conclusion This result suggests that the time difference does not imply high costs to the second stickleback male to ejaculate. Slow fertilization (and concomitant prolonged longevity of sperm may be the result of sexual conflict in which females aimed at complete fertilization and/or multiple paternity.

  8. Long-term effect on in vitro cloning efficiency after treatment of somatic cells with Xenopus egg extract in the pig.

    Science.gov (United States)

    Liu, Ying; Ostrup, Olga; Li, Rong; Li, Juan; Vajta, Gábor; Kragh, Peter M; Schmidt, Mette; Purup, Stig; Hyttel, Poul; Klærke, Dan; Callesen, Henrik

    2014-08-01

    In somatic cell nuclear transfer (SCNT), donor cell reprogramming is considered as a biologically important and vulnerable event. Various donor cell pre-treatments with Xenopus egg extracts can promote reprogramming. Here we investigated if the reprogramming effect of one treatment with Xenopus egg extract on donor cells was maintained for several cell passages. The extract treatment resulted in increased cell-colony formation from early passages in treated porcine fibroblasts (ExTES), and increased development of cloned embryos. Partial dedifferentiation was observed in ExTES cells, shown as a tendency towards upregulation of NANOG, c-MYC and KLF-4 and downregulation of DESMIM compared with ExTES at Passage 2. Compared with our routine SCNT, continuously increased development of cloned embryos was observed in the ExTES group, and ExTES cloned blastocysts displayed hypermethylated DNA patterns and hypermethylation of H3K4me3 and H3K27me3 in ICM compared with TE. All seven recipients became pregnant after transferral of ExTES cloned embryos and gave birth to 7-22 piglets per litter (average 12). In conclusion, our results demonstrate that one treatment of porcine fibroblasts with Xenopus egg extract can result in long-term increased ability of the cells to promote their in vitro function in subsequent SCNT. Finally these cells can also result in successful development of cloned embryos to term.

  9. Increased blastocyst formation of cloned porcine embryos produced with donor cells pre-treated with Xenopus egg extract and/or digitonin

    DEFF Research Database (Denmark)

    Liu, Ying; Østrup, Olga; Li, Juan

    2012-01-01

    from Xenopus laevis eggs. In Experiment 1, fetal fibroblasts were permeabilized by digitonin, incubated in egg extract and, after re-sealing of cell membranes, cultured for 3 or 5 days before use as donor cells in handmade cloning (HMC). Controls were produced by HMC with non-treated donor cells....... The blastocyst rate for reconstructed embryos increased significantly when digitonin-permeabilized, extract-treated cells were used after 5 days of culture after re-sealing. In Experiment 2, fetal and adult fibroblasts were treated with digitonin alone before re-sealing the cell membranes, then cultured for 3...... cells after pre-treatment with permeabilization/re-sealing and Xenopus egg extract. Interestingly, we observe a similar increase in cloning efficiency by permeabilization/re-sealing of donor cells without extract treatment that seems to depend on choice of donor cell type. Thus, pre-treatment of donor...

  10. Protein-Carbohydrate Interaction between Sperm and the Egg-Coating Envelope and Its Regulation by Dicalcin, a Xenopus laevis Zona Pellucida Protein-Associated Protein

    Directory of Open Access Journals (Sweden)

    Naofumi Miwa

    2015-05-01

    Full Text Available Protein-carbohydrate interaction regulates multiple important processes during fertilization, an essential biological event where individual gametes undergo intercellular recognition to fuse and generate a zygote. In the mammalian female reproductive tract, sperm temporarily adhere to the oviductal epithelium via the complementary interaction between carbohydrate-binding proteins on the sperm membrane and carbohydrates on the oviductal cells. After detachment from the oviductal epithelium at the appropriate time point following ovulation, sperm migrate and occasionally bind to the extracellular matrix, called the zona pellucida (ZP, which surrounds the egg, thereafter undergoing the exocytotic acrosomal reaction to penetrate the envelope and to reach the egg plasma membrane. This sperm-ZP interaction also involves the direct interaction between sperm carbohydrate-binding proteins and carbohydrates within the ZP, most of which have been conserved across divergent species from mammals to amphibians and echinoderms. This review focuses on the carbohydrate-mediated interaction of sperm with the female reproductive tract, mainly the interaction between sperm and the ZP, and introduces the fertilization-suppressive action of dicalcin, a Xenopus laevis ZP protein-associated protein. The action of dicalcin correlates significantly with a dicalcin-dependent change in the lectin-staining pattern within the ZP, suggesting a unique role of dicalcin as an inherent protein that is capable of regulating the affinity between the lectin and oligosaccharides attached on its target glycoprotein.

  11. 9 CFR 93.905 - Declaration and other documents for live fish, fertilized eggs, and gametes.

    Science.gov (United States)

    2010-01-01

    ... live fish, fertilized eggs, and gametes. 93.905 Section 93.905 Animals and Animal Products ANIMAL AND... for live fish, fertilized eggs, and gametes. (a) For all live fish, fertilized eggs, and gametes... fish, fertilized eggs, or gametes, the number, species, and the purpose of the importation, the name of...

  12. 9 CFR 93.903 - Import permits for live fish, fertilized eggs, and gametes.

    Science.gov (United States)

    2010-01-01

    ..., fertilized eggs, and gametes. 93.903 Section 93.903 Animals and Animal Products ANIMAL AND PLANT HEALTH... General Provisions for Svc-Regulated Fish Species § 93.903 Import permits for live fish, fertilized eggs, and gametes. (a) Live fish, fertilized eggs, or gametes of SVC-susceptible species imported into the...

  13. Fertility and hatchability of Japanese quail eggs under semi arid ...

    African Journals Online (AJOL)

    Some factors affecting the fertility and hatchability of Japanese quail eggs were studied at the Poultry Unit of the University of Maiduguri Livestock Teaching and Research Farm. A total of 1850 eggs were used for the study and data collected were subjected to Analysis of Variance. The mean values of fertility, hatchability of ...

  14. Metabolic Regulation of CaMKII Protein and Caspases in Xenopus laevis Egg Extracts*

    Science.gov (United States)

    McCoy, Francis; Darbandi, Rashid; Chen, Si-Ing; Eckard, Laura; Dodd, Keela; Jones, Kelly; Baucum, Anthony J.; Gibbons, Jennifer A.; Lin, Sue-Hwa; Colbran, Roger J.; Nutt, Leta K.

    2013-01-01

    The metabolism of the Xenopus laevis egg provides a cell survival signal. We found previously that increased carbon flux from glucose-6-phosphate (G6P) through the pentose phosphate pathway in egg extracts maintains NADPH levels and calcium/calmodulin regulated protein kinase II (CaMKII) activity to phosphorylate caspase 2 and suppress cell death pathways. Here we show that the addition of G6P to oocyte extracts inhibits the dephosphorylation/inactivation of CaMKII bound to caspase 2 by protein phosphatase 1. Thus, G6P sustains the phosphorylation of caspase 2 by CaMKII at Ser-135, preventing the induction of caspase 2-mediated apoptotic pathways. These findings expand our understanding of oocyte biology and clarify mechanisms underlying the metabolic regulation of CaMKII and apoptosis. Furthermore, these findings suggest novel approaches to disrupt the suppressive effects of the abnormal metabolism on cell death pathways. PMID:23400775

  15. Cadmium but not lead exposure affects Xenopus laevis fertilization and embryo cleavage

    Energy Technology Data Exchange (ETDEWEB)

    Slaby, Sylvain [Univ. Lille Nord de France, EA 4515 – LGCgE – Laboratoire Génie Civil et géo-Environnement, Université de Lille 1, Cité scientifique, SN3, F-59655 Villeneuve d’Ascq (France); Univ. Lille, CNRS, INRA, UMR 8576 – UGSF – Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille (France); Lemière, Sébastien [Univ. Lille Nord de France, EA 4515 – LGCgE – Laboratoire Génie Civil et géo-Environnement, Université de Lille 1, Cité scientifique, SN3, F-59655 Villeneuve d’Ascq (France); Hanotel, Julie; Lescuyer, Arlette [Univ. Lille, CNRS, INRA, UMR 8576 – UGSF – Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille (France); Demuynck, Sylvain [Univ. Lille Nord de France, EA 4515 – LGCgE – Laboratoire Génie Civil et géo-Environnement, Université de Lille 1, Cité scientifique, SN3, F-59655 Villeneuve d’Ascq (France); Bodart, Jean-François [Univ. Lille, CNRS, INRA, UMR 8576 – UGSF – Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille (France); and others

    2016-08-15

    Highlights: • First embryonic steps were studied. • Fertilization success was impacted by cadmium exposures. • Oocytes were most affected instead of spermatozoa by cadmium exposures. • First embryonic cleavages were slown down or stopped by cadmium exposures. • Lead exposures did not affected fertilization and segmentation. - Abstract: Among the toxicological and ecotoxicological studies, few have investigated the effects on germ cells, gametes or embryos, while an impact at these stages will result in serious damage at a population level. Thus, it appeared essential to characterize consequences of environmental contaminant exposures at these stages. Therefore, we proposed to assess the effects of exposure to cadmium and lead ions, alone or in a binary mixture, on early stages of Xenopus laevis life cycle. Fertilization and cell division during segmentation were the studied endpoints. Cadmium ion exposures decreased in the fertilization rates in a concentration-dependent manner, targeting mainly the oocytes. Exposure to this metal ions induced also delays or blockages in the embryonic development. For lead ion exposure, no such effect was observed. For the exposure to the mixture of the two metal ions, concerning the fertilization success, we observed results similar to those obtained with the highest cadmium ion concentration.

  16. Sea urchin egg fertilization and development

    Science.gov (United States)

    Young, R. S.

    1971-01-01

    The effects of subgravity (much less than unit gravity) on fertilization, cell division, differentiation, and growth of a relatively simple biological system (eggs of the sea urchin Arbacia punctulata) were considered. The experiment was flown on Gemini 3 and recovered as scheduled. However, the experiment objectives were not achieved, primarily for mechanical reasons.

  17. 9 CFR 93.902 - Ports designated for the importation of live fish, fertilized eggs, and gametes.

    Science.gov (United States)

    2010-01-01

    ... of live fish, fertilized eggs, and gametes. 93.902 Section 93.902 Animals and Animal Products ANIMAL... importation of live fish, fertilized eggs, and gametes. (a) The following ports are designated as ports of entry for live fish, fertilized eggs, and gametes of SVC-susceptible species imported under this subpart...

  18. A Biochemical Screen for Identification of Small-Molecule Regulators of the Wnt Pathway Using Xenopus Egg Extracts

    OpenAIRE

    Thorne, Curtis A.; Lafleur, Bonnie; Lewis, Michelle; Hanson, Alison J.; Jernigan, Kristin K.; Weaver, David C.; Huppert, Kari A.; Chen, Tony W.; Wichadiit, Chonlarat; Cselenyi, Christopher S.; Tahinci, Emilios; Meyers, Kelly C.; Waskow, Emily; Orton, Darren; Salic, Adrian

    2011-01-01

    Misregulation of the Wnt pathway has been shown to be responsible for a variety of human diseases, most notably cancers. Screens for inhibitors of this pathway have been performed almost exclusively using cultured mammalian cells or with purified proteins. We have previously developed a biochemical assay using Xenopus egg extracts to recapitulate key cytoplasmic events in the Wnt pathway. Using this biochemical system, we show that a recombinant form of the Wnt coreceptor, LRP6, regulates the...

  19. How does the Xenopus laevis embryonic cell cycle avoid spatial chaos?

    Science.gov (United States)

    Gelens, Lendert; Huang, Kerwyn Casey; Ferrell, James E.

    2015-01-01

    Summary Theoretical studies have shown that a deterministic biochemical oscillator can become chaotic when operating over a sufficiently large volume, and have suggested that the Xenopus laevis cell cycle oscillator operates close to such a chaotic regime. To experimentally test this hypothesis, we decreased the speed of the post-fertilization calcium wave, which had been predicted to generate chaos. However, cell divisions were found to develop normally and eggs developed into normal tadpoles. Motivated by these experiments, we carried out modeling studies to understand the prerequisites for the predicted spatial chaos. We showed that this type of spatial chaos requires oscillatory reaction dynamics with short pulse duration, and postulated that the mitotic exit in Xenopus laevis is likely slow enough to avoid chaos. In systems with shorter pulses, chaos may be an important hazard, as in cardiac arrhythmias, or a useful feature, as in the pigmentation of certain mollusk shells. PMID:26212326

  20. Mutant analysis of Cdt1's function in suppressing nascent strand elongation during DNA replication in Xenopus egg extracts.

    Science.gov (United States)

    Nakazaki, Yuta; Tsuyama, Takashi; Azuma, Yutaro; Takahashi, Mikiko; Tada, Shusuke

    2017-09-02

    The initiation of DNA replication is strictly regulated by multiple mechanisms to ensure precise duplication of chromosomes. In higher eukaryotes, activity of the Cdt1 protein is temporally regulated during the cell cycle, and deregulation of Cdt1 induces DNA re-replication. In previous studies, we showed that excess Cdt1 inhibits DNA replication by suppressing progression of replication forks in Xenopus egg extracts. Here, we investigated the functional regions of Cdt1 that are required for the inhibition of DNA replication. We constructed a series of N-terminally or C-terminally deleted mutants of Cdt1 and examined their inhibitory effects on DNA replication in Xenopus egg extracts. Our results showed that the region spanning amino acids (a. a.) 255-620 is required for efficient inhibition of DNA replication, and that, within this region, a. a. 255-289 have a critical role in inhibition. Moreover, one of the Cdt1 mutants, Cdt1 R285A, was compromised with respect to the licensing activity but still inhibited DNA replication. This result suggests that Cdt1 has an unforeseen function in the negative regulation of DNA replication, and that this function is located within a molecular region that is distinct from those required for the licensing activity. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Repair capacity of fertilized mouse eggs for X-ray damage induced in sperm and mature oocytes

    International Nuclear Information System (INIS)

    Matsuda, Yoichi; Tobari, Izuo

    1989-01-01

    To study the repair capacity of fertilized mouse eggs for X-ray damage induced in sperm and mature oocytes, the potentiating effects of 3 well-known repair inhibitors, arabinofuranosyl cytosine (ara-C), 3-aminobenzamide (3AB) and caffeine, on the frequency of induced chromosome aberrations were examined in eggs fertilized with X-irradiated sperm or in eggs irradiated with X-rays at the mature oocyte stage immediately before fertilization. Gametic treatment, fertilization and embryo culture wer carried out in vitro. Ara-C treatment was done only in the pre-DNA replication period, while treatment with 3AB and caffeine was continuous from fertilization to the first-cleavage metaphase. The induction of chromosome aberrations by exposing sperm or oocytes to X-rays was remarkably potentiated by post-treatment incubation in the presence of each of the 3 inhibitors. This result indicates the possibility that X-ray damage induced in sperm or oocytes is reparable in the fertilized eggs and that various types of repair processes are involved. (author). 39 refs.; 3 figs.; 5 tabs

  2. Fertilization Induces a Transient Exposure of Phosphatidylserine in Mouse Eggs

    Science.gov (United States)

    Curia, Claudio A.; Ernesto, Juan I.; Stein, Paula; Busso, Dolores; Schultz, Richard M.; Cuasnicu, Patricia S.; Cohen, Débora J.

    2013-01-01

    Phosphatidylserine (PS) is normally localized to the inner leaflet of the plasma membrane and the requirement of PS translocation to the outer leaflet in cellular processes other than apoptosis has been demonstrated recently. In this work we investigated the occurrence of PS mobilization in mouse eggs, which express flippase Atp8a1 and scramblases Plscr1 and 3, as determined by RT-PCR; these enzyme are responsible for PS distribution in cell membranes. We find a dramatic increase in binding of flouresceinated-Annexin-V, which specifically binds to PS, following fertilization or parthenogenetic activation induced by SrCl2 treatment. This increase was not observed when eggs were first treated with BAPTA-AM, indicating that an increase in intracellular Ca2+ concentration was required for PS exposure. Fluorescence was observed over the entire egg surface with the exception of the regions overlying the meiotic spindle and sperm entry site. PS exposure was also observed in activated eggs obtained from CaMKIIγ null females, which are unable to exit metaphase II arrest despite displaying Ca2+ spikes. In contrast, PS exposure was not observed in TPEN-activated eggs, which exit metaphase II arrest in the absence of Ca2+ release. PS exposure was also observed when eggs were activated with ethanol but not with a Ca2+ ionophore, suggesting that the Ca2+ source and concentration are relevant for PS exposure. Last, treatment with cytochalasin D, which disrupts microfilaments, or jasplakinolide, which stabilizes microfilaments, prior to egg activation showed that PS externalization is an actin-dependent process. Thus, the Ca2+ rise during egg activation results in a transient exposure of PS in fertilized eggs that is not associated with apoptosis. PMID:23951277

  3. Branching microtubule nucleation in Xenopus egg extracts mediated by augmin and TPX2

    Science.gov (United States)

    Petry, Sabine; Groen, Aaron C.; Ishihara, Keisuke; Mitchison, Timothy J.; Vale, Ronald D.

    2013-01-01

    Summary The microtubules that comprise mitotic spindles in animal cells are nucleated at centrosomes and by spindle assembly factors that are activated in the vicinity of chromatin. Indirect evidence also has suggested that microtubules might be nucleated from pre-existing microtubules throughout the spindle, but this process has not been observed directly. Here, we demonstrate microtubule nucleation from the sides of existing microtubules in meiotic Xenopus egg extracts. Daughter microtubules grow at a low branch angle and with the same polarity as mother filaments. Branching microtubule nucleation requires gamma-tubulin and augmin and is stimulated by GTP-bound Ran and its effector TPX2, factors previously implicated in chromatin-stimulated nucleation. Because of the rapid amplification of microtubule numbers and the preservation of microtubule polarity, microtubule-dependent microtubule nucleation is well suited for spindle assembly and maintenance. PMID:23415226

  4. Effects of salinity and sea salt type on egg activation, fertilization, buoyancy and early embryology of European eel, Anguilla anguilla

    DEFF Research Database (Denmark)

    Sørensen, Sune Riis; Butts, Ian; Munk, Peter

    2016-01-01

    sizes, while the remaining four salt types resulted in smaller eggs. All salt types except NaCl treatments led to high fertilization rates and had no effect on fertilization success as well as egg neutral buoyancies at 7 h post-fertilization. The study points to the importance of considering ionic...... and egg buoyancy. Egg diameter after activation, using natural seawater adjusted to different salinities, varied among female eels, but no consistent pattern emerged. Activation salinities between 30–40 practical salinity unit (psu) produced higher quality eggs and generally larger egg diameters. Chorion...

  5. The Expression of TALEN before Fertilization Provides a Rapid Knock-Out Phenotype in Xenopus laevis Founder Embryos.

    Science.gov (United States)

    Miyamoto, Kei; Suzuki, Ken-Ichi T; Suzuki, Miyuki; Sakane, Yuto; Sakuma, Tetsushi; Herberg, Sarah; Simeone, Angela; Simpson, David; Jullien, Jerome; Yamamoto, Takashi; Gurdon, J B

    2015-01-01

    Recent advances in genome editing using programmable nucleases have revolutionized gene targeting in various organisms. Successful gene knock-out has been shown in Xenopus, a widely used model organism, although a system enabling less mosaic knock-out in founder embryos (F0) needs to be explored in order to judge phenotypes in the F0 generation. Here, we injected modified highly active transcription activator-like effector nuclease (TALEN) mRNA to oocytes at the germinal vesicle (GV) stage, followed by in vitro maturation and intracytoplasmic sperm injection, to achieve a full knock-out in F0 embryos. Unlike conventional injection methods to fertilized embryos, the injection of TALEN mRNA into GV oocytes allows expression of nucleases before fertilization, enabling them to work from an earlier stage. Using this procedure, most of developed embryos showed full knock-out phenotypes of the pigmentation gene tyrosinase and/or embryonic lethal gene pax6 in the founder generation. In addition, our method permitted a large 1 kb deletion. Thus, we describe nearly complete gene knock-out phenotypes in Xenopus laevis F0 embryos. The presented method will help to accelerate the production of knock-out frogs since we can bypass an extra generation of about 1 year in Xenopus laevis. Meantime, our method provides a unique opportunity to rapidly test the developmental effects of disrupting those genes that do not permit growth to an adult able to reproduce. In addition, the protocol shown here is considerably less invasive than the previously used host transfer since our protocol does not require surgery. The experimental scheme presented is potentially applicable to other organisms such as mammals and fish to resolve common issues of mosaicism in founders.

  6. Development of a rapid and efficient microinjection technique for gene insertion into fertilized salmonid eggs

    Energy Technology Data Exchange (ETDEWEB)

    Chandler, D.P.; Welt, M.; Leung, F.C.

    1990-10-01

    An efficient one-step injection technique for gene insertion into fertilized rainbow trout (Oncorhynchus mykiss) eggs is described, and basic parameters affecting egg survival are reported. Freshly fertilized rainbow trout eggs were injected in the perivitelline space with a recombinant mouse metallothionein-genomic bovine growth hormone (bGH) DNA construct using a 30-gauge hypodermic needle and a standard microinjection system. Relative to control, site of injection and DNA concentration did not affect the egg survival, but injections later than 3--4 hours post fertilization were detrimental. The injection technique permitted treatment of 100 eggs/hr with survivals up to 100%, resulting in a 4% DNA uptake rate as indicated by DNA dot blot analysis. Positive dot blot results also indicated that the injected DNA is able to cross the vitelline membrane and persist for 50--60 days post hatching, obviating the need for direct injection into the germinal disk. Results are consistent with previous transgenic fish work, underscoring the usefulness of the technique for generating transgenic trout and salmonids. 24 refs., 6 figs., 3 tabs.

  7. Egg cell-secreted EC1 triggers sperm cell activation during double fertilization.

    Science.gov (United States)

    Sprunck, Stefanie; Rademacher, Svenja; Vogler, Frank; Gheyselinck, Jacqueline; Grossniklaus, Ueli; Dresselhaus, Thomas

    2012-11-23

    Double fertilization is the defining characteristic of flowering plants. However, the molecular mechanisms regulating the fusion of one sperm with the egg and the second sperm with the central cell are largely unknown. We show that gamete interactions in Arabidopsis depend on small cysteine-rich EC1 (EGG CELL 1) proteins accumulating in storage vesicles of the egg cell. Upon sperm arrival, EC1-containing vesicles are exocytosed. The sperm endomembrane system responds to exogenously applied EC1 peptides by redistributing the potential gamete fusogen HAP2/GCS1 (HAPLESS 2/GENERATIVE CELL SPECIFIC 1) to the cell surface. Furthermore, fertilization studies with ec1 quintuple mutants show that successful male-female gamete interactions are necessary to prevent multiple-sperm cell delivery. Our findings provide evidence that mutual gamete activation, regulated exocytosis, and sperm plasma membrane modifications govern flowering plant gamete interactions.

  8. Calculating the Degradation Rate of Individual Proteins Using Xenopus Extract Systems.

    Science.gov (United States)

    McDowell, Gary S; Philpott, Anna

    2018-05-16

    The Xenopus extract system has been used extensively as a simple, quick, and robust method for assessing the stability of proteins against proteasomal degradation. In this protocol, methods are provided for assessing the half-life of in vitro translated radiolabeled proteins using Xenopus egg or embryo extracts. © 2019 Cold Spring Harbor Laboratory Press.

  9. Process of egg formation in the female body cavity and fertilization in male eggs of Phytoseiulus persimilis (Acari: Phytoseiidae).

    Science.gov (United States)

    Toyoshima, S; Nakamura, M; Nagahama, Y; Amano, H

    2000-01-01

    The process of egg formation in the body cavity of a phytoseiid mite, Phytoseiulus persimilis, was observed to examine fertilization of male eggs. After insemination, one of the ova at the periphery of the ovary began to expand, taking up yolk. Two pronuclei appeared in the expanded egg, located dorsally in the ovary, and yolk granules were formed gradually. After the egg became filled with yolk granules the two pronuclei fused. The egg moved via the narrow entrance at the ventral region into the oviduct, where the eggshell was formed. When the eggshell was complete, and while embryogenesis proceeded, the egg was deposited. In the meantime some ova began to expand sequentially and two joining pronuclei appeared in expanding eggs. The joining pronuclei in the first egg proved male diploidy. This is additional evidence of pseudo-arrhenotoky in this phytoseiid mite species, since the first eggs developed into males.

  10. A general solution for optimal egg size during external fertilization, extended scope for intermediate optimal egg size and the introduction of Don Ottavio 'tango'

    NARCIS (Netherlands)

    Luttikhuizen, PC; Honkoop, PJC; Drent, J; van der Meer, J

    2004-01-01

    Egg sizes of marine invertebrates vary greatly, both within and between species. Among the proposed causes of this are a trade-off between egg size, egg number and survival probability of offspring, and a selection pressure exerted by sperm limitation during external fertilization. Although larger

  11. Dynein-Based Accumulation of Membranes Regulates Nuclear Expansion in Xenopus laevis Egg Extracts.

    Science.gov (United States)

    Hara, Yuki; Merten, Christoph A

    2015-06-08

    Nuclear size changes dynamically during development and has long been observed to correlate with the space surrounding the nucleus, as well as with the volume of the cell. Here we combine an in vitro cell-free system of Xenopus laevis egg extract with microfluidic devices to systematically analyze the effect of spatial constraints. The speed of nuclear expansion depended on the available space surrounding the nucleus up to a threshold volume in the nanoliter range, herein referred to as the nuclear domain. Under spatial constraints smaller than this nuclear domain, the size of microtubule-occupied space surrounding the nucleus turned out to be limiting for the accumulation of membranes around the nucleus via the motor protein dynein, therefore determining the speed of nuclear expansion. This mechanism explains how spatial information surrounding the nucleus, such as the positioning of the nucleus inside the cell, can control nuclear expansion. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. MCM interference during licensing of DNA replication in Xenopus egg extracts-Possible Role of a C-terminal region of MCM3.

    Science.gov (United States)

    Mimura, Satoru; Kubota, Yumiko; Takisawa, Haruhiko

    2018-01-01

    The minichromosome maintenance (MCM) complex, consisting of six subunits, Mcm2-7, is loaded onto replication origins through loading factors (origin recognition complex [ORC], Cdc6, and Cdt1) and forms an MCM double hexamer that licenses the initiation of DNA replication. Previous studies with Xenopus egg extracts showed that loading factors, especially Cdc6, dissociate from chromatin on MCM loading, but the molecular mechanism and physiological significance remain largely unknown. Using a cell-free system for MCM loading onto plasmid DNA in Xenopus egg extracts, we found that MCM loaded onto DNA prevents DNA binding of the loading factors ORC, Cdc6, and Cdt1. We further report that a peptide of the C-terminal region of MCM3 (MCM3-C), previously implicated in the initial association with ORC/Cdc6 in budding yeast, prevents ORC/Cdc6/Cdt1 binding to DNA in the absence of MCM loading. ATP-γ-S suppresses inhibitory activities of both the MCM loaded onto DNA and the MCM3-C peptide. Other soluble factors in the extract, but neither MCM nor Cdt1, are required for the activity. Conservation of the amino acid sequences of MCM3-C and its activity in vertebrates implies a novel negative autoregulatory mechanism that interferes with MCM loading in the vicinity of licensed origins to ensure proper origin licensing.

  13. Garlic essential oil increases rates of eggs fertilization and hatching of Rhamdia quelen larvae in an artificial incubation system

    Directory of Open Access Journals (Sweden)

    Nilton Garcia Marengoni

    Full Text Available ABSTRACT: The objective of this study was to evaluate the effects of garlic (Allium sativum essential oil on the rates of eggs fertilization and hatching and on the normal development of larvae of artificially incubated silver catfish (Rhamdia quelen. The experiment was carried out using a completely randomized experimental design with four treatments and five replicates. The treatments consisted of introducing garlic essential oil into the incubators, which used a closed system of water recirculation, at concentrations of 0, 1, 3, and 5mg L-1. Rates of eggs fertilization and larval hatching were determined at 10 and 24h after the beginning of eggs hydration. Morphology of larvae was assessed with the aid of a stereo microscope to determine the rate of normal development. The concentration of 5mg L-1 promoted better rates of eggs fertilization and larval hatching. Concentrations of garlic essential oil had a linear effect on eggs fertilization rates and larval hatching. Garlic oil did not influence the morphology of the larvae (P>0.05. It is recommended to use 5mg L-1 of garlic essential oil to promote better rates of eggs fertilization and larval hatching of artificially incubated silver catfish.

  14. Characterization of cell surface polypeptides of unfertilized, fertilized, and protease-treated zona-free mouse eggs

    International Nuclear Information System (INIS)

    Boldt, J.; Gunter, L.E.; Howe, A.M.

    1989-01-01

    The polypeptide composition of unfertilized, fertilized, and protease-treated zona-free mouse eggs was evaluated in this study. Zona-free eggs were radioiodinated by an Iodogen-catalyzed reaction. Light microscopic autoradiography of egg sections revealed that labeling was restricted to the cell surface. Labeled eggs were solubilized, and cell surface polypeptides were identified by one-dimensional SDS polyacrylamide gel electrophoresis and autoradiography. The unfertilized egg demonstrated 8-10 peptides that incorporated 125 I, with major bands observed at approximately 145-150, 94, and 23 kilodaltons (kD). Zona-free eggs fertilized in vitro and then radiolabeled demonstrated several new bands in comparison to unfertilized eggs, with a major band appearing at approximately 36 kD. Treatment of radiolabeled unfertilized eggs with either trypsin or chymotrypsin (1 mg/ml for 5-20 min) caused enzyme-specific modifications in labeled polypeptides. Trypsin (T) treatment resulted in time-dependant modification of the three major peptides at 145-150, 94, and 23 kD. Chymotrypsin (CT) treatment, in contrast, was associated with loss or modification of the 94 kD band, with no apparent effect on either the 145-150 or 23 kD band. Taken together with previous data indicating that T or CT egg treatment interferes with sperm-egg attachment and fusion, these results suggest a possible role for the 94 kD protein in sperm-egg interaction

  15. The effects of production water, WAF or CEWAF on the fertilization success of Atlantic cod eggs

    International Nuclear Information System (INIS)

    Burridge, L; Wong, D.; Trippel, E.

    2010-01-01

    Production water (PW), dispersed oil, and oil released from offshore oil and gas platforms can adversely affect fish populations. In this study, eggs from Atlantic cod were fertilized in the presence of PW collected from the Terra Nova and Hibernia offshore platforms in 2008. Cod eggs were also fertilized in the presence of the water accommodated fraction (WAF) and chemically enhanced water accommodated fraction (CEWAF) of Alaskan North Slope oil (ANS). Results of the study showed that exposure to the Terra Nova PW at 12 per cent (V/V) or higher resulted in significantly lower fertilization rates than those observed in controls. Exposure to Hibernia PW at 0.5 per cent (V/V) also resulted in significantly lower fertilization rates. The WAF of ANS up to 25 per cent (V/V) had no effect on fertilization rates. At 50 per cent (V/V), the fertilization rate for WAF ANS was much lower than rates observed in controls. The CEWAF significantly reduced fertilization at all tested concentrations.

  16. Autocatalytic microtubule nucleation determines the size and mass of Xenopus laevis egg extract spindles.

    Science.gov (United States)

    Decker, Franziska; Oriola, David; Dalton, Benjamin; Brugués, Jan

    2018-01-11

    Regulation of size and growth is a fundamental problem in biology. A prominent example is the formation of the mitotic spindle, where protein concentration gradients around chromosomes are thought to regulate spindle growth by controlling microtubule nucleation. Previous evidence suggests that microtubules nucleate throughout the spindle structure. However, the mechanisms underlying microtubule nucleation and its spatial regulation are still unclear. Here, we developed an assay based on laser ablation to directly probe microtubule nucleation events in Xenopus laevis egg extracts. Combining this method with theory and quantitative microscopy, we show that the size of a spindle is controlled by autocatalytic growth of microtubules, driven by microtubule-stimulated microtubule nucleation. The autocatalytic activity of this nucleation system is spatially regulated by the limiting amounts of active microtubule nucleators, which decrease with distance from the chromosomes. This mechanism provides an upper limit to spindle size even when resources are not limiting. © 2018, Decker et al.

  17. Egg fertility and hatchability in Avians broiler-breeder hens under ...

    African Journals Online (AJOL)

    Impact of temperature and humidity in different month of lay was evaluated on fertility and hatchability in broiler breeder hens reared in Sapele, Nigeria. Six million, six hundred and nineteen thousand, seven hundred and forty six eggs from flocks of Avians broiler-breeder hens reared between 2005 and 2006 in a farm ...

  18. Toxicity of nitrogenous fertilizers to eggs of snapping turtles (Chelydra serpentina) in field and laboratory exposures.

    Science.gov (United States)

    de Solla, Shane Raymond; Martin, Pamela Anne

    2007-09-01

    Many reptiles oviposit in soil of agricultural landscapes. We evaluated the toxicity of two commonly used nitrogenous fertilizers, urea and ammonium nitrate, on the survivorship of exposed snapping turtle (Chelydra serpentina) eggs. Eggs were incubated in a community garden plot in which urea was applied to the soil at realistic rates of up to 200 kg/ha in 2004, and ammonium nitrate was applied at rates of up to 2,000 kg/ha in 2005. Otherwise, the eggs were unmanipulated and were subject to ambient temperature and weather conditions. Eggs were also exposed in the laboratory in covered bins so as to minimize loss of nitrogenous compounds through volatilization or leaching from the soil. Neither urea nor ammonium nitrate had any impact on hatching success or development when exposed in the garden plot, despite overt toxicity of ammonium nitrate to endogenous plants. Both laboratory exposures resulted in reduced hatching success, lower body mass at hatching, and reduced posthatching survival compared to controls. The lack of toxicity of these fertilizers in the field was probably due to leaching in the soil and through atmospheric loss. In general, we conclude that nitrogenous fertilizers probably have little direct impacts on turtle eggs deposited in agricultural landscapes.

  19. Egg donation brokers: an analysis of agency versus in vitro fertilization clinic websites.

    Science.gov (United States)

    Holwell, Eve; Keehn, Jason; Leu, Cheng-shiun; Sauer, Mark V; Klitzman, Robert

    2014-01-01

    To compare websites of agencies that broker the services of women who provide human eggs for in vitro fertilization versus clinics that recruit egg providers. We examined 207 websites, of which 128 were egg provider agency 40%) or clinic (60%) websites that recruited providers online. We compared them regarding several variables related to adherence to American Society for Reproductive Medicine (ASRM) guidelines. According to their respective websites, agencies were more likely than clinics to mention ASRM guidelines, be located in the West/Pacific, indicate compensation, offer a fee range, set their minimum > $5,000, specify preferable traits, cap provider age at 31, require an education minimum, allow both parties to meet, discuss short-term risks, and not acknowledge a possible cancer risk. Only 25.5% of agencies and 19.5% of clinics mention psychological/emotional risks, and 11.8% and 5.2%, respectively, mention risk to future fertility. This research, the first to systematically compare several key aspects of egg provider agencies versus clinics, suggests it significant differences in adherence to guidelines, raising several concerns and suggesting needs for consideration of improved monitoring and regulation by ASRM or others.

  20. Hatching rate and growth rate of Nothobranchius guentheri fertilized eggs after space flight

    International Nuclear Information System (INIS)

    Guo Mingzhong; Zheng Leyun; Lin Guangji; Zhong Jianxing; Yang Huosheng; Zheng Yangfu

    2012-01-01

    Hatching, abnormal, growth and survival rate of the fertilized eggs of Nothobranchius guentheri were carried by Shenzhou 7 spacecraft were studied. The results indicated that the hatching and abnormal rate were no significant difference between the spaceflight group (99.3% and 16.8%) and ground group (97.2% and 10.4%); but the growth rate of male fish from spaceflight group was significant higher (0.094 g/d) than that of ground group (0.059 g/d), leading to the significant bigger of the male fish from spaceflight group. The survival rate of spaceflight group (66.7%) was higher than the ground group (47.9%). It was concluded that there was a higher growth and survival rate of Nothobranchius guentheri fertilized eggs after space flight. (authors)

  1. Effects of pre-incubation of eggs in fresh water and varying sperm concentration on fertilization rate in sterlet sturgeon, Acipenser ruthenus

    DEFF Research Database (Denmark)

    Siddique, Mohammad Abdul Momin; Butts, Ian; Psěnička, Martin

    2015-01-01

    , while it was significant at the 4300:1 and 430:1 ratios. The use of adequate experimental suboptimal sperm to egg ratio revealed a positive effect of pre-incubation time, such that at the 430:1 ratio, 0.5. min pre-incubation increased the fertilization rate than 10. min. At 0. min pre......-incubation the proportion of fertilized eggs increased at the 430,000:1 ratio, while at 1. min fertilization increased at the 4300:1 ratio. At the 10. min pre-incubation time, fertilization increased at the 43,000:1 ratio. Moreover, at the 0.5. min pre-incubation time, the 43,000:1 ratio increased the fertilization rate...... than the 430:1 ratio. Generally, for 430:1 ratio, the fertilization rate is lower than in control. Transmission electron microscopy showed that pre-incubation of eggs in water for

  2. Ubiquitin-mediated proteolysis in Xenopus extract.

    Science.gov (United States)

    McDowell, Gary S; Philpott, Anna

    2016-01-01

    The small protein modifier, ubiquitin, can be covalently attached to proteins in the process of ubiquitylation, resulting in a variety of functional outcomes. In particular, the most commonly-associated and well-studied fate for proteins modified with ubiquitin is their ultimate destruction: degradation by the 26S proteasome via the ubiquitin-proteasome system, or digestion in lysosomes by proteolytic enzymes. From the earliest days of ubiquitylation research, a reliable and versatile "cell-in-a-test-tube" system has been employed in the form of cytoplasmic extracts from the eggs and embryos of the frog Xenopus laevis. Biochemical studies of ubiquitin and protein degradation using this system have led to significant advances particularly in the study of ubiquitin-mediated proteolysis, while the versatility of Xenopus as a developmental model has allowed investigation of the in vivo consequences of ubiquitylation. Here we describe the use and history of Xenopus extract in the study of ubiquitin-mediated protein degradation, and highlight the versatility of this system that has been exploited to uncover mechanisms and consequences of ubiquitylation and proteolysis.

  3. Effect of mercuric chloride feeding on sexual maturity, egg production and fertility in Japanese quail

    Science.gov (United States)

    Hill, E.F.; Shaffner, C.S.

    1973-01-01

    Japanese quail (Coturnix c. japonica) were fed 0, 8, 16 or 32 p.p.m. of mercury as mercuric chloride from 3 days of age through 20 weeks of age. The onset of egg production generally occurred earlier for hens fed HgCl2. Average age in days at first oviposition for the control, 8 p.p.m., 16 p.p.m. and 32 p.p.m. was 48.4, 50.9, 46.9 and 44.0 respectively. The average rate of egg productivity from first oviposition to attainment of full growth (9 weeks of age) correlated positively with in increased dietary mercury (controls, 8 p.p.m., 16 p.p.m., 32 p.p.m. ? 75.2, 69.3, 86.1 and 93.3% respectively). By 20 weeks of age productivity was 81.0, 80.6, 87.5 and 92.9% for control, 8, 16 and 32 p.p.m. groups respectively. Fertility was depressed when hens were fed HgCl2. At 9 weeks of age average control fertility was 59% contrasted with 25% for the 32 p.p.m. group. At 12 weeks fertility increased to 89% and 57% for these groups. From this study it is apparent. that the onset and rate of egg production was stimulated by HgCl2, but fertility was adversely affected.

  4. Cold-shock eliminates female nucleus in fertilized eggs to induce androgenesis in the loach (Misgurnus anguillicaudatus, a teleost fish

    Directory of Open Access Journals (Sweden)

    Morishima Kagayaki

    2011-11-01

    Full Text Available Abstract Background Androgenesis (all-male inheritance is generally induced by means of irradiating the eggs to inactivate the maternal genome, followed by fertilization with normal sperm. In fish, the conventional technique for induced androgenesis has been applied for rapid fixation to traits, recovery of cryopreserved genotypes, sex-control, etc. A new method of androgenesis that eliminates the need to irradiate the egg was proposed using the loach, Misgurnus anguillicaudatus (a teleost fish. Results When the eggs of wild-type females were fertilized with sperm of albino or orange phenotype males and cold-shocked at 0 to 3°C for 60 min duration just after fertilization, generally more than 30% (with a peak of 100% of the hatched progeny were androgenotes. While a few of them were the normal diploid, most of them turned out to be abnormal haploid. All-male inheritance was verified by the expression of the recessive color trait (albino or orange and microsatellite genotypes comprising only paternally derived alleles. Nuclear behavior after the cold-shock treatment was traced by microscopic observation of DAPI (4'6-diamidino-2-phenylindole-stained samples and hematoxylin-eosin stained histological sections, and the extrusion of egg (maternal nucleus was observed in eggs treated in the optimum timing. Conclusion In this paper, we demonstrate that cold-shock treatment (at 0 and 3°C of loach eggs for 60 min just after fertilization successfully induces androgenetic haploid development. The most likely mechanism of cold-shock induced androgenesis is an elimination of the egg nucleus together along with the second polar body and subsequent development of a decondensed sperm nucleus or male pronucleus.

  5. In vitro fertilization and artificial activation of eggs of the direct-developing anuran Eleutherodactylus coqui

    Directory of Open Access Journals (Sweden)

    Toro Esteban

    2004-08-01

    Full Text Available Abstract Although much is known about the reproductive biology of pond-breeding frogs, there is comparatively little information about terrestrial-breeding anurans, a highly successful and diverse group. This study investigates the activation and in vitro fertilization of eggs of the Puerto Rican coqui frog obtained by hormonally induced ovulation. We report that spontaneous activation occurs in 34% of eggs, probably in response to mechanical stress during oviposition. Artificial activation, as evidenced by the slow block to polyspermy and the onset of zygote division, was elicited both by mechanical stimulation and calcium ionophore exposure in 64% and 83% of the cases, respectively. Finally, one in vitro fertilization protocol showed a 27% success rate, despite the fact that about one third of all unfertilized eggs obtained by hormone injection auto-activate. We expect these findings to aid in the conservation effort of Eleutherodactylus frogs, the largest vertebrate genus.

  6. Fertilization of sea urchin eggs and sperm motility are negatively impacted under low hypergravitational forces significant to space flight

    Science.gov (United States)

    Tash, J. S.; Kim, S.; Schuber, M.; Seibt, D.; Kinsey, W. H.

    2001-01-01

    Sperm and other flagellates swim faster in microgravity (microG) than in 1 G, raising the question of whether fertilization is altered under conditions of space travel. Such alterations have implications for reproduction of plant and animal food and for long-term space habitation by man. We previously demonstrated that microG accelerates protein phosphorylation during initiation of sperm motility but delays the sperm response to the egg chemotactic factor, speract. Thus sperm are sensitive to changes in gravitational force. New experiments using the NiZeMi centrifugal microscope examined whether low hypergravity (hyperG) causes effects opposite to microG on sperm motility, signal transduction, and fertilization. Sperm % motility and straight-line velocity were significantly inhibited by as little as 1.3 G. The phosphorylation states of FP130, an axonemal phosphoprotein, and FP160, a cAMP-dependent salt-extractable flagellar protein, both coupled to motility activation, showed a more rapid decline in hyperG. Most critically, hyperG caused an approximately 50% reduction in both the rate of sperm-egg binding and fertilization. The similar extent of inhibition of both fertilization parameters in hyperG suggests that the primary effect is on sperm rather than eggs. These results not only support our earlier microG data demonstrating that sperm are sensitive to small changes in gravitational forces but more importantly now show that this sensitivity affects the ability of sperm to fertilize eggs. Thus, more detailed studies on the impact of space flight on development should include studies of sperm function and fertilization.

  7. Xenopus laevis Kif18A is a highly processive kinesin required for meiotic spindle integrity

    Directory of Open Access Journals (Sweden)

    Martin M. Möckel

    2017-04-01

    Full Text Available The assembly and functionality of the mitotic spindle depends on the coordinated activities of microtubule-associated motor proteins of the dynein and kinesin superfamily. Our current understanding of the function of motor proteins is significantly shaped by studies using Xenopus laevis egg extract as its open structure allows complex experimental manipulations hardly feasible in other model systems. Yet, the Kinesin-8 orthologue of human Kif18A has not been described in Xenopus laevis so far. Here, we report the cloning and characterization of Xenopus laevis (Xl Kif18A. Xenopus Kif18A is expressed during oocyte maturation and its depletion from meiotic egg extract results in severe spindle defects. These defects can be rescued by wild-type Kif18A, but not Kif18A lacking motor activity or the C-terminus. Single-molecule microscopy assays revealed that Xl_Kif18A possesses high processivity, which depends on an additional C-terminal microtubule-binding site. Human tissue culture cells depleted of endogenous Kif18A display mitotic defects, which can be rescued by wild-type, but not tail-less Xl_Kif18A. Thus, Xl_Kif18A is the functional orthologue of human Kif18A whose activity is essential for the correct function of meiotic spindles in Xenopus oocytes.

  8. In vitro maintenance of spermatogenesis in Xenopus laevis testis explants cultured in serum-free media

    International Nuclear Information System (INIS)

    Risley, M.S.; Miller, A.; Bumcrot, D.A.

    1987-01-01

    Spermatogenesis has been maintained for extended periods in Xenopus laevis testis explants cultured in serum-free media supplemented with bovine serum albumin, insulin, transferrin, follicle-stimulating hormone, dihydrotestosterone, testosterone, retinol, ascorbate, and tocopherol. The organization of the testis fragments was maintained for 28 days, and all stages of development were present throughout the culture period. 3 H-Thymidine-labeled secondary (Type B) spermatogonia developed in 28 days into spermatids at the acrosomal vesicle stage whereas labeled zygotene spermatocytes became mature spermatids in 28 days. Spermatogonial proliferation also continued in vitro for 28 days. Germ cell differentiation was not dependent upon exogenous testosterone, ascorbate, or tocopherol since 3 H-labeled spermatogonia became mature spermatids in testes cultured 35 days in media lacking these supplements. Autoradiography demonstrated that 55% of the luminal sperm present in explants cultured 10 days had differentiated in vitro. Sperm from testes cultured 10-35 days were similar to sperm from freshly dissected testes with regard to motility and fecundity, and eggs fertilized with sperm from explant cultures developed normally into swimming tadpoles. The results demonstrate the feasibility of maintaining vertebrate spermatogenesis in culture and suggest that in vitro analysis of Xenopus spermatogenesis using defined media may provide important insights into the evolution of regulatory mechanisms in spermatogenesis

  9. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts.

    Science.gov (United States)

    Baur, Tina; Ramadan, Kristijan; Schlundt, Andreas; Kartenbeck, Jürgen; Meyer, Hemmo H

    2007-08-15

    Despite the progress in understanding nuclear envelope (NE) reformation after mitosis, it has remained unclear what drives the required membrane fusion and how exactly this is coordinated with nuclear pore complex (NPC) assembly. Here, we show that, like other intracellular fusion reactions, NE fusion in Xenopus laevis egg extracts is mediated by SNARE proteins that require activation by NSF. Antibodies against Xenopus NSF, depletion of NSF or the dominant-negative NSF(E329Q) variant specifically inhibited NE formation. Staging experiments further revealed that NSF was required until sealing of the envelope was completed. Moreover, excess exogenous alpha-SNAP that blocks SNARE function prevented membrane fusion and caused accumulation of non-flattened vesicles on the chromatin surface. Under these conditions, the nucleoporins Nup107 and gp210 were fully recruited, whereas assembly of FxFG-repeat-containing nucleoporins was blocked. Together, we define NSF- and SNARE-mediated membrane fusion events as essential steps during NE formation downstream of Nup107 recruitment, and upstream of membrane flattening and completion of NPC assembly.

  10. Effects of preincubation of eggs and activation medium on the percentage of eyed embryos in ide (Leuciscus idus), an externally fertilizing fish.

    Science.gov (United States)

    Siddique, Mohammad Abdul Momin; Linhart, Otomar; Krejszeff, Sławomir; Żarski, Daniel; Król, Jarosław; Butts, Ian Anthony Ernest

    2016-03-15

    Standardization of fertilization protocols is crucial for improving reproductive techniques for externally fertilizing fish in captive breeding. Therefore, the objectives of this study were to determine the effects of preincubation of eggs and activation medium on the percentage of eyed embryos for ide (Leuciscus idus). Pooled eggs from five females were preincubated in three different activating media for 0, 30, 60, 90, and 120 seconds and then fertilized by pooled sperm from five males. At the eyed-egg stage, the percentage of viable embryos was later calculated. Results showed that preincubation time was significant for the freshwater activation medium (P reproduction for this species. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Fate mapping of the silkworm, Bombyx mori, using localized UV irradiation of the egg at fertilization

    International Nuclear Information System (INIS)

    Myohara, Maroko

    1994-01-01

    Bombyx eggs at the fertilization stage (0-2 hours after oviposition) were irradiated with a scanning UV-laser microbeam (355 nm) over an area of about 1% of the total egg surface. In spite of absence of nuclei or cells at the irradiated sites, larvae from treated eggs showed localized cuticle defects in the integument. The location and frequency of the defects within the cuticular pattern correlated closely to the site of irradiation both in the anteroposterior and the dorsoventral direction. Based on the correlation, presumptive regions for each larval segment were located and a fate map of the Bombyx egg was established. (author)

  12. Deficient induction response in a Xenopus nucleocytoplasmic hybrid.

    Directory of Open Access Journals (Sweden)

    Patrick Narbonne

    2011-11-01

    Full Text Available Incompatibilities between the nucleus and the cytoplasm of sufficiently distant species result in developmental arrest of hybrid and nucleocytoplasmic hybrid (cybrid embryos. Several hypotheses have been proposed to explain their lethality, including problems in embryonic genome activation (EGA and/or nucleo-mitochondrial interactions. However, conclusive identification of the causes underlying developmental defects of cybrid embryos is still lacking. We show here that while over 80% of both Xenopus laevis and Xenopus (Silurana tropicalis same-species androgenetic haploids develop to the swimming tadpole stage, the androgenetic cybrids formed by the combination of X. laevis egg cytoplasm and X. tropicalis sperm nucleus invariably fail to gastrulate properly and never reach the swimming tadpole stage. In spite of this arrest, these cybrids show quantitatively normal EGA and energy levels at the stage where their initial gastrulation defects are manifested. The nucleocytoplasmic incompatibility between these two species instead results from a combination of factors, including a reduced emission of induction signal from the vegetal half, a decreased sensitivity of animal cells to induction signals, and differences in a key embryonic protein (Xbra concentration between the two species, together leading to inefficient induction and defective convergence-extension during gastrulation. Indeed, increased exposure to induction signals and/or Xbra signalling partially rescues the induction response in animal explants and whole cybrid embryos. Altogether, our study demonstrates that the egg cytoplasm of one species may not support the development promoted by the nucleus of another species, even if this nucleus does not interfere with the cytoplasmic/maternal functions of the egg, while the egg cytoplasm is also capable of activating the genome of that nucleus. Instead, our results provide evidence that inefficient signalling and differences in the

  13. Expression of multiple Src family kinases in sea urchin eggs and their function in Ca2+ release at fertilization.

    Science.gov (United States)

    Townley, Ian K; Schuyler, Erin; Parker-Gür, Michelle; Foltz, Kathy R

    2009-03-15

    Egg activation at fertilization in deuterostomes requires a rise in intracellular Ca(2+), which is released from the egg's endoplasmic reticulum. In sea urchins, a Src Family Kinase (SpSFK1) is necessary for the PLCgamma-mediated signaling event that initiates this Ca(2+) release (Giusti, A.F., O'Neill, F.J., Yamasu, K., Foltz, K.R. and Jaffe, L.A., 2003. Function of a sea urchin egg Src family kinase in initiating Ca2+ release at fertilization. Dev. Biol. 256, 367-378.). Annotation of the Strongylocentrotus purpuratus genome sequence led to the identification of additional, predicted SFKs (Bradham, C.A., Foltz, D.R., Beane, W.S., Amone, M.I., Rizzo, F., Coffman, J.A., Mushegian, A., Goel, M., Morales, J., Geneviere, A.M., Lapraz, F., Robertson, A.J., Kelkar, H., Loza-Coll, M., Townley, I.K., Raisch, M., Roux, M.M., Lepage, T., Gache, C., McClay, D.R., Manning, G., 2006. The sea urchin kinome: a first look. Dev. Biol. 300, 180-193.; Roux, M.M., Townley, I.K., Raisch, M., Reade, A., Bradham, C., Humphreys, G., Gunaratne, H.J., Killian, C.E., Moy, G., Su, Y.H., Ettensohn, C.A., Wilt, F., Vacquier, V.D., Burke, R.D., Wessel, G. and Foltz, K.R., 2006. A functional genomic and proteomic perspective of sea urchin calcium signaling and egg activation. Dev. Biol. 300, 416-433.). Here, we describe the cloning and characterization of these 4 additional SFKs and test their function during the initial Ca(2+) release at fertilization using the dominant-interfering microinjection method coupled with Ca(2+) recording. While two of the new SFKs (SpFrk and SpSFK3) are necessary for Ca(2+) release, SpSFK5 appears dispensable for early egg to embryo transition events. Interestingly, SpSFK7 may be involved in preventing precocious release of Ca(2+). Binding studies indicate that only SpSFK1 is capable of direct interaction with PLCgamma. Immunolocalization studies suggest that one or more SpSFK and PLCgamma are localized to the egg cortex and at the site of sperm-egg interaction

  14. Semen collection and fertility in naturally fertile sandhill cranes

    Science.gov (United States)

    Chen, G.; Gee, G.F.; Nicolich, Jane M.; Taylor, J.A.; Urbanek, R.P.; Stahlecker, D.W.

    1997-01-01

    Aviculturists often ask if semen collection will interfere with fertility in naturally fertile pairs of cranes. We used 12 naturally fertile Florida sandhill crane (Grus canadensis pratensis) pairs for this study, 6 control and 6 experimental. All pairs had produced fertile eggs in previous years and were in out-of-doors pens scattered throughout different pen complexes, within auditory range but physically isolated. Semen was collected on Tuesday mornings and Friday afternoons from 26 February 1993 to 4 June 1993. We used standard artificial insemination methods to collect and to evaluate the semen and spermatozoa. Semen collection did not affect semen quality or quantity. Semen volume, sperm density, sperm motility, sperm morphology, sperm live, sperm number per collection, and male response to semen collection exhibited significant daily variation (P semen collection began 13 days before the first egg in the experimental group, we observed no differences in the date of first egg laid or in fertility between experimental and control groups. Also, we observed no differences in the interval between clutches or in the percentage of broken eggs between experimental and control groups. Sires consistently producing better semen samples produced fewer fertile eggs than sires producing poorer semen samples (r = 0.60).

  15. Mlh1 is required for female fertility in Drosophila melanogaster: An outcome of effects on meiotic crossing over, ovarian follicles and egg activation.

    Science.gov (United States)

    Vimal, Divya; Kumar, Saurabh; Pandey, Ashutosh; Sharma, Divya; Saini, Sanjay; Gupta, Snigdha; Ravi Ram, Kristipati; Chowdhuri, Debapratim Kar

    2018-03-01

    Mismatch repair (MMR) system, a conserved DNA repair pathway, plays crucial role in DNA recombination and is involved in gametogenesis. The impact of alterations in MMR family of proteins (bacterial MutS and MutL homologues) on mammalian fertility is well documented. However, an insight to the role of MMR in reproduction of non-mammalian organisms is limited. Hence, in the present study, we analysed the impact of mlh1 (a MutL homologue) on meiotic crossing over/recombination and fertility in a genetically tractable model, Drosophila melanogaster. Using mlh1 e00130 hypomorphic allele, we report female specific adverse reproductive outcome for reduced mlh1 in Drosophila: mlh1 e00130 homozygous females had severely reduced fertility while males were fertile. Further, mlh1 e00130 females contained small ovaries with large number of early stages as well as significantly reduced mature oocytes, and laid fewer eggs, indicating discrepancies in egg production and ovulation. These observations contrast the sex independent and/or male specific sterility and normal follicular development as well as ovulation reported so far for MMR family proteins in mammals. However, analogous to the role(s) of mlh1 in meiotic crossing over and DNA repair processes underlying mammalian fertility, ovarian follicles from mlh1 e00130 females contained significantly increased DNA double strand breaks (DSBs) and reduced synaptonemal complex foci. In addition, large proportion of fertilized eggs display discrepancies in egg activation and fail to proceed beyond stage 5 of embryogenesis. Hence, reduction of the Mlh1 protein level leads to defective oocytes that fail to complete embryogenesis after fertilization thereby reducing female fertility. Copyright © 2017 Elsevier GmbH. All rights reserved.

  16. Does egg competition occur in marine broadcast-spawners?

    Science.gov (United States)

    Marshall, D J; Evans, J P

    2005-09-01

    When the availability of sperm limits female reproductive success, competition for sperm, may be an important broker of sexual selection. This is because sperm limitation can increase the variance in female reproductive success, resulting in strong selection on females to compete for limited fertilization opportunities. Sperm limitation is probably common in broadcast-spawning marine invertebrates, making these excellent candidates for investigating scramble competition between broods of eggs and its consequences for female reproductive success. Here, we report our findings from a series of experiments that investigate egg competition in the sessile, broadcast-spawning polychaete Galeolaria caespitosa. We initially tested whether the order in which eggs encounter sperm affects their fertilization success at two ecologically relevant current regimes. We used a split-clutch-split--ejaculate technique to compare the fertilization success of eggs from individual females that had either first access (competition-free treatment) or second access (egg competition treatment) to a batch of sperm. We found that fertilization success depended on the order in which eggs accessed sperm; eggs that were assigned to the competition-free treatment exhibited significantly higher fertilization rates than those assigned to the egg competition treatment at both current speeds. In subsequent experiments we found that prior exposure of sperm to eggs significantly reduced both the quantity and quality of sperm available to fertilize a second clutch of eggs, resulting in reductions in fertilization success at high and low sperm concentrations. These findings suggest that female traits that increase the likelihood of sperm-egg interactions (e.g. egg size) will respond to selection imposed by egg competition.

  17. The dynamics of plasma membrane PtdIns(4,5)P(2) at fertilization of mouse eggs.

    Science.gov (United States)

    Halet, Guillaume; Tunwell, Richard; Balla, Tamas; Swann, Karl; Carroll, John

    2002-05-15

    A series of intracellular Ca2+ oscillations are responsible for triggering egg activation and cortical granule exocytosis at fertilization in mammals. These Ca2+ oscillations are generated by an increase in inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)], which results from the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)]. Using confocal imaging to simultaneously monitor Ca2+ and plasma membrane PtdIns(4,5)P(2) in single living mouse eggs we have sought to establish the relationship between the kinetics of PtdIns(4,5)P(2) metabolism and the Ca2+ oscillations at fertilization. We report that there is no detectable net loss of plasma membrane PtdIns(4,5)P(2) either during the latent period or during the subsequent Ca2+ oscillations. When phosphatidylinositol 4-kinase is inhibited with micromolar wortmannin a limited decrease in plasma membrane PtdIns(4,5)P(2) is detected in half the eggs studied. Although we were unable to detect a widespread loss of PtdIns(4,5)P(2), we found that fertilization triggers a net increase in plasma membrane PtdIns(4,5)P(2) that is localized to the vegetal cortex. The fertilization-induced increase in PtdIns(4,5)P(2) follows the increase in Ca2+, is blocked by Ca2+ buffers and can be mimicked, albeit with slower kinetics, by photoreleasing Ins(1,4,5)P(3). Inhibition of Ca2+-dependent exocytosis of cortical granules, without interfering with Ca2+ transients, inhibits the PtdIns(4,5)P(2) increase. The increase appears to be due to de novo synthesis since it is inhibited by micromolar wortmannin. Finally, there is no increase in PtdIns(4,5)P(2) in immature oocytes that are not competent to extrude cortical granules. These studies suggest that fertilization does not deplete plasma membrane PtdIns(4,5)P(2) and that one of the pathways for increasing PtdIns(4,5)P(2) at fertilization is invoked by exocytosis of cortical granules.

  18. The timing of cortical granule fusion, content dispersal, and endocytosis during fertilization of the hamster egg: an electrophysiological and histochemical study.

    Science.gov (United States)

    Kline, D; Stewart-Savage, J

    1994-03-01

    To determine the temporal relationship between cortical granule exocytosis and the repetitive calcium transients, which are characteristic of mammalian fertilization, we monitored membrane addition from exocytosis during fertilization of hamster eggs. Continuous measurement of membrane capacitance by applying a 3.1-nA alternating current at 375 Hz showed addition of cortical granule membrane. Simultaneous measurement of membrane potential revealed each calcium transient by the appearance of transient hyperpolarizing responses due to calcium-activated potassium channels in the egg. The initial membrane capacitance of the eggs averaged 736 +/- 44 pF (mean +/- SD; n = 7) and an increase in capacitance of 61 +/- 19 pF occurred within 4 sec of the start of the first hyperpolarizing response (HR) after fertilization. Immediately after the first increase in capacitance there was a gradual decline in membrane capacitance in all eggs and in five/seven eggs the capacitance returned to the unfertilized level in 7.8 +/- 4.4 min. The gradual decline in capacitance after the first increase indicated endocytosis, which was confirmed by the internalization of fluorescently labeled dextran. Superimposed on the gradual decline in membrane capacitance were smaller increases in capacitance that occurred with the second and later HRs. The total increase in capacitance from the first three events averaged 72 +/- 19 pF, representing an average increase in capacitance of about 10% of the capacitance of the unfertilized egg. By labeling eggs before and after permeabilization with two different fluorochromes attached to Lens culinaris agglutinin, we demonstrate that the dispersal of the cortical granules contents does not occur immediately after exocytosis. Our results demonstrate that cortical granule exocytosis in hamster eggs is closely coupled to the periodic increases in calcium, that the contents of the cortical granules are slow to disperse, and that after exocytosis, the surface

  19. The effect of gamma irradiation on fertile eggs and hatched chick performance

    International Nuclear Information System (INIS)

    Mekkawy, S.H.

    1995-01-01

    Two handier fertile white arber acers eggs were exposed to gamma irradiation (Co 6 0) at the following levels 0, 100, 150 and 200 rad. They Were then hatched with the control eggs and hatchability, growth rate, feed consumption, feed conversion, carcass quality, carcass analysis and the blood analysis were determined. Hatchability was reduced gradually with increased radiation, but significantly, no differences in body weight between irradiated and control chicks were apparent at hatching time. At the end of the experimental 45 days, the mean heavight was that of the irradiation dose of 150 rad. The feed conversion efficiency was enhanced by the irradiation treatments, especially with the dose of 200 rad. There were no differences in the carcass quality and carcass composition between the treated and non-treated birds. The results demonstrated that, the irradiation treatments had no effects on the blood analysis of the birds. 7 tabs

  20. Observing dynamics of chromatin fibers in Xenopus egg extracts by single DNA manipulation using a transverse magnetic tweezer setup

    Science.gov (United States)

    Yan, Jie; Skoko, Dunja; Marko, John; Maresca, Tom; Heald, Rebecca

    2005-03-01

    We have studied assembly of chromatin on single DNAs using Xenopus egg extracts and a specially designed magnetic tweezer setup which generates controlled force in the focal plane of the objective, allowing us to visualize and measure DNA extension under a wide range of constant tensions. We found, in the absence of ATP, interphase extracts assembled nucleosomes against DNA tensions of up to 3.5 piconewtons (pN). We observed force-induced disassembly and opening-closing fluctuations indicating our experiments were in mechano-chemical equilibrium. We found that the ATP-depleted reaction can do mechanical work of 27 kcal/mol per nucleosome, providing a measurement of the free energy difference between core histone octamers on and off DNA. Addition of ATP leads to highly dynamic behavior: time courses show processive runs of assembly and disassembly of not observed in the -ATP case, with forces of 2 pN leading to nearly complete fiber disassembly. Our study shows that ATP hydrolysis plays a major role in nucleosome rearrangement and removal, and suggests that chromatin in vivo may be subject to continual assembly and disassembly.

  1. A NuRD Complex from Xenopus laevis Eggs Is Essential for DNA Replication during Early Embryogenesis

    Directory of Open Access Journals (Sweden)

    Christo P. Christov

    2018-02-01

    Full Text Available DNA replication in the embryo of Xenopus laevis changes dramatically at the mid-blastula transition (MBT, with Y RNA-independent random initiation switching to Y RNA-dependent initiation at specific origins. Here, we identify xNuRD, an MTA2-containing assemblage of the nucleosome remodeling and histone deacetylation complex NuRD, as an essential factor in pre-MBT Xenopus embryos that overcomes a functional requirement for Y RNAs during DNA replication. Human NuRD complexes have a different subunit composition than xNuRD and do not support Y RNA-independent initiation of DNA replication. Blocking or immunodepletion of xNuRD inhibits DNA replication initiation in isolated nuclei in vitro and causes inhibition of DNA synthesis, developmental delay, and embryonic lethality in early embryos. xNuRD activity declines after the MBT, coinciding with dissociation of the complex and emergence of Y RNA-dependent initiation. Our data thus reveal an essential role for a NuRD complex as a DNA replication factor during early Xenopus development.

  2. When Isolated at Full Receptivity, in Vitro Fertilized Wheat (Triticum aestivum, L. Egg Cells Reveal [Ca2+]cyt Oscillation of Intracellular Origin

    Directory of Open Access Journals (Sweden)

    Zsolt Pónya

    2014-12-01

    Full Text Available During in vitro fertilization of wheat (Triticum aestivum, L. in egg cells isolated at various developmental stages, changes in cytosolic free calcium ([Ca2+]cyt were observed. The dynamics of [Ca2+]cyt elevation varied, reflecting the difference in the developmental stage of the eggs used. [Ca2+]cyt oscillation was exclusively observed in fertile, mature egg cells fused with the sperm cell. To determine how [Ca2+]cyt oscillation in mature egg cells is generated, egg cells were incubated in thapsigargin, which proved to be a specific inhibitor of the endoplasmic reticulum (ER Ca2+-ATPase in wheat egg cells. In unfertilized egg cells, the addition of thapsigargin caused an abrupt transient increase in [Ca2+]cyt in the absence of extracellular Ca2+, suggesting that an influx pathway for Ca2+ is activated by thapsigargin. The [Ca2+]cyt oscillation seemed to require the filling of an intracellular calcium store for the onset of which, calcium influx through the plasma membrane appeared essential. This was demonstrated by omitting extracellular calcium from (or adding GdCl3 to the fusion medium, which prevented [Ca2+]cyt oscillation in mature egg cells fused with the sperm. Combined, these data permit the hypothesis that the first sperm-induced transient increase in [Ca2+]cyt depletes an intracellular Ca2+ store, triggering an increase in plasma membrane Ca2+ permeability, and this enhanced Ca2+ influx results in [Ca2+]cyt oscillation.

  3. Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks

    Energy Technology Data Exchange (ETDEWEB)

    Que, Emily L.; Bleher, Reiner; Duncan, Francesca E.; Kong, Betty Y.; Gleber, Sophie C.; Vogt, Stefan; Chen, Si; Garwin, Seth A.; Bayer, Amanda R.; Dravid, Vinayak P.; Woodruff, Teresa K.; O' Halloran, Thomas V.

    2014-12-15

    Fertilization of a mammalian egg initiates a series of 'zinc sparks' that are necessary to induce the egg-to-embryo transition. Despite the importance of these zinc-efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches that resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy-dispersive spectroscopy, X-ray fluorescence microscopy and three-dimensional elemental tomography for high-resolution elemental mapping. We show that the zinc spark arises from a system of thousands of zinc-loaded vesicles, each of which contains, on average, 10(6) zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. The discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes

  4. Effect of location of eggs in the incubator on hatchability of eggs from ...

    African Journals Online (AJOL)

    ... the hatchability of all eggs set and of fertile eggs was the highest in eggs placed in the front of the machine. Furthermore, early and middle stage deaths during incubation were lower in the front of the incubator compared to the back. The differences between eggs placed in the upper, middle and lower parts of the machine ...

  5. Fertility life table of Trichogramma pretiosum and Trichogramma acacioi on eggs of Anagasta kuehniella at different temperatures

    Directory of Open Access Journals (Sweden)

    Pratissoli Dirceu

    2004-01-01

    Full Text Available Species of the Trichogramma genus are among the most important ones for biological control. The objective of this research was to evaluate parasitism potential of two species of Trichogramma on eggs of Anagasta kuheniella through life fertility table, at temperatures between 15ºC and 35ºC. These species were collected in the State of Espírito Santo parasitising eggs of the avocado defoliator Nipteria panacea. Trichogramma pretiosum and T. acacioi showed adequate reproductive potential between 15ºC and 35ºC which indicates possibilities of using them in biological control programs in avocado plantations.

  6. Chromosomal analysis in mouse eggs fertilized in vitro with sperm exposed to ultraviolet light (UV) and methyl and ethyl methanesulfonate (MMS and EMS)

    International Nuclear Information System (INIS)

    Matsuda, Y.; Tobari, I.

    1988-01-01

    Chromosome aberrations were analyzed at the first-cleavage metaphase of mouse eggs fertilized in vitro with sperm exposed to ultraviolet light (UV) as well as to methyl and ethyl methanesulfonate (MMS and EMS). The frequencies of chromosome aberrations markedly increased with dose of UV as well as with concentration of MMS and EMS. In the UV-irradiation group, the frequency of chromosome-type aberrations was much higher than that of chromatid-type aberrations. About 90% of chromosome aberrations observed in the eggs following MMS and EMS treatment to sperm were chromosome type in which the frequency of chromosome fragments was the highest. The effects of UV on the induction of chromosome aberrations were clearly potentiated by post-treatment incubation of fertilized eggs in the presence of Ara-C or caffeine, but the effects of MMS and EMS were not pronounced by post-treatment of Ara-C or caffeine. The results indicate a possibility that UV damage induced in mouse sperm DNA is reparable in the eggs during the period between the entry of sperm into the egg cytoplasm and the first-cleavage metaphase. 35 refs.; 5 figs.; 4 tabs

  7. Lateral mobility of plasma membrane lipids in Xenopus eggs: Regional differences related to animal/vegetal polarity become extreme upon fertilization

    NARCIS (Netherlands)

    Bluemink, J.G.; Dictus, W.J.A.G.; Zoelen, E.J.J. van; Tetteroo, P.A.T.; Tertoolen, L.G.J.; Laat, S.W. de

    1984-01-01

    Regional differences in the lateral mobility properties of plasma membrane lipids have been studied in unfertilized and fertilizedxaqpus eggs by fluorescence photobleaching recovery (FPR) measurements. Out of a variety of commonly used lipid probes only the aminofluorescein-labeled fatty acids

  8. Effects of Hen Age and Egg Weight Class on the Hatchability of Free Range Indigenous Chicken Eggs

    Directory of Open Access Journals (Sweden)

    AM Abudabos

    Full Text Available ABSTRACT In total, 806 eggs of free-range Hassawi indigenous chickens were collected from local farm in Saudi Arabia. Eggs were weekly collected for 11 weeks. Initial egg weight (IEW was recorded, and eggs were graded into four classes (A: 35-40 g, B: 40-45 g, C: 45-50 g, and D: 50-55 g. Eggs were stored for seven days at 75-80% relative humidity and 14-16 C, after which egg weight losses (WL0 were calculated. During incubation, eggs were weighed on days 7 (W7 and 14 (W14, and egg weight losses on days 7 (WL7 and 14 (WL14, and total loss (WL0-14 were calculated. Hatchling weight (CW was measured. The proportion of CW relative to egg weight loss (WL on days0, 7 and 14 days of incubation (CW:WL0; CW:WL7 and CW:WL14, respectively, and break out analyses, fertility (F,total hatchability (HC and hatchability of fertile eggs (HF were also calculated. IEW decreased (p<0.05 with hen age. Stored egg weight (SEW were decreased as hen age increased (p<0.05. WL7, WL14 and WL0-14 showed significant differences (p<0.001 and increased up to first six-week of egg collection time. Hen age affected CW:WL before incubation, and on days 7 and 14 of incubation. Fertility (F was affected (p<0.05 in unpredicted way of increasing and decreasing by hen age. Egg weight class affected SEW, W7and W14 (p<0.001. Class D eggs were the highest weight. Class C eggs had highest HC. In summary, hatching eggs of Hassawi hens were affected by hen age and egg weight in randomly increase and decrease .

  9. Nuclear lamina and nuclear matrix organization in sperm pronuclei assembled in Xenopus egg extract.

    Science.gov (United States)

    Zhang, C; Jenkins, H; Goldberg, M W; Allen, T D; Hutchison, C J

    1996-09-01

    Nuclear lamina and matrices were prepared from sperm pronuclei assembled in Xenopus egg extracts using a fractionation and extraction procedure. Indirect immunofluorescence revealed that while chromatin was efficiently removed from nuclei during the extraction procedure, the distribution of lamins was unaffected. Consistent with this data, the amount of lamin B3, determined by immunoblotting, was not affected through the extraction procedure. Nuclear matrices were visualised in DGD sections by TEM. Within these sections filaments were observed both at the boundary of the nucleus (the lamina) and within the body of the nucleus (internal nuclear matrix filaments). To improve resolution, nuclear matrices were also prepared as whole mounts and viewed using field emission in lens scanning electron microscopy (FEISEM). This technique revealed two distinct networks of filaments. Filaments lying at the surface of nuclear matrices interconnected nuclear pores. These filaments were readily labelled with monoclonal anti-lamin B3 antibodies. Filaments lying within the body of the nuclear matrix were highly branched but were not readily labelled with antilamin B3 antibodies. Nuclear matrices were also prepared from sperm pronuclei assembled in lamin B3 depleted extracts. Using FEISEM, filaments were also detected in these preparations. However, these filaments were poorly organised and often appeared to aggregate. To confirm these results nuclear matrices were also observed as whole mounts using TEM. Nuclear matrices prepared from control nuclei contained a dense array of interconnected filaments. Many (but not all) of these filaments were labelled with anti-lamin B3 antibodies. In contrast, nuclear matrices prepared from "lamin depleted nuclei' contained poorly organised or aggregated filaments which were not specifically labelled with anti-lamin B3 antibodies.

  10. Have eggs. Will travel

    DEFF Research Database (Denmark)

    Kroløkke, Charlotte

    2015-01-01

    Feminist scholars have critically questioned the practices and ethics of reproductive mobility. While the reproductive mobility of fertility patients has been foregrounded, little is known of egg donor mobility including the experiences of travelling internationally to donate eggs. Based on written...... stories and photographic material provided by forty-two egg donors, this article uses feminist cluster analysis and the concept of eggpreneurship to illustrate how global egg donors negotiate reproductive agency and choice when they travel internationally to donate their eggs. In their stories, global egg...

  11. Humic Fertilizer and Vermicompost Applied to the Soil Can Positively Affect Population Growth Parameters of Trichogramma brassicae (Hymenoptera: Trichogrammatidae) on Eggs of Tuta absoluta (Lepidoptera: Gelechiidae).

    Science.gov (United States)

    Mohamadi, P; Razmjou, J; Naseri, B; Hassanpour, M

    2017-12-01

    The tomato leaf miner, Tuta absoluta (Meyrick), is a devastating pest of tomato worldwide. One of the control measures of T. absoluta is the use of biological control agents, such as Trichogramma wasps. Interactions between natural enemies and insect pests may be affected by application of fertilizers, because changes in plant quality through the fertilizer application may therefore affect herbivore characteristics and suitability of them to parasitism. Laboratory tests were carried out to evaluate the life table parameters of Trichogramma brassicae Bezdenko on T. absoluta eggs reared on tomato plants treated either with vermicompost (40%), humic fertilizer (2 g/kg soil), or control (suitable mixture of field soil and sand). Population growth parameters of T. brassicae were affected by fertilizer treatments. Significant differences were found for immature life period and total fecundity of T. brassicae on the treatments. Differences of intrinsic rate of natural increase (r m ), finite rate of increase (λ), net reproductive rate (R 0 ), mean generation time (T), and doubling time (DT) of T. brassicae among treatments were also significant. The lowest values of r m , λ, and R 0 were recorded for T. brassicae developed on T. absoluta eggs on control treatment, whereas the highest values of these parameters were observed on 2 g/kg humic fertilizer. Furthermore, T. brassicae had the shortest T and DT values on 2 g/kg humic fertilizer and 40% vermicompost treatments. Our results showed that application of humic fertilizer and vermicompost could positively affect population growth parameters of T. brassicae on eggs of T. absoluta fed on tomato plants.

  12. Social dominance molds cuticular and egg chemical blends in a paper wasp.

    Science.gov (United States)

    Dapporto, Leonardo; Romana Dani, Francesca; Turillazzi, Stefano

    2007-07-03

    Hamilton's theory [1] for the evolution of social behaviour predicts that helpers may renounce direct reproduction to help their more fertile kin. Intra-colony recognition among queens and helpers (subordinate queens or workers) is consequently a central issue in insect sociobiology. In social insects, cuticular hydrocarbons (CHCs) are involved in recognition, and egg-laying and non-egg-laying individuals often differ in CHC composition. These differences are assumed to be directly determined by fertility status [2,3]. In several ants and in Polistes wasps, when egg-layers disappear they are substituted by helpers, which develop their ovaries and become chemically similar to their former queens [2,3]. Sometimes helpers lay eggs in the presence of queens, which recognize and destroy the subordinates' eggs [4]. In ponerine ants, eggs often have the same chemical signature as the maternal cuticle [2]. If chemical signatures depend on fertility, egg-laying subordinates should match the queen's signature even when she is present, making egg recognition and differential oophagy impossible. In the study reported here, we experimentally separated fertility from dominance and analyzed the dynamics of hydrocarbon profiles of the cuticle of Polistes dominulus foundresses and the shell surface of their eggs. We have demonstrated that, contrary to the widely accepted view, dominance, rather than fertility, determines chemical signatures in Polistes wasps. This explains why queens can recognize their own eggs and police reproduction by subordinates if they become fertile and lay eggs.

  13. Maternal mRNAs of PEM and macho 1, the ascidian muscle determinant, associate and move with a rough endoplasmic reticulum network in the egg cortex.

    Science.gov (United States)

    Sardet, Christian; Nishida, Hiroki; Prodon, Francois; Sawada, Kaichiro

    2003-12-01

    Localization of maternal mRNAs in the egg cortex is an essential feature of polarity in embryos of Drosophila, Xenopus and ascidians. In ascidians, maternal mRNAs such as macho 1, a determinant of primary muscle-cell fate, belong to a class of postplasmic RNAs that are located along the animal-vegetal gradient in the egg cortex. Between fertilization and cleavage, these postplasmic RNAs relocate in two main phases. They further concentrate and segregate in small posterior blastomeres into a cortical structure, the centrosome-attracting body (CAB), which is responsible for unequal cleavages. By using high-resolution, fluorescent, in situ hybridization in eggs, zygotes and embryos of Halocynthia roretzi, we showed that macho 1 and HrPEM are localized on a reticulated structure situated within 2 mum of the surface of the unfertilized egg, and within 8 mum of the surface the vegetal region and then posterior region of the zygote. By isolating cortices from eggs and zygotes we demonstrated that this reticulated structure is a network of cortical rough endoplasmic reticulum (cER) that is tethered to the plasma membrane. The postplasmic RNAs macho 1 and HrPEM were located on the cER network and could be detached from it. We also show that macho 1 and HrPEM accumulated in the CAB and the cER network. We propose that these postplasmic RNAs relocalized after fertilization by following the microfilament- and microtubule-driven translocations of the cER network to the poles of the zygote. We also suggest that the RNAs segregate and concentrate in posterior blastomeres through compaction of the cER to form the CAB. A multimedia BioClip 'Polarity inside the egg cortex' tells the story and can be downloaded at www.bioclips.com/bioclip.html

  14. Campylobacter jejuni in commercial eggs.

    Science.gov (United States)

    Fonseca, Belchiolina Beatriz; Beletti, Marcelo Emílio; de Melo, Roberta Torres; Mendonça, Eliane Pereira; Coelho, Letícia Ríspoli; Nalevaiko, Priscila Christen; Rossi, Daise Aparecida

    2014-01-01

    This study evaluated the ability of Campylobacter jejuni to penetrate through the pores of the shells of commercial eggs and colonize the interior of these eggs, which may become a risk factor for human infection. Furthermore, this study assessed the survival and viability of the bacteria in commercial eggs. The eggs were placed in contact with wood shavings infected with C. jejuni to check the passage of the bacteria. In parallel, the bacteria were inoculated directly into the air chamber to assess the viability in the egg yolk. To determine whether the albumen and egg fertility interferes with the entry and survival of bacteria, we used varying concentrations of albumen and SPF and commercial eggs. C. jejuni was recovered in SPF eggs (fertile) after three hours in contact with contaminated wood shavings but not in infertile commercial eggs. The colonies isolated in the SPF eggs were identified by multiplex PCR and the similarity between strains verified by RAPD-PCR. The bacteria grew in different concentrations of albumen in commercial and SPF eggs. We did not find C. jejuni in commercial eggs inoculated directly into the air chamber, but the bacteria were viable during all periods tested in the wood shavings. This study shows that consumption of commercial eggs infected with C. jejuni does not represent a potential risk to human health.

  15. Avian Egg and Egg Coat.

    Science.gov (United States)

    Okumura, Hiroki

    2017-01-01

    An ovulated egg of vertebrates is surrounded by unique extracellular matrix, the egg coat or zona pellucida, playing important roles in fertilization and early development. The vertebrate egg coat is composed of two to six zona pellucida (ZP) glycoproteins that are characterized by the evolutionarily conserved ZP-domain module and classified into six subfamilies based on phylogenetic analyses. Interestingly, investigations of biochemical and functional features of the ZP glycoproteins show that the roles of each ZP-glycoprotein family member in the egg-coat formation and the egg-sperm interactions seemingly vary across vertebrates. This might be one reason why comprehensive understandings of the molecular basis of either architecture or physiological functions of egg coat still remain elusive despite more than 3 decades of intensive investigations. In this chapter, an overview of avian egg focusing on the oogenesis are provided in the first section, and unique features of avian egg coat, i.e., perivitelline layer, including the morphology, biogenesis pathway, and physiological functions are discussed mainly on chicken and quail in terms of the characteristics of ZP glycoproteins in the following sections. In addition, these features of avian egg coat are compared to mammalian zona pellucida, from the viewpoint that the structural and functional varieties of ZP glycoproteins might be associated with the evolutionary adaptation to their reproductive strategies. By comparing the egg coat of birds and mammals whose reproductive strategies are largely different, new insights into the molecular mechanisms of vertebrate egg-sperm interactions might be provided.

  16. Egg yolk plasma can replace egg yolk in stallion freezing extenders.

    Science.gov (United States)

    Pillet, E; Duchamp, G; Batellier, F; Beaumal, V; Anton, M; Desherces, S; Schmitt, E; Magistrini, M

    2011-01-01

    Hen egg yolk is normally used as a cryoprotective agent in semen freezing extenders, but its use has sanitary and practical disadvantages. Moreover the protection afforded by egg yolk has not yet been completely elucidated. The objective of this study was to compare the egg yolk plasma fraction to whole egg yolk in stallion freezing extender. Plasma contains mainly Low Density Lipoproteins (LDL), which are widely presumed to be the cryoprotective agent in egg yolk. Plasma can be produced on an industrial scale, sterilised by gamma-irradiation and incorporated in a ready-to-use extender (our ultimate objective). Plasma samples were subjected to different doses of gamma-irradiation (3, 5, 10 kGy) without dramatic chemical changes that may affect their cryoprotective properties. Stallion semen was frozen with whole egg yolk as a control and with sterilised egg yolk plasma. A fertility trial was conducted on a total of 70 mares' cycles. Fertility per cycle was 60% after insemination of semen frozen in our control extender containing egg yolk (EY), compared to 69% for the extender containing sterilised egg yolk plasma (EYP) (P > 0.05). Post-thaw motility and membrane integrity of spermatozoa were also analysed. Motility parameters were not significantly different between extenders except for the variable VAP (for EY versus EYP, VAP: 63 μm.s(-1) versus 59 μm.s(-1), a, b: P 0.05). Membrane integrity was better preserved in EY than in EYP but the difference between extenders was small (P < 0.05). Our results demonstrated that sterilised egg yolk plasma has the potential to replace egg yolk in stallion freezing extender. This experiment led to the development of a ready-to-use extender called INRA-Freeze(®) (IMV-Technologies, France). Copyright © 2011 Elsevier Inc. All rights reserved.

  17. Motility and fertilizing ability of cryopreserved Caspian brown trout (Salmo trutta caspius) sperm: Effect of post-thaw storage time and different sperm-to-egg ratios.

    Science.gov (United States)

    Golshahi, Karim; Shabani, Nariman; Aramli, Mohammad Sadegh; Noori, Elnaz

    2015-10-01

    This study was designed to test the effect of post-thaw storage time on sperm motility parameters of Caspian brown trout (n=7). Furthermore, we investigated the effect of sperm-to-egg ratios of 100,000:1, 300,000:1 and 600,000:1 on fertility of cryopreserved Caspian brown semen. Quality was assessed by measuring sperm motility parameters and fertilization rates at the eyed and hatching stages. The percentage of post-thawed sperm motility, curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH) were not affected by 60 min of storage, whereas a decrease in straight line velocity (VSL), average path velocity (VAP) and linearity (LIN) were found in cryopreserved semen. Thus, the cryopreserved sperm of Caspian brown trout could be stored up to 60 min without loss of the percentage of sperm motility. The fertilization rate was not affected by 60 min of post-thaw storage and was over 70% for sperm-to-egg ratios of both 300,000 and 600,000:1. To our knowledge, this study is the first to report the high post-thaw fertilization ability of Caspian brown trout semen at a sperm-to-egg ratio as low as 300,000:1. This procedure after scaling up can be recommended for routine Caspian brown trout sperm cryopreservation. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Fertilization Mechanisms in Flowering Plants.

    Science.gov (United States)

    Dresselhaus, Thomas; Sprunck, Stefanie; Wessel, Gary M

    2016-02-08

    Compared with the animal kingdom, fertilization is particularly complex in flowering plants (angiosperms). Sperm cells of angiosperms have lost their motility and require transportation as a passive cargo by the pollen tube cell to the egg apparatus (egg cell and accessory synergid cells). Sperm cell release from the pollen tube occurs after intensive communication between the pollen tube cell and the receptive synergid, culminating in the lysis of both interaction partners. Following release of the two sperm cells, they interact and fuse with two dimorphic female gametes (the egg and the central cell) forming the major seed components embryo and endosperm, respectively. This process is known as double fertilization. Here, we review the current understanding of the processes of sperm cell reception, gamete interaction, their pre-fertilization activation and fusion, as well as the mechanisms plants use to prevent the fusion of egg cells with multiple sperm cells. The role of Ca(2+) is highlighted in these various processes and comparisons are drawn between fertilization mechanisms in flowering plants and other eukaryotes, including mammals. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Effects of cadmium, estradiol-17beta and their interaction on gonadal condition and metamorphosis of male and female African clawed frog, Xenopus laevis

    Science.gov (United States)

    Sharma, Bibek; Patino, Reynaldo

    2010-01-01

    To assess interaction effects between cadmium (Cd, a putative xenoestrogen) and estradiol-17beta (E(2)) on sex differentiation and metamorphosis, Xenopus laevis were exposed to solvent-control (0.005% ethanol), Cd (10microgL(-1)), E(2) (1microgL(-1)), or Cd and E(2) (Cd+E(2)) in FETAX medium from fertilization to 75d postfertilization. Each treatment was applied to four aquaria, each with 30 fertilized eggs. Mortality was recorded and animals were sampled as they completed metamorphosis (Nieuwkoop and Faber stage 66). Gonadal sex of individuals (including >or= tadpoles NF stage 55 at day 75) was determined gross-morphologically and used to compute sex ratios. Time course and percent completion of metamorphosis, snout-vent length (SVL), hindlimb length (HLL) and weight were analyzed for each gender separately. Survival rates did not differ among treatments. The E(2) and Cd+E(2) treatments significantly skewed sex ratios towards females; however, no sex-ratio differences were observed between the control and Cd treatments or between the E(2) and Cd+E(2) treatments. Time course of metamorphosis was generally delayed and percent completion of metamorphosis was generally reduced in males and females exposed to Cd, E(2) or their combination compared to control animals. In males, but not females, the effect of Cd+E(2) was greater than that of individual chemicals. Weight at completion of metamorphosis was reduced only in females and only by the Cd+E(2) treatment. In conclusion, although Cd at an environmentally relevant concentration did not exhibit direct or indirect feminizing effects in Xenopus tadpoles, the metal and E(2) both had similar inhibitory effects on metamorphosis that were of greater magnitude in males than females.

  20. Effects of cadmium, estradiol-17β and their interaction on gonadal condition and metamorphosis of male and female African clawed frog, Xenopus laevis

    Science.gov (United States)

    Sharma, Bibek; Patino, Reynaldo

    2010-01-01

    To assess interaction effects between cadmium (Cd, a putative xenoestrogen) and estradiol-17?? (E2) on sex differentiation and metamorphosis, Xenopus laevis were exposed to solvent-control (0.005% ethanol), Cd (10 ??g L-1), E2 (1 ??g L-1), or Cd and E2 (Cd + E2) in FETAX medium from fertilization to 75 d postfertilization. Each treatment was applied to four aquaria, each with 30 fertilized eggs. Mortality was recorded and animals were sampled as they completed metamorphosis (Nieuwkoop and Faber stage 66). Gonadal sex of individuals (including tadpoles ???NF stage 55 at day 75) was determined gross-morphologically and used to compute sex ratios. Time course and percent completion of metamorphosis, snout-vent length (SVL), hindlimb length (HLL) and weight were analyzed for each gender separately. Survival rates did not differ among treatments. The E2 and Cd + E2 treatments significantly skewed sex ratios towards females; however, no sex-ratio differences were observed between the control and Cd treatments or between the E2 and Cd + E2 treatments. Time course of metamorphosis was generally delayed and percent completion of metamorphosis was generally reduced in males and females exposed to Cd, E2 or their combination compared to control animals. In males, but not females, the effect of Cd + E2 was greater than that of individual chemicals. Weight at completion of metamorphosis was reduced only in females and only by the Cd + E2 treatment. In conclusion, although Cd at an environmentally relevant concentration did not exhibit direct or indirect feminizing effects in Xenopus tadpoles, the metal and E2 both had similar inhibitory effects on metamorphosis that were of greater magnitude in males than females.

  1. Lateral mobility of plasma membrane lipids in Xenopus eggs: Regional differences related to animal/vegetal polarity become extreme upon fertilization

    OpenAIRE

    Bluemink, J.G.; Dictus, W.J.A.G.; Zoelen, E.J.J. van; Tetteroo, P.A.T.; Tertoolen, L.G.J.; Laat, S.W. de

    1984-01-01

    Regional differences in the lateral mobility properties of plasma membrane lipids have been studied in unfertilized and fertilizedxaqpus eggs by fluorescence photobleaching recovery (FPR) measurements. Out of a variety of commonly used lipid probes only the aminofluorescein-labeled fatty acids HEDAF (5-(N-hexadecanoyl)-aminofluorescein) and TEDAF (5-(N-tetradecanoyl)-aminofluorescein) appear to partition into the plasma membrane. Under all experimental conditions used these molecules show par...

  2. Selection responses for the number of fertile eggs of the Brown Tsaiya duck (Anas platyrhynchos after a single artificial insemination with pooled Muscovy (Cairina moschata semen

    Directory of Open Access Journals (Sweden)

    Tai Jui

    2002-09-01

    Full Text Available Abstract A seven-generation selection experiment comprising a selected (S and a control (C line was conducted with the objective of increasing the number of fertile eggs (F of the Brown Tsaiya duck after a single artificial insemination (AI with pooled Muscovy semen. Both lines consisted of about 20 males and 60 females since parents in each generation and each female duck was tested 3 times, at 26, 29 and 32 weeks of age. The fertile eggs were measured by candling at day 7 of incubation. The selection criterion in the S line was the BLUP animal model value for F. On average, 24.7% of the females and 15% of the males were selected. The direct responses to the selection for F, and correlated responses for the number of eggs set (Ie, the number of total dead embryos (M, the maximum duration of fertility (Dm and the number of hatched mule ducklings (H were measured by studying the differences across the generations of selection between the phenotypic value averages in the S and C lines. The predicted genetic responses were calculated by studying the differences between the S and C lines in averaged values of five traits of the BLUP animal model. The selection responses and the predicted responses showed similar trends. There was no genetic change for Ie. After seven generations of selection, the average selection responses per generation were 0.40, 0.33, 0.42, 0.41 genetic standard deviation units for F, M, Dm, and H respectively. Embryo viability was not impaired by this selection. For days 2–8 after AI, the fertility rates (F/Ie were 89.2% and 63.8%, the hatchability rates (H/F were 72.5% and 70.6%, and (H/Ie were 64.7% and 45.1% in the S and C lines respectively. It was concluded that upward selection on the number of fertile eggs after a single AI with pooled Muscovy semen may be effective in ducks to increase the duration of the fertile period and the fertility and hatchability rates with AI once a week instead of twice a week.

  3. EGG PRODUCTION AND HATCHABILITY OF LOCAL DUCKS UNDER SEMI INTENSIVE VS EXTENSIVE MANAGEMENTS

    Directory of Open Access Journals (Sweden)

    P. Widiyaningrum

    2016-06-01

    Full Text Available The study aimed to compare the egg production and hatchability of local ducks under different management systems, namely semi-intensive and extensive. The research conducted was an experimental one-way classification with two treatments. Each treatment used 15 male and 120 females (mating ratio 1:8, and ducks aged 13-14 months. Eggs production was recorded during three weeks period. About 300 eggs were selected from each treatment to be hatched. Fertility was observed at 4th days of hatching process using egg candler. Data of eggs production, eggs weight, fertility, and hatchability were analyzed using unpaired two sample Student’s t-test. The results showed that average of egg weight, fertility and hatchability were not different under two management system, but egg production in the semi-intensive maintenance was significantly higher (P<0.05 than those in the extensive system. Egg production in the semi-intensive was 12.3% higher than those in the extensive. In conclusion, the semi-intensive system that is applied in this study the number of eggs production but did not affect the average of egg weight, fertility and hatchability.

  4. Possible role of the 38 kDa protein, lacking in the gastrula-arrested Xenopus mutant, in gastrulation.

    Science.gov (United States)

    Tanaka, Tetsuya S; Ikenishi, Kohji

    2002-02-01

    An acidic, 38 kDa protein that is present in Xenopus wild-type embryos has been previously shown to be lacking in gastrula-arrested mutant embryos. To gain understanding of the role of this protein, its spatio-temporal distribution and involvement in gastrulation was investigated using the monoclonal antibody (9D10) against it. The protein was prominent in the cortical cytoplasm of cells facing the outside in the animal hemisphere of embryos until the gastrula stage, and in ciliated epithelial cells of embryos at stages later than the late neurula. When the 9D10 antibody was injected into fertilized wild-type eggs, they cleaved normally, but most of them had arrested development, always at the early stage of gastrulation, as in the mutant embryos. In contrast, the majority of the control antibody-injected eggs gastrulated normally and developed further. Cytoskeletal F-actin, which was mainly observed in the area beneath the plasma membrane facing the outside of the epithelial layer of not only the dorsal involuting marginal zone but also the dorsal, vegetal cell mass of the control antibody-injected embryos at the early gastrula stage, was scarcely recognized in the corresponding area of the 9D10 antibody-injected embryos. It is likely that the paucity of the F-actin caused by the 9D10 antibody inhibition of the 38 kDa protein might lead to a failure of cell movement in gastrulation, resulting in developmental arrest.

  5. Effect of Egg Orientation on Malposition in Hatching Eggs

    Directory of Open Access Journals (Sweden)

    İsmail Durmuş

    2015-09-01

    Full Text Available This research was conducted to determine the effects of orientation in the hatching eggs on hatching results and position disorders, so the eggs were placed in incubator for 3 groups as orientation of normal, horizontal and small end up. In this study, 1680 hatching eggs were divided into 3 groups with 7 replicates of 80 eggs in each. Hatchability of fertile eggs, malformation ratio, malposition ratio, weight loss ratio, chick quality, embryo death ratios in the early, middle and late stage of embriogenesis and mortality ratio were investigated in this research. There were significant differences among the groups in terms of egg weight loss ratio, embryo death in the late stage of embriogenesis, malposition ratio, hatching rate , whereas no significant differences among the groups in respect to malformation ratio, chick quality, embryo death in early and late embriogenesis. It was determined that the late embrionic mortality increased because of the position errors in case of hatching eggs orientation and the head of the embryo in small end up of eggs.

  6. effects of hatching egg weight and length of storage period

    African Journals Online (AJOL)

    metinpetek

    There were no significant effects of egg storage period on body weight and feed ... chicks from all groups were reared under the same growing conditions in battery ... on hatching time, apparent fertility, hatchability of fertile and total eggs data ...

  7. Eggs-ploiting women: a critical feminist analysis of the different principles in transplant and fertility tourism.

    Science.gov (United States)

    Pfeffer, Naomi

    2011-11-01

    Intergovernmental agencies have recognized that inconsistencies in the way that nation states regulate commerce in human kidneys lubricate transplant tourism, and have repeatedly exhorted recalcitrant governments of both organ-importing and organ-exporting nations to criminalize the exchange of cash for kidneys. Yet these same organizations have elected to remain silent on inconsistencies in the regulation of the trade in human eggs that lubricate fertility tourism. This article is a critical feminist analysis of this paradox. Sketches of the histories of regulation of the global markets in human kidneys and human eggs allow attribution of the different approaches to sales of kidneys and eggs to the triumph of neo-liberalism in the 1990s. Neo-liberalism supports the growth of the medical tourism industry and its niche market catering for infertility, and is responsible for exacerbating the relative disadvantage of poor and powerless women in destination countries, thereby creating the conditions for 'bioavailability', that is, the willingness to exchange body parts for cash. The paper identifies a disturbing correlation between deeply engrained conservative attitudes to women and a plentiful supply of eggs, and concludes by suggesting that what women need to lift themselves out of poverty and discrimination is secure and dignified work. Copyright © 2011 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  8. Repair in fertilized eggs of mice and its role in the production of chromosomal aberrations

    International Nuclear Information System (INIS)

    Generoso, W.M.

    1979-01-01

    The fertilized egg may influence the yield of dominant-lethal mutations produced from chemical treatment of male postmeiotic germ cells to a small or large extent depending upon the mutagen used and the competence of the egg to repair the premutational lesions induced. The strain of females has small influence on the yield of dominant-lethal mutations induced by TEM or EMS in spermatids and spermatozoa. On the contrary, it has large influence in the case of IMS. In addition to this difference, TEM and EMS induce high levels of heritable translocations at these germ cell stages while IMS is practically ineffective even though doses of these chemicals used produced comparable levels of dominant-lethal mutations. These differences between EMS and TEM on one hand and IMS on the other, were explained as a function of the types of chromosomal lesions present at the time of repair activity and whether or not chromosomal aberrations are already fixed at the time of postfertilization pronuclear DNA synthesis

  9. Effect of Number of Spermatozoa, Oviduct Condition and Timing of Artificial Insemination on Fertility and Fertile Period of Kampung Rooster Spermatozoa

    Directory of Open Access Journals (Sweden)

    DM Saleh

    2012-01-01

    Full Text Available Abstract. This study was carried out to determine the optimum fertility and fertile period using the number of spermatozoa, oviduct condition and timing of insemination of native rooster spermatozoa. Ninety six commercial Isa brown pullets and nine kampung roosters were used in this study in a 3×2×2 factorial arrangement with one bird in a cage constituting a unit. The factor levels were the number of spermatozoa (50, 100 and 150 million/0.1 ml, oviduct condition (hard-shelled eggs and free hard-shelled eggs, and timing of artificial insemination (in the morning, at 7 AM and in the afternoon, at 4 PM.  The results showed that among the treatments there was no significant interaction to fertility and fertile period. Insemination with 50 million sperm number seemed to be the same result with the other 2 treatments. Oviduct condition had a highly significant difference on fertility and fertile period percentage, and timing of insemination did not differ between morning and afternoon.  In conclusion, the only oviduct condition (free hard-shelled eggs was the best results for insemination in terms of fertility and fertile period of native roosters.  It is recommended that for the maximum fertility and fertile period, hens should be inseminated with 50 million spermatozoa, free of hard-shelled eggs and insemination performed in the morning or in the afternoon.   Keywords: timing of artificial insemination, fertility, fertile period, semen dose, oviduct condition Animal Production 14(1:32-36, January 2012

  10. Duplicate Abalone Egg Coat Proteins Bind Sperm Lysin Similarly, but Evolve Oppositely, Consistent with Molecular Mimicry at Fertilization

    Science.gov (United States)

    Aagaard, Jan E.; Springer, Stevan A.; Soelberg, Scott D.; Swanson, Willie J.

    2013-01-01

    Sperm and egg proteins constitute a remarkable paradigm in evolutionary biology: despite their fundamental role in mediating fertilization (suggesting stasis), some of these molecules are among the most rapidly evolving ones known, and their divergence can lead to reproductive isolation. Because of strong selection to maintain function among interbreeding individuals, interacting fertilization proteins should also exhibit a strong signal of correlated divergence among closely related species. We use evidence of such molecular co-evolution to target biochemical studies of fertilization in North Pacific abalone (Haliotis spp.), a model system of reproductive protein evolution. We test the evolutionary rates (d N/d S) of abalone sperm lysin and two duplicated egg coat proteins (VERL and VEZP14), and find a signal of co-evolution specific to ZP-N, a putative sperm binding motif previously identified by homology modeling. Positively selected residues in VERL and VEZP14 occur on the same face of the structural model, suggesting a common mode of interaction with sperm lysin. We test this computational prediction biochemically, confirming that the ZP-N motif is sufficient to bind lysin and that the affinities of VERL and VEZP14 are comparable. However, we also find that on phylogenetic lineages where lysin and VERL evolve rapidly, VEZP14 evolves slowly, and vice versa. We describe a model of sexual conflict that can recreate this pattern of anti-correlated evolution by assuming that VEZP14 acts as a VERL mimic, reducing the intensity of sexual conflict and slowing the co-evolution of lysin and VERL. PMID:23408913

  11. Duplicate abalone egg coat proteins bind sperm lysin similarly, but evolve oppositely, consistent with molecular mimicry at fertilization.

    Directory of Open Access Journals (Sweden)

    Jan E Aagaard

    Full Text Available Sperm and egg proteins constitute a remarkable paradigm in evolutionary biology: despite their fundamental role in mediating fertilization (suggesting stasis, some of these molecules are among the most rapidly evolving ones known, and their divergence can lead to reproductive isolation. Because of strong selection to maintain function among interbreeding individuals, interacting fertilization proteins should also exhibit a strong signal of correlated divergence among closely related species. We use evidence of such molecular co-evolution to target biochemical studies of fertilization in North Pacific abalone (Haliotis spp., a model system of reproductive protein evolution. We test the evolutionary rates (d(N/d(S of abalone sperm lysin and two duplicated egg coat proteins (VERL and VEZP14, and find a signal of co-evolution specific to ZP-N, a putative sperm binding motif previously identified by homology modeling. Positively selected residues in VERL and VEZP14 occur on the same face of the structural model, suggesting a common mode of interaction with sperm lysin. We test this computational prediction biochemically, confirming that the ZP-N motif is sufficient to bind lysin and that the affinities of VERL and VEZP14 are comparable. However, we also find that on phylogenetic lineages where lysin and VERL evolve rapidly, VEZP14 evolves slowly, and vice versa. We describe a model of sexual conflict that can recreate this pattern of anti-correlated evolution by assuming that VEZP14 acts as a VERL mimic, reducing the intensity of sexual conflict and slowing the co-evolution of lysin and VERL.

  12. Lateral mobility of plasma membrane lipids in dividing Xenopus eggs

    NARCIS (Netherlands)

    Laat, S.W. de; Tetteroo, P.A.T.; Bluemink, J.G.; Dictus, W.J.A.G.; Zoelen, E.J.J. van

    1984-01-01

    The lateral mobility of plasma membrane lipids was analyzed during first cleavage of Xaopus Levis eggs by fluorescence photobleaching recovery (FPR) measurements, using the lipid analogs 5-(N-hexadecanoyl)aminofluorescein (“HEDAF”) and 5-(N-tetradecanoyl)aminofluorescein (“TEDAF”) as probes. The

  13. Fertility results using bovine semen cryopreserved with extenders based on egg yolk and soy bean extract.

    Science.gov (United States)

    van Wagtendonk-de Leeuw, A M; Haring, R M; Kaal-Lansbergen, L M; den Daas, J H

    2000-07-01

    Semen extenders containing components such as egg yolk and skim milk are difficult to standardize and they introduce the risk of microbial contamination. A well-defined extender not originating from animal tissues would present a valuable contribution to the AI industry. We evaluated the fertility of bovine semen cryopreserved with 3 different extenders: 1) TRIS-Standard, prepared at 2 local AI laboratories, containing 20% (v/v) pasteurized egg yolk, 2) TRIS-Concentrate, prepared by adding 20% (v/v) pasteurized egg yolk and 1:5 (v/v) nonpyrogenic water, and 3) Biociphos Plus, a soybean extract containing extender, prepared by adding 1:5 nonpyrogenic water. Ejaculates of 4 Holstein bulls were split into 3 aliquots and cryopreserved with the 3 extenders. Prior to this study, the semen dose-response curve for each of the 4 bulls was developed in a field trial by freezing the semen and randomly distributing the straws throughout the Netherlands for insemination. Optimal semen doses were thus established to detect the effect of extenders on fertility, evaluated by 56-day non-return rate (NR56), and by the estimated conception rate and the calving rate, given a conception. We used the multiphasic model developed by Grossman et al. (7). A total of 22,246 first and second inseminations were recorded. The NR56 ranged among bulls from 67.0 to 70.1% for Tris-Standard, from 67.5 to 69.9% for Tris-Concentrate and from 60.2 to 66.7% for Biociphos Plus. No significant differences in NR56 were detected between Tris-Standard and Tris-Concentrate (P=0.54), whereas Biociphos Plus resulted in a significantly lower NR56 than Tris-Standard and Tris-Concentrate (P<0.05). Estimated conception rate was 72.1, 73.6 and 69.6% and estimated calving rate, given a conception was 80.6, 78.3 and 77.1 for Tris-Standard, Tris-Concentrate and Biociphos Plus, respectively. These results indicate that 1) semen extended with a custom made TRIS-Concentrate can be succesfully used in the field resulting

  14. Artificial activation of mature unfertilized eggs in the malaria vector mosquito, Anopheles stephensi (Diptera, Culicidae).

    Science.gov (United States)

    Yamamoto, Daisuke S; Hatakeyama, Masatsugu; Matsuoka, Hiroyuki

    2013-08-01

    In the past decade, many transgenic lines of mosquitoes have been generated and analyzed, whereas the maintenance of a large number of transgenic lines requires a great deal of effort and cost. In vitro fertilization by an injection of cryopreserved sperm into eggs has been proven to be effective for the maintenance of strains in mammals. The technique of artificial egg activation is a prerequisite for the establishment of in vitro fertilization by sperm injection. We demonstrated that artificial egg activation is feasible in the malaria vector mosquito, Anopheles stephensi (Diptera, Culicidae). Nearly 100% of eggs dissected from virgin females immersed in distilled water darkened, similar to normally oviposited fertilized eggs. It was revealed by the cytological examination of chromosomes that meiotic arrest was relieved in these eggs approximately 20 min after incubation in water. Biochemical examinations revealed that MAPK (mitogen-activated protein kinase)/ERK (extracellular signal-regulated protein kinase) and MEK (MAPK/ERK kinase) were dephosphorylated similar to that in fertilized eggs. These results indicate that dissected unfertilized eggs were activated in distilled water and started development. Injection of distilled water into body cavity of the virgin blood-fed females also induced activation of a portion of eggs in the ovaries. The technique of artificial egg activation is expected to contribute to the success of in vitro fertilization in A. stephensi.

  15. Calcium and Egg Activation in Drosophila

    Science.gov (United States)

    Sartain, Caroline V.; Wolfner, Mariana F.

    2012-01-01

    Summary In many animals, a rise in intracellular calcium levels is the trigger for egg activation, the process by which an arrested mature oocyte transitions to prepare for embryogenesis. In nearly all animals studied to date, this calcium rise, and thus egg activation, is triggered by the fertilizing sperm. However in the insects that have been examined, fertilization is not necessary to activate their oocytes. Rather, these insects’ eggs activate as they transit through the female’s reproductive tract, regardless of male contribution. Recent studies in Drosophila have shown that egg activation nevertheless requires calcium and that the downstream events and molecules of egg activation are also conserved, despite the difference in initial trigger. Genetic studies have uncovered essential roles for the calcium-dependent enzyme calcineurin and its regulator calcipressin, and have hinted at roles for calmodulin, in Drosophila egg activation. Physiological and in vitro studies have led to a model in which mechanical forces that impact the Drosophila oocyte as it moves through the reproductive tract triggers the influx of calcium from the external environment, thereby initiating egg activation. Future research will aim to test this model, as well as to determine the spatiotemporal dynamics of cytoplasmic calcium flux and mode of signal propagation in this unique system. PMID:23218670

  16. Ice, Ice, Baby? A Sociological Exploration of Social Egg Freezing

    OpenAIRE

    Baldwin, K.

    2016-01-01

    Social egg freezing is a fertility preservation strategy which enables women to preserve a number of healthy unfertilised eggs for potential future use when faced with the threat of age-related fertility decline. The overall aim of this thesis was to explore how women understand, construct and experience social egg freezing in the context of debates surrounding reproductive ‘choice’ and ‘delayed motherhood’. The study sought to provide insights into how women perceive the risks and benefi...

  17. Egg freezing and egg banking: empowerment and alienation in assisted reproduction.

    Science.gov (United States)

    Robertson, John A

    2014-06-01

    With the development of rapid freezing of human oocytes, many programs have reported IVF success rates comparable to those achieved with fresh eggs and thawed frozen embryos. Egg freezing is now gaining professional and regulatory acceptance as a safe and effective technique for women who wish to avoid discarding excess embryos, who face fertility-threatening medical treatments, or who want to preserve their eggs for use when they are better situated to have a family. This article focuses on the uses of and justification for egg freezing, the path to professional acceptance, the variability in success rates, and the controversy over freezing eggs for social rather than medical reasons. It also addresses the emergence of egg banking as a separate sector in the infertility industry, the regulatory issues that it poses, and its effect on egg donation. Key here is the legal control of stored eggs by banking women and their options when they wish to dispose of those eggs. The analysis is framed around empowerment and alienation. Egg freezing is generally empowering for women, but the donation or sale of unused eggs to infertile women, egg bankers, and researchers also raises issues of alienation.

  18. Fertilisation and hatching success of Atlantic cod (Gadus morhua) eggs when exposed to various concentrations of produced water

    International Nuclear Information System (INIS)

    Wong, D.; Lyons, M.; Burridge, L.; Lee, K.

    2010-01-01

    This study investigated the fertilization and hatching success of Atlantic cod (Gadus morhua) eggs that were exposed to various dilutions of produced water from a natural gas production platform. A control group of eggs was fertilized in sea water without any produced water. Another set of eggs were fertilized in sea water with 4 different concentrations of produced water and held their respective concentrations for 24 hours to evaluate the success of the fertilization. Viable fertilized eggs were transferred to 96 well plates containing fresh sea water and incubated in the dark at approximately 6 degrees C until hatch. Eggs began hatching about 14 days after fertilization (DPF), peaked at about 17 DPF, then finished hatching by 21 DPF. Analysis of PAH content in the diluted produced water showed that at 0.32 percent, 1.08 percent and 3.6 percent concentrations, the fertilization success was greater than or equal to that of the control group. However, for the 12 percent concentration, there were no viable fertilized eggs present after the 24 hour period. Survivability of eggs during sea water incubation was similar for the 0 percent, 0.32 percent and 1.08 percent treatment groups, but egg mortality increased for the 3.60 percent concentration group, to about 61.5 percent mean cumulative mortality at 6 DPF, then remained constant. The mean hatching success for all dilutions was presented.

  19. essential oil as hatching egg disinfectant

    African Journals Online (AJOL)

    STORAGESEVER

    2010-04-26

    Apr 26, 2010 ... disinfectant for hatching egg obtained from broiler breeder flock. Oregano essential ... contamination rate, hatchability of fertile egg, body weight at 21 and 42 days, body weight gain and total feed ... successful healthy hatchlings. Several ...... Insecticidal properties of essential plant oils against the mosquito.

  20. Albumin is synthesized in epididymis and aggregates in a high molecular mass glycoprotein complex involved in sperm-egg fertilization.

    Directory of Open Access Journals (Sweden)

    Kélen Fabíola Arroteia

    Full Text Available The epididymis has an important role in the maturation of sperm for fertilization, but little is known about the epididymal molecules involved in sperm modifications during this process. We have previously described the expression pattern for an antigen in epididymal epithelial cells that reacts with the monoclonal antibody (mAb TRA 54. Immunohistochemical and immunoblotting analyses suggest that the epitope of the epididymal antigen probably involves a sugar moiety that is released into the epididymal lumen in an androgen-dependent manner and subsequently binds to luminal sperm. Using column chromatography, SDS-PAGE with in situ digestion and mass spectrometry, we have identified the protein recognized by mAb TRA 54 in mouse epididymal epithelial cells. The ∼65 kDa protein is part of a high molecular mass complex (∼260 kDa that is also present in the sperm acrosomal vesicle and is completely released after the acrosomal reaction. The amino acid sequence of the protein corresponded to that of albumin. Immunoprecipitates with anti-albumin antibody contained the antigen recognized by mAb TRA 54, indicating that the epididymal molecule recognized by mAb TRA 54 is albumin. RT-PCR detected albumin mRNA in the epididymis and fertilization assays in vitro showed that the glycoprotein complex containing albumin was involved in the ability of sperm to recognize and penetrate the egg zona pellucida. Together, these results indicate that epididymal-derived albumin participates in the formation of a high molecular mass glycoprotein complex that has an important role in egg fertilization.

  1. Development of space-fertilized eggs and formation of primordial germ cells in the embryos of medaka fish

    Science.gov (United States)

    Ijiri, K.

    In the second International Microgravity Laboratory (IML-2) mission in 1994, four small Japanese killifish (Medaka, Oryzias latipes) made a space travel of 15 days aboard a space shuttle. These four adult Medaka fish successfully mated in space for the first time among vertebrate animals. Moreover, the eggs they laid developed normally, at least in their external appearance, hatching as fry (baby fish) in space. Fish mated and laid eggs every day during the first week. Near the end of the mission most of the eggs had a well-developed body with two pigmented eyes. In total, 43 eggs were laid (detected), out of which 8 fry hatched in space, as truly `space-originated' babies. A further 30 fry hatched within 3 days after landing. This is the normal hatching rate, compared with the ground-based data. Among the 8 space-originated fry, four were killed for histological sections, and germ cells at the gonadal region were counted for each fry. Their numbers were in the range of the germ cells of the normal control fry (ground-kept samples). Thus, as embryos developed normally in their external appearance, inside the embryos the formation of primordial germ cells took place normally in space, and their migration to the genital ridges was not hindered by microgravity. The two of the remaining space-originated fry have grown up and been creating their offspring in the laboratory. This proved that the primordial germ cells formed in space were also normal from a functional point of view. The four space-travelled adult fish re-started mating and laying eggs on the 7th day after landing and continued to do so every day afterward. Fertilization rate and hatchability of these eggs were as high as the eggs laid by the laboratory-kept fish. This fact implies that in gametogenesis of adult fish, there are no specific stages of germ cells extremely susceptible to microgravity.

  2. Behavior of centrosomes during fertilization and cell division in mouse oocytes and in sea urchin eggs

    Science.gov (United States)

    Schatten, Heide; Schatten, Gerald; Balczon, Ron; Simerly, Calvin; Mazia, Daniel

    1986-01-01

    The behavior of centrosomes during the stages of fertilization and cell division in mouse oocytes and in sea urchin eggs was monitored in an immunofluorescence microscope, using autoimmune centrosomal antiserum derived from a patient with scleroderma to label the centrosomal material. These observations showed that centrosomes reproduce during the interphase and aggregate and separate during cell mitosis. Results supported the hypothesis of Mazia (1984), who proposed that centrosomes are 'flexible bodies'. It was also found that, while the sea urchin centrosomes are paternally inherited as was initially proposed by Bovery (1904), the mouse centrosomes are of maternal origin.

  3. Life-history consequences of investment in free-spawned eggs and their accessory coats.

    Science.gov (United States)

    Podolsky, Robert D

    2004-05-01

    The optimal trade-off between offspring size and number can depend on details of the mode of reproduction or development. In marine organisms, broadcast spawning is widespread, and external coats are a common feature of spawned eggs. Egg jelly coats are thought to influence several aspects of fertilization and early development, including the size of the target for sperm, fertilization efficiency, egg suspension time, polyspermy, embryo survival, and fecundity. These costs and benefits of investment in jelly result in trade-offs that can influence optimal reproductive allocation and the evolution of egg size. I develop an optimization model that sequentially incorporates assumptions about the function of egg coats in fertilization. The model predicts large variation in coat size and limited variation in ovum size under a broad range of conditions. Heterogeneity among spawning events further limits the range of ovum sizes predicted to evolve under sperm limitation. In contrast, variation in larval mortality predicts a broad range of optimal ovum sizes that more closely reflects natural variation among broadcast-spawning invertebrates. By decoupling physical and energetic size, egg coats can enhance fertilization, maintain high fecundity, and buffer the evolution of ovum size from variation in spawning conditions.

  4. Probing the biology of cell boundary conditions through confinement of Xenopus cell-free cytoplasmic extracts.

    Science.gov (United States)

    Bermudez, Jessica G; Chen, Hui; Einstein, Lily C; Good, Matthew C

    2017-01-01

    Cell-free cytoplasmic extracts prepared from Xenopus eggs and embryos have for decades provided a biochemical system with which to interrogate complex cell biological processes in vitro. Recently, the application of microfabrication and microfluidic strategies in biology has narrowed the gap between in vitro and in vivo studies by enabling formation of cell-size compartments containing functional cytoplasm. These approaches provide numerous advantages over traditional biochemical experiments performed in a test tube. Most notably, the cell-free cytoplasm is confined using a two- or three-dimensional boundary, which mimics the natural configuration of a cell. This strategy enables characterization of the spatial organization of a cell, and the role that boundaries play in regulating intracellular assembly and function. In this review, we describe the marriage of Xenopus cell-free cytoplasm and confinement technologies to generate synthetic cell-like systems, the recent biological insights they have enabled, and the promise they hold for future scientific discovery. © 2017 Wiley Periodicals, Inc.

  5. Sterilization and lethal gamma radiation doses on adults and eggs of Sitotroga Cerealella (OLIVIER)

    International Nuclear Information System (INIS)

    Wiendl, F.M.; Bovi, O.A.; Arthur, V.

    1975-04-01

    The influence of lethal doses of radiation from a cobalt 60 gamma source on eggs, adults and fertitility of Sitotroga Cerealella (Olivier) is described. Eggs irradiated with a dose of 14 Krad still showed viability of 16.1%. On longevity doses up to 70 Krad were usually non lethal but some variation could be observed related to the larval diet. Females fertilized by males irradiated with a dose of 70 Krad produced 36% fertile eggs. When the females were irradiated with the same dose, their fertility dropped to 2.2% and when both sexes were irradiated with a 60 Krad dose, the fertility was 28.8%

  6. Young female cancer patients' experiences with fertility counselling and fertility preservation

    DEFF Research Database (Denmark)

    Hoeg, Didde; Schmidt, Lone; Tryde Macklon, Kirsten

    2016-01-01

    were aware that chemotherapy could destroy their eggs. The participants described how specialized fertility counselling and fertility preservation contributed to a belief in life after cancer, which gave them hope that they would survive their cancer disease. Further, the women described how......Introduction: Fertility counselling for young women newly diagnosed with cancer is an important field of preconceptional counselling. This qualitative, small-scale study explored how young women newly diagnosed with cancer experienced specialized fertility preservation counselling and treatment...... in the public Danish health care system. Methods: Semi-structured, in-depth interviews were conducted with five women below 40 years recently diagnosed with cancer. All women received fertility counselling by a fertility specialist at the Fertility Clinic, University Hospital of Copenhagen, Denmark before...

  7. Development of Atlantic salmon (Salmo salar) eggs and alevins under varied temperature regimes

    Energy Technology Data Exchange (ETDEWEB)

    Peterson, R H; Spinney, H C.E.; Sreedharan, A

    1977-01-01

    Atlantic salmon (Salmo salar) eggs and alevins were raised under conditions where the temperature was systematically varied either at fertilization, at the eyed egg stage, or at hatching. Mortality was more than 20% in eggs started immediately after fertilization at constant incubation temperatures <4/sup 0/C as compared with 5% or less at temperatures >4/sup 0/C. Alevins that eyed at 8/sup 0/C and higher were progressively smaller the higher the temperature. The optimum temperature from fertilization to eye pigmentation was near 6/sup 0/C. Eyed eggs reared at lower temperatures until hatching were larger than those hatched at higher temperatures. This size differential was maintained until the yolk was completely absorbed at all posthatching temperatures investigated. Sudden decreases in temperature at the eyed egg and hatching stages induced severe edema of the alevin yolk sac, resulting in slower growth and increased mortality.

  8. Activation of maternal centrosomes in unfertilized sea urchin eggs

    Science.gov (United States)

    Schatten, H.; Walter, M.; Biessmann, H.; Schatten, G.

    1992-01-01

    Centrosomes are undetectable in unfertilized sea urchin eggs, and normally the sperm introduces the cell's microtubule-organizing center (MTOC) at fertilization. However, artificial activation or parthenogenesis triggers microtubule assembly in the unfertilized egg, and this study explores the reappearance and behavior of the maternal centrosome. During activation with A23187 or ammonia, microtubules appear first at the cortex; centrosomal antigen is detected diffusely throughout the entire cytoplasm. Later, the centrosome becomes more distinct and organizes a radial microtubule shell, and eventually a compact centrosome at the egg center organizes a monaster. In these activated eggs, centrosomes undergo cycles of compaction and decompaction in synchrony with the chromatin, which also undergoes cycles of condensation and decondensation. Parthenogenetic activation with heavy water (50% D2O) or the microtubule-stabilizing drug taxol (10 microM) induces numerous centrosomal foci in the unfertilized sea urchin egg. Within 15 min after incubation in D2O, numerous fine centrosomal foci are detected, and they organize a connected network of numerous asters which fill the entire egg. Taxol induces over 100 centrosomal foci by 15 min after treatment, which organize a corresponding number of asters. The centrosomal material in either D2O- or taxol-treated eggs aggregates with time to form fewer but denser foci, resulting in fewer and larger asters. Fertilization of eggs pretreated with either D2O or taxol shows that the paternal centrosome is dominant over the maternal centrosome. The centrosomal material gradually becomes associated with the enlarged sperm aster. These experiments demonstrate that maternal centrosomal material is present in the unfertilized egg, likely as dispersed undetectable material, which can be activated without paternal contributions. At fertilization, paternal centrosomes become dominant over the maternal centrosomal material.

  9. Lateral mobility of plasma membrane lipids in dividing Xenopus eggs

    OpenAIRE

    Laat, S.W. de; Tetteroo, P.A.T.; Bluemink, J.G.; Dictus, W.J.A.G.; Zoelen, E.J.J. van

    1984-01-01

    The lateral mobility of plasma membrane lipids was analyzed during first cleavage of Xaopus Levis eggs by fluorescence photobleaching recovery (FPR) measurements, using the lipid analogs 5-(N-hexadecanoyl)aminofluorescein (“HEDAF”) and 5-(N-tetradecanoyl)aminofluorescein (“TEDAF”) as probes. The preexisting plasma membrane of the animal side showed an inhomogeneous, dotted fluorescence pattern after labeling and the lateral mobility of both probes used was below the detection limits of the FP...

  10. Selected egg parameters and hatching time of African catfish ...

    African Journals Online (AJOL)

    Egg weight, diameter and mean number per gram were determined for fertilized eggs from broodfish of Clarias gariepinus and Heterobranchus longifilis and F1, hybrid broodfish (C. gariepinus x H. longifilis). Mean egg weight was least in C. gariepinus (1.30 mg) followed by H. longifilis (1.80 mg) and highest in the F1 hybrid ...

  11. [Monitoring and SWOT analysis of Ascaris eggs pollution in soil of rural China].

    Science.gov (United States)

    Zhu, Hui-hui; Zhou, Chang-hai; Zang, Wei; Zhang, Xue-qiang; Chen, Ying-dan

    2014-06-01

    To understand the status of Ascaris eggs pollution in soil at national monitoring spots of soil-transmitted nematodiasis, so as to provide the evidence for making countermeasures and evaluating the control effect. Ten households were selected from each of the 22 national monitoring spots annually according to the National Surveillance Program of Soil-Transmitted Nematodiasis (Trial), and the soil samples from vegetable gardens, toilet periphery, courtyards and kitchens were collected and examined by using the modified floatation test with saturated sodium nitrate. Fertilized or unfertilized eggs as well as live or dead fertilized eggs were discriminated and identified. In addition, a SWOT analysis of monitoring of Ascaris eggs pollution in the soil of rural China was carried out. A total of 1 090 households were monitored in 22 monitoring spots from 2006 to 2010. The total detection rate of Ascaris eggs in the soil was 30.73%, and the detection rates of fertilized, unfertilized and live fertilized eggs were 13.21%, 26.42% and 20.28%, respectively. The total detection rates of Ascaris eggs in the vegetable garden, toilet periphery, courtyard and kitchen were 16.51%, 13.49%, 14.22% and 10.73% respectively. The SWOT analysis demonstrated that the monitoring work had both advantages and disadvantages, and was faced with opportunities as well as threats. The pollution status of Ascaris eggs in the soil is still quite severe at some national monitoring spots, and the counter-measures such as implementing hazard-free treatment of stool, improving water supply and sanitation and reforming environment should be taken to protect people from being infected.

  12. Targeted integration of genes in Xenopus tropicalis

    DEFF Research Database (Denmark)

    Shi, Zhaoying; Tian, Dandan; Xin, Huhu

    2017-01-01

    With the successful establishment of both targeted gene disruption and integration methods in the true diploid frog Xenopus tropicalis, this excellent vertebrate genetic model now is making a unique contribution to modelling human diseases. Here, we summarize our efforts on establishing homologous...... recombination-mediated targeted integration in Xenopus tropicalis, the usefulness, and limitation of targeted integration via the homology-independent strategy, and future directions on how to further improve targeted gene integration in Xenopus tropicalis....

  13. Ovarian hyperstimulation syndrome in gonadotropin-treated laboratory South African clawed frogs (Xenopus laevis).

    Science.gov (United States)

    Green, Sherril L; Parker, John; Davis, Corrine; Bouley, Donna M

    2007-05-01

    Ovarian hyperstimulation syndrome (OHS) is a rare but sometimes fatal iatrogenic complication of ovarian stimulation associated with the administration of exogenous gonadotropins to women undergoing treatment for infertility. Laboratory Xenopus spp are commonly treated with human chorionic gonadotropin (hCG) to stimulate ovulation and optimize the number of oocytes harvested for use in biomedical research. Here we report cases of OHS in 2 gonadotropin-treated laboratory Xenopus laevis. After receiving hCG, the frogs developed severe subcutaneous accumulation of fluid, coelomic distention, and whole-body edema and were unable to dive, although they continued to eat and swim. At postmortem examination, extensive subcutaneous edema was present; ascites and massive numbers of free-floating eggs were found in the coelomic cavity and in aberrant locations: around the heart-sac and adhered to the liver capsule. Whole-body edema, gross enlargement of the ovaries, ascites, and abdominal distention are findings comparable to those observed in women with OHS. The pathophysiology of OHS is thought to be related to hormonally induced disturbances of vasoactive mediators, one of which may be vascular endothelial growth factor secreted by theca and granulosa cells. We know of no other report describing OHSlike symptoms in gonadotropin-treated frogs, and the cases described here are 2 of the 3 we have observed at our respective institutions over the last 6 y. According to these results, OHS appears to be rare in gonadotropin-treated laboratory Xenopus. However, the condition should be included in the differential diagnosis for the bloated frog.

  14. Changes in X-ray sensitivity of mouse eggs from fertilization to the early pronuclear stage, and their repair capacity

    International Nuclear Information System (INIS)

    Matsuda, Yoichi; Utsugi-Takeuchi, Toyoko; Tobari, Izuo; Seki, Naohiko; Chiba Univ.

    1989-01-01

    The analysis of potentiation effects of 3-aminobenzamide and caffeine on the yield of X-ray-induced chromosome aberrations demonstrated that the increase of radiosensitivity and the decrease of chromosome-type exchange induction with pronuclear formation, may be closely correlated with alterations in chromatin configuration in the pronuclei and in repair capacity of fertilized eggs at the pre-DNA-synthetic stage. No evidence based on repair efficiency was found for the marked difference in radiosensitivity between male and female genomes during pronuclear formation. (author)

  15. PTK2b function during fertilization of the mouse oocyte

    International Nuclear Information System (INIS)

    Luo, Jinping; McGinnis, Lynda K.; Carlton, Carol; Beggs, Hilary E.; Kinsey, William H.

    2014-01-01

    Highlights: • PTK2b is expressed in oocytes and is activated following fertilization. • PTK2b suppression in oocytes prevents fertilization, but not parthenogenetic activation. • PTK2b suppression prevents the oocyte from fusing with or incorporating bound sperm. • PTK2b suppressed oocytes that fail to fertilize do not exhibit calcium oscillations. - Abstract: Fertilization triggers rapid changes in intracellular free calcium that serve to activate multiple signaling events critical to the initiation of successful development. Among the pathways downstream of the fertilization-induced calcium transient is the calcium-calmodulin dependent protein tyrosine kinase PTK2b or PYK2 kinase. PTK2b plays an important role in fertilization of the zebrafish oocyte and the objective of the present study was to establish whether PTK2b also functions in mammalian fertilization. PTK2b was activated during the first few hours after fertilization of the mouse oocyte during the period when anaphase resumption was underway and prior to the pronuclear stage. Suppression of PTK2b kinase activity in oocytes blocked sperm incorporation and egg activation although sperm-oocyte binding was not affected. Oocytes that failed to incorporate sperm after inhibitor treatment showed no evidence of a calcium transient and no evidence of anaphase resumption suggesting that egg activation did not occur. The results indicate that PTK2b functions during the sperm-egg fusion process or during the physical incorporation of sperm into the egg cytoplasm and is therefore critical for successful development

  16. PTK2b function during fertilization of the mouse oocyte

    Energy Technology Data Exchange (ETDEWEB)

    Luo, Jinping [Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160 (United States); McGinnis, Lynda K. [Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS 66160 (United States); Carlton, Carol [Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160 (United States); Beggs, Hilary E. [Department of Ophthalmology, University of California, San Francisco, CA (United States); Kinsey, William H., E-mail: wkinsey@kumc.edu [Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160 (United States)

    2014-08-01

    Highlights: • PTK2b is expressed in oocytes and is activated following fertilization. • PTK2b suppression in oocytes prevents fertilization, but not parthenogenetic activation. • PTK2b suppression prevents the oocyte from fusing with or incorporating bound sperm. • PTK2b suppressed oocytes that fail to fertilize do not exhibit calcium oscillations. - Abstract: Fertilization triggers rapid changes in intracellular free calcium that serve to activate multiple signaling events critical to the initiation of successful development. Among the pathways downstream of the fertilization-induced calcium transient is the calcium-calmodulin dependent protein tyrosine kinase PTK2b or PYK2 kinase. PTK2b plays an important role in fertilization of the zebrafish oocyte and the objective of the present study was to establish whether PTK2b also functions in mammalian fertilization. PTK2b was activated during the first few hours after fertilization of the mouse oocyte during the period when anaphase resumption was underway and prior to the pronuclear stage. Suppression of PTK2b kinase activity in oocytes blocked sperm incorporation and egg activation although sperm-oocyte binding was not affected. Oocytes that failed to incorporate sperm after inhibitor treatment showed no evidence of a calcium transient and no evidence of anaphase resumption suggesting that egg activation did not occur. The results indicate that PTK2b functions during the sperm-egg fusion process or during the physical incorporation of sperm into the egg cytoplasm and is therefore critical for successful development.

  17. Postmating sexual conflict and female control over fertilization during gamete interaction.

    Science.gov (United States)

    Firman, Renée C

    2018-06-01

    Males and females rarely have identical evolutionary interests over reproduction, and when the fitness of both sexes is dependent upon paternity outcomes, sexual conflict over fertilization is inevitable. In internal fertilizers, the female tract is a formidable selective force on the number and integrity of sperm that reach the egg. Selection on sperm quality is intensified when females mate multiply and rival males are forced to compete for fertilizations. While male adaptations to sperm competition have been well documented (e.g., increased sperm fertilizing capacity), much less attention has been given to the evolutionary consequences of postmating sexual conflict for egg form and function. Specifically, increased sperm competitiveness can be detrimental by giving rise to an elevation in reproductive failure resulting from polyspermy. Spanning literature on both internal and external fertilizers, in this review I discuss how females respond to sperm competition via fertilization barriers that mediate sperm entry. These findings, which align directly with sexual conflict theory, indicate that females have greater control over fertilization than has previously been appreciated. I then consider the implications of gametic sexual conflict in relation to the development of reproductive isolation and speculate on potential mechanisms accounting for "egg defensiveness." Finally, I discuss the functional significance of egg defensiveness for both the sexes, and sperm selection for females. © 2018 New York Academy of Sciences.

  18. Effect of Dietary L-ascorbic Acid (L-AA on Production Performance, Egg Quality Traits and Fertility in Japanese Quail ( at Low Ambient Temperature

    Directory of Open Access Journals (Sweden)

    N. Shit

    2012-07-01

    Full Text Available Environmental stress boosts the levels of stress hormones and accelerates energy expenditure which subsequently imbalance the body’s homeostasis. L-ascorbic acid (L-AA has been recognized to mitigate the negative impact of environmental stress on production performances in birds. The present investigation was carried out to elucidate the effect of different dietary levels of L-AA on production performance, egg quality traits and fertility in Japanese quail at low ambient temperature. Sixty matured females (15 wks were equally divided into three groups (20/group based on the different dietary levels of L-AA (0, 250 and 500 ppm and coupled with an equal number of males (1:1 obtained from the same hatch. They were managed in uniform husbandry conditions without restriction of feed and water at 14 h photo-schedule. Except for feed efficiency, body weight change, feed consumption and hen-day egg production were recorded highest in 500 ppm L-AA supplemented groups. Among the all egg quality traits studied, only specific gravity, shell weight and thickness differed significantly (p<0.05 in the present study. Fertility was improved significantly (p<0.01 to a dose dependent manner of L-AA. The findings of the present study concluded that dietary L-AA can be a caring management practice at least in part to alleviate the adverse effect of cold induced stress on production performance in Japanese quail.

  19. Three Cell Fusions during Double Fertilization.

    Science.gov (United States)

    Sprunck, Stefanie; Dresselhaus, Thomas

    2015-05-07

    Fertilization of both egg and central cell is a major distinguishing feature of flowering plants. Now, Maruyama et al. report a third cell fusion event between the persistent synergid and the fertilized central cell shortly after double fertilization in Arabidopsis. This causes rapid dilution of pollen tube attractant(s), preventing polytubey. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Running out of time: exploring women's motivations for social egg freezing.

    Science.gov (United States)

    Baldwin, Kylie; Culley, Lorraine; Hudson, Nicky; Mitchell, Helene

    2018-04-12

    Few qualitative studies have explored women's use of social egg freezing. Derived from an interview study of 31 participants, this article explores the motivations of women using this technology. Semi-structured interviews were conducted with 31 users of social egg freezing resident in UK (n = 23), USA (n = 7) and Norway (n = 1). Interviews were face to face (n = 16), through Skype and Facetime (n = 9) or by telephone (n = 6). Data were analyzed using interpretive thematic analysis. Women's use of egg freezing was shaped by fears of running out of time to form a conventional family, difficulties in finding a partner and concerns about "panic partnering", together with a desire to avoid future regrets and blame. For some women, use of egg freezing was influenced by recent fertility or health diagnoses as well as critical life events. A fifth of the participants also disclosed an underlying fertility or health issue as affecting their decision. The study provides new insights in to the complex motivations women have for banking eggs. It identifies how women's use of egg freezing was an attempt to "preserve fertility" in the absence of the particular set of "life conditions" they regarded as crucial for pursuing parenthood. It also demonstrates that few women were motivated by a desire to enhance their career and that the boundaries between egg freezing for medical and for social reasons may be more porous than first anticipated.

  1. Efficacy of egg-yolk citrate extender fortified with aqueous garlic ...

    African Journals Online (AJOL)

    An experiment was designed to harness the potential of aqueous fresh and dried garlic extracts supplement in egg yolk extender for rooster semen to improve fertility and hatchability of egg. Aqueous extract of fresh and dry garlic were obtained and supplemented into egg yolk-citrate buffer at 0,1, 2, 3, 4, 5, 6 and 7% for ...

  2. Trust women to choose: a response to John a Robertson's 'Egg freezing and Egg banking: empowerment and alienation in assisted reproduction'.

    Science.gov (United States)

    Goold, Imogen

    2017-12-01

    In 'Egg Freezing and Egg Banking: Empowerment and Alienation in Assisted Reproduction', John A Robertson responds to the American Society of Reproductive Medicine's statement that oocyte preservation should no longer be considered an experimental treatment. He explores the implications of this development, focusing on the potentially empowering impact of oocyte preservation as a means for women to preserve their fertility. He also engages with concerns about the possibility that such a development may raise issues of alienation. He highlights some of the potential problems that may emerge as women gain the capacity to store and either donate or sell any eggs they do not need for their own reproductive purposes. Much of his paper is valuable and considered, but in places, his views rest on assumptions about women's attitudes to their fertility, understanding of the technology, and relationship with their gametes that are open to dispute. This paper teases out some of these assumptions and puts pressure on them by drawing on the growing body of data about what women actually do think and feel about fertility issues. It focuses on two of his main concerns-that social egg freezing may give women a false sense of security and that women may be harmed if a market in eggs leads to their alienation from their gametes. Via this response to Robertson, I aim to redress the tendency often seen in discussions around women, infertility, aging, and empowerment to unquestioningly accept what I argue are stereotypes and assumptions about women's views and capacity to reason.

  3. Cryoprotective and contraceptive properties of egg yolk as an additive in rooster sperm diluents.

    Science.gov (United States)

    Santiago-Moreno, Julián; Castaño, Cristina; Toledano-Díaz, Adolfo; Coloma, Miguel A; López-Sebastián, Antonio; Prieto, María T; Campo, Jose L

    2012-12-01

    The addition of chicken egg yolk to semen extenders is thought to reduce the fertilizing potential of rooster spermatozoa--but not (or at least not as much) that of other avian species. The aim of the present study was to determine whether quail egg yolk, a novel extender additive, provides advantages over chicken egg yolk in the cryopreservation of rooster spermatozoa. Experiments were also performed to determine whether the harmful effect of egg yolk occurs during cryopreservation or during fertilization after artificial insemination. Heterospermic rooster semen samples were divided into aliquots and cooled in a polyvinylpyrrolidone-based medium containing 15% chicken egg yolk, 15% quail egg yolk or no egg yolk at all. The viability of spermatozoa of cooled samples (5 °C) without egg yolk were less viable (Prooster sperm cells against cold shock and during freezing and thawing, but exerts a contraceptive effect in the genital tract of the hen. Copyright © 2012 Elsevier Inc. All rights reserved.

  4. Effects of floor eggs on hatchability and later life performance in broiler chickens.

    Science.gov (United States)

    van den Brand, H; Sosef, M P; Lourens, A; van Harn, J

    2016-05-01

    Two experiments were conducted in which effects of floor eggs, washed floor eggs, and clean nest eggs were investigated on incubation characteristics and performance in later life of broiler chickens. In both experiments, a young and an older breeder flock were used in a 3×2 factorial design during incubation. In the second experiment, male and female chickens were reared separately until d 35 of age in floor pens. During this grow out trial, an extra group was created in which chickens obtained from clean nest eggs were mixed with chickens obtained from floor eggs, meaning that grow out period was set up as a 4×2×2 factorial design with 4 egg types, 2 breeder ages, and 2 sexes. In both experiments, fertility and hatchability of fertile eggs were lower in floor and washed eggs than in clean nest eggs (hatchability: experiment 1: 74.4 vs. 70.6 vs. 92.6% for floor eggs, washed floor eggs and clean nest eggs, respectively, P<0.001; experiment 2: 78.3 vs. 81.7 vs. 90.2%, respectively, P<0.001). In experiment 2, BW at d 0 of chickens obtained from clean nest eggs was higher than that of chickens from floor eggs and washed floor eggs (41.5 vs. 40.4 and 40.3 g, respectively; P<0.001). This difference disappeared during the grow out period and was absent at slaughter age at d 35 of age. Feed intake (FI), feed conversion ratio (FCR), and mortality during the grow out period were not affected by egg type. Incidence and severity of hock burns and footpad dermatitis were not affected by egg type or breeder age. Litter friability at d 35 of age tended to be lower in pens with chickens obtained from washed floor eggs compared to clean nest eggs. We conclude that incubation of floor eggs or washed floor eggs resulted in lower fertility and hatchability compared to clean nest eggs, but that performance during the grow out period was not affected. © 2016 Poultry Science Association Inc.

  5. The effect of long-term feeding of conjugated linoleic acid on fertility ...

    African Journals Online (AJOL)

    The objective of this study was to determine the effects of the long-term feeding of conjugated linoleic acid (CLA) on laying hen performance, egg fertility and hatchability of fertile eggs of Japanese quail (Coturnix coturnix japonica). One hundred and sixty 7-day old Japanese quail chicks were randomly assigned to four ...

  6. Stability of post-fertilization traveling waves

    Science.gov (United States)

    Flores, Gilberto; Plaza, Ramón G.

    This paper studies the stability of a family of traveling wave solutions to the system proposed by Lane et al. [D.C. Lane, J.D. Murray, V.S. Manoranjan, Analysis of wave phenomena in a morphogenetic mechanochemical model and an application to post-fertilization waves on eggs, IMA J. Math. Appl. Med. Biol. 4 (4) (1987) 309-331], to model a pair of mechanochemical phenomena known as post-fertilization waves on eggs. The waves consist of an elastic deformation pulse on the egg's surface, and a free calcium concentration front. The family is indexed by a coupling parameter measuring contraction stress effects on the calcium concentration. This work establishes the spectral, linear and nonlinear orbital stability of these post-fertilization waves for small values of the coupling parameter. The usual methods for the spectral and evolution equations cannot be applied because of the presence of mixed partial derivatives in the elastic equation. Nonetheless, exponential decay of the directly constructed semigroup on the complement of the zero eigenspace is established. We show that small perturbations of the waves yield solutions to the nonlinear equations decaying exponentially to a phase-modulated traveling wave.

  7. Unravelling anisogamy: egg size and ejaculate size mediate selection on morphology in free-swimming sperm.

    Science.gov (United States)

    Monro, Keyne; Marshall, Dustin J

    2016-07-13

    Gamete dimorphism (anisogamy) defines the sexes in most multicellular organisms. Theoretical explanations for its maintenance usually emphasize the size-related selection pressures of sperm competition and zygote survival, assuming that fertilization of all eggs precludes selection for phenotypes that enhance fertility. In external fertilizers, however, fertilization is often incomplete due to sperm limitation, and the risk of polyspermy weakens the advantage of high sperm numbers that is predicted to limit sperm size, allowing alternative selection pressures to target free-swimming sperm. We asked whether egg size and ejaculate size mediate selection on the free-swimming sperm of Galeolaria caespitosa, a marine tubeworm with external fertilization, by comparing relationships between sperm morphology and male fertility across manipulations of egg size and sperm density. Our results suggest that selection pressures exerted by these factors may aid the maintenance of anisogamy in external fertilizers by limiting the adaptive value of larger sperm in the absence of competition. In doing so, our study offers a more complete explanation for the stability of anisogamy across the range of sperm environments typical of this mating system and identifies new potential for the sexes to coevolve via mutual selection pressures exerted by gametes at fertilization. © 2016 The Author(s).

  8. Trust women to choose: a response to John a Robertson's ‘Egg freezing and Egg banking: empowerment and alienation in assisted reproduction’†

    Science.gov (United States)

    2017-01-01

    Abstract In ‘Egg Freezing and Egg Banking: Empowerment and Alienation in Assisted Reproduction’, John A Robertson responds to the American Society of Reproductive Medicine's statement that oocyte preservation should no longer be considered an experimental treatment. He explores the implications of this development, focusing on the potentially empowering impact of oocyte preservation as a means for women to preserve their fertility. He also engages with concerns about the possibility that such a development may raise issues of alienation. He highlights some of the potential problems that may emerge as women gain the capacity to store and either donate or sell any eggs they do not need for their own reproductive purposes. Much of his paper is valuable and considered, but in places, his views rest on assumptions about women's attitudes to their fertility, understanding of the technology, and relationship with their gametes that are open to dispute. This paper teases out some of these assumptions and puts pressure on them by drawing on the growing body of data about what women actually do think and feel about fertility issues. It focuses on two of his main concerns—that social egg freezing may give women a false sense of security and that women may be harmed if a market in eggs leads to their alienation from their gametes. Via this response to Robertson, I aim to redress the tendency often seen in discussions around women, infertility, aging, and empowerment to unquestioningly accept what I argue are stereotypes and assumptions about women's views and capacity to reason. PMID:29868183

  9. Successful pregnancy with donor eggs in-vitro fertilization after premature ovarian insufficiency in a tertiary hospital in a low-income setting: a case report

    Directory of Open Access Journals (Sweden)

    Thomas Obinchemti Egbe

    2016-11-01

    Full Text Available Abstract Background Premature Ovarian Insufficiency (POI is classically defined as 4–6 months of cessation of menses (amenorrhea in women under 40, associated with menopausal level of serum gonadotropins FSH > 40 IU/L and hypo-estrogenism and is also referred to as hypergonadotropic hypogonadism. This disorder can manifest as primary amenorrhea without the onset of menses (menarche, or as secondary amenorrhea after menarche and pubertal development. The diagnosis of this condition in Cameroon is sometimes difficult because of the high cost of hormonal assays and the few laboratories offering these services. Case presentation The patient was a 38-year-old G2P0020, blood group O Rh positive, genotype AA and BMI 19 kg/m2 who came to our service because of secondary amenorrhea and infertility of 2 years’ duration. She has a history of pulmonary tuberculosis that was treated in Cameroon. After laparoscopy and hormonal profile, the diagnosis of premature ovarian insufficiency was reached. The woman underwent a successful donor egg in-vitro fertilization cycle and delivered a female fetus. Two years later YE requested IVF with autologous eggs, which was not possible, and since then she has remained with one child. Conclusion The diagnosis of premature ovarian insufficiency is difficult in Cameroon because of the high cost of laboratory investigations and difficult access to the tests. In-vitro fertilization with donor egg is a better treatment option. Unfortunately, it is not accessible to most Cameroonians because of lack of technical ability and the existence of cultural and financial barriers.

  10. First production of larvae using cryopreserved sperm: Effects of preservation temperature and cryopreservation on European eel sperm fertilization capacity

    DEFF Research Database (Denmark)

    Asturiano, J.F.; Sørensen, Sune Riis; Perez, L.

    2016-01-01

    Sperm cryopreservation is a useful tool in captive fish reproduction management, that is to synchronize gamete production, especially in the case of species as the European eel, where the time of female spawning readiness is unpredictable. Several protocols to cryopreserve sperm of this species....... Fertilization of two egg batches was attempted. Diluted sperm caused a similar percentage of fertilized eggs and a similar number of embryos and larvae, independently of storage temperature (4 or 20°C). The cryopreserved sperm resulted in a lower percentage of fertilized eggs, but embryos developed and a few...... larvae ('cryolarvae') were obtained 55 h after fertilization in one of the two egg batches. This result evidences that the tested cryopreservation protocol is applicable for eel reproduction management, although improvements will be required to enhance fertilization success...

  11. Mature Oocyte Cryopreservation for Fertility Preservation.

    Science.gov (United States)

    Liang, Tina; Motan, Tarek

    2016-01-01

    In recent decades, advances in cancer treatment have led to a dramatic improvement in long term survival. This has led to an increasing focus on quality of life after surviving cancer treatment, with fertility being an important aspect. Given the known reproductive risks of cancer therapies, there has been a growing interest in the field of fertility preservation (also referred to as oncofertility). Mature oocyte cryopreservation is no longer considered experimental and has become a realistic option for reproductive aged women prior to undergoing cancer treatment. Additionally, as cryopreservation techniques improve, mature oocyte cryopreservation is increasing being marketed to healthy women without cancer wishing to delay child bearing, also termed "social egg freezing". This chapter provides a review of the current technology, use, and outcomes of mature oocyte cryopreservation. It also outlines the ethical debate surrounding social egg freezing and directions for future research in female fertility preservation.

  12. Identification, developmental expression and regulation of the Xenopus ortholog of human FANCG/XRCC9.

    Science.gov (United States)

    Stone, Stacie; Sobeck, Alexandra; van Kogelenberg, Margriet; de Graaf, Bendert; Joenje, Hans; Christian, Jan; Hoatlin, Maureen E

    2007-07-01

    Fanconi anemia (FA) is associated with variable developmental abnormalities, bone marrow failure and cancer susceptibility. FANCG/XRCC9 is member of the FA core complex, a group of proteins that control the monoubiquitylation of FANCD2, an event that plays a critical role in maintaining genomic stability. Here we report the identification of the Xenopus laevis ortholog of human FANCG (xFANCG), its expression during development, and its molecular interactions with a partner protein, xFANCA. The xFANCG protein sequence is 47% similar to its human ortholog, with highest conservation in the two putative N-terminal leucine zippers and the tetratricopeptide repeat (TPR) motifs. xFANCG is maternally and zygotically transcribed. Prior to the midblastula stage, a single xFANCG transcript is observed but two additional alternatively spliced mRNAs are detected after the midblastula transition. One of the variants is predicted to encode a novel isoform of xFANCG lacking exon 2. The mutual association between FANCG and FANCA required for their nuclear import is conserved in Xenopus egg extracts. Our data demonstrate that interactions between FANCA and FANCG occur at the earliest stage of vertebrate development and raise the possibility that functionally different isoforms of xFANCG may play a role in early development.

  13. Oocyte cryopreservation for donor egg banking.

    Science.gov (United States)

    Cobo, Ana; Remohí, José; Chang, Ching-Chien; Nagy, Zsolt Peter

    2011-09-01

    Oocyte donation is an efficient alternative to using own oocytes in IVF treatment for different indications. Unfortunately, 'traditional' (fresh) egg donations are challenged with inefficiency, difficulties of synchronization, very long waiting periods and lack of quarantine measures. Given the recent improvements in the efficiency of oocyte cryopreservation, it is reasonable to examine if egg donation through oocyte cryopreservation has merits. The objective of the current manuscript is to review existing literature on this topic and to report on the most recent outcomes from two established donor cryobank centres. Reports on egg donation using slow freezing are scarce and though results are encouraging, outcomes are not yet comparable to a fresh egg donation treatment. Vitrification on the other hand appears to provide high survival rates (90%) of donor oocytes and comparable fertilization, embryo development, implantation and pregnancy rates to traditional (fresh) egg donation. Besides the excellent outcomes, the ease of use for both donors and recipients, higher efficiency, lower cost and avoiding the problem of synchronization are all features associated with the benefit of a donor egg cryobank and makes it likely that this approach becomes the future standard of care. Oocyte donation is one of the last resorts in IVF treatment for couples challenged with infertility problems. However, traditional (fresh) egg donation, as it is performed today, is not very efficient, as typically all eggs from one donor are given to only one recipient, it is arduous as it requires an excellent synchronization between the donor and recipient and there are months or years of waiting time. Because of the development of an efficient oocyte cryopreservation technique, it is now possible to cryo-store donor (as well as non-donor) eggs, maintaining their viability and allowing their use whenever there is demand. Therefore, creating a donor oocyte cryobank would carry many advantages

  14. Confinement lowers fertility rate of helmeted guinea fowl ( Numida ...

    African Journals Online (AJOL)

    ... common game bird in Africa and there have been efforts to domesticate it for use as a source of human food. An important obstacle in successful domestication of guinea fowl is their low fertility rate.We studied the effects of semi-confinement on the fertility rates of helmeted guinea fowl by comparing egg fertility, hatch rate ...

  15. Effect of Some Egg Quality Traits on Hatching Results in Brown Pure Lines

    Directory of Open Access Journals (Sweden)

    İsmail Durmuş

    2016-09-01

    Full Text Available This study was conducted to determine the influence of eggshell thickness, albumen height, yolk height and haugh unit on hatching results. A total of 746 eggs, which were obtained and evaluated for egg quality parameters from 100 brown pure line hens of 42 weeks of age, were used. Eggs were collected from each hen for 12 days and individual incubation was carried out. Eggs were grouped based on quality parameters and evaluated accordingly. The findings suggested that hatchability, hatchability of fertile eggs, early, mid and late embryonic mortality did not differ in terms of haugh unit, albumen height, eggshell thickness and yolk height groups. However, early embryonic mortality was found different between the yolk height groups. No relationship was determined among albumen height, haugh unit, eggshell thickness and hatching results. There was no relation between yolk height and hatchability of fertile eggs, hatchability, mid and late embryonic mortalities but was a positive correlation with early embryonic mortality. The results of the present study demonstrate that egg quality parameters studied here had no influence on hatching results except that early embryonic mortality increased with the yolk height.

  16. 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.

  17. Novel genetic markers of the carbonic anhydrase II gene associated with egg production and reproduction traits in Tsaiya ducks.

    Science.gov (United States)

    Chang, M-T; Cheng, Y-S; Huang, M-C

    2013-02-01

    In our previous cDNA microarray study, we found that the carbonic anhydrase II (CA2) gene is one of the differentially expressed transcripts in the duck isthmus epithelium during egg formation period. The aim of this study was to identify the single-nucleotide polymorphisms (SNPs) in the CA2 gene of Tsaiya ducks. The relationship of SNP genotype with egg production and reproduction traits was also investigated. A total of 317 ducks from two lines, a control line with no selection and a selected line, were employed for testing. Three SNPs (C37T, A62G and A65G) in the 3'-untranslated region of the CA2 gene were found. SNP-trait association analysis showed that SNP C37T and A62G were associated with duck egg weight besides fertility. The ducks with the CT and AG genotypes had a 1.46 and 1.62 g/egg lower egg weight as compared with ducks with the CC and AA genotypes, respectively (p ducks with CT and AG genotypes had 5.20% and 4.22% higher fertility than those with CC and AA genotypes, respectively (p duck fertility, and the diplotype H1H4 was dominant for duck fertility. These findings might provide the basis for balanced selection and may be used in marker-assisted selection to improve egg weight and fertility simultaneously in the Tsaiya ducks. © 2012 Blackwell Verlag GmbH.

  18. Molecular basis of sperm capacitation acrosome reaction and interaction with eggs

    International Nuclear Information System (INIS)

    Sheikhnejad, G.

    1985-01-01

    A phospholipase C (PLC) which can hydrolyze 14 C-phosphatidylcholine was purified from bull seminal plasma. This PLC has an optimum at pH 7.2 and its PI was about 5.0. The enzyme was inhibited by EDTA, Cd 2+ , Pb 2+ , Ni 2+ , Fe 2+ , and Zn 2+ . PLC consists of two subunits one 69,000 and the other 55,000 daltons. The purified PLC was examined for induction of capacitation and acrosome reaction of guinea pig spermatozoa. Sperm were examined for the acrosome reaction 10 min after addition of 3.4 mM Ca 2+ . Fifty percent of the sperm underwent the acrosome reaction while the control had less than 5% acrosome reacted sperm. The antiserum to the inneracrosomal membrane isolated from sperm was labeled with FITC conjugated goat anti-guinea pig IgG. The conjugated antibody was used to localize sperm antigens. The antigens located on the IAM were only fluoresced when rabbit sperm were treated with methanol and/or MgCl 2 . Therefore anti-IAM antibody did not bind to the sperm plasma membrane. In vivo capacitated rabbit sperm were incubated with anti-IAM antibody (intact IgG and F(ab') 2 fragments) for 30 min prior to addition of rabbit eggs. After 24 h the eggs were examined for cleavage. The control eggs were fertilized (90%) while the antibody completely inhibited the fertilization of ova in vitro. The eggs incubated with antibody prior to the addition of sperm were still fertilizable. Thus, anti-IAM did not have any noticeable effect on the eggs. It was also shown that antibody inhibited fertilization of zona-free rabbit eggs in vitro as well

  19. Hyperinnervation improves Xenopus laevis limb regeneration.

    Science.gov (United States)

    Mitogawa, Kazumasa; Makanae, Aki; Satoh, Akira

    2018-01-15

    Xenopus laevis (an anuran amphibian) shows limb regeneration ability between that of urodele amphibians and that of amniotes. Xenopus frogs can initiate limb regeneration but fail to form patterned limbs. Regenerated limbs mainly consist of cone-shaped cartilage without any joints or branches. These pattern defects are thought to be caused by loss of proper expressions of patterning-related genes. This study shows that hyperinnervation surgery resulted in the induction of a branching regenerate. The hyperinnervated blastema allows the identification and functional analysis of the molecules controlling this patterning of limb regeneration. This paper focuses on the nerve affects to improve Xenopus limb patterning ability during regeneration. The nerve molecules, which regulate limb patterning, were also investigated. Blastemas grown in a hyperinnervated forelimb upregulate limb patterning-related genes (shh, lmx1b, and hoxa13). Nerves projecting their axons to limbs express some growth factors (bmp7, fgf2, fgf8, and shh). Inputs of these factors to a blastema upregulated some limb patterning-related genes and resulted in changes in the cartilage patterns in the regenerates. These results indicate that additional nerve factors enhance Xenopus limb patterning-related gene expressions and limb regeneration ability, and that bmp, fgf, and shh are candidate nerve substitute factors. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Effects of Dietary Vitamin E on Fertility Functions in Poultry Species

    Directory of Open Access Journals (Sweden)

    Deivendran Rengaraj

    2015-04-01

    Full Text Available Vitamin E is found in high quantities in vegetable oils. Although vitamin E has multiple functions in humans and animals, its key function is protecting cells from oxidative damage. Since its discovery, several studies have demonstrated that vitamin E deficiency causes impaired fertility in humans and lab animals. However, the effects of vitamin E deficiency or of its supplementation on the fertility of farm animals, particularly on poultry, are less well studied. Therefore, a comprehensive review of the effects of dietary vitamin E on the fertility of poultry species is needed in order to understand the beneficial role of vitamin E in the maintenance of sperm and egg qualities. Based on the observations reviewed here, we found that a moderate amount of vitamin E in poultry diet significantly protects semen/sperm qualities in male birds and egg qualities in female birds via decreasing the lipid peroxidation in semen/sperms and eggs. This review provides an overall understanding of the effects of dietary vitamin E on fertility functions in poultry species.

  1. In-Vitro Fertilization Practice: Awareness and Perceptions ...

    African Journals Online (AJOL)

    BACKGROUND: In-vitro fertilization(IVF) is a procedure that is one of the options in managing infertility. It involves fertilizing the egg from the female with the spermatozoa from the male outside the human body. It is a common practice in the developed world. However it is still a new phenomenon in the developing countries.

  2. A Unique Egg Cortical Granule Localization Motif Is Required for Ovastacin Sequestration to Prevent Premature ZP2 Cleavage and Ensure Female Fertility in Mice.

    Directory of Open Access Journals (Sweden)

    Bo Xiong

    2017-01-01

    Full Text Available Monospermic fertilization is mediated by the extracellular zona pellucida composed of ZP1, ZP2 and ZP3. Sperm bind to the N-terminus of ZP2 which is cleaved after fertilization by ovastacin (encoded by Astl exocytosed from egg cortical granules to prevent sperm binding. AstlNull mice lack the post-fertilization block to sperm binding and the ability to rescue this phenotype with AstlmCherry transgenic mice confirms the role of ovastacin in providing a definitive block to polyspermy. During oogenesis, endogenous ovastacin traffics through the endomembrane system prior to storage in peripherally located cortical granules. Deletion mutants of ovastacinmCherry expressed in growing oocytes define a unique 7 amino acid motif near its N-terminus that is necessary and sufficient for cortical granule localization. Deletion of the 7 amino acids by CRISPR/Cas9 at the endogenous locus (AstlΔ prevents cortical granule localization of ovastacin. The misdirected enzyme is present within the endomembrane system and ZP2 is prematurely cleaved. Sperm bind poorly to the zona pellucida of AstlΔ/Δ mice with partially cleaved ZP2 and female mice are sub-fertile.

  3. On the manganese content of the egg of Oryzias latipes and its change during the early development

    International Nuclear Information System (INIS)

    Hori, R.; Iwasaki, S.I.

    1976-01-01

    The content of manganese of the egg of Oryzias latipes was determined by atomic absorption spectrophotometry and activation analysis. The amount of manganese in the unfertilized egg was 0.51 mμg and increased remarkably in one hour after fertilization. Although the increasing rate became gradually lower in further stages, the content of manganese amounted to 2.61 mμg in the embryo after gastrulation. Tracer-experiments showed that the egg accumulated manganese positively from the surrounding medium during the short time after fertilization. (author)

  4. Fertilized eggs obtained from transplantation of frozen ovaries and parthenogenesis in combination with artificial insemination of frozen semen of the silkworm, Bombyx mori.

    Science.gov (United States)

    Mochida, Yuji; Takemura, Yoko; Kanda, Toshio; Horie, Yasuhiro

    2003-04-01

    A reliable method is reported for the long-term preservation of ovaries and spermatozoa of the silkworm (Bombyx mori). Three studies are presented. In the first, ovaries were removed from larvae at either 3rd, 4th, or 5th instar, cryopreserved, and stored in liquid nitrogen. Thawed ovaries were transplanted to surgically castrated female larvae at the same or a different developmental stage. The highest percentage of recipient females producing eggs resulted into either 3rd or 4th instar larvae (respectively, 22.1 and 8.7%). Similarly, the highest levels of other measurements of successful cryopreservation and transplanted ovary, and number of eggs laid, occurred with the same combination of donor and recipient developmental stages. Other combinations of ovary/recipient developmental stages yielded lower results. In the second experiment, semen was collected from male moths, cryopreserved, and then thawed semen was diluted with trypsin solution and artificially inseminated into females obtained from the best conditions of first experiment. A small percentage of inseminated moths laid eggs (8-10.3%) compared to that of controls (100%). In addition, the fertility of eggs from experimental moths was lower than that of control females (respectively, 40.3-88% and 97.5%). In the third experiment, eggs were surgically removed from ovarian tubules of moth following transplantation of thawed ovaries and subjected to parthenogenetic activation and artificial hatching. As expected, all resulting moths were female and, following natural mating or artificial insemination with thawed semen, yielded normal offspring at high rates.

  5. Artificial fertilization by intracytoplasmic sperm injection in a teleost fish, the medaka (Oryzias latipes).

    Science.gov (United States)

    Otani, Satoshi; Iwai, Toshiharu; Nakahata, Shingo; Sakai, Chiharu; Yamashita, Masakane

    2009-01-01

    Intracytoplasmic sperm injection (ICSI) is a technique that has been successfully used for assisting reproduction in mammals. However, this method is still not reliable in nonmammalian species, including teleosts. We succeeded in producing medaka individuals by ICSI with a rate of 13.4% (28 hatched embryos out of 209 eggs fertilized by ICSI), the best value reported so far in teleosts, including zebrafish and Nile tilapia. Although the technique was based on that developed for mammalian eggs, some critical modifications were made to adjust it to the medaka egg, which has a thick and hard envelope (the chorion) and a single sperm entry site (the micropyle). Medaka ICSI was performed by injecting a demembranated spermatozoon into an egg cytoplasm through the micropyle 10-15 sec after egg activation induced by a piezo-actuated vibration, the site and timing of sperm penetration being consistent with those in normal fertilization in medaka. To increase the efficiency of ICSI in medaka, we found that the fertilization by ICSI should precisely mimic the fertilization by insemination with intact sperm, both spatially and temporally. The success rate of ICSI was highly variable in batches of eggs (ranging from 0% to 56%), suggesting that the conditions of eggs are important factors in stabilizing the production of individuals by ICSI. The success in medaka ICSI provides a basis for future research to understand the basic mechanisms in gamete biology of teleosts as well as for development of new technology that can yield valuable applications in fisheries science.

  6. What to do now? How women with breast cancer make fertility preservation decisions.

    Science.gov (United States)

    Snyder, Karrie Ann; Tate, Alexandra Lee

    2013-07-01

    There has been increased attention paid to cancer-related infertility and fertility preservation. However, how cancer patients decide whether or not to pursue fertility preservation has not been fully examined. The data come from 34 interviews with women in the USA diagnosed with breast cancer prior to 40 years of age who contemplated fertility preservation prior to cancer treatment. Fully transcribed interviews were coded through a three-staged inductive process. Three sets of factors that shaped the decision-making process of the respondents regarding fertility preservation treatment options were identified: perceived benefits (e.g. ability to use 'younger' eggs in the future), inhibiting concerns (e.g. success rates) and influential relationships (e.g. physicians, parents and partners). Respondents saw their main fertility preservation decision as choosing whether or not to pursue egg/embryo banking. The decision-making process was complicated and included both health-related and personal considerations, with many respondents reporting a lack of support services for fertility issues. Findings suggest that greater attention needs to be placed on presenting patients with a wider range of options. Those who counsel patients regarding fertility preservation decisions should be aware of the influence of relationship dynamics, broader health care concerns, and fertility histories on these decisions. KEY MESSAGE POINTS: While fertility preservation has garnered greater attention, less is known about how cancer patients make fertility preservation decisions. Despite the range of choices for fertility preservation, respondents identified egg/embryo banking as their primary option. Many factors outside of cancer concerns inhibit and facilitate fertility preservation decisions including fertility history and family relationship dynamics.

  7. The histone H5 variant in Xenopus laevis

    NARCIS (Netherlands)

    Moorman, A. F.; de Boer, P. A.; Linders, M. T.; Charles, R.

    1984-01-01

    The presumptive histone H5 of Xenopus laevis has been characterized by SDS and acid-urea-Triton polyacrylamide gel electrophoresis and compared with chicken histone H5. Chicken H5 has a lower electrophoretic mobility compared to that of Xenopus H5 in both gel systems. It is shown, using a polyclonal

  8. Medical and social egg freezing

    DEFF Research Database (Denmark)

    Lallemant, Camille; Vassard, Ditte; Andersen, Anders Nyboe

    2016-01-01

    INTRODUCTION: Until recently, limited options for preserving fertility in order to delay childbearing were available. Although egg freezing and successful thawing is now possible, it remains unclear to what extent women are aware of the availability of this technique, their attitudes towards its...... use, or the circumstances under which this technique may be considered. MATERIAL AND METHODS: An online cross-sectional survey was designed to investigate knowledge and attitudes of women in Denmark and the UK on egg freezing and their potential intentions regarding the procedure. RESULTS: Data...... was collected from September 2012 to September 2013 and the responses of 973 women were analyzed. In total, 83% of women reported having heard of egg freezing, and nearly all considered it acceptable for medical indications, whilst 89% considered it acceptable for social reasons. Overall, 19% expressed active...

  9. Survival of Ascaris eggs and hygienic quality of human excreta in Vietnamese composting latrines

    DEFF Research Database (Denmark)

    Jensen, Peter K. M.; Phuc, Pham D.; Konradsen, Flemming

    2009-01-01

    this is the length of time that farmers have available to produce fertilizer between two cropping seasons. This study aimed to investigate whether hygienically safe fertilizer could be produced in the latrines within this period of time. Methods: By inoculating eggs of the helminth parasite indicator Ascaris suum...... from 9.4 to 11.6, a >99% die-off of eggs was obtained after 105 to 117 days of storage for all lime concentrations and 97% of eggs were nonviable after 88 days of storage. The most critical parameter found to determine the die-off process was the amount of ammonia (urine) in the excreta which indicates...... that longer storage periods are needed for parasite egg die-off if urine is separated from the excreta. Conclusion: By inactivating >99% of all A. suum eggs in human excreta during a storage period of only three months the commonly used Double Vault Composting (DVC) latrine, in which urine is not separated...

  10. The intimate genetics of Drosophila fertilization

    Science.gov (United States)

    Loppin, Benjamin; Dubruille, Raphaëlle; Horard, Béatrice

    2015-01-01

    The union of haploid gametes at fertilization initiates the formation of the diploid zygote in sexually reproducing animals. This founding event of embryogenesis includes several fascinating cellular and nuclear processes, such as sperm–egg cellular interactions, sperm chromatin remodelling, centrosome formation or pronuclear migration. In comparison with other aspects of development, the exploration of animal fertilization at the functional level has remained so far relatively limited, even in classical model organisms. Here, we have reviewed our current knowledge of fertilization in Drosophila melanogaster, with a special emphasis on the genes involved in the complex transformation of the fertilizing sperm nucleus into a replicated set of paternal chromosomes. PMID:26246493

  11. Oxygen requirements of separated hybrid catfish female Ictalurus punctatus male I. furcatus eggs

    Science.gov (United States)

    Channel catfish Ictalurus punctatus egg masses require ambient water with over 95% air saturation to maintain maximum oxygen consumption as they near hatch. Since hybrid catfish eggs (channel catfish ' X blue catfish I. furcatus ') are often kept separated after fertilization by the addition of full...

  12. Generation of eggs from mouse embryonic stem cells and induced pluripotent stem cells.

    Science.gov (United States)

    Hayashi, Katsuhiko; Saitou, Mitinori

    2013-08-01

    Oogenesis is an integrated process through which an egg acquires the potential for totipotency, a fundamental condition for creating new individuals. Reconstitution of oogenesis in a culture that generates eggs with proper function from pluripotent stem cells (PSCs) is therefore one of the key goals in basic biology as well as in reproductive medicine. Here we describe a stepwise protocol for the generation of eggs from mouse PSCs, such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). ESCs and iPSCs are first induced into primordial germ cell-like cells (PGCLCs) that are in turn aggregated with somatic cells of female embryonic gonads, the precursors for adult ovaries. Induction of PGCLCs followed by aggregation with the somatic cells takes up to 8 d. The aggregations are then transplanted under the ovarian bursa, in which PGCLCs grow into germinal vesicle (GV) oocytes in ∼1 month. The PGCLC-derived GV oocytes can be matured into eggs in 1 d by in vitro maturation (IVM), and they can be fertilized with spermatozoa by in vitro fertilization (IVF) to obtain healthy and fertile offspring. This method provides an initial step toward reconstitution of the entire process of oogenesis in vitro.

  13. Cortical mechanics and myosin-II abnormalities associated with post-ovulatory aging: implications for functional defects in aged eggs

    Science.gov (United States)

    Mackenzie, Amelia C.L.; Kyle, Diane D.; McGinnis, Lauren A.; Lee, Hyo J.; Aldana, Nathalia; Robinson, Douglas N.; Evans, Janice P.

    2016-01-01

    STUDY HYPOTHESIS Cellular aging of the egg following ovulation, also known as post-ovulatory aging, is associated with aberrant cortical mechanics and actomyosin cytoskeleton functions. STUDY FINDING Post-ovulatory aging is associated with dysfunction of non-muscle myosin-II, and pharmacologically induced myosin-II dysfunction produces some of the same deficiencies observed in aged eggs. WHAT IS KNOWN ALREADY Reproductive success is reduced with delayed fertilization and when copulation or insemination occurs at increased times after ovulation. Post-ovulatory aged eggs have several abnormalities in the plasma membrane and cortex, including reduced egg membrane receptivity to sperm, aberrant sperm-induced cortical remodeling and formation of fertilization cones at the site of sperm entry, and reduced ability to establish a membrane block to prevent polyspermic fertilization. STUDY DESIGN, SAMPLES/MATERIALS, METHODS Ovulated mouse eggs were collected at 21–22 h post-human chorionic gonadotrophin (hCG) (aged eggs) or at 13–14 h post-hCG (young eggs), or young eggs were treated with the myosin light chain kinase (MLCK) inhibitor ML-7, to test the hypothesis that disruption of myosin-II function could mimic some of the effects of post-ovulatory aging. Eggs were subjected to various analyses. Cytoskeletal proteins in eggs and parthenogenesis were assessed using fluorescence microscopy, with further analysis of cytoskeletal proteins in immunoblotting experiments. Cortical tension was measured through micropipette aspiration assays. Egg membrane receptivity to sperm was assessed in in vitro fertilization (IVF) assays. Membrane topography was examined by low-vacuum scanning electron microscopy (SEM). MAIN RESULTS AND THE ROLE OF CHANCE Aged eggs have decreased levels and abnormal localizations of phosphorylated myosin-II regulatory light chain (pMRLC; P = 0.0062). Cortical tension, which is mediated in part by myosin-II, is reduced in aged mouse eggs when compared with

  14. Cryopreservation of yamú (Brycon amazonicus) sperm for large scale fertilization

    DEFF Research Database (Denmark)

    Velasco-Santamaría, Yohana M.; Medina-Robles, Mauricio; Cruz-Casallas, Pablo E.

    2006-01-01

      To determine the effect of straw size and thawing temperature on cryopreserved sperm quality of yamú (Brycon amazonicus), ovulation and spermiation were induced in sexually mature broodstock using Carp Pituitary Extract. Sperm quality was evaluated by motility, activation time and fertility...... assays consisted of 40 g eggs inseminated with approximately 5.0 mL (ca. 75,000 motile spermatozoa/egg) of cryopreserved sperm in large straws thawed at 35 °C. The fertilization rate was estimated 6 h post-insemination. In all straws, postthaw motility was significantly lower than for fresh sperm (pb0.......05) to sperm frozen in 0.5-mL straws (48±2%, 51±2%, 52±2% and 54±3%, respectively). In large scale fertilization trials, fresh sperm showed a higher (pb0.05) fertilization rate (83±1%) than frozen-thawed sperm (68±1%). Although the fertility percentage with fresh sperm was significantly higher than with frozen...

  15. Cross-talk between free and bound spermatozoa to modulate initial sperm:egg ratios at the site of fertilization in the mammalian oviduct.

    Science.gov (United States)

    Hunter, R H F; Gadea, J

    2014-08-01

    This essay proposes that highly localized communication between free and bound spermatozoa in the caudal portion of the oviduct acts to regulate the numbers detaching from the epithelium and progressing to the site of fertilization close to the time of ovulation. Low initial sperm:egg ratios are essential for monospermic fertilization. Liberation of surface macromolecules and metabolic prompting from activated spermatozoa, together with altered patterns of sperm movement and dynamic differences in intracellular Ca(2+) ion status between neighboring sperm cells, would influence the progressive release of spermatozoa from the reservoir in the oviduct isthmus. Different intensities of preovulatory epithelial binding, reflecting a range of states in the sperm surface membranes and associated proteins, would provide a further explanation for a chronologically staggered periovulatory detachment of spermatozoa. Intimate sperm-sperm interactions within the confines of the oviduct isthmus offer a sensitive means of fine-tuning the vanguard of competent male gametes reaching the isthmo-ampullary junction. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Sperm depletion does not account for undeveloped eggs in blue tits Cyanistes caeruleus

    NARCIS (Netherlands)

    Schut, Elske; Vedder, Oscar; Komdeur, Jan; Magrath, Michael J. L.

    Hatching failure negatively impacts reproductive success in birds. One reason why eggs fail to hatch is that they are not fertilized, which may be because they receive insufficient sperm. In most passerines, copulation declines in frequency or ceases altogether after the laying of the first egg, so

  17. Downregulation of surface sodium pumps by endocytosis during meiotic maturation of Xenopus laevis oocytes

    International Nuclear Information System (INIS)

    Schmalzing, G.; Eckard, P.; Kroener, S.P.; Passow, H.

    1990-01-01

    During meiotic maturation, plasma membranes of Xenopus laevis oocytes completely lose the capacity to transport Na and K and to bind ouabain. To explore whether the downregulation might be due to an internalization of the sodium pump molecules, the intracellular binding of ouabain was determined. Selective permeabilization of the plasma membrane of mature oocytes (eggs) by digitonin almost failed to disclose ouabain binding sites. However, when the eggs were additionally treated with 0.02% sodium dodecyl sulfate (SDS) to permeabilize inner membranes, all sodium pumps present before maturation were recovered. Phosphorylation by [gamma-32P]ATP combined with SDS-polyacrylamide gel electrophoresis (PAGE) and autoradiography showed that sodium pumps were greatly reduced in isolated plasma membranes of eggs. According to sucrose gradient fractionation, maturation induced a shift of sodium pumps from the plasma membrane fraction to membranes of lower buoyant density with a protein composition different from that of the plasma membrane. Endocytosed sodium pumps identified on the sucrose gradient from [3H]ouabain bound to the cell surface before maturation could be phosphorylated with inorganic [32P]phosphate. The findings suggest that downregulation of sodium pumps during maturation is brought about by translocation of surface sodium pumps to an intracellular compartment, presumably endosomes. This contrasts the mechanism of downregulation of Na-dependent cotransport systems, the activities of which are reduced as a consequence of a maturation-induced depolarization of the membrane without a removal of the corresponding transporter from the plasma membrane

  18. Eleven generations of selection for the duration of fertility in the intergeneric crossbreeding of ducks

    Directory of Open Access Journals (Sweden)

    Liu Tai Jui-Jane

    2009-03-01

    Full Text Available Abstract A 12-generation selection experiment involving a selected line (S and a control line (C has been conducted since 1992 with the aim of increasing the number of fertile eggs laid by the Brown Tsaiya duck after a single artificial insemination (AI with pooled Muscovy semen. On average, 28.9% of the females and 17.05% of the males were selected. The selection responses and the predicted responses showed similar trends. The average predicted genetic responses per generation in genetic standard deviation units were 0.40 for the number of fertile eggs, 0.45 for the maximum duration of fertility, and 0.32 for the number of hatched mule ducklings' traits. The fertility rates for days 2–8 after AI were 89.14% in the S line and 61.46% in the C line. Embryo viability was not impaired by this selection. The largest increase in fertility rate per day after a single AI was observed from d5 to d11. In G12, the fertility rate in the selected line was 91% at d2, 94% at d3, 92% at days 3 and 4 then decreased to 81% at d8, 75% at d9, 58% at d10 and 42% at d11. In contrast, the fertility rate in the control line showed an abrupt decrease from d4 (74%. The same tendencies were observed for the evolution of hatchability according to the egg set rates. It was concluded that selection for the number of fertile eggs after a single AI with pooled Muscovy semen could effectively increase the duration of the fertile period in ducks and that research should now be focused on ways to improve the viability of the hybrid mule duck embryo.

  19. Diseases of amphibian eggs and embryos

    Science.gov (United States)

    Green, D.E.; Converse, K.A.; Majumdar, S.K.; Huffman, J.E.; Brenner, F.J.; Panah, A.I.

    2005-01-01

    Amphibians generally are prolific egg producers. In tropical and semi-tropical regions, deposition of eggs may occur year-round or may coincide with rainy seasons, while in temperate regions, deposition of eggs usually occurs immediately after emergence from hibernation. Numbers of eggs produced by each species may vary from a few dozen to thousands. Accordingly, some eggs may be infertile and wastage of embryos is to be expected. Fertility, viability and decomposition of eggs and embryos must be considered before it is assumed that diseases are present. An important consideration in the evaluation of egg masses is the fact that some will contain infertile and non-viable eggs. These infertile and nonviable eggs will undergo decomposition and they may appear similar to eggs that are infected by a pathogen. Evaluation of egg masses and embryos for the presence of disease may require repeated observations in a given breeding season as well as continued monitoring of egg masses during their growth and development and over successive breeding seasons. Amphibian eggs rarely are subjected to a comprehensive health (diagnostic) examination; hence, there is scant literature on the diseases of this life stage. Indeed, the eggs of some North American amphibians have yet to be described. Much basic physiology and normal biomedical baseline data on amphibian eggs is lacking. For example, it is known that the aquatic eggs of some species of shrimp quickly are coated by a protective and commensal bacterium that effectively impedes invasion of the eggs by other environmental organisms and potential pathogens. In the absence of this bacterium, shrimp eggs are rapidly killed by other bacteria and fungi (Green, 2001). The possibility that amphibian eggs also have important symbiotic or commensal bacteria needs to be investigated. Furthermore, the quantity and types of chemicals in the normal gelatinous capsules of amphibian eggs have scarcely been examined. Abnormalities of the

  20. Estimate the contribution of incubation parameters influence egg hatchability using multiple linear regression analysis.

    Science.gov (United States)

    Khalil, Mohamed H; Shebl, Mostafa K; Kosba, Mohamed A; El-Sabrout, Karim; Zaki, Nesma

    2016-08-01

    This research was conducted to determine the most affecting parameters on hatchability of indigenous and improved local chickens' eggs. Five parameters were studied (fertility, early and late embryonic mortalities, shape index, egg weight, and egg weight loss) on four strains, namely Fayoumi, Alexandria, Matrouh, and Montazah. Multiple linear regression was performed on the studied parameters to determine the most influencing one on hatchability. The results showed significant differences in commercial and scientific hatchability among strains. Alexandria strain has the highest significant commercial hatchability (80.70%). Regarding the studied strains, highly significant differences in hatching chick weight among strains were observed. Using multiple linear regression analysis, fertility made the greatest percent contribution (71.31%) to hatchability, and the lowest percent contributions were made by shape index and egg weight loss. A prediction of hatchability using multiple regression analysis could be a good tool to improve hatchability percentage in chickens.

  1. In p nduce protoc d spaw cols of wning hormo of Liza ones w egg a ...

    African Journals Online (AJOL)

    SAM

    environmental conditions and resistance to disease and stresses. Thin-lipped ... ship between egg quality and fatty acid content of eggs. Successful fertilization ...... mullet (Mugil cephalus L.) with fractional salmon pituitary extract. J. Fish Biol.

  2. Analysis of limited fertility in intracytoplasmic sperm injection of sperm obtained by electroejaculation

    Science.gov (United States)

    Nakamura, Yoshihiro; Kitamura, Masaya; Nishimura, Kenji; Tsujimura, Akira; Takeyama, Masami; Kondoh, Nobuyuki; Miyazaki, Kazunori; Okuyama, Akihiko

    2004-01-01

    Background and Aims:  We correlated findings in semen from patients with ejaculatory dysfunction with results of in vitro fertilization using their electroejaculated sperm. Methods and Results:  Electroejaculation was carried out in six patients with the above‐mentioned criteria for a total of eight times. Sperm was obtained in six attempts. Intracytoplasmic injection of these sperm was performed in 156 eggs. Sixty‐seven eggs were fertilized; most of these were injected with motile sperm. Two women became pregnant, both after injection with motile sperm. As previously reported, electroejaculated sperm showed low motility and a low fertilization rate, but even motile sperm had a low fertilization rate. Conclusion:  The results of the present study suggest the importance in fertilization of undetermined factors in addition to sperm motility. (Reprod Med Biol 2004; 3: 9–12) PMID:29662380

  3. Caracterização do microclima dos diferentes layouts de caixas no transporte de ovos férteis Characterization of microclimate in different layout of boxes during transport of fertile eggs

    Directory of Open Access Journals (Sweden)

    Aérica C. Nazareno

    2013-03-01

    Full Text Available Quando as condições microclimáticas e de transporte são combinadas inadequadamente, os componentes dos ovos férteis podem ser danificados. Objetivou-se, com este trabalho, caracterizar e analisar a dependência espacial dos perfis microclimáticos no transporte de ovos férteis em diferentes layouts de caixas. A pesquisa foi conduzida na empresa integradora avícola, em Mogi Mirim, SP, por meio do acompanhamento de cinco carregamentos. Utilizou-se um caminhão climatizado do tipo baú, com capacidade para 592 caixas de ovos e se avaliaram dois layouts da carga (caixas superiores e inferiores de 9 pilhas com seis caixas de ovos férteis. Um microprocessador foi instalado dentro das caixas de ovos, totalizando 18 microprocessadores. As variáveis meteorológicas analisadas foram: temperatura (ºC umidade relativa (% e entalpia específica (kJ kg-1 ar seco. O delineamento experimental adotado foi o inteiramente casualizado com parcelas subdividas e realizada a análise geoestatística (krigagem ordinária. No layout das caixas superiores foram registrados valores elevados de temperatura e baixos valores de umidade relativa e entalpia específica. As regiões de piores condições microclimáticas nos dois layouts se situaram nas partes central e traseira do caminhão, durante o transporte dos ovos férteis.The fertile egg components may be damaged when the microclimatic conditions and transportation characteristics are combined inadequately. Thus, the aim of this study was to characterize and to assess the spatial dependence of microclimatic profiles during transport of fertile eggs with different layout of boxes. A trial was conducted in a commercial poultry industry in Mogi Mirim in the State of São Paulo, Brazil, through monitoring five loads of hatching eggs. An air conditioned truck was used, with maximum capacity of 592 egg boxes. Two layout of boxes were assessed in the present study. One data logger was installed inside the egg boxes

  4. Indian egg donors’ characteristics, motivations and feelings towards the recipient and resultant child

    Directory of Open Access Journals (Sweden)

    V. Jadva

    2015-12-01

    Full Text Available This is the first study to examine characteristics, motivations and experiences of Indian egg donors. In-depth interviews were conducted with 25 egg donors who had donated during the previous 8 months at a fertility clinic in Mumbai. The semi-structured interviews were conducted in Hindi and English. In addition to demographic information, data were collected on donors’ motivations for donating, with whom they had discussed donation, and feelings towards the recipients. The response rate was 66%. All participants were literate and had attended school. Twenty (80% egg donors had children and five (20% did not. The most common motivation (19, 76% for donating was financial need. Egg donors had discussed their donation with their husband or with close family/friends, with almost all mentioning that wider society would disapprove. The majority (20, 80% had no information about the recipients and 11 (44% preferred not to. The findings highlight the similarities and differences between egg donors from India and those from other countries and that egg donors are of a more varied demographic background than surrogates in India. Given that India has been a popular destination for fertility treatment, the findings have important implications for regulation and practice within India and internationally.

  5. Indian egg donors' characteristics, motivations and feelings towards the recipient and resultant child.

    Science.gov (United States)

    Jadva, V; Lamba, N; Kadam, K; Golombok, S

    2015-12-01

    This is the first study to examine characteristics, motivations and experiences of Indian egg donors. In-depth interviews were conducted with 25 egg donors who had donated during the previous 8 months at a fertility clinic in Mumbai. The semi-structured interviews were conducted in Hindi and English. In addition to demographic information, data were collected on donors' motivations for donating, with whom they had discussed donation, and feelings towards the recipients. The response rate was 66%. All participants were literate and had attended school. Twenty (80%) egg donors had children and five (20%) did not. The most common motivation (19, 76%) for donating was financial need. Egg donors had discussed their donation with their husband or with close family/friends, with almost all mentioning that wider society would disapprove. The majority (20, 80%) had no information about the recipients and 11 (44%) preferred not to. The findings highlight the similarities and differences between egg donors from India and those from other countries and that egg donors are of a more varied demographic background than surrogates in India. Given that India has been a popular destination for fertility treatment, the findings have important implications for regulation and practice within India and internationally.

  6. Age- and size-dependent mating performance and fertility in a pelagic copepod, Temora longicornis

    DEFF Research Database (Denmark)

    Sichlau, Mie Hylstofte; Kiørboe, Thomas

    2011-01-01

    Prepress abstract: In many species, size and age have been shown to be strong determinants of the reproductive success for both sexes. Here we examine age- and size dependent reproductive performance (egg- and sperm production, mating success) in a pelagic copepod. Compared to smaller males, larger...... males produce larger spermatophores containing more spermatozoa, and fertilize a larger fraction of available females. Females mating with large males produce more offspring than those mating with small males. Similarly, large females have higher egg production rates as well as a higher life-time egg...... fertilize females for only about eight days after they mature. The strong size- and age-dependent fertility observed in this species is conducive to the development of sexual selection via mate choice for young and large partners, as has been shown in one other copepod species...

  7. Amphibian fertilization and development in microgravity

    Science.gov (United States)

    Souza, Kenneth A.

    1993-01-01

    During the year before launch, female frogs will be tested every 3 months for the quantity and quality of eggs produced. Two weeks or more prior to launch, male and female frogs will be transported to the John F. Kennedy Space Center (KSC). During the few weeks before launch, groups will be periodically tested for egg quality to assure that the frogs have adapted to the KSC laboratory environment. About 27 hours before launch, four females will be placed in a damp foam-lined box, called the Adult Frog Box (AFB), through which 100 cc/min of air wil be circulated. The AFB will be lowered into the Spacelab and loaded into the Frog Environmental Unit (FEU) during the final pre-launch preparations. A sperm suspension, for use in flight to fertilize the eggs, will also be prepared and loaded during the pre-launch period. The sperm suspension, together with a kit of syringes containing Human Chorionic Gonadotropin (HCG), will be stored in a refrigerator aboard the shuttle until needed in flight. On the first day of flight, the AFB will be transferred from the FEU to the General Purpose Work Station (GPWS), which is a type of glove box specially designed to allow the crew to use chemicals and biological materials during the flight without contaminating the shuttle/Spacelab environment. Inside the GPWS the four adult frogs will be injected with the HCG hormone and returned to the FEU. Approximately 16 hours after injection, ovulation should have taken place and 15 to 20 eggs from each frog will be placed on each of two egg baskets and covered with sperm for 10 minutes. The egg baskets are inserted into acrylic egg chambers and 50 ml of 'pond water' (20 percent strength Modified Ringers solution (is added. One of the chambers from each frog will be placed on a centrifuge within the FEU and rotated to simulate normal terrestrial gravity (1 g). The remaining chambers are incubated under microgravity conditions within the FEU. Forty minutes after fertilization, the four

  8. X-ray- and mitomycin C (MMC)-induced chromosome aberrations in spermiogenic germ cells and the repair capacity of mouse eggs for the X-ray and MMC damage

    International Nuclear Information System (INIS)

    Matsuda, Yoichi; Utsugi-Takeuchi, Toyoko; Tobari, Izuo; Seki, Naohiko; Chiba Univ.

    1989-01-01

    Chromosome aberrations induced at the first-cleavage metaphase of eggs fertilized with sperm recovered from spermiogenic cells which had been X-irradiated and treated with mitomycin C (MMC) at various stages were observed using in vitro fertilization and embryo culture technique. Furthermore, the repair capacity of the fertilized eggs for X-ray- and MMC-induced DNA damage which was induced in the spermiogenic cells and retained in the sperm until fertilization was investigated by analysis of the potentiation effects of 2 repair inhibitors, 3-aminobenzamide (3AB) and caffeine on the yield of chromosome aberrations. The frequency of chromosome aberrations observed in the eggs fertilized with sperm recovered from speratozoa to late spermatid stage (0-8 days after X-irradiation). The induced chromosome aberrations followed by chromosome exchange through all the spermiogenic stages. The results suggest that the large amount of DNA lesions induced in spermiogenic cells by X-rays and MMC persist as reparable damage until sperm maturation and are effectively repaired in the cytoplasm of the fertilized eggs. (author). 29 refs.; 2 figs.; 2 tabs

  9. The beginning of a seed: regulatory mechanisms of double fertilization.

    Science.gov (United States)

    Bleckmann, Andrea; Alter, Svenja; Dresselhaus, Thomas

    2014-01-01

    THE LAUNCH OF SEED DEVELOPMENT IN FLOWERING PLANTS (ANGIOSPERMS) IS INITIATED BY THE PROCESS OF DOUBLE FERTILIZATION: two male gametes (sperm cells) fuse with two female gametes (egg and central cell) to form the precursor cells of the two major seed components, the embryo and endosperm, respectively. The immobile sperm cells are delivered by the pollen tube toward the ovule harboring the female gametophyte by species-specific pollen tube guidance and attraction mechanisms. After pollen tube burst inside the female gametophyte, the two sperm cells fuse with the egg and central cell initiating seed development. The fertilized central cell forms the endosperm while the fertilized egg cell, the zygote, will form the actual embryo and suspensor. The latter structure connects the embryo with the sporophytic maternal tissues of the developing seed. The underlying mechanisms of double fertilization are tightly regulated to ensure delivery of functional sperm cells and the formation of both, a functional zygote and endosperm. In this review we will discuss the current state of knowledge about the processes of directed pollen tube growth and its communication with the synergid cells resulting in pollen tube burst, the interaction of the four gametes leading to cell fusion and finally discuss mechanisms how flowering plants prevent multiple sperm cell entry (polyspermy) to maximize their reproductive success.

  10. Naturally Occurring Egg Drop Syndrome Infection in Turkeys

    Directory of Open Access Journals (Sweden)

    Z. Biđin

    2007-01-01

    Full Text Available A decrease in the egg quality, production, fertility and hatchability without serious clinical signs of illness was recorded in turkey fl ocks in Croatia at the beginning of 2002. It was assumed that the egg drop syndrome virus might be one of the etiological agents responsible for the abnormalities in the egg production. The systematic serological monitoring, using a haemagglutination inhibition test, showed that the antibodies to the egg drop syndrome virus existed in 94.4 and 55.1% of the sera analysed in 2002 and 2003, respectively. The haemagglutination inhibition titres ranged from 16 to 128. The sera samples were randomly collected from 11 - to 46-week-old hens from the affected fl ocks. The serological evidence of the egg drop syndrome virus infection was confirmed by detection of the presence of the virus genome in the turkey sera by the polymerase chain reaction. Vaccination of the 18- and 25-week-old turkey hens against the egg drop syndrome virus started in March 2003. After this period, the presence of antibodies to the egg drop syndrome virus (the haemagglutination inhibition titres between 16 and 256 was found in 96.7% of the analysed sera, while the egg production reached normal or higher values for the Nicholas hybrid line of turkeys.

  11. Effects of host genetics and environment on egg-associated microbiotas in brown trout (Salmo trutta).

    Science.gov (United States)

    Wilkins, Laetitia G E; Fumagalli, Luca; Wedekind, Claus

    2016-10-01

    Recent studies found fish egg-specific bacterial communities that changed over the course of embryogenesis, suggesting an interaction between the developing host and its microbiota. Indeed, single-strain infections demonstrated that the virulence of opportunistic bacteria is influenced by environmental factors and host immune genes. However, the interplay between a fish embryo host and its microbiota has not been studied yet at the community level. To test whether host genetics affects the assemblage of egg-associated bacteria, adult brown trout (Salmo trutta) were sampled from a natural population. Their gametes were used for full-factorial in vitro fertilizations to separate sire from dam effects. In total, 2520 embryos were singly raised under experimental conditions that differently support microbial growth. High-throughput 16S rRNA amplicon sequencing was applied to characterize bacterial communities on milt and fertilized eggs across treatments. Dam and sire identity influenced embryo mortality, time until hatching and composition of egg-associated microbiotas, but no link between bacterial communities on milt and on fertilized eggs could be found. Elevated resources increased embryo mortality and modified bacterial communities with a shift in their putative functional potential. Resource availability did not significantly affect any parental effects on embryo performance. Sire identity affected bacterial diversity that turned out to be a significant predictor of hatching time: embryos associated with high bacterial diversity hatched later. We conclude that both host genetics and the availability of resources define diversity and composition of egg-associated bacterial communities that then affect the life history of their hosts. © 2016 John Wiley & Sons Ltd.

  12. Standardization of fertilization protocols for the European eel, Anguilla anguilla

    DEFF Research Database (Denmark)

    Butts, Ian; Sørensen, Sune Riis; Politis, Sebastian Nikitas

    2014-01-01

    of this research were to provide a rapid, accurate and precise method to quantify sperm density by examining the relationship between sperm density and absorbance by use of a spectrophotometer, determine the optimal number of sperm required to fertilize eggs in a controlled setting, and explore how long eggs...... are receptive to fertilization post-stripping. Mean sperm density and absorbance at 350nm were 1.54e+10±4.95e+9sperm/mL and 1.91±0.22nm, respectively. Regression analysis demonstrated a highly significant positive relationship between sperm density and absorbance using a spectrophotometer at 350nm (R2=0.94, p

  13. Xenopus: An Emerging Model for Studying Congenital Heart Disease

    Science.gov (United States)

    Kaltenbrun, Erin; Tandon, Panna; Amin, Nirav M.; Waldron, Lauren; Showell, Chris; Conlon, Frank L.

    2011-01-01

    Congenital heart defects affect nearly 1% of all newborns and are a significant cause of infant death. Clinical studies have identified a number of congenital heart syndromes associated with mutations in genes that are involved in the complex process of cardiogenesis. The African clawed frog, Xenopus, has been instrumental in studies of vertebrate heart development and provides a valuable tool to investigate the molecular mechanisms underlying human congenital heart diseases. In this review, we discuss the methodologies that make Xenopus an ideal model system to investigate heart development and disease. We also outline congenital heart conditions linked to cardiac genes that have been well-studied in Xenopus and describe some emerging technologies that will further aid in the study of these complex syndromes. PMID:21538812

  14. EGG QUALITY AND HATCHABILITY OF In situ - REARED KEDU AND CEMANI HENS FED DIET OF FARMER FORMULATION SUPPLEMENTED WITH VITAMIN E

    Directory of Open Access Journals (Sweden)

    H.I. Wahyuni

    2014-10-01

    Full Text Available The aim of study was to evaluate the effect of dietary vitamin E supplementation in Kedu andCemani hens reraed in situ and given farmer-formulated diet on egg performances. A total of 120 femaleand 24 male birds were equally divided into two groups of Kedu and Cemani, with average body weightof 1890+216.79 and 1830+396.23 g, respectively. Basal ration was based on the diet formulated by thefarmer (R1 consisting of corn (30%, rice bran (50%, protein concentrate (15%, and premix (5%.Dietary treatments evaluated were R1 (without additional vitamin E, and the other three were theinclusion of DL α-tocopheryl acetate into the R1 diet up to 2 (R2, 4 (R3 and 6 IU (R4. The experimentwas arranged in a Randomized Block Design with 4 treatments, and 2 different groups of hen (Kedu andCemani were assigned as block. Parameters observed were feed consumption, hen day production, feedconversion ratio, egg quality, fertility, and hatchability. The result showed that vitamin Esupplementation did not affect whatever parameters, except egg fertility. Egg hatchability between Keduand Cemani hens was significantly different (p<0.05. Trend comparison test indicated significantlylinear (p<0.05 for fertility and hatchability. In conclusion, vitamin E supplementation up to 6 IUincreased linearly egg fertility and hatchability of about 5-8%, and egg hatchability of Kedu hens washigher (11% than that of Cemani.

  15. The unexpected teratogenicity of RXR antagonist UVI3003 via activation of PPARγ in Xenopus tropicalis

    International Nuclear Information System (INIS)

    Zhu, Jingmin; Janesick, Amanda; Wu, Lijiao; Hu, Lingling; Tang, Weiyi; Blumberg, Bruce; Shi, Huahong

    2017-01-01

    The RXR agonist (triphenyltin, TPT) and the RXR antagonist (UVI3003) both show teratogenicity and, unexpectedly, induce similar malformations in Xenopus tropicalis embryos. In the present study, we exposed X. tropicalis embryos to UVI3003 in seven specific developmental windows and identified changes in gene expression. We further measured the ability of UVI3003 to activate Xenopus RXRα (xRXRα) and PPARγ (xPPARγ) in vitro and in vivo. We found that UVI3003 activated xPPARγ either in Cos7 cells (in vitro) or Xenopus embryos (in vivo). UVI3003 did not significantly activate human or mouse PPARγ in vitro; therefore, the activation of Xenopus PPARγ by UVI3003 is novel. The ability of UVI3003 to activate xPPARγ explains why UVI3003 and TPT yield similar phenotypes in Xenopus embryos. Our results indicate that activating PPARγ leads to teratogenic effects in Xenopus embryos. More generally, we infer that chemicals known to specifically modulate mammalian nuclear hormone receptors cannot be assumed to have the same activity in non-mammalian species, such as Xenopus. Rather they must be tested for activity and specificity on receptors of the species in question to avoid making inappropriate conclusions. - Highlights: • UVI3003 is a RXRs antagonist and shows teratogenicity to Xenopus embryos. • UVI3003 activated xPPARγ either in Cos7 cells or Xenopus embryos. • UVI3003 did not activate human or mouse PPARγ in Cos7 cells. • Activating PPARγ leads to teratogenic effects in Xenopus embryos.

  16. Localisation of RNAs into the germ plasm of vitellogenic Xenopus oocytes.

    Directory of Open Access Journals (Sweden)

    Sarbjit Nijjar

    Full Text Available We have studied the localisation of mRNAs in full-grown Xenopus laevis oocytes by injecting fluorescent RNAs, followed by confocal microscopy of the oocyte cortex. Concentrating on RNA encoding the Xenopus Nanos homologue, nanos1 (formerly Xcat2, we find that it consistently localised into aggregated germ plasm ribonucleoprotein (RNP particles, independently of cytoskeletal integrity. This implies that a diffusion/entrapment-mediated mechanism is active, as previously reported for previtellogenic oocytes. Sometimes this was accompanied by localisation into scattered particles of the "late", Vg1/VegT pathway; occasionally only late pathway localisation was seen. The Xpat RNA behaved in an identical fashion and for neither RNA was the localisation changed by any culture conditions tested. The identity of the labelled RNP aggregates as definitive germ plasm was confirmed by their inclusion of abundant mitochondria and co-localisation with the germ plasm protein Hermes. Further, the nanos1/Hermes RNP particles are interspersed with those containing the germ plasm protein Xpat. These aggregates may be followed into the germ plasm of unfertilized eggs, but with a notable reduction in its quantity, both in terms of injected molecules and endogenous structures. Our results conflict with previous reports that there is no RNA localisation in large oocytes, and that during mid-oogenesis even germ plasm RNAs localise exclusively by the late pathway. We find that in mid oogenesis nanos1 RNA also localises to germ plasm but also by the late pathway. Late pathway RNAs, Vg1 and VegT, also may localise into germ plasm. Our results support the view that mechanistically the two modes of localisation are extremely similar, and that in an injection experiment RNAs might utilise either pathway, the distinction in fates being very subtle and subject to variation. We discuss these results in relation to their biological significance and the results of others.

  17. An evaluation of soybean lecithin as an alternative to avian egg yolk in the cryopreservation of fish sperm.

    Science.gov (United States)

    Yildiz, Cengiz; Bozkurt, Yusuf; Yavas, Ilker

    2013-08-01

    Plant-derived lecithin has been used as a more sanitary alternative to avian egg yolk in livestock sperm cryopreservation protocols but its efficacy for cryopreserving fish sperm has not previously been tested comparatively. Here various concentrations of soybean lecithin were evaluated for the cryopreservation of carp (Cyprinus carpio) sperm. Sexually mature fish were induced to spermiation and ovulation with ovopel. The extenders were prepared by using 300 mM glucose, 10% DMSO, supplemented with different ratios of lecithin (5%, 10%, 15%, and 20%) and 10% egg yolk (control I). Negative control was made without egg yolk and soybean lecithin (control II). The pooled semen was diluted separately at ratio of 1:3 (v/v) by using egg yolk and soybean-based extenders. Diluted semen placed into 0.25 ml straws were equilibrated at 4 °C for 15 min and frozen in liquid nitrogen vapor. Fertilization was conducted using a ratio of 1 × 10(5)spermatozoa/egg. Supplementation of 10% lecithin to extender showed the best cryoprotective effect for sperm motility and duration of motility against freezing damage compared to 15%, 20% and control II groups (plecithin provided a greater result in terms of fertilization success when compared to extenders containing 20% lecithin or control II (plecithin has a similar cryoprotective actions with conventional egg yolk-based extender against freezing damages and fertilization. Therefore, soybean lecithin is a suitable alternative to avian egg yolk for the cryopreservation of fish sperm. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. [Controlling effect of antagonist bioorganic fertilizer on tomato root-knot nematode].

    Science.gov (United States)

    Zhu, Zhen; Chen, Fang; Xiao, Tong-jian; Wang, Xiao-hui; Ran, Wei; Yang, Xing-ming; Shen, Qi-rong

    2011-04-01

    Indoor in vitro culture experiment and greenhouse pot experiment were conducted to evaluate the capabilities of three bacterial strains XZ-173 (Bacillus amyloliquefaciens), SL-25 (B. gibsonii), and KS-62 (Paenibacillus polymyxa) that can hydrolyze collagen protein in controlling tomato root-knot nematode. In the in vitro culture experiment, suspensions of XZ-173, SL-25, and KS-62 induced a mortality rate of 75.9%, 66.7%, and 50.0% to the second-stage junior nematode within 24 h, and decreased the egg hatching rate to 17.8%, 28.9% and 37.6% after 7-day incubation, respectively, in contrast to the 17.4% mortality rate and 53.6% egg hatching rate in the control (sterilized water). In the greenhouse pot experiment, the bioorganic fertilizer mixed with equal parts of fermented XZ-173, SL-25, and KS-62 gained the best result, with the root-knot nematode population in rhizosphere soil decreased by 84.0% as compared with the control. The bioorganic fertilizer also decreased the numbers of galls and eggs on tomato roots significantly, and increased the underground and aboveground biomass of tomato. Therefore, antagonist bioorganic fertilizer has promising potential in controlling root-knot nematode.

  19. Using fertile couples as embryo donors: An ethical dilemma.

    Science.gov (United States)

    Alizadeh, Leila; Omani Samani, Reza

    2014-03-01

    The use of donated embryos has offered hope for infertile couples who have no other means to have children. In Iran, fertility centers use fertile couples as embryo donors. In this paper, the advantages and disadvantages of this procedure will be discussed. We conclude that embryo-donation should be performed with frozen embryos thus preventing healthy donors from being harmed by fertility drugs. There must be guidelines for choosing the appropriate donor families. In countries where commercial egg donation is acceptable, fertile couples can be procured as embryo donors thus fulfilling the possible shortage of good quality embryos. Using frozen embryos seems to have less ethical, religious and legal problems when compared to the use of fertile embryo donors.

  20. The Effects of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD on the Mortality and Growth of Two Amphibian Species (Xenopus laevis and Pseudacris triseriata

    Directory of Open Access Journals (Sweden)

    Alex Collier

    2008-12-01

    Full Text Available We observed a slight drop in the growth of Xenopus laevis and Pseudacris triseriata larvae following acute exposure (24-48 h during egg development to three concentrations of TCDD (0.3, 3.0, 30.0 μg/l. Our exposure protocol was modeled on a previous investigation that was designed to mimic the effects of maternal deposition of TCDD. The doses selected were consistent with known rates of maternal transfer between mother and egg using actual adult body burdens from contaminated habitats. Egg and embryonic mortality immediately following exposure increased only among 48 h X. laevis treatments. Control P. triseriata and X. laevis completed metamorphosis more quickly than TCDDtreated animals. The snout-vent length of recently transformed P. triseriata did not differ between treatments although controls were heavier than high-dosed animals. Likewise, the snout-vent length and weight of transformed X. laevis did not differ between control and TCDD treatments. These findings provide additional evidence that amphibians, including P. triseriata and X. laevis are relatively insensitive to acute exposure to TCDD during egg and embryonic development. Although the concentrations selected for this study were relatively high, they were not inconsistent with our current understanding of bioaccumulation via maternal transfer.

  1. Egg quality traits and predictors of embryo and fry viability in red snapper Lutjanus campechanus

    Directory of Open Access Journals (Sweden)

    Agnès Bardon-Albaret

    2017-08-01

    Full Text Available The quality of red snapper eggs is highly variable and unpredictable in aquaculture, leading to high mortality during early larval rearing. In this work, the viability of red snapper eggs was monitored from fertilization until unfed larvae expired because of exhaustion of vitelline reserves to determine egg quality traits in this species. The spawns were obtained via strip spawning wild-caught females following hormonal induction with chorionic gonadotropin. Females were induced immediately after capture (wild group, n = 17 or held captive for the entire maturation period prior to induction (captive group, n = 7. Candidate predictors of egg quality measured on the female parent at the time of induction or on the spawn at ovulation were evaluated using correlation and multiple regression analysis. The fertilization rate, the hatching rate, and the duration of survival of unfed larvae post hatch were weakly correlated to each other (-0.23 < r < −0.08, revealing occurrence of distinct and independent components of egg quality. Spawns from captive females were characterized by a longer latency interval between hormonal induction and ovulation, lower fecundity, and lower hatching rates, as compared to those from wild females. Among the wild brood fish, a positive correlation was observed between the age of the female and the hatching rate. The best model optimized during stepwise multiple regression analysis of hatching rate data only explained 34% of the variance for this trait and no model could be optimized for the prediction of fertilization rate or the duration of survival post hatch. These results highlight the need to develop alternative egg quality measures to predict the viability of fry with confidence.

  2. Sperm swimming velocity predicts competitive fertilization success in the green swordtail Xiphophorus helleri.

    Directory of Open Access Journals (Sweden)

    Clelia Gasparini

    Full Text Available Sperm competition is expected to favour the evolution of traits that influence the performance of sperm when they compete to fertilize a female's eggs. While there is considerable evidence that selection favours increases in sperm numbers, much less is known about how sperm quality contributes towards competitive fertilization success. Here, we determine whether variation in sperm quality influences competitive fertilization success in the green swordtail Xiphophorus helleri, a highly promiscuous livebearing fish. We use artificial insemination as a method of controlled sperm delivery and show that sperm swimming velocity is the primary determinant of fertilization success when ejaculates from two males compete to fertilize a female's eggs. By contrast, we found no evidence that sperm length had any effect on siring success. We also found no evidence that pre- and postcopulatory sexual traits were phenotypically integrated in this species, suggesting that the previous observation that reproductive skew favours males with high mating rates is unlikely to be due to any direct association between sperm quality and male sexual ornamentation.

  3. Evolution of egg coats: linking molecular biology and ecology.

    Science.gov (United States)

    Shu, Longfei; Suter, Marc J-F; Räsänen, Katja

    2015-08-01

    One central goal of evolutionary biology is to explain how biological diversity emerges and is maintained in nature. Given the complexity of the phenotype and the multifaceted nature of inheritance, modern evolutionary ecological studies rely heavily on the use of molecular tools. Here, we show how molecular tools help to gain insight into the role of egg coats (i.e. the extracellular structures surrounding eggs and embryos) in evolutionary diversification. Egg coats are maternally derived structures that have many biological functions from mediating fertilization to protecting the embryo from environmental hazards. They show great molecular, structural and functional diversity across species, but intraspecific variability and the role of ecology in egg coat evolution have largely been overlooked. Given that much of the variation that influences egg coat function is ultimately determined by their molecular phenotype, cutting-edge molecular tools (e.g. proteomics, glycomics and transcriptomics), combined with functional assays, are needed for rigorous inferences on their evolutionary ecology. Here, we identify key research areas and highlight emerging molecular techniques that can increase our understanding of the role of egg coats in the evolution of biological diversity, from adaptation to speciation. © 2015 John Wiley & Sons Ltd.

  4. Utilization of laserpuncture induction as spawning stimulation in catfish (Clarias spp. crossbreeding toward egg quality

    Directory of Open Access Journals (Sweden)

    Pungky S.W. Kusuma

    2015-12-01

    Full Text Available The induction of laserpuncture on the reproductive acupoint of catfish can accelerate gonadotropin hormone formation from the pituitary especially gonadotropin II (GTH-II which has a role in the final stage of oocyte maturation, ovulation and spawning stimulation. The purpose of this study is to evaluate the effects of laserpuncture induction toward the egg quality from crossbreeding catfish male var. Paiton and female var. Sangkuriang. The egg quality was measured by the following parameters: fertilization rate (FR, egg hatching rate (HR, and larvae survival rate (SR. The research treatments were conducted using two levels along with eight repetitions. The results show that the crossbreed catfish using laserpuncture induction affected the parameters by increasing the mean value of fertilization rate, egg hatching rate and larvae survival rate significantly (P < 0.05 compared with mean value of fish without induction. This study concluded that laserpuncture induction on the crossbreeding between broodstock of male catfish var. Paiton and female var. Sangkuriang will increase FR, HR and SR.

  5. Comparing sugar type supplementation for cryopreservation of boar semen in egg yolk based extender.

    Science.gov (United States)

    Malo, C; Gil, L; Gonzalez, N; Cano, R; de Blas, I; Espinosa, E

    2010-08-01

    Cryopreservation of boar semen is still considered suboptimal due to lower fertility when compared to fresh semen. The aim of this study was to evaluate the effects of the addition of different sugars (lactose, trehalose and glucose) on boar spermatozoa cryopreserved in an egg yolk based extender. Ejaculates were collected from a boar previously selected and semen samples were processed using the straw freezing procedure. In experiment 1, subsamples of semen were frozen in three different extenders: recommended lactose egg yolk extender (LEY); trehalose egg yolk extender (TEY) and glucose egg yolk extender (GEY). Sperm quality was assessed for motility, viability, acrosome integrity and hypoosmotic swelling test response upon collection, after freezing and thawing and then every hour for 3h. Results showed that total motility at 1 and 3h, progressive motility at 3h, positive hypoosmotic response at 2 and 3h and acrosome integrity at all times were significantly improved when trehalose was added to the extender. In experiment 2, sugar influence was also demonstrated in vitro fertilization. A total of 1691 oocytes were in vitro matured and inseminated with frozen-thawed sperm at 2000:1 sperm:oocyte ratio and coincubated for 6h. Presumptive zygotes were cultured in NCSU-23 medium to assess fertilization parameters and embryo development. Both penetration and monospermy rates were significantly higher for trehalose frozen semen. A significant increase was observed in efficiency and blastocyst formation rates from TEY to the other groups. Our results demonstrated that trehalose extender enhances spermatozoa viability and its in vitro fertilization parameters in boar ejaculates with good sperm freezability. Further studies are necessary to assess the impact of sugars on the entire population. (c) 2010 Elsevier Inc. All rights reserved.

  6. “It’s a Trash”: Poetic Responses to the Experiences of a Mexican Egg Donor

    Directory of Open Access Journals (Sweden)

    Heather L. Walmsley

    2017-03-01

    Full Text Available This paper explores the use of found poetry as interpretive and aesthetic inquiry into the meaning and experience of reproductive tourism. The context is an ethnographic study of transnational egg donation, focusing upon the fertility services industry in Cancun, Mexico. Our source is an audio-recorded interview conducted with Maria, a young Mexican woman who struggles to maintain her integrity as a single mother donating eggs to a fertility clinic. Drawing upon Maria’s story, we experiment with three forms of found poetry as a method for listening deeply to her voice. In this paper, we share our research process, poems, and reflections.

  7. Effects of inverting the position of layers eggs during storage on hatchery performance parameters

    Directory of Open Access Journals (Sweden)

    JCS de Lima

    2012-12-01

    Full Text Available Storing hatchable eggs is a common practice in commercial hatcheries. However, storage time may negative effects on several performance parameters. An experiment was carried out to evaluate inverting egg position during storage of eggs laid by young and old layer breeders. Fertile eggs of 32 and 58-week-old breeders were stored for seven, 14, and 21 days at 18ºC ± 2ºC and 80% ± 10% relative humidity (RU. The following parameters were evaluated: egg weight loss, hatchability and hatchling weight, and embryodiagnosis results. Eggs stored with the small end up lost less weight during storage compared with the control eggs. Storing eggs for 14 days with the small end up reduced early embryo mortality, improving hatchability. In addition, hatchling weight increased. These results show that the detrimental effects of long storage periods may be alleviated when eggs are stored with the small end up to 14 days of storage.

  8. Influences of DMP on the fertilization process and subsequent embryogenesis of abalone (Haliotis diversicolor supertexta by gametes exposure.

    Directory of Open Access Journals (Sweden)

    Jin Zhou

    Full Text Available Di-methyl phthalate (DMP, a typical endocrine disrupting chemical (EDC, is ubiquitously distributed in aquatic environments; yet studies regarding its impact on gametes and the resulting effects on embryogenesis in marine gastropods are relatively scarce. In this study, the influences of DMP on the gametes and subsequent developmental process of abalone (Haliotis diversicolor supertexta, a representative marine benthic gastropod were assessed. Newborn abalone eggs and sperm were exposed separately to different DMP concentrations (1, 10 or 100 ppb for 60 min. At the end-point of exposure, the DMP-treated eggs and sperm were collected for analysis of their ultra-structures, ATPase activities and total lipid levels, and the fertilized gametes (embryos were collected to monitor related reproductive parameters (fertilization rate, abnormal development rate and hatching success rate. Treatment with DMP did not significantly alter the structure or total lipid content of eggs at any of the doses tested. Hatching failures and morphological abnormalities were only observed with the highest dose of DMP (100 ppb. However, DMP exposure did suppress sperm ATPase activities and affect the morphological character of their mitochondria. DMP-treated sperm exhibited dose-dependent decreases in fertilization efficiency, morphogenesis and hatchability. Relatively obvious toxicological effects were observed when both sperm and eggs were exposed to DMP. Furthermore, RT-PCR results indicate that treatment of gametes with DMP changed the expression patterns of physiologically-regulated genes (cyp3a, 17β-HSD-11 and 17β-HSD-12 in subsequent embryogenesis. Taken together, this study proofed that pre-fertilization exposure of abalone eggs, sperm or both to DMP adversely affects the fertilization process and subsequent embryogenesis.

  9. EGG YOLK AND LDL: POSSIBILITIES FOR ARTIFICIAL INSEMINATION IN EQUINES

    Directory of Open Access Journals (Sweden)

    Igor F. Canisso

    2008-12-01

    Full Text Available The world horse industry exerts an important role as a job and income generation source. Reproductive technologies arises as an important tool in the service of world equine growth. Artificial insemination (AI is perhaps the biotechnology with greater impact on equine breeding; a stallion can leave hundreds of offsprings over his reproductive life if AI is efficiently used. In some countries, egg yolk is frequently used as part of equine seminal extenders. The egg yolk provides the spermatozoa “resistance factors’’ when it is added. The protective fraction of the egg yolk probably is the low density lipoproteins (LDL. Several studies have reported successful results with the addition and replacement of egg yolk by LDL. There are many citations about the use of egg yolk in seminal extenders for stallion’s cooled and frozen semen, and in the equine reproduction practice. The egg yolk dilutors are used with good fertility results. New research is needed for the better understanding of the protective effects of egg yolk and the LDL for stallion semen. The LDL would be a great solution for dilutors to artificial insemination in horse. This review discusses the use and the advantages of egg yolk and LDL as constituents of equine semen extenders.

  10. Ocean acidification hampers sperm-egg collisions, gamete fusion, and generation of Ca2+ oscillations of a broadcast spawning bivalve, Tegillarca granosa.

    Science.gov (United States)

    Shi, Wei; Han, Yu; Guo, Cheng; Zhao, Xinguo; Liu, Saixi; Su, Wenhao; Wang, Yichen; Zha, Shanjie; Chai, Xueliang; Liu, Guangxu

    2017-09-01

    Although the effect of ocean acidification on fertilization success of marine organisms is increasingly well documented, the underlying mechanisms are not completely understood. The fertilization success of broadcast spawning invertebrates depends on successful sperm-egg collisions, gamete fusion, and standard generation of Ca 2+ oscillations. Therefore, the realistic effects of future ocean pCO 2 levels on these specific aspects of fertilization of Tegillarca granosa were investigated in the present study through sperm velocity trials, fertilization kinetics model analysis, and intracellular Ca 2+ assays, respectively. Results obtained indicated that ocean acidification significantly reduced the fertilization success of T. granosa, which could be accountable by (i) decreased sperm velocity hence reducing the probability for sperm-egg collisions; (ii) lowered probability of gamete fusion for each gamete collision event; and (iii) disrupted intracellular Ca 2+ oscillations. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. The morphology and attachment of Protopolystoma xenopodis (Monogenea: Polystomatidae infecting the African clawed frog Xenopus laevis

    Directory of Open Access Journals (Sweden)

    Theunissen Maxine

    2014-01-01

    Full Text Available The African clawed frog Xenopus laevis (Anura: Pipidae is host to more than 25 parasite genera encompassing most of the parasitic invertebrate groups. Protopolystoma xenopodis Price, 1943 (Monogenea: Polystomatidae is one of two monogeneans infecting X. laevis. This study focussed on the external morphology of different developmental stages using scanning electron microscopy, histology and light microscopy. Eggs are released continuously and are washed out when the frog urinates. After successful development, an active swimming oncomiracidium leaves the egg capsule and locates a potential post-metamorphic clawed frog. The oncomiracidium migrates to the kidney where it attaches and starts to feed on blood. The parasite then migrates to the urinary bladder where it reaches maturity. Eggs are fusiform, about 300 μm long, with a smooth surface and are operculated. Oncomiracidia are elongated and cylindrical in shape, with an oval posterior cup-shaped haptor that bears a total of 20 sclerites; 16 marginal hooklets used for attachment to the kidney of the host and two pairs of hamulus primordia. Cilia from the 64 ciliated cells enable the oncomiracidium to swim for up to 24 h when the cilia subsequently curl up, become non-functional and are shed from the body. The tegument between the ciliated cells bears a series of sensory papillae. The body of the mature parasite is elongated and pyriform and possesses an opisthaptor armed with three pairs of suckers and two pairs of falciform hooks to ensure a firm grip on the flexible internal surface of the urinary bladder.

  12. Relationship between cell cycle stage in the fertilized egg of mice and repair capacity for X-ray-induced damage in the sperm

    International Nuclear Information System (INIS)

    Matsuda, Y.; Maemori, M.; Tobari, I.

    1989-01-01

    The potentiation effects of 3-aminobenzamide, caffeine, hydroxyurea and arabinofuranosyl cytosine on the yield of X-ray-induced chromosome aberrations of mouse sperm were examined at the first-cleavage metaphase, to clarify a correlation between chromosome aberrations and cell cycle dependency of repair capacity of the fertilized egg. The result provided evidence that there are two major types of DNA damage in X-irradiated sperm: (1) short-lived DNA lesions; the lesions are subject to repair inhibitions by agents added in G 1 and are converted into chromosome-type aberrations during G 1 , and (2) long-lived DNA lesions; the lesions persist until S phase and repair of the lesions is inhibited by caffeine, hydroxyurea and arabinofuranosyl cytosine in G 2 . (author)

  13. Relationship between cell cycle stage in the fertilized egg of mice and repair capacity for X-ray-induced damage in the sperm

    Energy Technology Data Exchange (ETDEWEB)

    Matsuda, Y.; Maemori, M.; Tobari, I. (National Inst. of Radiological Sciences, Chiba (Japan))

    1989-09-01

    The potentiation effects of 3-aminobenzamide, caffeine, hydroxyurea and arabinofuranosyl cytosine on the yield of X-ray-induced chromosome aberrations of mouse sperm were examined at the first-cleavage metaphase, to clarify a correlation between chromosome aberrations and cell cycle dependency of repair capacity of the fertilized egg. The result provided evidence that there are two major types of DNA damage in X-irradiated sperm: (1) short-lived DNA lesions; the lesions are subject to repair inhibitions by agents added in G{sub 1} and are converted into chromosome-type aberrations during G{sub 1}, and (2) long-lived DNA lesions; the lesions persist until S phase and repair of the lesions is inhibited by caffeine, hydroxyurea and arabinofuranosyl cytosine in G {sub 2}. (author).

  14. A comparison of fatty acid composition and quality aspects of eggs and larvae from cultured and wild broodstock of common sole ( Solea solea L . )

    DEFF Research Database (Denmark)

    Lund, Ivar; Steenfeldt, Svend Jørgen; Suhr, Karin

    2008-01-01

    compared with respect to fatty acid (FA) composition, egg size, fertilization rate and hatching rate. Based on a multivariate analysis of the FA profiles, it was possible to discriminate between culture and wild inheritance. Eggs from cultured broodstock had high levels of C20:1(n-9), C18:2(n-6) and C18......:3(n-3), whereas eggs from wild fish had high levels of C16:1(n-7), C20:4(n-6) and C20:5(n-3). Differences in FA profiles were most likely related to dietary differences. Fertilization and hatching rates were generally low and lowest in eggs from cultured broodstock, but not related to FA composition...

  15. Comparative Analysis of Cartilage Marker Gene Expression Patterns during Axolotl and Xenopus Limb Regeneration.

    Directory of Open Access Journals (Sweden)

    Kazumasa Mitogawa

    Full Text Available Axolotls (Ambystoma mexicanum can completely regenerate lost limbs, whereas Xenopus laevis frogs cannot. During limb regeneration, a blastema is first formed at the amputation plane. It is thought that this regeneration blastema forms a limb by mechanisms similar to those of a developing embryonic limb bud. Furthermore, Xenopus laevis frogs can form a blastema after amputation; however, the blastema results in a terminal cone-shaped cartilaginous structure called a "spike." The causes of this patterning defect in Xenopus frog limb regeneration were explored. We hypothesized that differences in chondrogenesis may underlie the patterning defect. Thus, we focused on chondrogenesis. Chondrogenesis marker genes, type I and type II collagen, were compared in regenerative and nonregenerative environments. There were marked differences between axolotls and Xenopus in the expression pattern of these chondrogenesis-associated genes. The relative deficit in the chondrogenic capacity of Xenopus blastema cells may account for the absence of total limb regenerative capacity.

  16. Conservation and divergence of ADAM family proteins in the Xenopus genome

    Directory of Open Access Journals (Sweden)

    Shah Anoop

    2010-07-01

    Full Text Available Abstract Background Members of the disintegrin metalloproteinase (ADAM family play important roles in cellular and developmental processes through their functions as proteases and/or binding partners for other proteins. The amphibian Xenopus has long been used as a model for early vertebrate development, but genome-wide analyses for large gene families were not possible until the recent completion of the X. tropicalis genome sequence and the availability of large scale expression sequence tag (EST databases. In this study we carried out a systematic analysis of the X. tropicalis genome and uncovered several interesting features of ADAM genes in this species. Results Based on the X. tropicalis genome sequence and EST databases, we identified Xenopus orthologues of mammalian ADAMs and obtained full-length cDNA clones for these genes. The deduced protein sequences, synteny and exon-intron boundaries are conserved between most human and X. tropicalis orthologues. The alternative splicing patterns of certain Xenopus ADAM genes, such as adams 22 and 28, are similar to those of their mammalian orthologues. However, we were unable to identify an orthologue for ADAM7 or 8. The Xenopus orthologue of ADAM15, an active metalloproteinase in mammals, does not contain the conserved zinc-binding motif and is hence considered proteolytically inactive. We also found evidence for gain of ADAM genes in Xenopus as compared to other species. There is a homologue of ADAM10 in Xenopus that is missing in most mammals. Furthermore, a single scaffold of X. tropicalis genome contains four genes encoding ADAM28 homologues, suggesting genome duplication in this region. Conclusions Our genome-wide analysis of ADAM genes in X. tropicalis revealed both conservation and evolutionary divergence of these genes in this amphibian species. On the one hand, all ADAMs implicated in normal development and health in other species are conserved in X. tropicalis. On the other hand, some

  17. Musashi and Plasticity of Xenopus and Axolotl Spinal Cord Ependymal Cells

    Science.gov (United States)

    Chernoff, Ellen A. G.; Sato, Kazuna; Salfity, Hai V. N.; Sarria, Deborah A.; Belecky-Adams, Teri

    2018-01-01

    The differentiated state of spinal cord ependymal cells in regeneration-competent amphibians varies between a constitutively active state in what is essentially a developing organism, the tadpole of the frog Xenopus laevis, and a quiescent, activatable state in a slowly growing adult salamander Ambystoma mexicanum, the Axolotl. Ependymal cells are epithelial in intact spinal cord of all vertebrates. After transection, body region ependymal epithelium in both Xenopus and the Axolotl disorganizes for regenerative outgrowth (gap replacement). Injury-reactive ependymal cells serve as a stem/progenitor cell population in regeneration and reconstruct the central canal. Expression patterns of mRNA and protein for the stem/progenitor cell-maintenance Notch signaling pathway mRNA-binding protein Musashi (msi) change with life stage and regeneration competence. Msi-1 is missing (immunohistochemistry), or at very low levels (polymerase chain reaction, PCR), in both intact regeneration-competent adult Axolotl cord and intact non-regeneration-competent Xenopus tadpole (Nieuwkoop and Faber stage 62+, NF 62+). The critical correlation for successful regeneration is msi-1 expression/upregulation after injury in the ependymal outgrowth and stump-region ependymal cells. msi-1 and msi-2 isoforms were cloned for the Axolotl as well as previously unknown isoforms of Xenopus msi-2. Intact Xenopus spinal cord ependymal cells show a loss of msi-1 expression between regeneration-competent (NF 50–53) and non-regenerating stages (NF 62+) and in post-metamorphosis froglets, while msi-2 displays a lower molecular weight isoform in non-regenerating cord. In the Axolotl, embryos and juveniles maintain Msi-1 expression in the intact cord. In the adult Axolotl, Msi-1 is absent, but upregulates after injury. Msi-2 levels are more variable among Axolotl life stages: rising between late tailbud embryos and juveniles and decreasing in adult cord. Cultures of regeneration-competent Xenopus tadpole

  18. Musashi and Plasticity of Xenopus and Axolotl Spinal Cord Ependymal Cells

    Directory of Open Access Journals (Sweden)

    Ellen A. G. Chernoff

    2018-02-01

    Full Text Available The differentiated state of spinal cord ependymal cells in regeneration-competent amphibians varies between a constitutively active state in what is essentially a developing organism, the tadpole of the frog Xenopus laevis, and a quiescent, activatable state in a slowly growing adult salamander Ambystoma mexicanum, the Axolotl. Ependymal cells are epithelial in intact spinal cord of all vertebrates. After transection, body region ependymal epithelium in both Xenopus and the Axolotl disorganizes for regenerative outgrowth (gap replacement. Injury-reactive ependymal cells serve as a stem/progenitor cell population in regeneration and reconstruct the central canal. Expression patterns of mRNA and protein for the stem/progenitor cell-maintenance Notch signaling pathway mRNA-binding protein Musashi (msi change with life stage and regeneration competence. Msi-1 is missing (immunohistochemistry, or at very low levels (polymerase chain reaction, PCR, in both intact regeneration-competent adult Axolotl cord and intact non-regeneration-competent Xenopus tadpole (Nieuwkoop and Faber stage 62+, NF 62+. The critical correlation for successful regeneration is msi-1 expression/upregulation after injury in the ependymal outgrowth and stump-region ependymal cells. msi-1 and msi-2 isoforms were cloned for the Axolotl as well as previously unknown isoforms of Xenopus msi-2. Intact Xenopus spinal cord ependymal cells show a loss of msi-1 expression between regeneration-competent (NF 50–53 and non-regenerating stages (NF 62+ and in post-metamorphosis froglets, while msi-2 displays a lower molecular weight isoform in non-regenerating cord. In the Axolotl, embryos and juveniles maintain Msi-1 expression in the intact cord. In the adult Axolotl, Msi-1 is absent, but upregulates after injury. Msi-2 levels are more variable among Axolotl life stages: rising between late tailbud embryos and juveniles and decreasing in adult cord. Cultures of regeneration

  19. Normal development of hamster and rabbit eggs fertilized by spermatozoa labelled with the fluorescent thiol alkylating agent, monobromobimane

    International Nuclear Information System (INIS)

    Fleming, A.D.; Cummins, J.M.; Kuehl, T.J.; Seidel, G.E. Jr.; Yanagimachi, R.

    1986-01-01

    Cauda epididymal spermatozoa of the golden hamster were labelled with the thiol-alkylating reagent, monobromobimane (MB). Female hamsters underwent uterine insemination with labelled spermatozoa at laparotomy under metofane anesthesia. All 12 females examined between 5 and 54 h postinsemination yielded a total of 83/100 (83%) eggs in the process of fertilization or embryos. Under ultraviolet (UV) exposure all exhibited a fluorescent tail which, in the 4- and 8-cell embryos, could be seen to be fraying or disintegrating. As cleavage progressed, labelled tail components came to be restricted among the blastomeres such that at the 4- and 8-cell stage the tail could be seen in only one to three blastomeres. To study complete development and pregnancy another 12 females received uterine insemination. After recovery from anesthesia (approximately equal to 4 h) these females were mated with a vasectomized male bearing a dominant genetic marker (black eyes) to allow unequivocal determination of paternity in the fetuses and young produced. Seven became pregnant with one female losing her pregnancy about Day 7 of gestation. Two females sacrificed on Day 13 produced 17 normal fetuses and one resorption. Four females delivered 16 young, all normal at birth and in subsequent growth and fertility. In addition, insemination of female rabbits with MB-labelled spermatozoa yielded normal embryos (50/52 96%) from 3 does on Day 2 and (38/64 60%) from 4 does on Days 4 or 5. Two normal litters (9 bunnies) have delivered from 3 does allowed to carry to term

  20. Effects on the Growth and Survival of Eggs and Embryos of the California Grunion (Leuresthes tenuis) Exposed to Trace Levels of Tributyltin.

    Science.gov (United States)

    1985-05-01

    postfertilization exposure of grunion eggs to low doses of TBT has no effect on hatch success or notochord length at time of hatch. 2. No = exposure of...grunion eggs and sperm during fertilization and sub- sequent low-level dose incubation has no effect on fertilization, hatch suc- cess, or notochord ...clean glass vials for stor- -.- - age. Ten randomly selected larvae from each replicate were also measured for notochord length and examined for

  1. Next generation sequencing and comparative analyses of Xenopus mitogenomes

    Directory of Open Access Journals (Sweden)

    Lloyd Rhiannon E

    2012-09-01

    Full Text Available Abstract Background Mitochondrial genomes comprise a small but critical component of the total DNA in eukaryotic organisms. They encode several key proteins for the cell’s major energy producing apparatus, the mitochondrial respiratory chain. Additonally, their nucleotide and amino acid sequences are of great utility as markers for systematics, molecular ecology and forensics. Their characterization through nucleotide sequencing is a fundamental starting point in mitogenomics. Methods to amplify complete mitochondrial genomes rapidly and efficiently from microgram quantities of tissue of single individuals are, however, not always available. Here we validate two approaches, which combine long-PCR with Roche 454 pyrosequencing technology, to obtain two complete mitochondrial genomes from individual amphibian species. Results We obtained two new xenopus frogs (Xenopus borealis and X. victorianus complete mitochondrial genome sequences by means of long-PCR followed by 454 of individual genomes (approach 1 or of multiple pooled genomes (approach 2, the mean depth of coverage per nucleotide was 9823 and 186, respectively. We also characterised and compared the new mitogenomes against their sister taxa; X. laevis and Silurana tropicalis, two of the most intensely studied amphibians. Our results demonstrate how our approaches can be used to obtain complete amphibian mitogenomes with depths of coverage that far surpass traditional primer-walking strategies, at either the same cost or less. Our results also demonstrate: that the size, gene content and order are the same among xenopus mitogenomes and that S. tropicalis form a separate clade to the other xenopus, among which X. laevis and X. victorianus were most closely related. Nucleotide and amino acid diversity was found to vary across the xenopus mitogenomes, with the greatest diversity observed in the Complex 1 gene nad4l and the least diversity observed in Complex 4 genes (cox1-3. All protein

  2. Ultrastructural dynamics of human reproduction, from ovulation to fertilization and early embryo development.

    Science.gov (United States)

    Familiari, Giuseppe; Heyn, Rosemarie; Relucenti, Michela; Nottola, Stefania A; Sathananthan, A Henry

    2006-01-01

    This study describes the updated, fine structure of human gametes, the human fertilization process, and human embryos, mainly derived from assisted reproductive technology (ART). As clearly shown, the ultrastructure of human reproduction is a peculiar multistep process, which differs in part from that of other mammalian models, having some unique features. Particular attention has been devoted to the (1) sperm ultrastructure, likely "Tygerberg (Kruger) strict morphology criteria"; (2) mature oocyte, in which the MII spindle is barrel shaped, anastral, and lacking centrioles; (3) three-dimensional microarchitecture of the zona pellucida with its unique supramolecular filamentous organization; (4) sperm-egg interactions with the peculiarity of the sperm centrosome that activates the egg and organizes the sperm aster and mitotic spindles of the embryo; and (5) presence of viable cumulus cells whose metabolic activity is closely related to egg and embryo behavior in in vitro as well as in vivo conditions, in a sort of extraovarian "microfollicular unit." Even if the ultrastructural morphodynamic features of human fertilization are well understood, our knowledge about in vivo fertilization is still very limited and the complex sequence of in vivo biological steps involved in human reproduction is only partially reproduced in current ART procedures.

  3. Uptake and elimination of aromatic hydrocarbons a chlorinated biphenyl in eggs and larvae of cod Gadus morhua

    OpenAIRE

    Solbakken, Jan Erik; Tilseth, Snorre; Palmork, Karsten H.

    1984-01-01

    Artificially fertilized eggs and newly hatched larvae of cod Gadus rnorhua L. were exposed to 14C labelled naphthalene, phenanthrene, benzo(a)pyrene and 2. 4. 5. 2', 4', 5'-hexachlorobiphenyl (PCB) for 24 h and thereafter transferred to clean seawater. Radioactivity in eggs and larvae was measured at different times during exposure and after transfer to clean seawater. Maximum accumulation was found with phenanthrene, whereas naphthalene accumulated only slightly in both eggs a...

  4. Effects of gamma irradiation on the egg stage of the meal moth, Pyralis Farinalis L

    International Nuclear Information System (INIS)

    Gharib, O.H.; Abdelkawy, F.K.

    1993-01-01

    Two days-old eggs of the meal moth, Pyralis Farinalis L. Were exposed to different doses of gamma radiation ranged from 10 to 150 gray. Reduction in egg hatch was highly correlated with the given dose. At control, 83.2% of the eggs hatched and 75.6% of the eggs reached adult stage, whereas egg hatch was reduced to 21.8 , 16.1 and 5.8% after exposures to 100,120 and 150 gray, respectively. No adults emerged after egg irradiation to 150 gray. Gamma irradiation had almost slight effect on larval pupal duration without significant differences. Irradiation of eggs could alter the reproductive ability of the emerged moths. The doses 10, 20 and 40 gray delivered to the egg stage reduced egg production and fertility of the emerging moths. The greatest reduction was obtained when both sexes had been irradiated at 40 gray and paired together. Females were more radiosensitive than males. 2 tab

  5. On Reproductive Work in Spain: Transnational Adoption, Egg Donation, Surrogacy.

    Science.gov (United States)

    Marre, Diana; San Román, Beatriz; Guerra, Diana

    2018-01-01

    Spain's plummeting fertility since the late twentieth century may seem to reflect a waning desire for children. Nevertheless, reproductive disappointments resulting from gender inequalities cause many Spanish women to postpone motherhood and experience age-related fertility problems. For them, creating a family often becomes possible only through the reproductive labor of other women. Our analysis of transnational adoption, egg donation, and surrogacy in Spain shows how anonymity and altruism play out in these three strategies, with implications for the valuation of women's reproductive work and relationships among reproductive providers, intermediaries, recipients, and the resulting children.

  6. A new large egg type from the marine live feed calanoid copepod Acartia tonsa (Dana)-Perspectives for selective breeding of designer feed for hatcheries

    DEFF Research Database (Denmark)

    Hammervold, Stian Halsen; Glud, Ronnie N.; Evjemo, Jan Ove

    2015-01-01

    correspondingly large, and the copepods remained large when developing into adulthood. The fact that copepods hatched from large eggs were fertile indicates no chromosomal abnormalities and suggests that this egg type represents the upper tail end of eggs in a classical normal distribution of size vs. frequency...

  7. Feeding broiler breeder males. 4. Deficient feed allocation reduces fertility and broiler progeny body weight.

    Science.gov (United States)

    Romero-Sanchez, H; Plumstead, P W; Leksrisompong, N; Brannan, K E; Brake, J

    2008-04-01

    Two experiments were conducted to evaluate the effects of male broiler breeder feed intake on broiler progeny performance. In experiment 1, a low cumulative nutrition program supplied 29,580 kcal of ME and 1,470 g of CP, whereas a high cumulative nutrition program supplied 33,500 kcal of ME and 1,730 g of CP to photostimulation at 21 wk of age. Two diets (HiDiet and LoDiet) were formulated, and a single feeding program was used to achieve the selected nutrient intakes. The HiDiet group of males in experiment 1 achieved greater BW and exhibited lower fertility when fed as the LoDiet males from the onset of egg production. The HiDiet breeder males subsequently produced male broilers from eggs laid at 29 wk of age that exhibited lower BW at 42 d. This was due to the heaviest 50% of the breeder males in this treatment not gaining BW consistently due to less-than-adequate ME intake relative to their greater BW requirements. Two feeding programs during the production period (constant or increasing) were compared in experiment 2. Broilers were hatched from eggs laid at 32 and 48 wk of age to evaluate the vertical effect of male treatments on progeny performance. No difference in fertility or broiler performance was found at 32 wk. However, the constant feeding program produced lower fertility from 36 to 55 wk of age, and this resulted in a lower male and female broiler progeny BW at 42 d of age from eggs collected at 48 wk of age. Adequate breeder male feed allocation during the production period improved fertility and favorably affected broiler progeny performance in both experiments. However, broiler progeny effects were observed only when there were differences in fertility, which suggests that the males with the greatest genetic potential were not mating at these times.

  8. Live Birth After In-Vitro Fertilization (IVF) in a 53 Year Old Woman: A ...

    African Journals Online (AJOL)

    TNHJOURNALPH

    It has adverse social and psychological implications especially for ... resulting in lower fertilization cleavage and implantation rates. ... on fertility. However even amongst women who receive donated eggs,pregnancy ratesare lower in women aged 40 years and above[5,6]. Advanced maternal age is a risk factor for almost all ...

  9. Histone titration against the genome sets the DNA-to-cytoplasm threshold for the Xenopus midblastula transition

    Science.gov (United States)

    Amodeo, Amanda A.; Jukam, David; Straight, Aaron F.; Skotheim, Jan M.

    2015-01-01

    During early development, animal embryos depend on maternally deposited RNA until zygotic genes become transcriptionally active. Before this maternal-to-zygotic transition, many species execute rapid and synchronous cell divisions without growth phases or cell cycle checkpoints. The coordinated onset of transcription, cell cycle lengthening, and cell cycle checkpoints comprise the midblastula transition (MBT). A long-standing model in the frog, Xenopus laevis, posits that MBT timing is controlled by a maternally loaded inhibitory factor that is titrated against the exponentially increasing amount of DNA. To identify MBT regulators, we developed an assay using Xenopus egg extract that recapitulates the activation of transcription only above the DNA-to-cytoplasm ratio found in embryos at the MBT. We used this system to biochemically purify factors responsible for inhibiting transcription below the threshold DNA-to-cytoplasm ratio. This unbiased approach identified histones H3 and H4 as concentration-dependent inhibitory factors. Addition or depletion of H3/H4 from the extract quantitatively shifted the amount of DNA required for transcriptional activation in vitro. Moreover, reduction of H3 protein in embryos induced premature transcriptional activation and cell cycle lengthening, and the addition of H3/H4 shortened post-MBT cell cycles. Our observations support a model for MBT regulation by DNA-based titration and suggest that depletion of free histones regulates the MBT. More broadly, our work shows how a constant concentration DNA binding molecule can effectively measure the amount of cytoplasm per genome to coordinate division, growth, and development. PMID:25713373

  10. OCT imaging of craniofacial anatomy in xenopus embryos (Conference Presentation)

    Science.gov (United States)

    Deniz, Engin; Jonas, Stephan M.; Griffin, John; Hooper, Michael C.; Choma, Michael A.; Khokha, Mustafa K.

    2016-03-01

    The etiology of craniofacial defects is incompletely understood. The ability to obtain large amounts of gene sequence data from families affected by craniofacial defects is opening up new ways to understand molecular genetic etiological factors. One important link between gene sequence data and clinical relevance is biological research into candidate genes and molecular pathways. We present our recent research using OCT as a nondestructive phenotyping modality of craniofacial morphology in Xenopus embryos, an important animal model for biological research in gene and pathway discovery. We define 2D and 3D scanning protocols for a standardized approach to craniofacial imaging in Xenopus embryos. We define standard views and planar reconstructions for visualizing normal anatomy and landmarks. We compare these views and reconstructions to traditional histopathology using alcian blue staining. In addition to being 3D, nondestructive, and having much faster throughout, OCT can identify craniofacial features that are lost during traditional histopathological preparation. We also identify quantitative morphometric parameters to define normative craniofacial anatomy. We also note that craniofacial and cardiac defects are not infrequently present in the same patient (e.g velocardiofacial syndrome). Given that OCT excels at certain aspects of cardiac imaging in Xenopus embryos, our work highlights the potential of using OCT and Xenopus to study molecular genetic factors that impact both cardiac and craniofacial development.

  11. Marine gametes in a changing ocean: Impacts of climate change stressors on fecundity and the egg.

    Science.gov (United States)

    Foo, Shawna A; Byrne, Maria

    2017-07-01

    In marine invertebrates, the environmental history of the mother can influence fecundity and egg size. Acclimation of females in climate change stressors, increased temperature and low pH, results in a decrease in egg number and size in many taxa, with the exception of cephalopods, where eggs increase in size. With respect to spawned eggs, near future levels of ocean acidification can interfere with the egg's block to polyspermy and intracellular pH. Reduction of the extracellular egg jelly coat seen in low pH conditions has implications for impaired egg function and fertilization. Some fast generation species (e.g. copepods, polychaetes) have shown restoration of female reproductive output after several generations in treatments. It will be important to determine if the changes to egg number and size induced by exposure to climate change stressors are heritable. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Skin wound healing in different aged Xenopus laevis.

    Science.gov (United States)

    Bertolotti, Evelina; Malagoli, Davide; Franchini, Antonella

    2013-08-01

    Xenopus froglets can perfectly heal skin wounds without scarring. To explore whether this capacity is maintained as development proceeds, we examined the cellular responses during the repair of skin injury in 8- and 15-month-old Xenopus laevis. The morphology and sequence of healing phases (i.e., inflammation, new tissue formation, and remodeling) were independent of age, while the timing was delayed in older frogs. At the beginning of postinjury, wound re-epithelialization occurred in form of a thin epithelium followed by a multilayered epidermis containing cells with apoptotic patterns and keratinocytes stained by anti-inducible nitric oxide synthase (iNOS) antibody. The inflammatory response, early activated by recruitment of blood cells immunoreactive to anti-tumor necrosis factor (TNF)-α, iNOS, transforming growth factor (TGF)-β1, and matrix metalloproteinase (MMP)-9, persisted over time. The dermis repaired by a granulation tissue with extensive angiogenesis, inflammatory cells, fibroblasts, and anti-α-SMA positive myofibroblasts. As the healing progressed, wounded areas displayed vascular regression, decrease in cellularity, and rearrangement of provisional matrix. The epidermis restored to a prewound morphology while granulation tissue was replaced by a fibrous tissue in a scar-like pattern. The quantitative PCR analysis demonstrated an up-regulated expression of Xenopus suppressor of cytokine signaling 3 (XSOCS-3) and Xenopus transforming growth factor-β2 (XTGF-β2) soon after wounding and peak levels were detected when granulation tissue was well developed with a large number of inflammatory cells. The findings indicate that X. laevis skin wound healing occurred by a combination of regeneration (in epidermis) and repair (in dermis) and, in contrast to froglet scarless wound healing, the growth to a more mature adult stage is associated with a decrease in regenerative capacity with scar-like tissue formation. Copyright © 2013 Wiley Periodicals, Inc.

  13. Biology and fertility life table of Agrotis ipsilon on artificial diet

    International Nuclear Information System (INIS)

    Bento, Flavia de Moura Manoel; Fortes, Priscila; Zerio, Neide Graciano; Parra, Jose Roberto Postali

    2007-01-01

    The objective of this work was to develop an artificial diet to rear Agrotis ipsilon in laboratory using biological parameters and fertility life table. The artificial diet was prepared with bean, casein, soybean protein, yeast and wheat germ as protein sources. The biological aspects duration and viability of larval and pupal stages, pupal weight, sex ratio, life span of adults, preoviposition period, egg laying capacity and fertility life table were evaluated. Six larval instars were observed comprising larval duration of 28,4 days and 93% of viability mean pupal duration of 12,4 days and viability of 96%. The total viability of the life cycle was 72%. The pupae weight was 387 mg for males and 484 mg for females. The sex ratio was 0,46 and the preoviposition period lasted one day and egg laying was 1,806 eggs per female. The net reproductive rate per generation and increase finite rate were 616,9 and 1,14, respectively. Artificial diet is adequate for rearing A. ipsilon in laboratory. (author)

  14. Sperm storage and duration of fertility in female ostriches ( Struthio ...

    African Journals Online (AJOL)

    , the duration of sperm storage and the fertile period following separation of sexes were investigated by egg break-out and by counting the sperm in the perivitelline membrane (spermOPVL) above the germinal disc (GD) region. Fertilisation ...

  15. Fertility signals in the bumblebee Bombus terrestris (Hymenoptera: Apidae)

    Science.gov (United States)

    Sramkova, A.; Schulz, C.; Twele, R.; Francke, W.; Ayasse, M.

    2008-06-01

    In eusocial Hymenoptera, queen control over workers is probably inseparable from the mechanism of queen recognition. In primitively eusocial bumblebees ( Bombus), worker reproduction is controlled not only by the presence or absence of a dominant queen but also by other dominant workers. Furthermore, it was shown that the queen dominance is maintained by pheromonal cues. We investigated whether there is a similar odor signal released by egg-laying queens and workers that may have a function as a fertility signal. We collected cuticular surface extracts from nest-searching and breeding Bombus terrestris queens and workers that were characterized by their ovarian stages. In chemical analyses, we identified 61 compounds consisting of aldehydes, alkanes, alkenes, and fatty acid esters. Nest-searching queens and all groups of breeding females differed significantly in their odor bouquets. Furthermore, workers before the competition point (time point of colony development where workers start to develop ovaries and lay eggs) differed largely from queens and all other groups of workers. Breeding queens showed a unique bouquet of chemical compounds and certain queen-specific compounds, and the differences toward workers decrease with an increasing development of the workers’ ovaries, hinting the presence of a reliable fertility signal. Among the worker groups, the smallest differences were found after the competition point. Egg-laying females contained higher total amounts of chemical compounds and of relative proportions of wax-type esters and aldehydes than nest-searching queens and workers before the competition point. Therefore, these compounds may have a function as a fertility signal present in queens and workers.

  16. Polystyrene nanoparticles affect Xenopus laevis development

    Energy Technology Data Exchange (ETDEWEB)

    Tussellino, Margherita; Ronca, Raffaele [University of Naples Federico II, Department of Biology (Italy); Formiggini, Fabio [Italian Institute of Technology, Center for Advanced Biomaterials for Health Care IIT@CRIB (Italy); Marco, Nadia De [University of Naples Federico II, Department of Biology (Italy); Fusco, Sabato; Netti, Paolo Antonio [Italian Institute of Technology, Center for Advanced Biomaterials for Health Care IIT@CRIB (Italy); Carotenuto, Rosa, E-mail: rosa.carotenuto@unina.it [University of Naples Federico II, Department of Biology (Italy)

    2015-02-15

    Exposing living organisms to nanoparticulates is potentially hazardous, in particular when it takes place during embryogenesis. In this investigation, we have studied the effects of 50-nm-uncoated polystyrene nanoparticles (PSNPs) as a model to investigate the suitability of their possible future employments. We have used the standardized Frog Embryo Teratogenesis Assay-Xenopus test during the early stages of larval development of Xenopus laevis, and we have employed either contact exposure or microinjections. We found that the embryos mortality rate is dose dependent and that the survived embryos showed high percentage of malformations. They display disorders in pigmentation distribution, malformations of the head, gut and tail, edema in the anterior ventral region, and a shorter body length compared with sibling untreated embryos. Moreover, these embryos grow more slowly than the untreated embryos. Expressions of the mesoderm markers, bra (T-box Brachyury gene), myod1 (myogenic differentiation1), and of neural crest marker sox9 (sex SRY (determining region Y-box 9) transcription factor sox9), are modified. Confocal microscopy showed that the nanoparticles are localized in the cytoplasm, in the nucleus, and in the periphery of the digestive gut cells. Our data suggest that PSNPs are toxic and show a potential teratogenic effect for Xenopus larvae. We hypothesize that these effects may be due either to the amount of NPs that penetrate into the cells and/or to the “corona” effect caused by the interaction of PSNPs with cytoplasm components. The three endpoints of our study, i.e., mortality, malformations, and growth inhibition, suggest that the tests we used may be a powerful and flexible bioassay in evaluating pollutants in aquatic embryos.

  17. Polystyrene nanoparticles affect Xenopus laevis development

    International Nuclear Information System (INIS)

    Tussellino, Margherita; Ronca, Raffaele; Formiggini, Fabio; Marco, Nadia De; Fusco, Sabato; Netti, Paolo Antonio; Carotenuto, Rosa

    2015-01-01

    Exposing living organisms to nanoparticulates is potentially hazardous, in particular when it takes place during embryogenesis. In this investigation, we have studied the effects of 50-nm-uncoated polystyrene nanoparticles (PSNPs) as a model to investigate the suitability of their possible future employments. We have used the standardized Frog Embryo Teratogenesis Assay-Xenopus test during the early stages of larval development of Xenopus laevis, and we have employed either contact exposure or microinjections. We found that the embryos mortality rate is dose dependent and that the survived embryos showed high percentage of malformations. They display disorders in pigmentation distribution, malformations of the head, gut and tail, edema in the anterior ventral region, and a shorter body length compared with sibling untreated embryos. Moreover, these embryos grow more slowly than the untreated embryos. Expressions of the mesoderm markers, bra (T-box Brachyury gene), myod1 (myogenic differentiation1), and of neural crest marker sox9 (sex SRY (determining region Y-box 9) transcription factor sox9), are modified. Confocal microscopy showed that the nanoparticles are localized in the cytoplasm, in the nucleus, and in the periphery of the digestive gut cells. Our data suggest that PSNPs are toxic and show a potential teratogenic effect for Xenopus larvae. We hypothesize that these effects may be due either to the amount of NPs that penetrate into the cells and/or to the “corona” effect caused by the interaction of PSNPs with cytoplasm components. The three endpoints of our study, i.e., mortality, malformations, and growth inhibition, suggest that the tests we used may be a powerful and flexible bioassay in evaluating pollutants in aquatic embryos

  18. Laboratory technique for obtaining fathead minnow eggs for use in toxicity experiments

    International Nuclear Information System (INIS)

    Till, J.E.

    1977-01-01

    A laboratory technique for collecting freshly fertilized eggs of the fathead minnow (Pimephales promelas) has been developed for use in toxicity testing of radioactive materials or other environmental pollutants. The fathead minnow egg was selected as a test organism because the period of embryogenesis is relatively short, the number of eggs per spawning is sufficiently large to provide a test group and a control group, and the size of the parent fish is compatible with laboratory facilities. Under a controlled laboratory environment, eggs are available all year. Nesting sites were constructed by cutting plastic pipe in half and covering the inside of the pipe with 1-mm-thick translucent polyethylene. The polyethylene with the eggs attached was removed from the shelter and cut into two pieces, each piece containing approximately the same number of eggs. One piece of polyethylene was floated in the test solution, and the other in spring water as a control. Hatching occurs in 168 h at 24 0 C. Toxicity of the test solution may be scored on the percentage of eggs hatching, the number of abnormal larvae produced, and larval survival

  19. Bisphenol A accumulation in eggs disrupts the endocrine regulation of growth in rainbow trout larvae

    Energy Technology Data Exchange (ETDEWEB)

    Birceanu, Oana; Servos, Mark R.; Vijayan, Mathilakath M., E-mail: matt.vijayan@ucalgary.ca

    2015-04-15

    Highlights: • BPA in eggs reduces growth and increases food conversion ratio in trout larvae. • BPA in eggs disrupts larval transcript abundance of genes involved in GH/IGF axis. • BPA in eggs disrupts thyroid hormone receptor mRNA levels. • BPA in eggs consistently suppressed IGF-1rb mRNA levels during early development. - Abstract: Bisphenol A (BPA), a monomer used in the production of plastics and epoxy resins, is ubiquitously present in the aquatic environment. BPA is considered a weak estrogen in fish, but the effects of this chemical on early developmental events are far from clear. We tested the hypothesis that BPA accumulation in eggs, mimicking maternal transfer, disrupts growth hormone/insulin-like growth factor (GH/IGF) axis function, leading to defects in larval growth in rainbow trout. Trout oocytes were exposed to 0 (control), 0.3, 3, and 30 μg ml{sup −1} BPA for 3 h, which led to an accumulation of around 0, 1, 4 and 40 ng BPA per egg, respectively. All treatment groups were fertilized with clean milt and reared in clean water for the rest of the experiment. The embryo BPA content declined over time in all groups and was completely eliminated by 42 days post-fertilization (dpf). Hatchlings from BPA accumulated eggs had higher water content and reduced total energy levels prior to first feed. There was an overall reduction in the specific growth rate and food conversion ratio in larvae reared from BPA-laden eggs. BPA accumulation disrupted the mRNA abundance of genes involved in GH/IGF axis function, including GH isoforms and their receptors, IGF-1 and -2 and IGF receptors, in a life stage-dependent manner. Also, there was a temporal disruption in the mRNA levels of thyroid hormone receptors in the larvae raised from BPA-laden eggs. Altogether, BPA accumulation in eggs, mimicking maternal transfer, affects larval growth and the mode of action involves disruption of genes involved in the GH/IGF and thyroid axes function in trout.

  20. Bisphenol A accumulation in eggs disrupts the endocrine regulation of growth in rainbow trout larvae

    International Nuclear Information System (INIS)

    Birceanu, Oana; Servos, Mark R.; Vijayan, Mathilakath M.

    2015-01-01

    Highlights: • BPA in eggs reduces growth and increases food conversion ratio in trout larvae. • BPA in eggs disrupts larval transcript abundance of genes involved in GH/IGF axis. • BPA in eggs disrupts thyroid hormone receptor mRNA levels. • BPA in eggs consistently suppressed IGF-1rb mRNA levels during early development. - Abstract: Bisphenol A (BPA), a monomer used in the production of plastics and epoxy resins, is ubiquitously present in the aquatic environment. BPA is considered a weak estrogen in fish, but the effects of this chemical on early developmental events are far from clear. We tested the hypothesis that BPA accumulation in eggs, mimicking maternal transfer, disrupts growth hormone/insulin-like growth factor (GH/IGF) axis function, leading to defects in larval growth in rainbow trout. Trout oocytes were exposed to 0 (control), 0.3, 3, and 30 μg ml −1 BPA for 3 h, which led to an accumulation of around 0, 1, 4 and 40 ng BPA per egg, respectively. All treatment groups were fertilized with clean milt and reared in clean water for the rest of the experiment. The embryo BPA content declined over time in all groups and was completely eliminated by 42 days post-fertilization (dpf). Hatchlings from BPA accumulated eggs had higher water content and reduced total energy levels prior to first feed. There was an overall reduction in the specific growth rate and food conversion ratio in larvae reared from BPA-laden eggs. BPA accumulation disrupted the mRNA abundance of genes involved in GH/IGF axis function, including GH isoforms and their receptors, IGF-1 and -2 and IGF receptors, in a life stage-dependent manner. Also, there was a temporal disruption in the mRNA levels of thyroid hormone receptors in the larvae raised from BPA-laden eggs. Altogether, BPA accumulation in eggs, mimicking maternal transfer, affects larval growth and the mode of action involves disruption of genes involved in the GH/IGF and thyroid axes function in trout

  1. Effect of gamma-radiated males on egg production in Acarus siro

    International Nuclear Information System (INIS)

    Szlendak, E.; Boczek, J.; Bruce, W.; Davis, R.

    1985-01-01

    Mating of unirradiated Acarus siro L. females with normal and gamma irradiated males was studied. Females usually laid eggs throughout their life span if they were mated at least once a week with normal males. Females paired with treated males contained spermatophores but did not lay any eggs. Previously mated fertile females of A. siro subsequently mated with irradiated, sterilized males ceased egg-laying during the next 1-2 weeks. There was no significant difference when compared to the untreated control in the number of eggs laid when females were mated for one week with irradiated males and then with normal males. Weekly alternation of mating with irradiated and normal males reduced significantly the number of eggs laid by the females and significantly reduced their longevity. Because this mite species mates many times during its life span, it is doubtful that it could be eradicated solely by the introduction of sterile males into a natural population. However, the presence of such males would decrease the number of eggs laid by normal females and it would lead to the decrease in the number of offspring in the next generation

  2. Radiosensitivity of eggs of the rice moth, corcyra cephalonica (STAINT)

    International Nuclear Information System (INIS)

    Abdel-Salam, K.A.

    1991-01-01

    The sensitivity of 1-day-old eggs of Coreyra cephalonica staint., to ionizing radiation has been investigated in view of producing sterile male adults which might help in possible future application of sterile-male technique to control this species. The irradiation effects on subsequent developmental stages were evaluated. A dose of 5 krad was found to prevent hatching of eggs while a dose of 4 krad was the highest dose that allowed an adequate percentage of the irradiated eggs (28.23) to complete development to the adult stage. The mating percentage and mating frequency of resultant adults appeared to be unaffected and the percentage of multiple mating was variable. Egg production and fertility of resulting adults were significantly decreased and all effects of the treatment were more severe for pairings of irradiated males with irradiated females than pairings of either sex with unirradiated partner. However, the results show that a dose of irradiation low enough to permit adequate numbers of adults to develop from irradiated eggs ( 4 krad ) is insufficient to induce high levels of sterility. Thus, it could be concluded that irradiation of eggs for mass production of either sterile or partially sterile adults was found not to be feasible.4 tab., 1 fig

  3. ADAM2 interactions with mouse eggs and cell lines expressing α4/α9 (ITGA4/ITGA9 integrins: implications for integrin-based adhesion and fertilization.

    Directory of Open Access Journals (Sweden)

    Ulyana V Desiderio

    2010-10-01

    Full Text Available Integrins are heterodimeric cell adhesion molecules, with 18 α (ITGA and eight β (ITGB subunits forming 24 heterodimers classified into five families. Certain integrins, especially the α(4/α(9 (ITGA4/ITGA9 family, interact with members of the ADAM (a disintegrin and metalloprotease family. ADAM2 is among the better characterized and also of interest because of its role in sperm function. Having shown that ITGA9 on mouse eggs participates in mouse sperm-egg interactions, we sought to characterize ITGA4/ITGA9-ADAM2 interactions.An anti-β(1/ITGB1 function-blocking antibody that reduces sperm-egg binding significantly inhibited ADAM2 binding to mouse eggs. Analysis of integrin subunit expression indicates that mouse eggs could express at least ten different integrins, five in the RGD-binding family, two in the laminin-binding family, two in the collagen-binding family, and ITGA9-ITGB1. Adhesion assays to characterize ADAM2 interactions with ITGA4/ITGA9 family members produced the surprising result that RPMI 8866 cell adhesion to ADAM2 was inhibited by an anti-ITGA9 antibody, noteworthy because ITGA9 has only been reported to dimerize with ITGB1, and RPMI 8866 cells lack detectable ITGB1. Antibody and siRNA studies demonstrate that ITGB7 is the β subunit contributing to RPMI 8866 adhesion to ADAM2.These data indicate that a novel integrin α-β combination, ITGA9-ITGB7 (α(9β(7, in RPMI 8866 cells functions as a binding partner for ADAM2. ITGA9 had previously only been reported to dimerize with ITGB1. Although ITGA9-ITGB7 is unlikely to be a widely expressed integrin and appears to be the result of "compensatory dimerization" occurring in the context of little/no ITGB1 expression, the data indicate that ITGA9-ITGB7 functions as an ADAM binding partner in certain cellular contexts, with implications for mammalian fertilization and integrin function.

  4. Highly efficient gene knockout by injection of TALEN mRNAs into oocytes and host transfer in Xenopus laevis

    Directory of Open Access Journals (Sweden)

    Keisuke Nakajima

    2015-01-01

    Full Text Available Zinc-finger nucleases, transcription activator-like effector nucleases (TALENs and the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins system are potentially powerful tools for producing tailor-made knockout animals. However, their mutagenic activity is not high enough to induce mutations at all loci of a target gene throughout an entire tadpole. In this study, we present a highly efficient method for introducing gene modifications at almost all target sequences in randomly selected embryos. The gene modification activity of TALEN is enhanced by adopting the host-transfer technique. In our method, the efficiency is further improved by injecting TALEN mRNAs fused to the 3′UTR of the Xenopus DEADSouth gene into oocytes, which are then transferred into a host female frog, where they are ovulated and fertilized. The addition of the 3′UTR of the DEADSouth gene promotes mRNA translation in the oocytes and increases the expression of TALEN proteins to near-maximal levels three hours post fertilization (hpf. In contrast, TALEN mRNAs without this 3′UTR are translated infrequently in oocytes. Our data suggest that genomic DNA is more sensitive to TALEN proteins from fertilization to the midblastula (MBT stage. Our method works by increasing the levels of TALEN proteins during the pre-MBT stages.

  5. EBF proteins participate in transcriptional regulation of Xenopus muscle development.

    Science.gov (United States)

    Green, Yangsook Song; Vetter, Monica L

    2011-10-01

    EBF proteins have diverse functions in the development of multiple lineages, including neurons, B cells and adipocytes. During Drosophila muscle development EBF proteins are expressed in muscle progenitors and are required for muscle cell differentiation, but there is no known function of EBF proteins in vertebrate muscle development. In this study, we examine the expression of ebf genes in Xenopus muscle tissue and show that EBF activity is necessary for aspects of Xenopus skeletal muscle development, including somite organization, migration of hypaxial muscle anlagen toward the ventral abdomen, and development of jaw muscle. From a microarray screen, we have identified multiple candidate targets of EBF activity with known roles in muscle development. The candidate targets we have verified are MYOD, MYF5, M-Cadherin and SEB-4. In vivo overexpression of the ebf2 and ebf3 genes leads to ectopic expression of these candidate targets, and knockdown of EBF activity causes downregulation of the endogenous expression of the candidate targets. Furthermore, we found that MYOD and MYF5 are likely to be direct targets. Finally we show that MYOD can upregulate the expression of ebf genes, indicating the presence of a positive feedback loop between EBF and MYOD that we find to be important for maintenance of MYOD expression in Xenopus. These results suggest that EBF activity is important for both stabilizing commitment and driving aspects of differentiation in Xenopus muscle cells. Copyright © 2010 Elsevier Inc. All rights reserved.

  6. Evidence for participation of GCS1 in fertilization of the starlet sea anemone Nematostella vectensis: Implication of a common mechanism of sperm–egg fusion in plants and animals

    Energy Technology Data Exchange (ETDEWEB)

    Ebchuqin, Eerdundagula; Yokota, Naoto; Yamada, Lixy [Sugashima Marine Biological Laboratory, Graduate School of Science, Nagoya University, Sugashima, Toba 517-0004 (Japan); Yasuoka, Yuuri [Marine Genomics Unit, Okinawa Institute of Science and Technology, Onna, Okinawa 904-0495 (Japan); Akasaka, Mari [Laboratory of Molecular Genetics, Graduate School of Science, Nagoya University, Chikusa-ku, Furo-cho, Nagoya 464-8602 (Japan); Arakawa, Mio; Deguchi, Ryusaku [Department of Biology, Miyagi University of Education, Sendai, Miyagi 980-0845 (Japan); Mori, Toshiyuki [Waseda Institute for Advanced Study, Waseda University, Shinjuku, Tokyo 169-8050 (Japan); Sawada, Hitoshi, E-mail: hsawada@bio.nagoya-u.ac.jp [Sugashima Marine Biological Laboratory, Graduate School of Science, Nagoya University, Sugashima, Toba 517-0004 (Japan)

    2014-09-05

    Highlights: • GCS1 is a sperm transmembrane protein that is essential for gamete fusion in flowering plants. • The GCS1 gene is present not only in angiosperms but also in unicellular organisms and animals. • NvGCS1 gene is expressed in the testis and GCS1 protein exists in sperm of a sea anemone. • Anti-GCS1 antibodies inhibited the fertilization, showing the participation in fertilization. - Abstract: It has been reported that GCS1 (Generative Cell Specific 1) is a transmembrane protein that is exclusively expressed in sperm cells and is essential for gamete fusion in flowering plants. The GCS1 gene is present not only in angiosperms but also in unicellular organisms and animals, implying the occurrence of a common or ancestral mechanism of GCS1-mediated gamete fusion. In order to elucidate the common mechanism, we investigated the role of GCS1 in animal fertilization using a sea anemone (Cnidaria), Nematostella vectensis. Although the existence of the GCS1 gene in N. vectensis has been reported, the expression of GCS1 in sperm and the role of GCS1 in fertilization are not known. In this study, we showed that the GCS1 gene is expressed in the testis and that GCS1 protein exists in sperm by in situ hybridization and proteomic analysis, respectively. Then we made four peptide antibodies against the N-terminal extracellular region of NvGCS1. These antibodies specifically reacted to NvGCS1 among sperm proteins on the basis of Western analysis and potently inhibited fertilization in a concentration-dependent manner. These results indicate that sperm GCS1 plays a pivotal role in fertilization, most probably in sperm–egg fusion, in a starlet sea anemone, suggesting a common gamete-fusion mechanism shared by eukaryotic organisms.

  7. Efficacy and toxicity of iodine disinfection of Atlantic salmon eggs

    Science.gov (United States)

    Chalupnicki, M.A.; Ketola, H.G.; Starliper, C.E.; Gallagher, D.

    2011-01-01

    Recent interest in the restoration of Atlantic salmon Salmo salar in the Great Lakes has given rise to new culture techniques and management programs designed to reduce pathogen transmission while stabilizing and enhancing wild populations. We examined the toxicity of iodine to Atlantic salmon eggs and its effectiveness as a disinfectant against bacteria on egg surfaces. We spawned and fertilized eight gravid Atlantic salmon from Cayuga Lake, New York, and exposed their eggs to 10 concentrations of iodine (5, 10, 50, 75, 100, 500, 750, 1,000, 5,000, and 7,500 mg/L) for 30 min during water hardening. An additional subsample of unfertilized eggs was also exposed to some of the same concentrations of iodine (5, 10, 50, 75, and 100 mg/L) to determine the efficiency of disinfection. Viable eggs were only obtained from four females. Survival of eggs to the eyed stage and hatch tended to be reduced at iodine concentrations of 50 and 75 mg/L and was significantly reduced at concentrations of 100 mg/L iodine or more. We calculated the concentrations of iodine that killed 50% of the Atlantic salmon eggs at eye-up and hatch to be 175 and 85 mg/L, respectively. Aeromonas veronii, A. schubertii, A. hydrophila, A. caviae, Plesiomonas shiggeloides, and Citrobacter spp. were the predominant bacteria present on the surface of green eggs and were significantly reduced by an iodine immersion. The use of iodine as a disinfectant on Atlantic salmon eggs was effective at low concentrations (50–75 mg/L), for which toxicity to Atlantic salmon was minimal.

  8. Young female cancer patients' experiences with fertility counselling and fertility preservation-a qualitative small-scale study within the Danish health care setting.

    Science.gov (United States)

    Hoeg, Didde; Schmidt, Lone; Macklon, Kirsten T

    2016-07-14

    Fertility counselling for young women newly diagnosed with cancer is an important field of preconceptional counselling. This qualitative, small-scale study explored how young women newly diagnosed with cancer experienced specialized fertility preservation counselling and treatment in the public Danish health care system. Semi-structured, in-depth interviews were conducted with five women below 40 years recently diagnosed with cancer. All women received fertility counselling by a fertility specialist at the Fertility Clinic, University Hospital of Copenhagen, Denmark before initiation of cancer treatment. Participants were interviewed at a place chosen by them, and interviews were recorded and transcribed verbatim. Data were analysed using systematic text condensation developed by Malterud and inspired by Giorgi's phenomenological analysis. None of the participants were aware that chemotherapy could destroy their eggs. The participants described how specialized fertility counselling and fertility preservation contributed to a belief in life after cancer, which gave them hope that they would survive their cancer disease. Further, the women described how the possibility of fertility preservation removed a huge concern and enabled them to concentrate on their cancer treatment and on getting better. Overall, the specialized fertility counselling and treatment to preserve fertility was highly valued. The women felt it gave them a choice about their future fertility. The fertility expert presented the various fertility-preserving scenarios, and the women were content that they had an actual choice.

  9. DNA methylation profiles correlated to striped bass sperm fertility

    Science.gov (United States)

    Striped bass (Morone saxatilis) spermatozoa are used to fertilize in vitro the eggs of white bass (Morone chrysops) to produce the preferred hybrid for the striped bass aquaculture industry. Currently, only one source of domestic striped bass juveniles are available to growers that are not obtained ...

  10. Chemical surface disinfection of eggs of Baltic cod, Gadus morhua L

    DEFF Research Database (Denmark)

    Overton, J L; Bruun, Morten Sichlau; Dalsgaard, Inger

    2010-01-01

    The effect of two disinfectants on eggs and larvae of Baltic cod, Gadus morhua, was investigated. The eggs were disinfected for 10 min using various concentrations of either glutaraldehyde (100, 200, 400, 600 and 800 mg L−1) or iodophor (10, 50, 100 and 150 mg L−1), 1–4-days post......-fertilization. Bactericidal effect of disinfection, survival to hatching, hatching success and larval abnormalities were assessed. Larval survival was recorded at 5-, 10- and 15-days post-hatch (dph). Although Baltic cod eggs have an unusually thin chorion, they could tolerate surface disinfection. A reduction in bacterial...... growth was observed with increased concentrations of disinfectant (3.0 × 107–1.6 × 101 CFU mL−1). Abnormalities in newly hatched larvae were not related to disinfection. Survival of the yolk sac larvae was significantly better for eggs treated with 400 mg L−1 glutaraldehyde for 10 min at 10 and 15 dph...

  11. Effects of caffeine on cleavage delay of sea urchin eggs induced by ethidium bromide or puromycin

    International Nuclear Information System (INIS)

    Kimura, Hiroshi

    1975-01-01

    The mitotic delay was induced when sea urchin eggs were treated with either ethidium bromide or puromycin, as observed with eggs fertilized with x- or UV-irradiated sperm. Treatment of these eggs with caffeine during the period of early prophase block resulted in the reduction of the mitotic delay. Protein synthesis of these eggs was not affected by x-irradiation but inhibited by ethidium bromide or puromycin. Caffeine was almost ineffective in changing the protein synthesis of eggs inseminated with x-irradiated sperm or treated with ethidium bromide. These facts mean that additive synthesis of protein is not required for the reduction by caffeine of the mitotic delay. Some role of protein synthesis in the reduction by caffeine of the cleavage delay is not excluded for puromycin treated eggs, since caffeine counteracted the inhibitory effect of puromycin on protein synthesis. (author)

  12. Isthmin is a novel secreted protein expressed as part of the Fgf-8 synexpression group in the Xenopus midbrain-hindbrain organizer.

    Science.gov (United States)

    Pera, Edgar M; Kim, James I; Martinez, Sarah L; Brechner, Mariel; Li, Su Yu; Wessely, Oliver; De Robertis, E M

    2002-08-01

    Patterning of the central nervous system is regulated by a signaling center located at the midbrain-hindbrain boundary (MHB), or isthmus organizer. Fibroblast growth factors secreted from the MHB are required and sufficient to direct the ordered growth and regionalization of the midbrain and anterior hindbrain. In an unbiased secretion cloning screen of Xenopus gastrula embryos we identified a novel gene, which we designated as Isthmin (xIsm) due to its prominent expression at the MHB. xIsm encodes a secreted protein of 449 amino acids containing one copy of the thrombospondin type 1 repeat (TSR). We also found orthologous Isthmin genes in human (hIsm) and mouse (mIsm), as well as a gene encoding an Isthmin-like human unknown protein (hIsm-l). The conservation of a unique carboxy-terminal region between hIsm and hIsm-l suggests that Isthmin is the founding member of a new family of secreted proteins. xIsm was strongly expressed maternally in the Xenopus egg and showed zygotic expression in the ventral blastopore lip, notochord, and MHB. Additional expression domains were detected in neural crest, ear vesicle, and developing blood islands. Interestingly, xIsm was co-expressed with Fibroblast growth factor-8 (xFgf-8) at multiple sites including the MHB, indicating that these two genes are part of a synexpression group which also includes sprouty and sef homologs.

  13. Energy source in the developing eggs of the Indian horseshoe crab, Tachypleus gigas (Muller)

    Digital Repository Service at National Institute of Oceanography (India)

    Chatterji, A.; Aguiar, Q.; Saldanha, C.

    Wet weight, dry weight, water content, ash weight, soluble and insoluble proteins, carbohydrates, lipids, and glycogen were determined from 0 to 40th day after fertilization of the developing eggs of the Indian horseshoe crab, Tachypleus gigas...

  14. Zygotic Genome Activation Occurs Shortly after Fertilization in Maize.

    Science.gov (United States)

    Chen, Junyi; Strieder, Nicholas; Krohn, Nadia G; Cyprys, Philipp; Sprunck, Stefanie; Engelmann, Julia C; Dresselhaus, Thomas

    2017-09-01

    The formation of a zygote via the fusion of an egg and sperm cell and its subsequent asymmetric division herald the start of the plant's life cycle. Zygotic genome activation (ZGA) is thought to occur gradually, with the initial steps of zygote and embryo development being primarily maternally controlled, and subsequent steps being governed by the zygotic genome. Here, using maize ( Zea mays ) as a model plant system, we determined the timing of zygote development and generated RNA-seq transcriptome profiles of gametes, zygotes, and apical and basal daughter cells. ZGA occurs shortly after fertilization and involves ∼10% of the genome being activated in a highly dynamic pattern. In particular, genes encoding transcriptional regulators of various families are activated shortly after fertilization. Further analyses suggested that chromatin assembly is strongly modified after fertilization, that the egg cell is primed to activate the translational machinery, and that hormones likely play a minor role in the initial steps of early embryo development in maize. Our findings provide important insights into gamete and zygote activity in plants, and our RNA-seq transcriptome profiles represent a comprehensive, unique RNA-seq data set that can be used by the research community. © 2017 American Society of Plant Biologists. All rights reserved.

  15. Eggs containing larvae of Enterobius vermicularis in vaginal smear

    Directory of Open Access Journals (Sweden)

    Jyothi B Shetty

    2012-01-01

    Full Text Available Enterobius vermicularis also known commonly as pinworm is the most common intestinal parasite. It is a nematode that inhabits the human terminal ileum, colon and appendix. The fertilized female migrates to the perianal area where eggs are deposited but occasionally introduces itself into adjacent orifices, most commonly the female genitourinary tract. Thus the eggs can be seen in the vaginal smear as a result of contamination. We report a case wherein the patient presented with signs and symptoms of vulvovaginitis. In her vaginal smear there were eggs of Enterobius vermicularis which showed a coiled larva within it. In the background there were plenty of acute inflammatory cells. This patient responded favorably to antihelminthics. We report this case to highlight the morphology of the parasite and also to emphasize that such findings should not be neglected. Timely reporting and appropriate treatment of such cases will prevent further complications of this parasite including endometritis, salphingitis and peritonitis.

  16. IVF versus ICSI for the fertilization of in-vitro matured human oocytes.

    Science.gov (United States)

    Walls, M; Junk, S; Ryan, J P; Hart, R

    2012-12-01

    Traditional dogma suggests that intracytoplasmic sperm injection (ICSI) should be performed to ensure successful oocyte fertilization in an in-vitro maturation (IVM) cycle. This study postulated that there would be no difference in the fertilization rate when ICSI was compared with IVF. This hypothesis was tested in a randomized trial of IVF versus ICSI in IVM. A total of 150 immature oocytes were collected in eight cycles of IVM for patients diagnosed with polycystic ovarian syndrome (PCOS). Patients were primed with minimal FSH before transvaginal oocyte aspiration. Sibling oocytes were inseminated by 50% IVF and 50% ICSI. There was no significant difference in fertilization, useable or total blastocyst development between the two insemination technique groups. Clinical pregnancy results for combined fresh and cryopreserved transfers were identical between the two insemination techniques with a total of two fresh and five cryopreserved IVF-inseminated embryos resulting in three clinical pregnancies (42.9%) and five fresh and two cryopreserved ICSI-derived embryos resulting in three clinical pregnancies (42.9%). This research has shown IVF to be a legitimate fertilization technique for IVM oocytes in PCOS patients and provides a greater awareness of the use of a fertilization method previously not utilized with IVM. In-vitro maturation (IVM) is an alternative treatment method to traditional IVF. Due to the minimal use of stimulating hormones in this treatment, IVM has a lower risk of ovarian hyperstimulation syndrome, it can be used for fertility preservation in cancer patients and it is more cost conservative. Early research into the effects of IVM showed a hardening effect on the membrane surrounding the egg (the zona pellucida). It was initially believed that, to overcome this hardening in order to allow the egg to be fertilized, spermatozoa would need to be injected into the egg using intracytoplasmic sperm injection. Due to recent advances in hormonal

  17. ZP Domain Proteins in the Abalone Egg Coat Include a Paralog of VERL under Positive Selection That Binds Lysin and 18-kDa Sperm Proteins

    Science.gov (United States)

    Aagaard, Jan E.; Vacquier, Victor D.; MacCoss, Michael J.; Swanson, Willie J.

    2010-01-01

    Identifying fertilization molecules is key to our understanding of reproductive biology, yet only a few examples of interacting sperm and egg proteins are known. One of the best characterized comes from the invertebrate archeogastropod abalone (Haliotis spp.), where sperm lysin mediates passage through the protective egg vitelline envelope (VE) by binding to the VE protein vitelline envelope receptor for lysin (VERL). Rapid adaptive divergence of abalone lysin and VERL are an example of positive selection on interacting fertilization proteins contributing to reproductive isolation. Previously, we characterized a subset of the abalone VE proteins that share a structural feature, the zona pellucida (ZP) domain, which is common to VERL and the egg envelopes of vertebrates. Here, we use additional expressed sequence tag sequencing and shotgun proteomics to characterize this family of proteins in the abalone egg VE. We expand 3-fold the number of known ZP domain proteins present within the VE (now 30 in total) and identify a paralog of VERL (vitelline envelope zona pellucida domain protein [VEZP] 14) that contains a putative lysin-binding motif. We find that, like VERL, the divergence of VEZP14 among abalone species is driven by positive selection on the lysin-binding motif alone and that these paralogous egg VE proteins bind a similar set of sperm proteins including a rapidly evolving 18-kDa paralog of lysin, which may mediate sperm–egg fusion. This work identifies an egg coat paralog of VERL under positive selection and the candidate sperm proteins with which it may interact during abalone fertilization. PMID:19767347

  18. A Female Identity Switch Helps Keep Only One Egg in the Basket

    NARCIS (Netherlands)

    Weijers, Dolf

    2016-01-01

    The flowering plant female gametophyte carries two gametes, an egg cell and a central cell, whose double fertilization gives rise to embryo and endosperm, respectively. In this issue of Developmental Cell, Yuan et al. (2016) identify the protein CKI1 as a key switch that controls the differential

  19. Collection of gametes from live axolotl, Ambystoma mexicanum, and standardization of in vitro fertilization.

    Science.gov (United States)

    Mansour, N; Lahnsteiner, F; Patzner, R A

    2011-01-15

    This study established the first protocol for collection of gametes from live axolotl, Ambystoma mexicanum, by gentle abdominal massage and in vitro fertilization. To stimulate spermiation and ovulation, human chorionic gonadotrophin (hCG) and Ovopel pellets, which are commercially used to stimulate spawning in fish, were tested. The hCG was more effective than Ovopel pellets and yielded a higher semen volume in the injected males and a shorter response time in the females. Collected semen by this method was already motile and fertile. Fertile eggs could be collected in 3-4 successive collection times after the female has started the typical spawning behaviour. The fertilization condition that yielded the highest hatching rate was mixing semen with eggs before the addition of a fertilization saline solution (20 mmol/l NaCl, 1 mmol/l KCl, 1 mmol/l Mg(2)SO(4), 1 mmol Ca(2)Cl, 3 mmol NaHCO(3), 10 mmol/l Tris, pH 8.5 - Osmolality = 65 mosmol/kg). When the pH of the fertilization solution was increased to ≥ 10, the hatching rate was significantly increased. The use of fertilization solutions with osmolalities of ≥ 150 and ≥ 182 were accompanied with a significant decrease in hatching rates and the appearance of deformed larvae, respectively. In conclusion, a reliable protocol for gamete collection from live axolotl is established as a laboratory model of in vitro fertilization for urodele amphibians. This protocol may be transferable to endangered urodeles. Copyright © 2011 Elsevier Inc. All rights reserved.

  20. Effect of cobalt 60 gamma-ray irradiation on the hatching process of chicken eggs

    International Nuclear Information System (INIS)

    Ohkuma, Yoshikazu

    1981-01-01

    An experiment on fertilized chicken eggs was carried out to determine the effects of 60 Co irradiation on the embryos, their fatality, and growth impairment or deformity, in particular. The experimental groups, consisting of 10 eggs each, recieved a 60 Co irradiation of 50 to 2,000 rads on any one day between day 0 and day 20 of incubation. The larger the irradiation dose, the greater was the number of dead embryos. The fatality was higher in the groups receiving irradiation in the earlier stage (1st week). The resultant death was a chronic one. The irradiation also caused body weight decrease and growth impairment. A decrease in the brain and liver weights was noted, suggesting insufficiency in these organs. Deformity occurred in 4%, most of which involved impairments of skeletal growth, of the bones of the extremities and the bill, in particular. Administration of the SH amino acid, cysteine tended to alleviate the adverse effects of the 60 Co irradiation. These results for fertilized chicken eggs suggest the possibility of abortion and the occurrence of deformities in human fetuses if they should be subjected to 60 Co irradiation. (author)

  1. Possible natural hybridization of two morphologically distinct species of Acropora (Cnidaria, Scleractinia in the Pacific: fertilization and larval survival rates.

    Directory of Open Access Journals (Sweden)

    Naoko Isomura

    Full Text Available Natural hybridization of corals in the Indo-Pacific has been considered rather rare. However, field studies have observed many corals with intermediate interspecific or unusual morphologies. Given that the existence of F1 hybrids with intermediate interspecific morphologies has been proven in the Caribbean, hybrids may also inhabit the Indo-Pacific and occur more frequently than expected. In this study, we focused on two morphologically different species, Acropora florida and A. intermedia, and performed crossing experiments at Akajima Island, Japan. Results showed that these species could hybridize in both directions via eggs and sperm, but that fertilization rates significantly differed according to which species provided eggs. These results are similar to those reported from the Caribbean. Although all embryos developed normally to the planular larval stage, the developmental processes of some hybrid embryos were delayed by approximately 1 h compared with conspecific embryos, suggesting that fertilization occurred 1 h later in interspecific crosses than in intraspecific crosses. More successful hybridization could occur under conditions with low numbers of conspecific colonies. Additionally, a comparison of survival rates between hybrid and intraspecific larvae revealed that intra- and interspecific larvae produced from eggs of A. florida survived for significantly longer than those produced from eggs of A. intermedia. Considering these data, under specific conditions, hybrids can be expected to be produced and survive in nature in the Pacific. Furthermore, we identified one colony with intermediate morphology between A. florida and A. intermedia in the field. This colony was fertilized only by eggs of A. florida, with high fertilization rates, suggesting that this colony would be a hybrid of these two species and might be backcrossed.

  2. Young female cancer patients’ experiences with fertility counselling and fertility preservation—a qualitative small-scale study within the Danish health care setting

    Science.gov (United States)

    Hoeg, Didde; Schmidt, Lone; Macklon, Kirsten T.

    2016-01-01

    Introduction Fertility counselling for young women newly diagnosed with cancer is an important field of preconceptional counselling. This qualitative, small-scale study explored how young women newly diagnosed with cancer experienced specialized fertility preservation counselling and treatment in the public Danish health care system. Methods Semi-structured, in-depth interviews were conducted with five women below 40 years recently diagnosed with cancer. All women received fertility counselling by a fertility specialist at the Fertility Clinic, University Hospital of Copenhagen, Denmark before initiation of cancer treatment. Participants were interviewed at a place chosen by them, and interviews were recorded and transcribed verbatim. Data were analysed using systematic text condensation developed by Malterud and inspired by Giorgi’s phenomenological analysis. Results None of the participants were aware that chemotherapy could destroy their eggs. The participants described how specialized fertility counselling and fertility preservation contributed to a belief in life after cancer, which gave them hope that they would survive their cancer disease. Further, the women described how the possibility of fertility preservation removed a huge concern and enabled them to concentrate on their cancer treatment and on getting better. Conclusion Overall, the specialized fertility counselling and treatment to preserve fertility was highly valued. The women felt it gave them a choice about their future fertility. The fertility expert presented the various fertility-preserving scenarios, and the women were content that they had an actual choice. PMID:27413812

  3. Factors related to shell deaths during artificial incubation of ostrich eggs

    Directory of Open Access Journals (Sweden)

    Z. Brand

    2007-06-01

    Full Text Available he ostrich industry experiences a high rate of embryonic mortalities during artificial incubation of eggs. Embryonic deaths were studied from data recorded on 37 740 fertile eggs incubated artificially during the 1998-2005 breeding seasons. Roughly 10 000 eggs that sustained embryonic mortalities were classified according to the stage and nature of death, i.e. before 21 days of incubation, after 21 days of incubation, deaths after pipping and rotten eggs. Although infection may have played a role in ~1300 rotten eggs, no detailed knowledge of the pathogens involved was available. The remainder of deaths could not be related to pathogens and the deaths were thus generally referred to as non-infectious. The overall level of embryonic mortality in all the eggs studied was 28.5 %. Overall embryonic mortality was affected by incubator, with higher levels (57.0 % found in eggs incubated in an African Incubator(R and also in eggs that were transferred between incubators during incubation (38.1 %. Overall embryonic mortality also increased in eggs produced by older females. Eggs produced in the autumn had the highest level of embryonic mortality at 53.6 %, whereas eggs produced in the winter had a marginally higher level of embryonic mortalities of 29.2 % compared with eggs produced during summer (27.4 %. Eggs produced by South African (SA Black males crossed to Zimbabwean Blue females had high levels of embryonic losses of 45.7 %. The embryonic mortality of eggs produced by SA Blacks or Zimbabwean Blue breeding birds subjected to pure breeding was similar at ~33-34 %, but embryonic mortality was improved in eggs produced by Zimbabwean Blue males crossed to SA Black females (27 %. Embryonic mortality was increased in eggs that were set directly (32.0 % or subjected to longer than 6 days of storage (43.5 %. Embryonic mortality was affected by year. The results that were obtained will assist in determining non-infectious factors that have a negative

  4. Fertility preservation in Turner syndrome.

    Science.gov (United States)

    Grynberg, Michaël; Bidet, Maud; Benard, Julie; Poulain, Marine; Sonigo, Charlotte; Cédrin-Durnerin, Isabelle; Polak, Michel

    2016-01-01

    Premature ovarian insufficiency is a relatively rare condition that can appear early in life. In a non-negligible number of cases the ovarian dysfunction results from genetic diseases. Turner syndrome (TS), the most common sex chromosome abnormality in females, is associated with an inevitable premature exhaustion of the follicular stockpile. The possible or probable infertility is a major concern for TS patients and their parents, and physicians are often asked about possible options to preserve fertility. Unfortunately, there are no recommendations on fertility preservation in this group. The severely reduced follicle pool even during prepubertal life represents the major limit for fertility preservation and is the root of numerous questions regarding the competence of gametes or ovarian tissue crybanked. In addition, patients suffering from TS show higher than usual rates of spontaneous abortion, fetal anomaly, and maternal morbidity and mortality, which should be considered at the time of fertility preservation and before reutilization of the cryopreserved gametes. Apart from fulfillment of the desire of becoming genetic parents, TS patients may be potential candidates for egg donation, gestational surrogacy, and adoption. The present review discusses the different options for preserving female fertility in TS and the ethical questions raised by these approaches. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  5. 7 CFR 1250.306 - Commercial eggs or eggs.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false Commercial eggs or eggs. 1250.306 Section 1250.306... AGREEMENTS AND ORDERS; MISCELLANEOUS COMMODITIES), DEPARTMENT OF AGRICULTURE EGG RESEARCH AND PROMOTION Egg Research and Promotion Order Definitions § 1250.306 Commercial eggs or eggs. Commercial eggs or eggs means...

  6. Fertilization capacity with rainbow trout DNA-damaged sperm and embryo developmental success.

    Science.gov (United States)

    Pérez-Cerezales, S; Martínez-Páramo, S; Beirão, J; Herráez, M P

    2010-06-01

    Mammalian spermatozoa undergo a strong selection process along the female tract to guarantee fertilization by good quality cells, but risks of fertilization with DNA-damaged spermatozoa have been reported. In contrast, most external fertilizers such as fish seem to have weaker selection procedures. This fact, together with their high prolificacy and external embryo development, indicates that fish could be useful for the study of the effects of sperm DNA damage on embryo development. We cryopreserved sperm from rainbow trout using egg yolk and low-density lipoprotein as additives to promote different rates of DNA damage. DNA fragmentation and oxidization were analyzed using comet assay with and without digestion with restriction enzymes, and fertilization trials were performed. Some embryo batches were treated with 3-aminobenzamide (3AB) to inhibit DNA repair by the poly (ADP-ribose) polymerase, which is an enzyme of the base excision repair pathway. Results showed that all the spermatozoa cryopreserved with egg yolk carried more than 10% fragmented DNA, maintaining fertilization rates of 61.1+/-2.3 but a high rate of abortions, especially during gastrulation, and only 14.5+/-4.4 hatching success. Furthermore, after 3AB treatment, hatching dropped to 3.2+/-2.2, showing that at least 10% DNA fragmentation was repaired. We conclude that trout sperm maintains its ability to fertilize in spite of having DNA damage, but that embryo survival is affected. Damage is partially repaired by the oocyte during the first cleavage. Important advantages of using rainbow trout for the study of processes related to DNA damage and repair during development have been reported.

  7. Induced ovulation and egg deposition in the direct developing anuran Eleutherodactylus coqui

    Directory of Open Access Journals (Sweden)

    Estrada Alberto R

    2004-01-01

    Full Text Available Abstract This study investigates ovulation and egg deposition behaviors in the anuran Eleutherodactylus coqui from Puerto Rico in response to stimulation with gonadotropin and gonadotropin releasing hormones. Five hormones were tested by injection over a range of doses, including mammalian LHRH, avian LHRH, fish LHRH, D-Ala6, des-Gly10 ethylamide LHRH and hCG. We report a low level of ovulation and egg deposition in response to all hormones, with the most complete and consistent results from the non-natural D-Ala6, des-Gly10 ethylamide LHRH derivative. To confirm the viability of eggs produced in this manner we performed in vitro fertilization experiments that resulted in the development of normal frogs. Reproductive behaviors in E. coqui are apparently not controlled by a mammalian form of LHRH as reported in other common laboratory anuran species. D-Ala6, des-Gly10 ethylamide LHRH induces ovulation and deposition of mature and fertilizable eggs in E. coqui.

  8. Effects of toxic compounds in Montipora capitata on exogenous and endogenous zooxanthellae performance and fertilization success.

    Directory of Open Access Journals (Sweden)

    Mary Hagedorn

    Full Text Available Studies have identified chemicals within the stony coral genus Montipora that have significant biological activities. For example, Montiporic acids A and B and other compounds have been isolated from the adult tissue and eggs of Montipora spp. and have displayed antimicrobial activity and cytotoxicity in cultured cells. The ecological role of these toxic compounds is currently unclear. This study examines the role these toxins play in reproduction. Toxins were found in the eggs and larvae of the coral Montipora capitata. Releasing these toxins by crushing both the eggs and larvae resulted in irreversible inhibition of photosynthesis in endogenous and exogenous zooxanthellae within minutes. Moreover, these toxins were stable, as frozen storage of eggs and larvae did not affect toxicity. Photosynthetic competency of Porites compressa zooxanthellae treated with either frozen or fresh, crushed eggs was inhibited similarly (P > 0.05, ANCOVA. Addition of toxic eggs plugs to live P. compressa fragments caused complete tissue necrosis under the exposed area on the fragments within 1 week. Small volumes of M. capitata crushed eggs added to sperm suspensions reduced in vitro fertilization success by killing the sperm. After 30 min, untreated sperm maintained 90 ± 1.9% SEM motility while those treated with crushed eggs were rendered immotile, 4 ± 1.4% SEM. Flow cytometry indicated membrane disruption of the immotile sperm. Fertilization success using untreated sperm was 79 ± 4% SEM, whereas the success rate dropped significantly after exposure to the crushed eggs, 1.3 ± 0% SEM. Unlike the eggs and the larvae, M. capitata sperm did not reduce the photosynthetic competency of P. compressa zooxanthellae, suggesting the sperm was nontoxic. The identity of the toxins, cellular mechanism of action, advantage of the toxins for M. capitata and their role on the reef are still unknown.

  9. Effects of toxic compounds in Montipora capitata on exogenous and endogenous zooxanthellae performance and fertilization success.

    Science.gov (United States)

    Hagedorn, Mary; Farrell, Ann; Carter, Virginia; Zuchowicz, Nikolas; Johnston, Erika; Padilla-Gamiño, Jacqueline; Gunasekera, Sarath; Paul, Valerie

    2015-01-01

    Studies have identified chemicals within the stony coral genus Montipora that have significant biological activities. For example, Montiporic acids A and B and other compounds have been isolated from the adult tissue and eggs of Montipora spp. and have displayed antimicrobial activity and cytotoxicity in cultured cells. The ecological role of these toxic compounds is currently unclear. This study examines the role these toxins play in reproduction. Toxins were found in the eggs and larvae of the coral Montipora capitata. Releasing these toxins by crushing both the eggs and larvae resulted in irreversible inhibition of photosynthesis in endogenous and exogenous zooxanthellae within minutes. Moreover, these toxins were stable, as frozen storage of eggs and larvae did not affect toxicity. Photosynthetic competency of Porites compressa zooxanthellae treated with either frozen or fresh, crushed eggs was inhibited similarly (P > 0.05, ANCOVA). Addition of toxic eggs plugs to live P. compressa fragments caused complete tissue necrosis under the exposed area on the fragments within 1 week. Small volumes of M. capitata crushed eggs added to sperm suspensions reduced in vitro fertilization success by killing the sperm. After 30 min, untreated sperm maintained 90 ± 1.9% SEM motility while those treated with crushed eggs were rendered immotile, 4 ± 1.4% SEM. Flow cytometry indicated membrane disruption of the immotile sperm. Fertilization success using untreated sperm was 79 ± 4% SEM, whereas the success rate dropped significantly after exposure to the crushed eggs, 1.3 ± 0% SEM. Unlike the eggs and the larvae, M. capitata sperm did not reduce the photosynthetic competency of P. compressa zooxanthellae, suggesting the sperm was nontoxic. The identity of the toxins, cellular mechanism of action, advantage of the toxins for M. capitata and their role on the reef are still unknown.

  10. Scrambled or bisected mouse eggs and the basis of patterning in mammals.

    Science.gov (United States)

    Gardner, R L

    1999-04-01

    Several findings challenge the notion that specification of cell types and embryonic axes in mammals are rooted entirely in the temporal and spatial relations between cleaving blastomeres. They raise the question as to whether, as in most non-mammalian species, these processes depend on information already present in the egg. However, experiments designed to investigate this possibility directly by perturbing the organization of the zygote or, very recently, by deleting one or other of its polar regions [M. Zernicka-Goetz. Fertile offspring derived from mammalian eggs lacking either animal or vegetal poles. Development 1998;125:4803-4808 (Ref. 1)], have been interpreted to mean that such a role for the egg can be discounted. This conclusion seems premature in view of continuing uncertainty regarding the developmental potential of individual blastomeres in mammals.

  11. Reproductive broodstock performance and egg quality of wild-caught and first-generation domesticated Seriola rivoliana reared under same culture conditions

    Directory of Open Access Journals (Sweden)

    Marcos F Quiñones-Arreola

    2015-11-01

    Full Text Available Almaco jack, Seriola rivoliana as well as some related species is of great interest in marine fish aquaculture. However, there are few studies about their reproduction in captivity. In this research work, reproductive performance and egg quality in two groups of adult Seriola rivoliana, caught in the wild and domesticated-F1 analyzed and compared, reared under optimal maturation conditions in a commercial private Laboratory. A total of 28 wild adult (>5 kg were caught at La Paz Bay, Baja California Sur, Mexico, and 30 adult domesticated-F1 broodstock (>5 kg, were obtained from an original stock of 1,000 juveniles (3.5 g body weight produced at Kona Blue (Hawaii, USA sea farm. Fishes were transported to the Rancheros del Mar commercial private hatchery, where they were grown to adult size. Both groups were evaluated during eight months (May to December 2012 and compared in terms of reproduction performance (total number of spawning events, monthly spawning frequency, total number of eggs, total number of eggs per mL, and fertilization rate, egg biochemical composition (total proteins, total lipids, total carbohydrates, and triacylglycerides and egg diameter. Results indicated that wild caught broostock showed a better reproductive performance in terms of fertilization rate, total number of spawning, monthly spawning frequency and total number of eggs produced. However, biochemical composition and egg diameter did not show statistical differences (P < 0.05 between two groups. The reproductive performance of broodstock and quality of eggs analyzed in this study are important traits to improve the aquaculture management of this species.

  12. Blood vessels segmentation of hatching eggs based on fully convolutional networks

    Science.gov (United States)

    Geng, Lei; Qiu, Ling; Wu, Jun; Xiao, Zhitao

    2018-04-01

    FCN, trained end-to-end, pixels-to-pixels, predict result of each pixel. It has been widely used for semantic segmentation. In order to realize the blood vessels segmentation of hatching eggs, a method based on FCN is proposed in this paper. The training datasets are composed of patches extracted from very few images to augment data. The network combines with lower layer and deconvolution to enables precise segmentation. The proposed method frees from the problem that training deep networks need large scale samples. Experimental results on hatching eggs demonstrate that this method can yield more accurate segmentation outputs than previous researches. It provides a convenient reference for fertility detection subsequently.

  13. Evaluation of developmental toxicity and teratogenicity of diclofenac using Xenopus embryos.

    Science.gov (United States)

    Chae, Jeong-Pil; Park, Mi Seon; Hwang, Yoo-Seok; Min, Byung-Hwa; Kim, Sang-Hyun; Lee, Hyun-Shik; Park, Mae-Ja

    2015-02-01

    Diclofenac is a non-steroidal anti-inflammatory drug (NSAID) with analgesic and anti-pyretic properties. This compound is therefore used to treat pain, inflammatory disorders, and dysmenorrhea. Due to its multimodal mechanism of action and ability to penetrate placenta, diclofenac is known to have undesirable side effects including teratogenicity. However, limited data exist on its teratogenicity, and a detailed investigation regarding harmful effects of this drug during embryogenesis is warranted. Here, we analyzed the developmental toxic effects of diclofenac using Xenopus embryos according to the Frog Embryo Teratogenesis Assay-Xenopus (FETAX) protocol. Diclofenac treatment exerted a teratogenic effect on Xenopus embryos with a teratogenic index (TI) value of 2.64 TI; if this value is higher than 1.2, the cut-off value indicative of toxicity. In particular, mortality of embryos treated with diclofenac increased in a concentration-dependent manner and a broad spectrum of malformations such as shortening and kinking of the axis, abdominal bulging, and prominent blister formation, was observed. The shape and length of internal organs also differed compared to the control group embryos and show developmental retardation on histological label. However, the expression of major tissue-specific markers did not change when analyzed by reverse transcription-polymerase chain reaction (RT-PCR). In conclusion, diclofenac treatment can promote teratogenicity that results in morphological anomalies, but not disrupt the developmental tissue arrangement during Xenopus embryogenesis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Does magnesium hardness in hatching waters affect the fertilization and hatching success of hybrid catfish eggs

    Science.gov (United States)

    Embryonic development is deemed to be the most sensitive stage in the life cycle of a teleost. As egg development takes outside the fish’s body, water hardness is one abioitic parameter, suggested to have a major effect on egg development and embryo survival. Ca2+ and Mg2+ contribute to water har...

  15. Effects of dietary fatty acids on the production and quality of eggs and larvae of Atlantic cod (Gadus morhua L.)

    DEFF Research Database (Denmark)

    Røjbek, Maria; Støttrup, Josianne; Jacobsen, Charlotte

    2014-01-01

    Cultivated Atlantic cod (Gadus morhua) entering their first year of gamete maturation were fed diets with different levels of arachidonic acid (ARA) and eicosapentaenoic acid (EPA) for 6.5 months prior to commencement of spawning. Gravid females were stripped three times: at the beginning, peak......–173% higher in eggs than in diets. Cod fed the diet with the lowest EPA/ARA ratio had the greatest egg production. Eggs from fish on a diet with high ARA level had significantly higher fertilization and hatching success than those fed low levels of ARA. This diet produced on average 71 viable eggs g 1 female...

  16. Relationships of chemical composition, quantity of milt to fertility and ...

    African Journals Online (AJOL)

    African Journal of Food, Agriculture, Nutrition and Development ... determining the right quantity of milt that can be used to fertilize certain quantity of eggs. ... quantity of milt used for artificial reproduction in fish significantly affect water quality used ... is the major organic substance that supplies the spermatozoa with energy.

  17. In-cell NMR spectroscopy of proteins inside Xenopus laevis oocytes

    International Nuclear Information System (INIS)

    Sakai, Tomomi; Tochio, Hidehito; Tenno, Takeshi; Ito, Yutaka; Kokubo, Tetsuro; Hiroaki, Hidekazu; Shirakawa, Masahiro

    2006-01-01

    In-cell NMR is an application of solution NMR that enables the investigation of protein conformations inside living cells. We have measured in-cell NMR spectra in oocytes from the African clawed frog Xenopus laevis. 15 N-labeled ubiquitin, its derivatives and calmodulin were injected into Xenopus oocytes and two-dimensional 1 H- 15 N correlation spectra of the proteins were obtained. While the spectrum of wild-type ubiquitin in oocytes had rather fewer cross-peaks compared to its in vitro spectrum, ubiquitin derivatives that are presumably unable to bind to ubiquitin-interacting proteins gave a markedly larger number of cross-peaks. This observation suggests that protein-protein interactions between ubiquitin and ubiquitin-interacting proteins may cause NMR signal broadening, and hence spoil the quality of the in-cell HSQC spectra. In addition, we observed the maturation of ubiquitin precursor derivative in living oocytes using the in-cell NMR technique. This process was partly inhibited by pre-addition of ubiquitin aldehyde, a specific inhibitor for ubiquitin C-terminal hydrolase (UCH). Our work demonstrates the potential usefulness of in-cell NMR with Xenopus oocytes for the investigation of protein conformations and functions under intracellular environmental conditions

  18. The Genome of the Western Clawed Frog Xenopus tropicalis

    Energy Technology Data Exchange (ETDEWEB)

    Hellsten, Uffe; Harland, Richard M.; Gilchrist, Michael J.; Hendrix, David; Jurka, Jerzy; Kapitonov, Vladimir; Ovcharenko, Ivan; Putnam, Nicholas H.; Shu, Shengqiang; Taher, Leila; Blitz, Ira L.; Blumberg, Bruce; Dichmann, Darwin S.; Dubchak, Inna; Amaya, Enrique; Detter, John C.; Fletcher, Russell; Gerhard, Daniela S.; Goodstein, David; Graves, Tina; Grigoriev, Igor V.; Grimwood, Jane; Kawashima, Takeshi; Lindquist, Erika; Lucas, Susan M.; Mead, Paul E.; Mitros, Therese; Ogino, Hajime; Ohta, Yuko; Poliakov, Alexander V.; Pollet, Nicolas; Robert, Jacques; Salamov, Asaf; Sater, Amy K.; Schmutz, Jeremy; Terry, Astrid; Vize, Peter D.; Warren, Wesley C.; Wells, Dan; Wills, Andrea; Wilson, Richard K.; Zimmerman, Lyle B.; Zorn, Aaron M.; Grainger, Robert; Grammer, Timothy; Khokha, Mustafa K.; Richardson, Paul M.; Rokhsar, Daniel S.

    2009-10-01

    The western clawed frog Xenopus tropicalis is an important model for vertebrate development that combines experimental advantages of the African clawed frog Xenopus laevis with more tractable genetics. Here we present a draft genome sequence assembly of X. tropicalis. This genome encodes over 20,000 protein-coding genes, including orthologs of at least 1,700 human disease genes. Over a million expressed sequence tags validated the annotation. More than one-third of the genome consists of transposable elements, with unusually prevalent DNA transposons. Like other tetrapods, the genome contains gene deserts enriched for conserved non-coding elements. The genome exhibits remarkable shared synteny with human and chicken over major parts of large chromosomes, broken by lineage-specific chromosome fusions and fissions, mainly in the mammalian lineage.

  19. Crystallization and X-ray analysis of the salmon-egg lectin SEL24K

    Energy Technology Data Exchange (ETDEWEB)

    Murata, Kenji [Department of Animal Science, University of California, Davis 95616 (United States); Fisher, Andrew J. [Department of Chemistry, University of California, Davis 95616 (United States); Hedrick, Jerry L., E-mail: jlhedrick@ucdavis.edu [Department of Animal Science, University of California, Davis 95616 (United States)

    2007-05-01

    The 24 kDa egg lectin of Chinook salmon (Oncorhynchus tshawytscha) was purified by affinity chromatography from salmon eggs and crystallized by the hanging-drop vapor-diffusion method using 15/4 EO/OH (pentaerythritol ethoxylate) as a precipitant. The 24 kDa egg lectin of Chinook salmon (Oncorhynchus tshawytscha) is released from the egg during the cortical reaction. The lectin functions in blocking polyspermy during the fertilization process. The egg lectin was purified by affinity chromatography from salmon eggs and crystallized by the hanging-drop vapor-diffusion method using 15/4 EO/OH (pentaerythritol ethoxylate) as a precipitant. The crystal diffracted synchrotron-radiation X-rays to 1.63 Å resolution. The crystal belongs to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 93.0, b = 73.6, c = 113.6 Å, α = 90, β = 92.82, γ = 90°. The crystal is likely to contain eight molecules in the asymmetric unit (V{sub M} = 2.3 Å{sup 3} Da{sup −1}), corresponding to a solvent content of 45.5%. A self-rotation function suggests an arrangement with 222 point symmetry within the asymmetric unit.

  20. Activation of eggs of the frog, Rana nigromaculata Hallowell, after localized UV-irradiation

    International Nuclear Information System (INIS)

    Sambuichi, Hajime

    1981-01-01

    To analyze the mode of transmission of the activating stimulus, the change in time course of the 2nd maturation division was examined in eggs of the frog, Rana nigromaculata, which were activated by insemination or by pricking after UV-irradiation at either the animal or the vegetal hemisphere. Irradiation of the animal (UAF) or the vegetal (UVF) hemisphere before insemination retarded the 2nd maturation division in comparison with that in normally fertilized eggs (CF). When the eggs were pricked at the irradiated surface (UAP and UVP, respectively), however, the 2nd maturation division progressed faster than that of CF. The comparison of the velocity of the 2nd maturation division between UAF and UVF, and also between UAP and UVP, showed that the cortical reaction in the vegetal hemisphere was affected by UV-irradiation more severely than that in the animal hemisphere. The results suggested that the quality of the granules or the constituent within the egg cortex might vary according to the regional difference in the egg surface. (author)

  1. It's what's inside that counts: egg contaminant concentrations are influenced by estimates of egg density, egg volume, and fresh egg mass.

    Science.gov (United States)

    Herzog, Mark P; Ackerman, Joshua T; Eagles-Smith, Collin A; Hartman, C Alex

    2016-05-01

    In egg contaminant studies, it is necessary to calculate egg contaminant concentrations on a fresh wet weight basis and this requires accurate estimates of egg density and egg volume. We show that the inclusion or exclusion of the eggshell can influence egg contaminant concentrations, and we provide estimates of egg density (both with and without the eggshell) and egg-shape coefficients (used to estimate egg volume from egg morphometrics) for American avocet (Recurvirostra americana), black-necked stilt (Himantopus mexicanus), and Forster's tern (Sterna forsteri). Egg densities (g/cm(3)) estimated for whole eggs (1.056 ± 0.003) were higher than egg densities estimated for egg contents (1.024 ± 0.001), and were 1.059 ± 0.001 and 1.025 ± 0.001 for avocets, 1.056 ± 0.001 and 1.023 ± 0.001 for stilts, and 1.053 ± 0.002 and 1.025 ± 0.002 for terns. The egg-shape coefficients for egg volume (K v ) and egg mass (K w ) also differed depending on whether the eggshell was included (K v  = 0.491 ± 0.001; K w  = 0.518 ± 0.001) or excluded (K v  = 0.493 ± 0.001; K w  = 0.505 ± 0.001), and varied among species. Although egg contaminant concentrations are rarely meant to include the eggshell, we show that the typical inclusion of the eggshell in egg density and egg volume estimates results in egg contaminant concentrations being underestimated by 6-13 %. Our results demonstrate that the inclusion of the eggshell significantly influences estimates of egg density, egg volume, and fresh egg mass, which leads to egg contaminant concentrations that are biased low. We suggest that egg contaminant concentrations be calculated on a fresh wet weight basis using only internal egg-content densities, volumes, and masses appropriate for the species. For the three waterbirds in our study, these corrected coefficients are 1.024 ± 0.001 for egg density, 0.493 ± 0.001 for K v , and 0.505 ± 0.001 for K w .

  2. Irradiation of liquid egg, frozen egg, powdered egg, egg yolk and white of egg: reducing the population of Salmonella enteritidis and sensory aspects and physico-chemical

    International Nuclear Information System (INIS)

    Froehlich, Angela

    2004-01-01

    Eggs and their products have been incriminated in foodborne disease outbreaks due to Salmonella enteritidis contamination. Irradiation is a food preservation technology that could be applied to minimize the problem. The aim of this study was to determine the effect of irradiation in liquid and frozen egg as well as in powdered egg, egg yolk and egg white spiked with Salmonella enteritidis. Spiked samples of liquid egg, egg white and egg yolk were exposed to 0,5; 1,0; 1,5; 2,0; 2,5; 3,0 kGy and spiked samples of frozen and powdered egg were exposed to 0,5; 1,0; 1,5; 2,0; 2,5; 3,0; 3,5 e 4,0 kGy. Raw odour, cooked odour and taste of non inoculated and irradiated samples and non irradiated samples of egg and egg products were analysed by a trained penal. Viscosity and lipid oxidation (malonaldehyde concentration) were also determined. Doses of 2,0; 3,0; 3,5; 3,0 e 3,5 kGy reduced in 5 log the population of S. Enteritidis in liquid and frozen egg, powdered egg yolk, egg white and egg, respectively, with moderate alterations in relation to non irradiated samples detected by the trained penal. Viscosity and lipid oxidation in the powdered products, however, showed more intense alterations. Therefore, irradiation can be considered a feasible process for liquid and frozen egg while when applied to powdered products it should be considered the type of food product to which they will be added due to alterations in viscosity. (author)

  3. A role for carbohydrate recognition in mammalian sperm-egg binding

    International Nuclear Information System (INIS)

    Clark, Gary F.

    2014-01-01

    Highlights: • Mammalian sperm-egg binding as a carbohydrate dependent species recognition event. • The role of carbohydrate recognition in human, mouse and pig sperm-egg binding. • Historical perspective and future directions for research focused on gamete binding. - Abstract: Mammalian fertilization usually requires three sequential cell–cell interactions: (i) initial binding of sperm to the specialized extracellular matrix coating the egg known as the zona pellucida (ZP); (ii) binding of sperm to the ZP via the inner acrosomal membrane that is exposed following the induction of acrosomal exocytosis; and (iii) adhesion of acrosome-reacted sperm to the plasma membrane of the egg cell, enabling subsequent fusion of these gametes. The focus of this review is on the initial binding of intact sperm to the mammalian ZP. Evidence collected over the past fifty years has confirmed that this interaction relies primarily on the recognition of carbohydrate sequences presented on the ZP by lectin-like egg binding proteins located on the plasma membrane of sperm. There is also evidence that the same carbohydrate sequences that mediate binding also function as ligands for lectins on lymphocytes that can inactivate immune responses, likely protecting the egg and the developing embryo up to the stage of blastocyst hatching. The literature related to initial sperm-ZP binding in the three major mammalian models (human, mouse and pig) is discussed. Historical perspectives and future directions for research related to this aspect of gamete adhesion are also presented

  4. A role for carbohydrate recognition in mammalian sperm-egg binding

    Energy Technology Data Exchange (ETDEWEB)

    Clark, Gary F., E-mail: clarkgf@health.missouri.edu

    2014-08-01

    Highlights: • Mammalian sperm-egg binding as a carbohydrate dependent species recognition event. • The role of carbohydrate recognition in human, mouse and pig sperm-egg binding. • Historical perspective and future directions for research focused on gamete binding. - Abstract: Mammalian fertilization usually requires three sequential cell–cell interactions: (i) initial binding of sperm to the specialized extracellular matrix coating the egg known as the zona pellucida (ZP); (ii) binding of sperm to the ZP via the inner acrosomal membrane that is exposed following the induction of acrosomal exocytosis; and (iii) adhesion of acrosome-reacted sperm to the plasma membrane of the egg cell, enabling subsequent fusion of these gametes. The focus of this review is on the initial binding of intact sperm to the mammalian ZP. Evidence collected over the past fifty years has confirmed that this interaction relies primarily on the recognition of carbohydrate sequences presented on the ZP by lectin-like egg binding proteins located on the plasma membrane of sperm. There is also evidence that the same carbohydrate sequences that mediate binding also function as ligands for lectins on lymphocytes that can inactivate immune responses, likely protecting the egg and the developing embryo up to the stage of blastocyst hatching. The literature related to initial sperm-ZP binding in the three major mammalian models (human, mouse and pig) is discussed. Historical perspectives and future directions for research related to this aspect of gamete adhesion are also presented.

  5. Interactions among motility, fertilizing ability, and testosterone binding on spermatozoa of bonnet monkey (Macaca radiata).

    Science.gov (United States)

    Warikoo, P K; Majumdar, S S; Allag, I S; Das, R P; Roy, S

    1986-01-01

    Fresh ejaculates of bonnet monkeys were separated into fractions rich with highly motile and sluggishly motile spermatozoa. The motility, ability to fertilize zona-free hamster eggs, and distribution of testosterone-binding sites on spermatozoa were assessed to determine the relation between these sperm functions. Two parameters of objective assessment of motility--velocity and degree of flagellar bending--were significantly correlated with the ability to form pronuclei in zona-free hamster eggs. Only spermatozoa with good motility could form pronuclei, which might be important for assessment of the fertilizing ability. The motility was directly related to the distribution of testosterone-binding sites; the fraction having mostly motile spermatozoa was distributed over the sperm surface. The technique is simple and may be used to evaluate semen of nonhuman primates.

  6. Selection for reproductive traits in white egg stock using independent culling levels

    Directory of Open Access Journals (Sweden)

    GS Schmidt

    2005-12-01

    Full Text Available The objective of this study was to report the response to selection for fertility and hatchability in white egg layers strains, based on sire and dam performance, and the effects of inbreeding on these traits. Two selected white egg strains (CC and DD from Embrapa´s poultry program under multiple trait selection were compared to a control strain (CCc. The control strain was established by randomly mating one male from each sire family to a non-related female from each dam family and was used to monitor genetic change in the selected lines. CC and DD were selected using family and individual information on hen-day egg production up to 40 weeks of age, egg weight, sexual maturity and 16-week-old body weight over five generations. Independent culling levels (ICL were also used during population expansion to eliminate sires and dams with low fertility (FERT and hatchability (HATC. Initially, FERT and HATC were over 90.0% in both selected and control strains. FERT and HATC of the selected and the control strains were compared during the last generation. FERT averages were 93.8, 93.8 and 94.4%, and HATCH averages were 93.2, 91.6 and 93.1% for CC, DD and CCc respectively. FERT and HATCH means were not different among strains. Estimated inbreeding increased at a rate of 0.4% per generation in all strains. Selection using ICL was able to effectively maintain the high initial FERT and HATC levels, and provide potential for high selection intensity in other traits. Inbreeding depression was not observed for any trait, indicating that selection compensated for any negative effects of inbreeding.

  7. Cracking the egg: An insight into egg hypersensitivity.

    Science.gov (United States)

    Dhanapala, Pathum; De Silva, Chamika; Doran, Tim; Suphioglu, Cenk

    2015-08-01

    Hypersensitivity to the chicken egg is a widespread disorder mainly affecting 1-2% of children worldwide. It is the second most common food allergy in children, next to cow's milk allergy. Egg allergy is mainly caused by hypersensitivity to four allergens found in the egg white; ovomucoid, ovalbumin, ovotransferrin and lysozyme. However, some research suggests the involvement of allergens exclusively found in the egg yolk such as chicken serum albumin and YGP42, which may play a crucial role in the overall reaction. In egg allergic individuals, these allergens cause conditions such as itching, atopic dermatitis, bronchial asthma, vomiting, rhinitis, conjunctivitis, laryngeal oedema and chronic urticaria, and anaphylaxis. Currently there is no permanent cure for egg allergy. Upon positive diagnosis for egg allergy, strict dietary avoidance of eggs and products containing traces of eggs is the most effective way of avoiding future hypersensitivity reactions. However, it is difficult to fully avoid eggs since they are found in a range of processed food products. An understanding of the mechanisms of allergic reactions, egg allergens and their prevalence, egg allergy diagnosis and current treatment strategies are important for future studies. This review addresses these topics and discusses both egg white and egg yolk allergy as a whole. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Food restriction but not fish oil increases fertility in hens: role of RARRES2?

    Science.gov (United States)

    Mellouk, Namya; Ramé, Christelle; Delaveau, Joël; Rat, Christophe; Marchand, Maxime; Mercerand, Frédéric; Travel, Angélique; Brionne, Aurélien; Chartrin, Pascal; Ma, Linlin; Froment, Pascal; Dupont, Joëlle

    2018-04-01

    Overfed hens selected for their rapid growth become fatter and develop reproductive disorders. Herein, we aimed to demonstrate that food restriction leading to a weight reduction and/or a supplementation with fish oil may be effective in preventing reproductive disorders through the regulation of adipokine expression in broiler hens. This study included four groups of food restricted (Rt) or ad libitum hens (Ad, feeding at a rate 1.7 times greater than Rt hens) supplemented or unsupplemented with fish oil (1%). The Rt diet significantly increased plasma chemerin (RARRES2) levels during the laying period, delayed sexual maturity by one week and improved egg quality and fertility. These effects were associated with higher progesterone production in response to IGF1 (or LH) in cultured granulosa cells and in vivo egg yolk, as compared with Ad hens. Fish oil supplementation had similar effects to the Rt diet on progesterone ( P  food restriction but not fish oil supplementation improved fertility, and this was associated with variations in RARRES2 plasma and ovarian expression in hens. © 2018 Society for Reproduction and Fertility.

  9. Efeito do chumbo sobre a fertilização artificial e incubação de ovos de jundiá cinza (Rhamdia quelen = The effect of lead on artificial fertilization and incubation of gray jundiá (Rhamdia quelen eggs

    Directory of Open Access Journals (Sweden)

    Cleonice Cristina Hilbig

    2008-04-01

    Full Text Available A poluição no meio aquático afeta o potencial reprodutivo de várias espécies de peixes. Diante disso, o presente experimento foi conduzido com o objetivo de avaliar os efeitos causados pela contaminação da água utilizada no processo de fertilização artificial e incubação inicial de ovos de jundiá cinza (Rhamdia quelen por chumbo. Foram realizados dois ensaios, os quais foram distribuídos em delineamento experimental inteiramente casualizado, compostos por seis tratamentos, com quatro e seis repetições, para o primeiro eo segundo ensaio, respectivamente. O primeiro ensaio consistiu na contaminação da água utilizada no processo de fertilização artificial, com os níveis de 0,00; 0,01; 0,02; 0,03; 0,04 e0,05 mg de PB L-1, e incubação dos ovos em água limpa. O segundo ensaio utilizou os níveis de contaminação de 0,00; 0,15; 0,30; 0,45; 0,60 e 0,75 mg de PB L -1 de água utilizada nos processos de fertilização e de incubação dos ovos. Observou-se que apenas no segundo ensaio houve efeito (p Aquatic environmental pollution affects the reproductive potential of various species of fish. The present experiment was carried out to evaluate the effect of lead contamination of the water used in artificial fertilization and initial egg incubation of gray jundiá (Rhamdia quelen. Two assays were carried out distributed inrandomly experimental designs, composed by six treatments, with four and six replications, for the first and second assays, respectively. The first assay consisted of the contamination of artificial fertilization water with 0.00; 0.01; 0.02; 0.03; 0.04 and 0.05 mg of Pb L-1 and egg incubation in pure water. The second assay consisted of the contamination of artificial fertilization water and egg incubation water with 0.00; 0.15; 0.30; 0.45; 0.60 and 0.75 mg ofPb L-1. Only the second assay was affected (p < 0.05 by the treatments, showing a “plateau” tendency until 0.25 mg of Pb L-1. A linear reduction of the

  10. Expression of the adhesion G protein-coupled receptor A2 (adgra2) during Xenopus laevis development.

    Science.gov (United States)

    Seigfried, Franziska A; Dietmann, Petra; Kühl, Michael; Kühl, Susanne J

    2018-06-01

    The adhesion G protein-coupled receptor A2 (Adgra2) is a seven transmembrane receptor that has been described to be a regulator for angiogenesis in mice. Furthermore, the zebrafish ouchless mutant is unable to develop dorsal root ganglia through a disrupted trafficking of Adgra2. Besides RNA sequencing data, nothing is reported about Adgra2 in the south African crawled frog Xenopus laevis. In this study, we investigated for the first time the spatio-temporal expression of adgra2 during early Xenopus embryogenesis in detail. In silico approaches showed that the genomic adgra2 region as well as the Adgra2 protein sequence is highly conserved among different species including Xenopus. RT-PCR experiments confirmed that embryonic adgra2 expression is primarily detected at the beginning of neurulation and is then present throughout the whole Xenopus embryogenesis until stage 42. Whole mount in situ hybridization approaches visualized adgra2 expression in many tissues during Xenopus embryogenesis such as the cardiovascular system including the heart, the migrating neural crest cells and the developing eye including the periocular mesenchyme. Our results indicate a role of Adgra2 for embryogenesis and are a good starting point for further functional studies during early vertebrate development. Copyright © 2018 Elsevier B.V. All rights reserved.

  11. Enteropathogenic bacterial contamination of a latosol following application of organic fertilizer

    Directory of Open Access Journals (Sweden)

    Pedro Alexandre Escosteguy

    2015-10-01

    Full Text Available Poultry manure is used as fertilizer in natura, but little is known about whether it contaminates the soil with pathogenic organisms. The aim of this study was to assess the effects of organic, organomineral and mineral fertilizers on soil contamination by enteric pathogens, using poultry manure as the organic fertilizer. Manure was applied in field experiments at rates of 7.0 ton. ha-1 (maize crop, 2008/2009, 8.0 ton. ha-1 (wheat crop, 2009 and 14 ton. ha-1 (maize crop, 2010/2011. Organomineral fertilizer was applied at the same rates but was comprised of 50% manure and 50% mineral fertilizer. At 30 and 70 days after fertilization, the organic fertilizer and the upper 0-5 cm layer of the soil were tested for the presence of helminth eggs and larvae and enteropathogenic bacteria. Fecal and non-fecal coliforms (Escherichia coli and Clostridium perfringes were found in the organic fertilizer, but neither Salmonella spp. nor enteroparasites were detected. The population of enteropathogenic bacteria in the soil was similar among the treatments for all crops at both evaluation times. The population of thermotolerant coliforms in the organic fertilizer was larger than the maximum level allowed in Brazil, but neither the organic or nor the organomineral fertilizer contaminated the soil.

  12. Phosphorylation of both nucleoplasmin domains is required for activation of its chromatin decondensation activity

    DEFF Research Database (Denmark)

    Bañuelos, Sonia; Omaetxebarria, Miren J; Ramos, Isbaal

    2007-01-01

    Nucleoplasmin (NP) is a histone chaperone involved in nucleosome assembly, chromatin decondensation at fertilization, and apoptosis. To carry out these activities NP has to interact with different types of histones, an interaction that is regulated by phosphorylation. Here we have identified...... are found at the tail domain, flanking the nuclear localization signal. Phosphorylation-mimicking mutations render a recombinant protein as active in chromatin decondensation as hyperphosphorylated NP isolated from Xenopus laevis eggs. Comparison of mutants in which the core and tail domains of the protein...... were independently or simultaneously "activated" indicates that activation or phosphorylation of both protein domains is required for NP to efficiently extract linker-type histones from chromatin....

  13. Evidence of Biased Advertising in the Case of Social Egg Freezing.

    Science.gov (United States)

    Barbey, Christopher

    2017-11-01

    Oocyte cryopreservation, or 'egg freezing,' is the practice of preserving unfertilised oocytes for later fertilisation. This practice allows women to extend their reproductive years. In 2014, Facebook and Apple announced that they would subsidise their female employees' elective - or 'social' - use of egg freezing so that these women can more easily reconcile the demands of career and family life. This announcement engendered controversy and moral debate. Given that social egg freezing (SEF) is becoming more popular, ethical and empirical analyses are warranted. Here, I utilise content analysis to examine media messages in advertising for SEF. I conclude that many fertility clinics engage in biased advertising - i.e. they advertise the service persuasively, not informatively, emphasising indirect benefits while minimising risks and the low chance of successfully bringing a child to term. As advertising for medical services has been shown to influence the use of those services, advertising for SEF should emphasise clear and easily interpretable statistics about success rate, usage rate, cost, and risk.

  14. A sensory bias has triggered the evolution of egg-spots in cichlid fishes.

    Directory of Open Access Journals (Sweden)

    Bernd Egger

    Full Text Available Although, generally, the origin of sex-limited traits remains elusive, the sensory exploitation hypothesis provides an explanation for the evolution of male sexual signals. Anal fin egg-spots are such a male sexual signal and a key characteristic of the most species-rich group of cichlid fishes, the haplochromines. Males of about 1500 mouth-brooding species utilize these conspicuous egg-dummies during courtship--apparently to attract females and to maximize fertilization success. Here we test the hypothesis that the evolution of haplochromine egg-spots was triggered by a pre-existing bias for eggs or egg-like coloration. To this end, we performed mate-choice experiments in the basal haplochromine Pseudocrenilabrus multicolor, which manifests the plesiomorphic character-state of an egg-spot-less anal fin. Experiments using computer-animated photographs of males indeed revealed that females prefer images of males with virtual ('in-silico' egg-spots over images showing unaltered males. In addition, we tested for color preferences (outside a mating context in a phylogenetically representative set of East African cichlids. We uncovered a strong preference for yellow, orange or reddish spots in all haplochromines tested and, importantly, also in most other species representing more basal lines. This pre-existing female sensory bias points towards high-quality (carotenoids-enriched food suggesting that it is adaptive.

  15. Identification of genes associated with regenerative success of Xenopus laevis hindlimbs

    Directory of Open Access Journals (Sweden)

    Barker Donna

    2008-06-01

    Full Text Available Abstract Background Epimorphic regeneration is the process by which complete regeneration of a complex structure such as a limb occurs through production of a proliferating blastema. This type of regeneration is rare among vertebrates but does occur in the African clawed frog Xenopus laevis, traditionally a model organism for the study of early development. Xenopus tadpoles can regenerate their tails, limb buds and the lens of the eye, although the ability of the latter two organs to regenerate diminishes with advancing developmental stage. Using a heat shock inducible transgene that remains silent unless activated, we have established a stable line of transgenic Xenopus (strain N1 in which the BMP inhibitor Noggin can be over-expressed at any time during development. Activation of this transgene blocks regeneration of the tail and limb of Xenopus tadpoles. Results In the current study, we have taken advantage of the N1 transgenic line to directly compare morphology and gene expression in same stage regenerating vs. BMP signalling deficient non-regenerating hindlimb buds. The wound epithelium of N1 transgenic hindlimb buds, which forms over the cut surface of the limb bud after amputation, does not transition normally into the distal thickened apical epithelial cap. Instead, a basement membrane and dermis form, indicative of mature skin. Furthermore, the underlying mesenchyme remains rounded and does not expand to form a cone shaped blastema, a normal feature of successful regeneration. Using Affymetrix Gene Chip analysis, we have identified genes linked to regenerative success downstream of BMP signalling, including the BMP inhibitor Gremlin and the stress protein Hsp60 (no blastema in zebrafish. Gene Ontology analysis showed that genes involved in embryonic development and growth are significantly over-represented in regenerating early hindlimb buds and that successful regeneration in the Xenopus hindlimb correlates with the induction of

  16. Soya-lecithin in extender improves the freezability and fertility of buffalo (Bubalus bubalis) bull spermatozoa.

    Science.gov (United States)

    Akhter, S; Ansari, M S; Andrabi, S M H; Rakha, B A; Ullah, N; Khalid, M

    2012-10-01

    Egg yolk is routinely used as a cryoprotectant in semen extenders. However, it may contain cryoprotective antagonists, and there are hygienic risks associated with its use. Proteins of plant origin, like soya-lecithin, lack these hazards. The aim of this study was to use soya-lecithin as a cryoprotectant in extender and to investigate its effects on in vitro quality and in vivo fertility of buffalo semen. Semen from three buffalo bulls was frozen in tris-citric extender containing 5.0%, 10% or 15% soya-lecithin or 20% egg yolk. Sperm motility, plasma membrane integrity and viability were assessed post-dilution, pre-freezing and post-thaw. In Post-dilution and pre-freezing, the values for motility, plasma membrane integrity and viability remained higher (p ≤ 0.05) in extenders containing 10% soya-lecithin and control compared with extender containing 5% and 15% soya-lecithin. However, motility, plasma membrane integrity and viability were higher (p lecithin compared with control and extenders containing 5% and 15% soya-lecithin. Semen from two buffalo bulls was frozen in tris-citric extender containing either 10% soya-lecithin or 20% egg yolk. Higher (p lecithin (56%) compared with 20% egg yolk (41.5%). The results suggest that 10% soya-lecithin in extender improves the freezability and fertility of buffalo bull spermatozoa and can be used as an alternate to egg yolk in cryopreservation of buffalo semen. © 2011 Blackwell Verlag GmbH.

  17. Sexual reproduction of Nausithoe aurea (Scyphozoa, Coronatae. Gametogenesis, egg release, embryonic development, and gastrulation

    Directory of Open Access Journals (Sweden)

    André C. Morandini

    2001-06-01

    Full Text Available The structure of the ovaries and testes of Nausithoe aurea, reared in the laboratory, is described to update the knowledge of coronate scyphomedusae gametogenesis and early development. The testis is similar to those of other scyphozoans. The organization of the ovary agrees with the description for other coronates, with free oocytes in the mesoglea. The oocytes develop in a limited region of the gastrodermis, and a maturation gradient is observed from this point on. Egg release, embryonic development, and gastrulation mode of Nausithoe aurea are also described. Egg production was continuous for 55 days, and the output of released eggs oscillated without observed cue. Cleavage was holoblastic and adequal, but after the 8-cell stage, the cleavage became pseudospiral. Gastrulation occurred through multipolar ingression and began 24 hours after fertilization.

  18. Polarity and reorganization of the endoplasmic reticulum during fertilization and ooplasmic segregation in the ascidian egg

    OpenAIRE

    1993-01-01

    During the first cell cycle of the ascidian egg, two phases of ooplasmic segregation create distinct cytoplasmic domains that are crucial for later development. We recently defined a domain enriched in ER in the vegetal region of Phallusia mammillata eggs. To explore the possible physiological and developmental function of this ER domain, we here investigate its organization and fate by labeling the ER network in vivo with DiIC16(3), and observing its distribution before and after fertilizati...

  19. Developing Xenopus Embryos Recover by Compacting and Expelling Single-Wall Carbon Nanotubes

    Science.gov (United States)

    Holt, Brian D.; Shawky, Joseph H.; Dahl, Kris Noel; Davidson, Lance A.; Islam, Mohammad F.

    2015-01-01

    Single-wall carbon nanotubes are high aspect ratio nanomaterials that are being developed for use in materials, technological and biological applications due to their high mechanical stiffness, optical properties, and chemical inertness. Because of their prevalence, it is inevitable that biological systems will be exposed to nanotubes, yet studies of the effects of nanotubes on developing embryos have been inconclusive and are lacking for single-wall carbon nanotubes exposed to the widely studied model organism Xenopus laevis (African clawed frog). Microinjection of experimental substances into the Xenopus embryo is a standard technique for toxicology studies and cellular lineage tracing. Here we report the surprising finding that superficial (12.5 ± 7.5 μm below the membrane) microinjection of nanotubes dispersed with Pluronic F127 into one-to-two cell Xenopus embryos resulted in the formation and expulsion of compacted, nanotube-filled, punctate masses, at the blastula to mid-gastrula developmental stages, which we call “boluses”. Such expulsion of microinjected materials by Xenopus embryos has not been reported before and is dramatically different from the typical distribution of the materials throughout the progeny of the microinjected cells. Previous studies of microinjections of nanomaterials such as nanodiamonds, quantum dots or spherical nanoparticles report that nanomaterials often induce toxicity and remain localized within the embryos. In contrast, our results demonstrate an active recovery pathway for embryos after exposure to Pluronic F127-coated nanotubes, which we speculate is due to a combined effect of the membrane activity of the dispersing agent, Pluronic F127, and the large aspect ratio of nanotubes. PMID:26153061

  20. Developing Xenopus embryos recover by compacting and expelling single wall carbon nanotubes.

    Science.gov (United States)

    Holt, Brian D; Shawky, Joseph H; Dahl, Kris Noel; Davidson, Lance A; Islam, Mohammad F

    2016-04-01

    Single wall carbon nanotubes are high aspect ratio nanomaterials being developed for use in materials, technological and biological applications due to their high mechanical stiffness, optical properties and chemical inertness. Because of their prevalence, it is inevitable that biological systems will be exposed to nanotubes, yet studies of the effects of nanotubes on developing embryos have been inconclusive and are lacking for single wall carbon nanotubes exposed to the widely studied model organism Xenopus laevis (African clawed frog). Microinjection of experimental substances into the Xenopus embryo is a standard technique for toxicology studies and cellular lineage tracing. Here we report the surprising finding that superficial (12.5 ± 7.5 µm below the membrane) microinjection of nanotubes dispersed with Pluronic F127 into one- to two-cell Xenopus embryos resulted in the formation and expulsion of compacted, nanotube-filled, punctate masses, at the blastula to mid-gastrula developmental stages, which we call "boluses." Such expulsion of microinjected materials by Xenopus embryos has not been reported before and is dramatically different from the typical distribution of the materials throughout the progeny of the microinjected cells. Previous studies of microinjections of nanomaterials such as nanodiamonds, quantum dots or spherical nanoparticles report that nanomaterials often induce toxicity and remain localized within the embryos. In contrast, our results demonstrate an active recovery pathway for embryos after exposure to Pluronic F127-coated nanotubes, which we speculate is due to a combined effect of the membrane activity of the dispersing agent, Pluronic F127, and the large aspect ratio of nanotubes. Copyright © 2015 John Wiley & Sons, Ltd.

  1. Apoptosis regulates notochord development in Xenopus

    OpenAIRE

    Malikova, Marina; Van Stry, Melanie; Symes, Karen

    2007-01-01

    The notochord is the defining characteristic of the chordate embryo, and plays critical roles as a signaling center and as the primitive skeleton. In this study we show that early notochord development in Xenopus embryos is regulated by apoptosis. We find apoptotic cells in the notochord beginning at the neural groove stage and increasing in number as the embryo develops. These dying cells are distributed in an anterior to posterior pattern that is correlated with notochord extension through ...

  2. Viability and fertility of cooled equine semen diluted with skimmed milk or glycine egg yolk-based extenders

    Directory of Open Access Journals (Sweden)

    Guilherme Pugliesi

    2012-12-01

    Full Text Available Two semen extenders were compared for their ability to maintain viability of horse semen during 24 hours of cold preservation, and for the pregnancy rate after artificial insemination. In the experiment 1, five ejaculates from three stallions were split-diluted in either a skimmed milk-based extender (Kenney extender or a glycine egg yolk-based extender (Foote extender and cooled at 6-8 ºC for 24 hours. Semen samples stored in Kenney extender for 24 hours had higher motility and spermatic vigor compared with those stored in Foote extender. However, samples stored in Foote extender had higher number of reactive sperm by hypoosmotic test and greater viability by epifluorescence test compared with those in Kenney extender. In the experiment 2, 17 and 23 ejaculates from two stallions were split-diluted with Kenney extender and Foote extender. The sperm concentration in each extender was adjusted to 500 million viable sperms per insemination dose. Semen was cooled to 6-8 ºC and stored for 24 hours. Seventy-four cycles of crossbred mares were inseminated with either semen diluted in Kenney extender or semen diluted in Foote extender. The pregnancy rate was higher from semen diluted in Kenney extender than that from semen in Foote extender (0.553 vs. 0.306. The Kenney extender is effective in preserving the motility, vigor and fertility of stallion semen after 24 hours of cold storage, whereas the Foote extender is not acceptable.

  3. Xenopus iaevis (Anura: Pipidae) Mating systems - A preliminary ...

    African Journals Online (AJOL)

    was inhibited if the frogs were injected with saline to simulate ripe ovaries. Russel (1954) described a low frequency tremor given by males when in amplexus with females. This behaviour was also noted by Grimm (1952). Although many authors have provided verbal or phonetic descriptions of the calls of Xenopus laevis, ...

  4. Comparative biology of sperm factors and fertilization-induced calcium signals across the animal kingdom.

    Science.gov (United States)

    Kashir, Junaid; Deguchi, Ryusaku; Jones, Celine; Coward, Kevin; Stricker, Stephen A

    2013-10-01

    Fertilization causes mature oocytes or eggs to increase their concentrations of intracellular calcium ions (Ca²⁺) in all animals that have been examined, and such Ca²⁺ elevations, in turn, provide key activating signals that are required for non-parthenogenetic development. Several lines of evidence indicate that the Ca²⁺ transients produced during fertilization in mammals and other taxa are triggered by soluble factors that sperm deliver into oocytes after gamete fusion. Thus, for a broad-based analysis of Ca²⁺ dynamics during fertilization in animals, this article begins by summarizing data on soluble sperm factors in non-mammalian species, and subsequently reviews various topics related to a sperm-specific phospholipase C, called PLCζ, which is believed to be the predominant activator of mammalian oocytes. After characterizing initiation processes that involve sperm factors or alternative triggering mechanisms, the spatiotemporal patterns of Ca²⁺ signals in fertilized oocytes or eggs are compared in a taxon-by-taxon manner, and broadly classified as either a single major transient or a series of repetitive oscillations. Both solitary and oscillatory types of fertilization-induced Ca²⁺ signals are typically propagated as global waves that depend on Ca²⁺ release from the endoplasmic reticulum in response to increased concentrations of inositol 1,4,5-trisphosphate (IP₃). Thus, for taxa where relevant data are available, upstream pathways that elevate intraoocytic IP3 levels during fertilization are described, while other less-common modes of producing Ca²⁺ transients are also examined. In addition, the importance of fertilization-induced Ca²⁺ signals for activating development is underscored by noting some major downstream effects of these signals in various animals. © 2013 Wiley Periodicals, Inc.

  5. Milt-Egg Ratio in Artificial Fertilization of Pangasiid Catfish Injected by Gonadotropin Releasing Hormone-Analog (GnRH-a and Domperidone Mixture

    Directory of Open Access Journals (Sweden)

    J. Subagja

    2007-08-01

    Full Text Available Study on the level of gonadotropin hormone treatments combined with latency time to induce ovulation in Pangasius djambal was conducted in the Research Instalation of Germ Plasm, Cijeruk, Bogor. This study was carried out to investigate the effect of GnRH-a and domperidone mixture on the milt production and fertilization rates of Pangasius djambal, and to determine the optimal milt-egg ratio required for artificial fertilization. Different doses of hormone i.e: 0,3; 0,5 and 0,7 ml kg"1 body weight combined with latency time of 12, 24 and 48 h after inducing hormone were applied to increase milt-production. Milt dilution was 10"1, 10~2, 10"3, 10"4, 10"5, 10"6 and 10"7and evaluated for hatching rate and normality of larvae. The results showed that mean milt production was 4,3 ml/kg body weight, and there was interaction between hormone dose of 0,5 ml/kg of body weight and latency time 12 and 24 h that giving hatching rate of 77 to 83% ( p Key words : Fertilization, milt production, domperidone, Pangasius djambal   ABSTRAK Suatu studi penyuntikan hormon gonadotropin dengan perbedaan dosis dan waktu laten terhadap spesies Pangasius djambal telah dilakukan di Instalasi Riset Plasma Nutfah Air Tawar, Cijeruk, Bogor. Tujuan penelitian ini adalah untuk mengetahui pengaruh hormon GnRH-a dan domperidon terhadap produktivitas semen ikan jantan dan viabilitasnya pada pembuahan buatan. Tujuan lainnya untuk menentukan perbandingan optimal antara jumlah spermatozoa dengan telur dalam fertilisasi buatan. Dosis hormon perlakuan untuk peningkatan produksi semen yaitu 0,3; 0,5 dan 0,7 ml.kg"1 bobot badan yang di kombinasikan dengan waktu inkubasi jantan 12, 24 dan 24 jam setelah penyuntikan hormon. Semen diencerkan mulai dari 10"', 10"2, 10"3, 10"4, 10~5, 10'6 dan 10"7, dan dilakukan pembuahan terhadap telur. Daya tetas dan abnormalitas larva dievaluasi. Hasil analisis menunjukan produksi semen rata-rata 4,3 ml/kg bobot badan, ada interaksi antara dosis hormon 0

  6. HPLC Quantification of astaxanthin and canthaxanthin in Salmonidae eggs.

    Science.gov (United States)

    Tzanova, Milena; Argirova, Mariana; Atanasov, Vasil

    2017-04-01

    Astaxanthin and canthaxanthin are naturally occurring antioxidants referred to as xanthophylls. They are used as food additives in fish farms to improve the organoleptic qualities of salmonid products and to prevent reproductive diseases. This study reports the development and single-laboratory validation of a rapid method for quantification of astaxanthin and canthaxanthin in eggs of rainbow trout (Oncorhynchus mykiss) and brook trout (Salvelinus fontinalis М.). An advantage of the proposed method is the perfect combination of selective extraction of the xanthophylls and analysis of the extract by high-performance liquid chromatography and photodiode array detection. The method validation was carried out in terms of linearity, accuracy, precision, recovery and limits of detection and quantification. The method was applied for simultaneous quantification of the two xanthophylls in eggs of rainbow trout and brook trout after their selective extraction. The results show that astaxanthin accumulations in salmonid fish eggs are larger than those of canthaxanthin. As the levels of these two xanthophylls affect fish fertility, this method can be used to improve the nutritional quality and to minimize the occurrence of the M74 syndrome in fish populations. Copyright © 2016 John Wiley & Sons, Ltd.

  7. Proteomics analysis of egg white proteins from different egg varieties.

    Science.gov (United States)

    Wang, Jiapei; Liang, Yue; Omana, Dileep A; Kav, Nat N V; Wu, Jianping

    2012-01-11

    The market of specialty eggs, such as omega-3-enriched eggs, organic eggs, and free-range eggs, is continuously growing. The nutritional composition of egg yolk can be manipulated by feed diet; however, it is not known if there is any difference in the composition of egg white proteins among different egg varieties. The purpose of the study was to compare the egg white proteins among six different egg varieties using proteomics analysis. Egg white proteins were analyzed using two-dimensional gel electrophoresis (2-DE), and 89 protein spots were subjected to LC-MS/MS. A total of 23 proteins, belonging to Gallus gallus , were identified from 72 detected protein spots. A quiescence-specific protein precursor in egg white was identified for the first time in this study. Significant differences in the abundant levels of 19 proteins (from 65 protein spots) were observed among six egg varieties. Four proteins, ovalbumin-related protein Y, cystatin, avidin, and albumin precursor, were not different among these six egg varieties. These findings suggest that the abundance, but not the composition, of egg white proteins varied among the egg varieties.

  8. Method of Euthanasia Influences the Oocyte Fertilization Rate with Fresh Mouse Sperm

    Science.gov (United States)

    Hazzard, Karen C; Watkins-Chow, Dawn E; Garrett, Lisa J

    2014-01-01

    In vitro fertilization (IVF) is used to produce mouse embryos for a variety of reasons. We evaluated the effect of the method of euthanasia on the fertilization rate in 2 different IVF protocols. Oocytes collected from C57BL/6J female mice euthanized by CO2 inhalation or cervical dislocation were used in IVF with fresh sperm from either wild-type or genetically engineered C57BL/6J. Compared with CO2 inhalation, cervical dislocation improved the resulting rate of fertilization by 18% in an IVF method using Cook media and by 13% in an IVF method using methyl-B cyclodextrin and reduced glutathione. The lower fertilization rate due to euthanasia by CO2 inhalation was accompanied by changes in blood pH and body temperature despite efforts to minimize temperature drops. In our hands, euthanasia by cervical dislocation improved fertilization rates and consequently reduced the number of egg-donor mice required. PMID:25650969

  9. Method of euthanasia influences the oocyte fertilization rate with fresh mouse sperm.

    Science.gov (United States)

    Hazzard, Karen C; Watkins-Chow, Dawn E; Garrett, Lisa J

    2014-11-01

    In vitro fertilization (IVF) is used to produce mouse embryos for a variety of reasons. We evaluated the effect of the method of euthanasia on the fertilization rate in 2 different IVF protocols. Oocytes collected from C57BL/6J female mice euthanized by CO2 inhalation or cervical dislocation were used in IVF with fresh sperm from either wild-type or genetically engineered C57BL/6J. Compared with CO2 inhalation, cervical dislocation improved the resulting rate of fertilization by 18% in an IVF method using Cook media and by 13% in an IVF method using methyl-B cyclodextrin and reduced glutathione. The lower fertilization rate due to euthanasia by CO2 inhalation was accompanied by changes in blood pH and body temperature despite efforts to minimize temperature drops. In our hands, euthanasia by cervical dislocation improved fertilization rates and consequently reduced the number of egg-donor mice required.

  10. A marker of animal-vegetal polarity in the egg of the sea urchin Paracentrotus lividus. The pigment band.

    Science.gov (United States)

    Sardet, C; Chang, P

    1985-09-01

    We have examined the subequatorial accumulation of pigment granules (the so-called 'pigment band') in the egg of the sea urchin Paracentrotus lividus, which constitutes an unambiguous marker of animal-vegetal polarity. Most of the reddish pigment granules are situated at the periphery of the egg. They exhibit occasional saltatory movements and can aggregate into large patches. Pigment granules are retained as a band in the isolated cortex when the egg surface complex is isolated by shearing eggs attached to polylysine-coated surfaces with calcium-free isotonic solutions. Pigment granules remain as the main vesicular component of fertilized egg cortices or of unfertilized egg cortices perfused with calcium to provoke cortical granule exocytosis. They may be anchored to the isolated cortex through associations with the plasma membrane and with an extensive subsurface network of rough endoplasmic reticulum (rough ER). Pigment granules contain antimonate-precipitable calcium and, in this respect and many others, resemble acidic vesicles recently identified in the cortex of unpigmented sea urchin eggs. We discuss the similarities observed between granules and acidic vesicles in various urchin egg species and their possible functions.

  11. Genetic characterization of egg weight, egg production and age at first egg in quails

    NARCIS (Netherlands)

    Marubayashi Hidalgo, A.; Martins, E.N.; Santos, A.L.; Quadros, T.C.O.; Ton, A.P.S.; Teixeira, R.

    2011-01-01

    The objective of this research was to estimate genetic parameters for the traits egg weight, egg production in 189 days and age at first egg in three laying quails and one meat line of quails. Data was analyzed by Bayesian procedures using Gibbs sampling. The heritability estimates for egg weight,

  12. A transgenic Xenopus laevis reporter model to study lymphangiogenesis

    Directory of Open Access Journals (Sweden)

    Annelii Ny

    2013-07-01

    The importance of the blood- and lymph vessels in the transport of essential fluids, gases, macromolecules and cells in vertebrates warrants optimal insight into the regulatory mechanisms underlying their development. Mouse and zebrafish models of lymphatic development are instrumental for gene discovery and gene characterization but are challenging for certain aspects, e.g. no direct accessibility of embryonic stages, or non-straightforward visualization of early lymphatic sprouting, respectively. We previously demonstrated that the Xenopus tadpole is a valuable model to study the processes of lymphatic development. However, a fluorescent Xenopus reporter directly visualizing the lymph vessels was lacking. Here, we created transgenic Tg(Flk1:eGFP Xenopus laevis reporter lines expressing green fluorescent protein (GFP in blood- and lymph vessels driven by the Flk1 (VEGFR-2 promoter. We also established a high-resolution fluorescent dye labeling technique selectively and persistently visualizing lymphatic endothelial cells, even in conditions of impaired lymph vessel formation or drainage function upon silencing of lymphangiogenic factors. Next, we applied the model to dynamically document blood and lymphatic sprouting and patterning of the initially avascular tadpole fin. Furthermore, quantifiable models of spontaneous or induced lymphatic sprouting into the tadpole fin were developed for dynamic analysis of loss-of-function and gain-of-function phenotypes using pharmacologic or genetic manipulation. Together with angiography and lymphangiography to assess functionality, Tg(Flk1:eGFP reporter tadpoles readily allowed detailed lymphatic phenotyping of live tadpoles by fluorescence microscopy. The Tg(Flk1:eGFP tadpoles represent a versatile model for functional lymph/angiogenomics and drug screening.

  13. Growth inhibition of Staphylococcus aureus and escherichia coli strains by neutralizing IgY antibodies from ostrich egg yolk

    Directory of Open Access Journals (Sweden)

    Fernando Luiz Tobias

    2012-06-01

    Full Text Available Ostrich raising around the world have some key factors and farming profit depend largely on information and ability of farmers to rear these animals. Non fertilized eggs from ostriches are discharged in the reproduction season. Staphylococcus aureus and Escherichia coli are microorganisms involved in animal and human diseases. In order to optimize the use of sub products of ostrich raising, non fertilized eggs of four selected birds were utilized for development of polyclonal IgY antibodies. The birds were immunized (200ug/animal with purified recombinant staphylococcal enterotoxin C (recSEC and synthetic recRAP, both derived from S. aureus, and recBFPA and recEspB involved in E. coli pathogenicity, diluted in FCA injected in the braquial muscle. Two subsequent immunization steps with 21 days intervals were repeated in 0,85% saline in FIA. Blood and eggs samples were collected before and after immunization steps. Egg yolk immunoglobulins were purified by precipitation with 19% sodium sulfate and 20% ammonium sulphate methodologies. Purified IgY 50µL aliquots were incubated in 850µL BHI broth containing 50µL inoculums of five strains of S. aureus and five strains of E.coli during four hours at 37ºC. Growth inhibition was evaluated followed by photometry reading (DO550nm. Egg yolk IgY preparation from hiperimmunized birds contained antibodies that inhibited significantly (p<0,05 growth of strains tested. Potential use of ostrich IgY polyclonal antibodies as a diagnostic and therapeutic tool is proposed for diseased animals.

  14. Genetic Compatibility Underlies Benefits of Mate Choice in an External Fertilizer.

    Science.gov (United States)

    Aguirre, J David; Blows, Mark W; Marshall, Dustin J

    2016-05-01

    Mate choice is a common feature of sexually reproducing species. In sessile or sedentary external fertilizers, however, direct interactions between reproductive partners are minimal, and instead mate recognition and choice must occur at the level of gametes. It is common for some sperm and egg combinations to have higher fertilization success than others, but it remains unclear whether differences in fertilization reflect gamete-level mate choice (GMC) for paternal quality or parental compatibility. Here, we examine the mechanisms underlying GMC in an externally fertilizing ascidian. A manipulative mate-choice assay confirmed that offspring viability was greater in clutches where we allowed GMC than in clutches where we precluded GMC. A complementary quantitative genetic experiment then revealed that paternal quality effects were generally weaker than parental compatibility effects, particularly for the trait combination underlying the benefits of GMC. Overall, our data suggest that gametes that are more compatible at fertilization produce more viable offspring than gametes that are less compatible at fertilization. Therefore, although the regalia we typically associate with sexual selection are absent in external fertilizers, mechanisms that allow females to bias fertilization in favor of some males over others produce significant fitness benefits in organisms reproducing via the ancestral strategy.

  15. Immunologic changes in children with egg allergy ingesting extensively heated egg.

    Science.gov (United States)

    Lemon-Mulé, Heather; Sampson, Hugh A; Sicherer, Scott H; Shreffler, Wayne G; Noone, Sally; Nowak-Wegrzyn, Anna

    2008-11-01

    Prior studies have suggested that heated egg might be tolerated by some children with egg allergy. We sought to confirm tolerance of heated egg in a subset of children with egg allergy, to evaluate clinical and immunologic predictors of heated egg tolerance, to characterize immunologic changes associated with continued ingestion of heated egg, and to determine whether a diet incorporating heated egg is well tolerated. Subjects with documented IgE-mediated egg allergy underwent physician-supervised oral food challenges to extensively heated egg (in the form of a muffin and a waffle), with tolerant subjects also undergoing regular egg challenges (in a form of scrambled egg or French toast). Heated egg-tolerant subjects incorporated heated egg into their diets. Skin prick test wheal diameters and egg white, ovalbumin, and ovomucoid IgE levels, as well as ovalbumin and ovomucoid IgG4 levels, were measured at baseline for all subjects and at 3, 6, and 12 months for those tolerant of heated egg. Sixty-four of 117 subjects tolerated heated egg, 23 tolerated regular egg, and 27 reacted to heated egg. Heated egg-reactive subjects had larger skin test wheals and greater egg white-specific, ovalbumin-specific, and ovomucoid-specific IgE levels compared with heated egg- and egg-tolerant subjects. Continued ingestion of heated egg was associated with decreased skin test wheal diameters and ovalbumin-specific IgE levels and increased ovalbumin-specific and ovomucoid-specific IgG4 levels. The majority of subjects with egg allergy were tolerant of heated egg. Continued ingestion of heated egg was well tolerated and associated with immunologic changes that paralleled the changes observed with the development of clinical tolerance to regular egg.

  16. Patterns of globalized reproduction: Egg cells regulation in Israel and Austria.

    Science.gov (United States)

    Shalev, Carmel; Werner-Felmayer, Gabriele

    2012-04-18

    Since the successful introduction of in vitro fertilization in 1978, medically assisted reproduction (MAR) has proliferated in multiple clinical innovations. Consequently, egg cells have become an object of demand for both infertility treatment and stem cell research, and this raises complex legal, ethical, social and economic issues.In this paper we compare how the procurement and use of human egg cells is regulated in two countries: Israel and Austria. Israel is known for its scientific leadership, generous public funding, high utilization and liberal regulation of assisted reproductive technology (ART). Austria lies at the other extreme of the regulatory spectrum in terms of restrictions on reproductive interventions.In both countries, however, there is a constant increase in the use of the technology, and recent legal developments make egg cells more accessible. Also, in both countries the scarcity of egg cells in concert with the rising demand for donations has led to the emergence of cross-border markets and global 'reproductive tourism' practices. In Israel, in particular, a scandal known as the 'eggs affair' was followed by regulation that allowed egg cell donations from outside the country under certain conditions.Cross-border markets are developed by medical entrepreneurs, driven by global economic gaps, made possible by trans-national regulatory lacunae and find expression as consumer demand. The transnational practice of egg cell donations indicates the emergence of a global public health issue, but there is a general lack of medical and epidemiological data on its efficacy and safety. We conclude that there is need for harmonisation of domestic laws and formulation of new instruments for international governance.

  17. Patterns of globalized reproduction: Egg cells regulation in Israel and Austria

    Directory of Open Access Journals (Sweden)

    Shalev Carmel

    2012-04-01

    Full Text Available Abstract Since the successful introduction of in vitro fertilization in 1978, medically assisted reproduction (MAR has proliferated in multiple clinical innovations. Consequently, egg cells have become an object of demand for both infertility treatment and stem cell research, and this raises complex legal, ethical, social and economic issues. In this paper we compare how the procurement and use of human egg cells is regulated in two countries: Israel and Austria. Israel is known for its scientific leadership, generous public funding, high utilization and liberal regulation of assisted reproductive technology (ART. Austria lies at the other extreme of the regulatory spectrum in terms of restrictions on reproductive interventions. In both countries, however, there is a constant increase in the use of the technology, and recent legal developments make egg cells more accessible. Also, in both countries the scarcity of egg cells in concert with the rising demand for donations has led to the emergence of cross-border markets and global 'reproductive tourism' practices. In Israel, in particular, a scandal known as the 'eggs affair' was followed by regulation that allowed egg cell donations from outside the country under certain conditions. Cross-border markets are developed by medical entrepreneurs, driven by global economic gaps, made possible by trans-national regulatory lacunae and find expression as consumer demand. The transnational practice of egg cell donations indicates the emergence of a global public health issue, but there is a general lack of medical and epidemiological data on its efficacy and safety. We conclude that there is need for harmonisation of domestic laws and formulation of new instruments for international governance.

  18. Regulation of ALF promoter activity in Xenopus oocytes.

    Directory of Open Access Journals (Sweden)

    Dan Li

    Full Text Available BACKGROUND: In this report we evaluate the use of Xenopus laevis oocytes as a matched germ cell system for characterizing the organization and transcriptional activity of a germ cell-specific X. laevis promoter. PRINCIPAL FINDINGS: The promoter from the ALF transcription factor gene was cloned from X. laevis genomic DNA using a PCR-based genomic walking approach. The endogenous ALF gene was characterized by RACE and RT-PCR for transcription start site usage, and by sodium bisulfite sequencing to determine its methylation status in somatic and oocyte tissues. Homology between the X. laevis ALF promoter sequence and those from human, chimpanzee, macaque, mouse, rat, cow, pig, horse, dog, chicken and X. tropicalis was relatively low, making it difficult to use such comparisons to identify putative regulatory elements. However, microinjected promoter constructs were very active in oocytes and the minimal promoter could be narrowed by PCR-mediated deletion to a region as short as 63 base pairs. Additional experiments using a series of site-specific promoter mutants identified two cis-elements within the 63 base pair minimal promoter that were critical for activity. Both elements (A and B were specifically recognized by proteins present in crude oocyte extracts based on oligonucleotide competition assays. The activity of promoter constructs in oocytes and in transfected somatic Xenopus XLK-WG kidney epithelial cells was quite different, indicating that the two cell types are not functionally equivalent and are not interchangeable as assay systems. CONCLUSIONS: Overall the results provide the first detailed characterization of the organization of a germ cell-specific Xenopus promoter and demonstrate the feasibility of using immature frog oocytes as an assay system for dissecting the biochemistry of germ cell gene regulation.

  19. Characterization of Cer-1 cis-regulatory region during early Xenopus development.

    Science.gov (United States)

    Silva, Ana Cristina; Filipe, Mário; Steinbeisser, Herbert; Belo, José António

    2011-05-01

    Cerberus-related molecules are well-known Wnt, Nodal, and BMP inhibitors that have been implicated in different processes including anterior–posterior patterning and left–right asymmetry. In both mouse and frog, two Cerberus-related genes have been isolated, mCer-1 and mCer-2, and Xcer and Xcoco, respectively. Until now, little is known about the mechanisms involved in their transcriptional regulation. Here, we report a heterologous analysis of the mouse Cerberus-1 gene upstream regulatory regions, responsible for its expression in the visceral endodermal cells. Our analysis showed that the consensus sequences for a TATA, CAAT, or GC boxes were absent but a TGTGG sequence was present at position -172 to -168 bp, relative to the ATG. Using a series of deletion constructs and transient expression in Xenopus embryos, we found that a fragment of 1.4 kb of Cer-1 promoter sequence could reproduce the endogenous expression pattern of Xenopus cerberus. A 0.7-kb mcer-1 upstream region was able to drive reporter expression to the involuting mesendodermal cells, while further deletions abolished reporter gene expression. Our results suggest that although no sequence similarity was found between mouse and Xenopus cerberus cis-regulatory regions, the signaling cascades regulating cerberus expression, during gastrulation, is conserved.

  20. Animal egg as evolutionary innovation: a solution to the "embryonic hourglass" puzzle.

    Science.gov (United States)

    Newman, Stuart A

    2011-11-15

    The evolutionary origin of the egg stage of animal development presents several difficulties for conventional developmental and evolutionary narratives. If the egg's internal organization represents a template for key features of the developed organism, why can taxa within a given phylum exhibit very different egg types, pass through a common intermediate morphology (the so-called "phylotypic stage"), only to diverge again, thus exemplifying the embryonic "hourglass"? Moreover, if different egg types typically represent adaptations to different environmental conditions, why do birds and mammals, for example, have such vastly different eggs with respect to size, shape, and postfertilization dynamics, whereas all these features are more similar for ascidians and mammals? Here, I consider the possibility that different body plans had their origin in self-organizing physical processes in ancient clusters of cells, and suggest that eggs represented a set of independent evolutionary innovations subsequently inserted into the developmental trajectories of such aggregates. I first describe how "dynamical patterning modules" (DPMs) associations between components of the metazoan developmental-genetic toolkit and certain physical processes and effects may have organized primitive animal body plans independently of an egg stage. Next, I describe how adaptive specialization of cells released from such aggregates could have become "proto-eggs," which regenerated the parental cell clusters by cleavage, conserving the characteristic DPMs available to a lineage. Then, I show how known processes of cytoplasmic reorganization following fertilization are often based on spontaneous, self-organizing physical effects ("egg-patterning processes": EPPs). I suggest that rather than acting as developmental blueprints or prepatterns, the EPPs refine the phylotypic body plans determined by the DPMs by setting the boundary and initial conditions under which these multicellular patterning

  1. The cellular distribution of histone H5 in embryonic and adult tissues of Xenopus laevis and chicken

    NARCIS (Netherlands)

    Moorman, A. F.; de Boer, P. A.; Lamers, W. H.; Charles, R.

    1986-01-01

    The cellular distribution of histone H5 in embryonic and adult tissues of Xenopus laevis and chicken has been established with monoclonal antibodies to histone H5. Both in Xenopus and in chicken, the protein has presumably a more widespread cellular distribution than hitherto expected but is absent

  2. 9 CFR 590.905 - Importation of restricted eggs or eggs containing more restricted eggs than permitted in the...

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Importation of restricted eggs or eggs containing more restricted eggs than permitted in the official standards for U.S. Consumer Grade B. 590.905... EGG PRODUCTS INSPECTION INSPECTION OF EGGS AND EGG PRODUCTS (EGG PRODUCTS INSPECTION ACT) Imports...

  3. Effektive representasjoner? Forventninger til og bekymringer for forskning på befruktede egg

    Directory of Open Access Journals (Sweden)

    Marie Auensen Antonsen

    2011-05-01

    ,kontrovers, representasjoner, Mehmet-sakenEnglish abstract: Effective representations? Concerns and expectationsregarding research on fertilized eggsSince the 1980s, there have been several debates and controversies concerningthe regulation of medical biotechnology. In this article, we analyze three suchinstances in Norway, mainly the debates concerning research on fertilizedeggs. Analyzing the debates surrounding the regulations of 1994, 2003/2004and 2008, we investigate how different stakeholders strived to mobilize whatwe call different framings of the controversy, utilizing the different scientificand political representations of the fertilized egg.Framing involves creating baselines for rhetorical and political stratagems.Two framings appear more pronounced than others in the Norwegian debate.On one hand, we find attempts of framing the controversy in a risk centricframework, which mostly emphasizes the potential negative effects of medicalbiotechnology as it focuses on the universal reverence for the unborn life. Onthe other hand, we see attempts to mobilize a positive frame of expectationthat emphasizes on the hope of finding new and effective treatment options forthe severely ill.The ban against research on fertilized eggs that was passed in Norway in1987 was repeated in 1994 and 2003. However, with the changes introducedto the law in 2008, there was a noticeable shift in Norwegian legislation, asresearch on fertilized eggs became permissible if specific conditions were met.We argue that the turning point in this regulative change was the Mehmentcase, a particular case that entailed the so-called «overflowing» (Callon 1998in both the negative and the positive frame. The Mehmet case exposed a moregeneral democratic and political dilemma. In our opinion, it illustrates howvulnerable can regulation that is based on general ethical principles be in theface of what appears to the laity as unreasonable. Thus, we argue that thisactualizes the need for more hybridized ways to view the

  4. Development of eggs and larvae of Emmelichthys nitidus (Percoidei: Emmelichthyidae) in south-eastern Australia, including a temperature-dependent egg incubation model

    Science.gov (United States)

    Neira, Francisco J.; Keane, John P.; Lyle, Jeremy M.; Tracey, Sean R.

    2008-08-01

    Reared eggs and field-collected material were employed to describe the development of the pelagic eggs and larvae of Emmelichthys nitidus (Emmelichthyidae), a small (36 cm TL) mid-water schooling species common in shelf waters of temperate Australia. Hydrated oocytes from adults trawled from spawning grounds off eastern Tasmania were fertilized and reared to the yolk-sac larval stage, and the data employed to build a temperature-dependent egg incubation model. Embryogenesis lasted 96, 84 and 54 h at mean temperatures of 13.1, 14.4 and 16.5 °C respectively, and was divided into seven stages based on extent of epiboly until blastopore closure (stages I-III) and embryo growth (stages IV-VII). Eggs (1.00-1.05 mm diameter) are spherical with a smooth chorion, small perivitelline space and prominent, unsegmented yolk with a single, posteriorly-located oil globule (0.18-0.20 mm diameter) that becomes pigmented from stage III. Embryos have two distinct snout melanophores, and a paired melanophore row laterally along the trunk and tail. Morphological identification of eggs collected during surveys in October 2005 and 2006 was validated using quantitative PCR amplification of the mtDNA d-loop gene region unique to E. nitidus, producing an 80-100% agreement across all seven stages. Newly-emerged larvae (1.9-3.3 mm) possess a prominent yolk sac with the posteriorly-located, pigmented oil globule, mouth not yet functional and unpigmented eyes. Notochord flexion occurs between 5 and 8 mm while fins are formed by 12 mm. Larvae examined (3.3-17.4 mm) are lightly pigmented and possess percoid features such as an elongate to moderate body, coiled, triangular-shaped gut, preopercular spines and 24-25 myomeres; two prominent pigment patches opposite each other dorsally and ventrally along the tail are diagnostic. Variability of mean egg ages ( y) by temperature ( t) and stage ( i) was best described by the deterministic stage-to-age model y = 35.911exp[-(0.155 t + 0.262 i)] i2

  5. Effect of seminal plasma and egg yolk concentration on freezability of goat semen

    Directory of Open Access Journals (Sweden)

    Valéria da Silva Ferreira

    2014-10-01

    Full Text Available The objective of this study was to evaluate the effects of egg yolk and seminal plasma on the viability of cryopreserved goat semen. To this end, four fertile Saanen bucks, aged between 10 months and 1 year, and weighing 18 to 25 kg, were used. Semen was collected from each buck by the artificial vagina method at the end of breeding season (June-July. The extender used was the yolk citrate, which was split into two equal aliquots: 5% egg yolk (2.5 mL egg yolk: 47.5 mL citrate solution were added to one of the samples and 10% egg yolk (5.0 mL egg yolk: 45.0 mL citrate solution were added to another. The sperm motility and vigor after thawing and post thermal resistance test (TRT were evaluated and the data were subjected to analysis of variance and means were compared by the F test at 5.0% probability. The observed values for motility and vigor after thawing and post thermal resistance test (TRT, fast and slow, according to the presence of seminal plasma and egg yolk percentage were: 5% egg yolk with plasma (25.0% and 3.3; 1.60% and 0.7; 12.36% and 1.6, respectively; 5% egg yolk without plasma (23.61% and 3.1; 1.25% and 0.2; 9.93% and 1.3, respectively; 10% egg yolk with plasma (30.8% and 3.3; 4.4% and 1.9; 19.5% and 2.7, respectively; and 10% egg yolk without plasma (13.4% and 2.5; 4.1% and 0.5; 17.0% and 1.0, respectively. There were significant differences between the analyzed data in relation to semen with or without plasma at different percentages of egg yolk, and the group that presented the best results was 10% egg yolk citrate in extender with plasma. The presence of seminal plasma and higher concentration of egg yolk in extender provide a higher viability of cryopreserved goat semen.

  6. An analysis of radiation-induced damage in the spider mite. Relationship between mortality of haploid and diploid eggs in two successive generations

    International Nuclear Information System (INIS)

    Leenhouts, H.P.; Chadwick, K.H.

    1976-01-01

    Unfertilized females of the spider mite (Tetranychus urticae) produce only haploid eggs which develop into a haploid male. Fertilized females produce both haploid eggs (unfertilized), which develop into males, and diploid eggs (fertilized), which develop into females. Radiobiological experiments performed by A.M. Feldmann (Association Euratom-ITAL) made data available on the radiation-induced mortality of haploid and diploid eggs in the F 1 and F 2 generation following irradiation of either males or females with X rays or fast neutrons. The data have been analysed using the molecular theory of cell survival where it is assumed that DNA double strand breaks, induced randomly in the cell, are the critical radiation-induced lesions, which lead to cell death. Theoretical relationships are derived for the dose dependence of hatchability in haploid and diploid eggs in the first and second generations expressed as a function of the radiation damage in the parental genome. These theoretical relationships can be used to derive the inter-relationship between the different hatchabilities, and the results from the spider mite have been analysed using these considerations. It is concluded that the radiation-induced genetic damage arises from one type of initial lesion. The eventual radiobiological implications of this analysis are discussed, expecially with respect to the transmittance of radiation-induced genetic damage after low-level radiation. (author)

  7. Egg size and offspring quality: a meta-analysis in birds.

    Science.gov (United States)

    Krist, Miloš

    2011-08-01

    Parents affect offspring fitness by propagule size and quality, selection of oviposition site, quality of incubation, feeding of dependent young, and their defence against predators and parasites. Despite many case studies on each of these topics, this knowledge has not been rigorously integrated into individual parental care traits for any taxon. Consequently, we lack a comprehensive, quantitative assessment of how parental care modifies offspring phenotypes. This meta-analysis of 283 studies with 1805 correlations between egg size and offspring quality in birds is intended to fill this gap. The large sample size enabled testing of how the magnitude of the relationship between egg size and offspring quality depends on a number of variables. Egg size was positively related to nearly all studied offspring traits across all stages of the offspring life cycle. Not surprisingly, the relationship was strongest at hatching but persisted until the post-fledging stage. Morphological traits were the most closely related to egg size but significant relationships were also found with hatching success, chick survival, and growth rate. Non-significant effect sizes were found for egg fertility, chick immunity, behaviour, and life-history or sexual traits. Effect size did not depend on whether chicks were raised by their natural parents or were cross-fostered to other territories. Effect size did not depend on species-specific traits such as developmental mode, clutch size, and relative size of the egg, but was larger if tested in captive compared to wild populations and between rather than within broods. In sum, published studies support the view that egg size affects juvenile survival. There are very few studies that tested the relationship between egg size and the fecundity component of offspring fitness, and no studies on offspring survival as adults or on global fitness. More data are also needed for the relationships between egg size and offspring behavioural and

  8. Egg Allergy

    Science.gov (United States)

    ... Staying Safe Videos for Educators Search English Español Egg Allergy KidsHealth / For Teens / Egg Allergy What's in ... but it's worth it. What Happens With an Egg Allergy? Eggs aren't bad. But when you' ...

  9. A model for investigating developmental eye repair in Xenopus laevis.

    Science.gov (United States)

    Kha, Cindy X; Son, Philip H; Lauper, Julia; Tseng, Kelly Ai-Sun

    2018-04-01

    Vertebrate eye development is complex and requires early interactions between neuroectoderm and surface ectoderm during embryogenesis. In the African clawed frog, Xenopus laevis, individual eye tissues such as the retina and lens can undergo regeneration. However, it has been reported that removal of either the specified eye field at the neurula stage or the eye during tadpole stage does not induce replacement. Here we describe a model for investigating Xenopus developmental eye repair. We found that tailbud embryos can readily regrow eyes after surgical removal of over 83% of the specified eye and lens tissues. The regrown eye reached a comparable size to the contralateral control by 5 days and overall animal development was normal. It contained the expected complement of eye cell types (including the pigmented epithelium, retina and lens), and is connected to the brain. Our data also demonstrate that apoptosis, an early mechanism that regulates appendage regeneration, is also required for eye regrowth. Treatment with apoptosis inhibitors (M50054 or NS3694) blocked eye regrowth by inhibiting caspase activation. Together, our findings indicate that frog embryos can undergo successful eye repair after considerable tissue loss and reveals a required role for apoptosis in this process. Furthermore, this Xenopus model allows for rapid comparisons of productive eye repair and developmental pathways. It can also facilitate the molecular dissection of signaling mechanisms necessary for initiating repair. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

  10. Changes in the number of eggs loaded in Pantala flavescens females with age from mass flights (Odonata: Libellulidae).

    Science.gov (United States)

    Ichikawa, Yuta; Watanabe, Mamoru

    2014-11-01

    The wandering glider dragonfly Pantala flavescens migrates to Japan every spring, where the population increases until autumn, in which mass flights often occur, followed by death in the winter. There have been no reports to date on the maturation process of this species throughout its lifespan in Japan. We collected females from mass flights when the flight height was low, and classified them into seven age stages by examining their wing condition. Very few females of the older stage were collected from the mass flights. The wing condition corresponded with the change in body color and with the egg production process in the ovaries. While pre-reproductive-stage females did not release eggs when treated with our artificial oviposition technique, each reproductive-stage female released about 640 eggs. Nearly all eggs released were fertilized. The ovaries developed with the stage, and reproductive-stage females had about 1100 ovarioles. The estimated maximum fecundity was about 29,000 eggs. The lifetime number of eggs laid of P. flavescens should be revealed by dissection.

  11. For Whom the Clock Ticks: Reproductive Ageing and Egg Freezing in Dutch and British News Media

    Directory of Open Access Journals (Sweden)

    Lucy van de Wiel

    2014-01-01

    Full Text Available The last century saw struggles for women's reproductive choices both to avoid childbearing (i.e. abortion, contraception and to achieve it (i.e. IVF, artificial insemination. Now, after the turn of the millennium, these two approaches to regulate reproduction are combined in oocyte cryopreservation (OC, or egg freezing. With it, a new reproductive question has emerged as egg freezing simultaneously represents an active choice not to have children at present and a commitment to a future, possibly assisted, reproduction. Women's usage of OC to preserve fertility is itself an act of refusing current childbearing, thus calling into question an easy distinction between reproductive and non-reproductive behaviour. In this article, I discuss the representation of this new choice by using a selection of Dutch and UK news media pieces, focusing specifically on the implications of egg freezing for conceptualisations of the female reproductive body as site of a gendered politics of ageing. Firstly, I address the rhetorical divisions between 'medical' and 'social' motivations for egg freezing and then I argue that the media narratives around these divisions create new subject positions related to reproductive identity through which medical authority becomes extended into new parts of social life. Secondly, I address the discursive construction of the decision to use egg freezing technology in relation to the notion of the 'biological clock' and the reconceptualisations of age-specific (non-reproductive bodies, which focus on the ovum as the locus of fertility. The news coverage of OC thus reveals a gendered politics of ageing, predicated on reproductive ability as the organising principle for the temporal structuring of life, which not only interpellates (potentially infertile women who desire to reproduce, but also has an impact on the wider public.

  12. Hormonal priming, induction of ovulation and in-vitro fertilization of the endangered Wyoming toad (Bufo baxteri

    Directory of Open Access Journals (Sweden)

    Seratt Jessica

    2006-06-01

    Full Text Available Abstract The endangered Wyoming toad (Bufo baxteri is the subject of an extensive captive breeding and reintroduction program. Wyoming toads in captivity rarely ovulate spontaneously and hormonal induction is used to ovulate females or to stimulate spermiation in males. With hormonal induction, ovulation is unreliable and egg numbers are low. The sequential administration of anovulatory doses of hormones (priming has increased egg numbers and quality in both anurans and fish. Consequently, we tested the efficacy of a combination of human Chorionic Gonadotrophin (hCG and Luteinizing Hormone Releasing Hormone analogue (LHRHa administered as one dose, or two or three sequential doses to Bufo baxteri on egg numbers, fertilization and early embryo development. Spawning toads deposited eggs into Simplified Amphibian Ringers (SAR solution to enable controlled in-vitro fertilization (IVF with sperm from hormonally induced male toads. Unprimed females receiving a single mixed normally ovulatory dose of 500 IU hCG plus 4 micrograms of LHRHa produced no eggs. Whereas females primed with this dose and an anovulatory dose (100 IU hCG and 0.8 micrograms of LHRHa of the same hormones, or primed only with an anovulatory dose, spawned after then receiving an ovulatory dose. Higher total egg numbers were produced with two primings than with one priming. Moreover, two primings produced significantly more eggs from each individual female than one priming. The cleavage rate of eggs was not found to differ between one or two primings. Nevertheless, embryo development with eggs from two primings gave a significantly greater percentage neurulation and swim-up than those from one priming. Of the male toads receiving a single dose of 300 IU hCG, 80% produced spermic urine with the greatest sperm concentration 7 hours post-administration (PA. However, peak sperm motility (95% was achieved at 5 hours PA and remained relatively constant until declining 20 hours PA. In

  13. Presence of helminth eggs in wastewater sludge; Presencia de huevos de helmintos en fangos de depuracion de aguas residuales

    Energy Technology Data Exchange (ETDEWEB)

    GeovanPerez Ortiz, O. G.; Gomez Vera, D. [Estacion Depuradora de Aguas Residuales. Alboraia. Valencia (Spain); Faubell Brell, M. [Universidad de Valencia (Spain); Morenilla Martinez, J. J.; Bernacer Bonora, I. [Entididad Publica de Saneamiento. Valencia (Spain); Bernabeu Adrian, A. [S. A. Agricultures de la Vega. Valencia (Spain); Amores Blasco, S. [Depuracion de Aguas del Mediterraneo. Valencia (Spain)

    2000-07-01

    Land application is one of the best options for the elimination of wastewater sludge, because sludge is useful as conditioner and fertilizer in soils. However, land application represents a sanitary hazards due to the sludge's content in pathogenic agents. Helminth eggs are among these agents, and their hardness and resistance to adverse environmental conditions allows them to survive in soils for months. With this in mind, we have launched a study of the presence of helminth eggs in the sludge of a wastewater treatment plant in Alboraia (Valencia). The results show a concentration of helminth eggs in digestion sludge of 27.000 eggs/kg dry matter. We discuss the potential sanitary relevant of these results and the need to continue with this evaluation program. (Author) 20 refs.

  14. Exposing broiler eggs to green, red and white light during incubation.

    Science.gov (United States)

    Archer, G S

    2017-07-01

    Previous work has shown that exposing broiler eggs to white light during incubation can improve hatchability and post-hatch animal welfare. It was hypothesized that due to how different wavelengths of light can affect avian physiology differently, and how pigmented eggshells filter light that different monochromatic wavelengths would have differential effects on hatchability and post-hatch animal welfare indicators. To determine, we incubated chicken eggs (n=6912) under either no light (dark), green light, red light or white light; the light level was 250 lux. White and red light were observed to increase hatch of fertile (P0.05). Fear response of during isolation and tonic immobility was reduced (P0.05) from dark incubated broilers. All light incubated broilers had lower (Phatchery efficiency and post-hatch animal welfare at the same time.

  15. IDENTIFICATION AND MOLECULAR CLONING OF XENOPUS LAEVIS SP22, A PROTEIN ASSOCIATED WITH FERTILIZATION IN MAMMALS

    Science.gov (United States)

    ABSTRACTSP22 is a protein that has been characterized in rats where it has been related with fertility. SP22 homologues have been studied in mouse and man and a definitive role for the protein has not been assigned yet. By means of a polyclonal IgG to recombinant rat SP22...

  16. Medical and social egg freezing: internet-based survey of knowledge and attitudes among women in Denmark and the UK.

    Science.gov (United States)

    Lallemant, Camille; Vassard, Ditte; Nyboe Andersen, Anders; Schmidt, Lone; Macklon, Nick

    2016-12-01

    Until recently, limited options for preserving fertility in order to delay childbearing were available. Although egg freezing and successful thawing is now possible, it remains unclear to what extent women are aware of the availability of this technique, their attitudes towards its use, or the circumstances under which this technique may be considered. An online cross-sectional survey was designed to investigate knowledge and attitudes of women in Denmark and the UK on egg freezing and their potential intentions regarding the procedure. Data was collected from September 2012 to September 2013 and the responses of 973 women were analyzed. In total, 83% of women reported having heard of egg freezing, and nearly all considered it acceptable for medical indications, whilst 89% considered it acceptable for social reasons. Overall, 19% expressed active interest in the procedure, and 27% expressed possible interest. Key factors found to positively influence attitudes to accepting the procedure were reassurance that it would not affect future fertility and greater than 50% chance of achieving a live birth. Characteristics significantly associated with intention to freeze eggs were being single, age under 35 years, childlessness, and a history of infertility. In this group, risk and cost were less important considerations. This study indicates that there is widespread awareness and support of the availability of eggs freezing for reproductive planning. Reassurance regarding its efficacy appears more important than its potential adverse effects on their health or that of future children, or the costs of the procedure. © 2016 Nordic Federation of Societies of Obstetrics and Gynecology.

  17. Toxicity of road deicing salt (NaCl) and copper (Cu) to fertilization and early developmental stages of Atlantic salmon (Salmo salar).

    Science.gov (United States)

    Mahrosh, Urma; Kleiven, Merethe; Meland, Sondre; Rosseland, Bjørn Olav; Salbu, Brit; Teien, Hans-Christian

    2014-09-15

    In many countries, salting of ice or snow covered roads may affect aquatic organisms in the catchment directly or indirectly by mobilization of toxic metals. We studied the toxicity of road deicing salt and copper (Cu) on the vulnerable early life stages of Atlantic salmon (Salmo salar), from fertilization till hatching. Controlled episodic exposure to road salt (≥ 5,000 mg/L) during fertilization resulted in reduced swelling and less percent egg survival. Exposure to Cu both during and post fertilization caused delayed hatching. Larval deformities were, however found as an additional effect, when eggs were exposed to high salt concentration (≥ 5,000 mg/L) mixed with Cu (10 μg Cu/L) during fertilization. Thus, it appears that the sensitivity of early developmental stages of Atlantic salmon increased when exposed to these stressors, and road salt application during spawning can pose threat to Atlantic salmon in water bodies receiving road runoff. The study gives insight on assessment and management of risks on Atlantic salmon population posed by road related hazardous chemicals. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Effects of Environmental Enrichment on the Fertility and Fecundity of Zebrafish (Danio rerio).

    Science.gov (United States)

    Wafer, Lemnique N; Jensen, V Behrana; Whitney, Jesse C; Gomez, Thomas H; Flores, Rene; Goodwin, Bradford S

    2016-01-01

    Zebrafish (Danio rerio) are a popular vertebrate model in biomedical research, but information describing the effects of environmental enrichment on fertility and fecundity of zebrafish is sparse. In the current study, 18 breeding pairs were placed in divided 1.5-L breeding tanks containing 1 of 3 enrichment conditions: plastic grass (n = 6), plastic leaves (n = 6), or no enrichment (n = 6, control). The pairs were allowed to spawn for 3 h the next day, after which eggs were counted and breeding pairs were returned to holding tanks for use in subsequent sessions. Spawning sessions were repeated at 7-d intervals until the completion of 9 trials, with pairs rotating to a different condition at each interval. Total egg count (mean ± SEM) after 3 h was greater for zebrafish spawning in the grass environment (48.0 ± 7.7 eggs) than in the leaf or control environments (29.4 ± 5.3 and 20.4 ± 3.7 eggs, respectively). An interaction emerged between enrichment type and the age of the spawning pair on the number of fry at 6 d postfertilization (dpf). Initially, more fry were obtained from 110- and 160-dpf pairs with the grass enrichment, but from 173- and 180-dpf pairs there were more obtained with leaf enrichment than grass. A separate experiment showed that enrichment type did not have an effect on fry survivability. Overall, our data indicates that, under certain conditions, zebrafish fertility and fecundity are greater in a breeding tank containing environmental enrichment than in a bare tank.

  19. 7 CFR 57.905 - Importation of restricted eggs or eggs containing more restricted eggs than permitted in the...

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Importation of restricted eggs or eggs containing more restricted eggs than permitted in the official standards for U.S. Consumer Grade B. 57.905 Section 57.905... AGRICULTURAL MARKETING ACT OF 1946 AND THE EGG PRODUCTS INSPECTION ACT (CONTINUED) INSPECTION OF EGGS (EGG...

  20. Host-defense and trefoil factor family peptides in skin secretions of the Mawa clawed frog Xenopus boumbaensis (Pipidae).

    Science.gov (United States)

    Conlon, J Michael; Mechkarska, Milena; Kolodziejek, Jolanta; Leprince, Jérôme; Coquet, Laurent; Jouenne, Thierry; Vaudry, Hubert; Nowotny, Norbert; King, Jay D

    2015-10-01

    Peptidomic analysis of norepinephrine-stimulated skin secretions from the octoploid Mawa clawed frog Xenopus boumbaensis Loumont, 1983 led to the identification and characterization of 15 host-defense peptides belonging to the magainin (two peptides), peptide glycine-leucine-amide (PGLa; three peptides), xenopsin precursor fragment (XPF; three peptides), caerulein precursor fragment (CPF; two peptides), and caerulein precursor fragment-related peptide (CPF-RP; five peptides) families. In addition, caerulein and three peptides with structural similarity to the trefoil factor family (TFF) peptides, xP2 and xP4 from Xenopus laevis were also present in the secretions. Consistent with data from comparisons of the nucleotides sequence of mitochondrial and nuclear genes, the primary structures of the peptides suggest a close phylogenetic relationship between X. boumbaensis and the octoploid frogs Xenopus amieti and Xenopus andrei. As the three species occupy disjunct ranges within Cameroon, it is suggested that they diverged from a common ancestor by allopatric speciation. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Vocal competition in male Xenopus laevis frogs

    OpenAIRE

    Tobias, Martha L.; Corke, Anna; Korsh, Jeremy; Yin, David; Kelley, Darcy B.

    2010-01-01

    Male Xenopus laevis frogs produce underwater advertisement calls that attract gravid females and suppress calling by male competitors. Here we explore whether groups of males establish vocal ranks and whether auditory cues alone suffice for vocal suppression. Tests of male–male pairs within assigned groups reveal linear vocal dominance relations, in which each male has a defined rank. Both the duration over which males interact, as well as the number of competitive opportunities, affect linea...

  2. Dealing with multicollinearity in predicting egg components from egg weight and egg dimension

    Directory of Open Access Journals (Sweden)

    Tarek M. Shafey

    2014-10-01

    Full Text Available Measurements of 174 eggs from meat-type breeder flock (Ross at 36 weeks of age were used to study the problem of multicollinearity (MC instability in the estimation of egg components of yolk weight (YKWT, albumen weight (ALBWT and eggshell weight (SHWT. Egg weight (EGWT, egg shape index (ESI=egg width (EGWD*100/egg length (EGL and their interaction (EGWTESI were used in the context of un-centred vs centred data and principal components regression (PCR models. The pairwise phenotypic correlations, variance inflation factor (VIF, eigenvalues, condition index (CI, and variance proportions were examined. Egg weight had positive correlations with EGWD and EGL (r=0.56 and 0.50, respectively; P<0.0001 and EGL had a negative correlation with ESI (r=-0.79; P<0.0001. The highest correlation was observed between EGWT and ALBWT (r=0.94; P<0.0001, while the lowest was between EGWD and SHWT (r=0.33; P<0.0001. Multicollinearity problems were found in EGWT, ESI and their interaction as shown by VIF (>10, eigenvalues (near zero, CI (>30 and high corresponding proportions of variance of EGWT, ESI and EGWTESI with respect to EGWTESI. Results from this study suggest that mean centring and PCR were appropriate to overcome the MC instability in the estimation of egg components from EGWT and ESI. These methods improved the meaning of intercept values and produced much lower standard error values for regression coefficients than those from un-centred data.

  3. Cell cycle in egg cell and its progression during zygotic development in rice.

    Science.gov (United States)

    Sukawa, Yumiko; Okamoto, Takashi

    2018-03-01

    Rice egg is arrested at G1 phase probably by OsKRP2. After fusion with sperm, karyogamy, OsWEE1-mediated parental DNA integrity in zygote nucleus, zygote progresses cell cycle to produce two-celled embryo. In angiosperms, female and male gametes exist in gametophytes after the complementation of meiosis and the progression of nuclear/cell division of the haploid cell. Within the embryo sac, the egg cell is specially differentiated for fertilization and subsequent embryogenesis, and cellular programs for embryonic development, such as restarting the cell cycle and de novo gene expression, are halted. There is only limited knowledge about how the cell cycle in egg cells restarts toward zygotic division, although the conversion of the cell cycle from a quiescent and arrested state to an active state is the most evident transition of cell status from egg cell to zygote. This is partly due to the difficulty in direct access and analysis of egg cells, zygotes and early embryos, which are deeply embedded in ovaries. In this study, precise relative DNA amounts in the nuclei of egg cells, developing zygotes and cells of early embryos were measured, and the cell cycle of a rice egg cell was estimated as the G1 phase with a 1C DNA level. In addition, increases in DNA content in zygote nuclei via karyogamy and DNA replication were also detectable according to progression of the cell cycle. In addition, expression profiles for cell cycle-related genes in egg cells and zygotes were also addressed, and it was suggested that OsKRP2 and OsWEE1 function in the inhibition of cell cycle progression in egg cells and in checkpoint of parental DNA integrity in zygote nucleus, respectively.

  4. Identification of a Paralog-Specific Notch1 Intracellular Domain Degron

    OpenAIRE

    Broadus, Matthew R.; Chen, Tony W.; Neitzel, Leif R.; Ng, Victoria H.; Jodoin, Jeanne; Lee, Laura A.; Salic, Adrian; Robbins, David J.; Capobianco, Anthony J.; Patton, James G.; Huppert, Stacey S.; Lee, Ethan

    2016-01-01

    Upon Notch pathway activation, the receptor is cleaved to release the Notch intracellular domain (NICD), which translocates to the nucleus to activate gene transcription. Using Xenopus egg extracts, we have identified a Notch1-specific destruction signal (N1-Box). We show that mutations in the N1-Box inhibit NICD1 degradation and that the N1-Box is transferable for the promotion of degradation of heterologous proteins in Xenopus egg extracts and in cultured human cells. Mutation of the N1-Box...

  5. Fertility Preservation for Pediatric Patients: Current State and Future Possibilities.

    Science.gov (United States)

    Johnson, Emilie K; Finlayson, Courtney; Rowell, Erin E; Gosiengfiao, Yasmin; Pavone, Mary Ellen; Lockart, Barbara; Orwig, Kyle E; Brannigan, Robert E; Woodruff, Teresa K

    2017-07-01

    This review provides an overview of pediatric fertility preservation. Topics covered include the patient populations who could benefit, the current state of fertility preservation options and research, and considerations related to ethics and program development. A broad Embase® and PubMed® search was performed to identify publications discussing investigational, clinical, ethical and health care delivery issues related to pediatric fertility preservation. Relevant publications were reviewed and summarized. Populations who could benefit from fertility preservation in childhood/adolescence include oncology patients, patients with nononcologic conditions requiring gonadotoxic chemotherapy, patients with differences/disorders of sex development and transgender individuals. Peripubertal and postpubertal fertility preservation options are well established and include cryopreservation of oocytes, embryos or sperm. Prepubertal fertility preservation is experimental. Multiple lines of active research aim to develop technologies that will enable immature eggs and sperm to be matured and used to produce a biological child in the future. Ethical challenges include the need for parental proxy decision making and the fact that fertility preservation procedures can be considered not medically necessary. Successful multidisciplinary fertility preservation care teams emphasize partnerships with adult colleagues, prioritize timely consultations and use standardized referral processes. Some aspects of fertility preservation are not covered by insurance and out-of-pocket costs can be prohibitive. Pediatric fertility preservation is an emerging, evolving field. Fertility preservation options for prepubertal patients with fertility altering conditions such as cancer and differences/disorders of sex development are currently limited. However, multiple lines of active research hold promise for the future. Key considerations include establishing a multidisciplinary team to provide

  6. Egg production of turbot, Scophthalmus maximus, in the Baltic Sea

    Science.gov (United States)

    Nissling, Anders; Florin, Ann-Britt; Thorsen, Anders; Bergström, Ulf

    2013-11-01

    In the brackish water Baltic Sea turbot spawn at ~ 6-9 psu along the coast and on offshore banks in ICES SD 24-29, with salinity influencing the reproductive success. The potential fecundity (the stock of vitellogenic oocytes in the pre-spawning ovary), egg size (diameter and dry weight of artificially fertilized 1-day-old eggs) and gonad dry weight were assessed for fish sampled in SD 25 and SD 28. Multiple regression analysis identified somatic weight, or total length in combination with Fulton's condition factor, as main predictors of fecundity and gonad dry weight with stage of maturity (oocyte packing density or leading cohort) as an additional predictor. For egg size, somatic weight was identified as main predictor while otolith weight (proxy for age) was an additional predictor. Univariate analysis using GLM revealed significantly higher fecundity and gonad dry weight for turbot from SD 28 (3378-3474 oocytes/g somatic weight) compared to those from SD 25 (2343 oocytes/g somatic weight), with no difference in egg size (1.05 ± 0.03 mm diameter and 46.8 ± 6.5 μg dry weight; mean ± sd). The difference in egg production matched egg survival probabilities in relation to salinity conditions suggesting selection for higher fecundity as a consequence of poorer reproductive success at lower salinities. This supports the hypothesis of higher size-specific fecundity towards the limit of the distribution of a species as an adaptation to harsher environmental conditions and lower offspring survival probabilities. Within SD 28 comparisons were made between two major fishing areas targeting spawning aggregations and a marine protected area without fishing. The outcome was inconclusive and is discussed with respect to potential fishery induced effects, effects of the salinity gradient, effects of specific year-classes, and effects of maturation status of sampled fish.

  7. Effects of Ascorbic Acid Injection in Incubated Eggs Submitted to Heat Stress on Incubation Parameters and Chick Quality

    Directory of Open Access Journals (Sweden)

    S Sgavioli

    2015-06-01

    Full Text Available Dose-dependent positive effects on hatchability and hatchling weight have been attributed to ascorbic acid (AA when eggs were submitted or not to intermittent heat stress during incubation. Fertile breeder (Cobb(r eggs were used to determine if the pre-incubation injection of AA in ovo affects the incubation and hatchling quality of egg incubated under thermoneutral or intermittent heat stress conditions. Eggs were not injected or injected with 0, 2,4, or 6% AA/100µL water and incubated at continuous thermoneutral (37.5ºC or hot (39.0ºC temperature. Eggshell temperature (EST increased in the second half of the incubation period in all experimental groups. The EST of non-injected eggs and of those injected with water was higher when incubated at 39°C than at 37.5°C, but EST was not different among eggs injected with AA. Egg mass loss and eggshell conductance were higher in the eggs incubated at 39°C than at 37.5°C.Hatchability was lower in the eggs injected with AA. Liver and yolk sac weights were higher, whereas heart and liver weights were lower in hatchlings from eggs incubated at 39°C; however, hatchling weight was not affected by incubation temperature. The results showed that AA doses affected egg conductive heat loss and hatchability, and that they did not minimize the effects of high incubation temperature on liver and heart development.

  8. Phenotypic correlations between egg weight and some egg quality ...

    African Journals Online (AJOL)

    Eggs were examined for both internal and external egg quality traits.Data obtained were subjected to one-way analysis of variance using the general linear procedure of SAS (2012). Differences in means were ranked using the Duncan's multiple Range test. Phenotypic correlations between egg weight and other egg quality ...

  9. Tumor immunology viewed from alternative animal models—the Xenopus story

    Science.gov (United States)

    Banach, Maureen; Robert, Jacques

    2017-01-01

    a) Purpose of review Nonmammalian comparative animal models are important not only to gain fundamental evolutionary understanding of the complex interactions of tumors with the immune system, but also to better predict the applicability of novel immunotherapeutic approaches to humans. After reviewing recent advances in developing alternative models, we focus on the amphibian Xenopus laevis and its usefulness in deciphering the perplexing roles of MHC class I-like molecules and innate (i)T cells in tumor immunity. b) Recent findings Experiments using MHC-defined inbred and cloned animals, tumor cell lines, effective reagents, sequenced genomes, and adapted gene editing techniques in Xenopus, have revealed that the critical involvement of class I-like molecules and iT cells in tumor immunity has been conserved during evolution. c) Summary Comparative studies with the X. laevis tumor immunity model can contribute to the development of better and more efficient cancer immunotherapies. PMID:28944105

  10. Chromosome Banding in Amphibia. XXXII. The Genus Xenopus (Anura, Pipidae).

    Science.gov (United States)

    Schmid, Michael; Steinlein, Claus

    2015-01-01

    Mitotic chromosomes of 16 species of the frog genus Xenopus were prepared from kidney and lung cell cultures. In the chromosomes of 7 species, high-resolution replication banding patterns could be induced by treating the cultures with 5-bromodeoxyuridine (BrdU) and deoxythymidine (dT) in succession, and in 6 of these species the BrdU/dT-banded chromosomes could be arranged into karyotypes. In the 3 species of the clade with 2n = 20 and 4n = 40 chromosomes (X. tropicalis, X. epitropicalis, X. new tetraploid 1), as well as in the 3 species with 4n = 36 chromosomes (X. laevis, X. borealis, X. muelleri), the BrdU/dT-banded karyotypes show a high degree of homoeology, though differences were detected between these groups. Translocations, inversions, insertions or sex-specific replication bands were not observed. Minor replication asynchronies found between chromosomes probably involve heterochromatic regions. BrdU/dT replication banding of Xenopus chromosomes provides the landmarks necessary for the exact physical mapping of genes and repetitive sequences. FISH with an X. laevis 5S rDNA probe detected multiple hybridization sites at or near the long-arm telomeric regions in most chromosomes of X. laevis and X. borealis, whereas in X. muelleri, the 5S rDNA sequences are located exclusively at the long-arm telomeres of a single chromosome pair. Staining with the AT base pair-specific fluorochrome quinacrine mustard revealed brightly fluorescing heterochromatic regions in the majority of X. borealis chromosomes which are absent in other Xenopus species.

  11. Egg temperature and embryonic metabolism of A- and B-eggs of ...

    African Journals Online (AJOL)

    Macaroni and rockhopper penguins lay two eggs but rear only one chick to independence. The eggs are markedly dimorphic in size and, although the smaller A-egg is laid several days before the B-egg, in nests where both eggs are incubated, the B-egg always hatches first. Incubation temperatures and embryonic oxygen ...

  12. [IVF and endometriosis, oocyte donation and fertility preservation].

    Science.gov (United States)

    d'Argent, Emmanuelle Mathieu; Antoine, Jean-Marie

    2017-12-01

    Endometriosis is a common condition, causing pain and infertility. In infertile woman with superficial peritoneal endometriosis and patent tubes, laparoscopy is recommended, followed by ovarian stimulation alone or in combination with intrauterine inseminations. In case of ovarian or deep endometriosis, the indications of surgery and assisted reproductive technologies remain to be defined precisely. In vitro fertilization is generally proposed after the failure of up to three inseminations, directly for ovarian or deep endometriosis, or in case of an associated factor of infertility, mainly male. Before ovarian stimulation in view to in vitro fertilization, a pretreatment by GnRH agonist for 2 to 6 months or combined contraceptive for 6 to 8 weeks would improve the pregnancy rate. Egg donation is effective in patients with advanced ovarian failure or lack of ovarian response to stimulation. Fertility preservation, especially by oocytes vitrified, must be proposed preventively to women with endometriosis at risk of ovarian failure, without close wish to be pregnant. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  13. Rapid Evolution of Assortative Fertilization between Recently Allopatric Species of Drosophila.

    Science.gov (United States)

    Ahmed-Braimah, Yasir H; McAllister, Bryant F

    2012-01-01

    The virilis group of Drosophila represents a relatively unexplored but potentially useful model to investigate the genetics of speciation. Good resolution of phylogenetic relationships and the ability to obtain fertile hybrid offspring make the group especially promising for analysis of genetic changes underlying reproductive isolation separate from hybrid sterility and inviability. Phylogenetic analyses reveal a close relationship between the sister species, Drosophila americana and D. novamexicana, yet excepting their contemporary allopatric distributions, factors that contribute to reproductive isolation between this species pair remain uncharacterized. A previous report has shown reduced progeny numbers in laboratory crosses between the two species, especially when female D. novamexicana are crossed with male D. americana. We show that the hatch rate of eggs produced from heterospecific matings is reduced relative to conspecific matings. Failure of eggs to hatch, and consequent reduction in hybrid progeny number, is caused by low fertilization success of heterospecific sperm, thus representing a postmating, prezygotic incompatibility. Following insemination, storage and motility of heterospecific sperm is visibly compromised in female D. novamexicana. Our results provide evidence for a mechanism of reproductive isolation that is seldom reported for Drosophila species, and indicate the rapid evolution of postmating, prezygotic reproductive barriers in allopatry.

  14. Egg CD9 protein tides correlated with sperm oscillations tune the gamete fusion ability in mammal.

    Science.gov (United States)

    Ravaux, Benjamin; Favier, Sophie; Perez, Eric; Gourier, Christine

    2018-01-23

    Mammalian fertilization involves membrane events -adhesion, fusion, sperm engulfment, membrane block to polyspermy- whose causes remain largely unknown. Recently, specific oscillations of the sperm in contact with the egg were shown to be necessary for fusion. Using a microfluidic chip to impose the venue for the encounter of two gametes allowed real-time observation of the membrane remodelling occurring at the sperm/egg interface. The spatiotemporal mapping of egg CD9 revealed that this protein concentrates at the egg/sperm interface as a result of sperm oscillations, until a CD9-rich platform is nucleated on which fusion immediately takes place. Within 2 to 5 minutes after fusion, most of the CD9 leaves the egg for the external aqueous medium. Then an egg membrane wave engulfs the sperm head in approximately 25 minutes. These results show that sperm oscillations initiate the CD9 recruitment that causes gamete fusion after which CD9 and associated proteins leave the membrane in a process likely to contribute to block polyspermy. They highlight that the gamete fusion story in mammals is an unexpected interplay between mechanical constraints and proteins. © The Author(s) (2018). Published by Oxford University Press on behalf of Journal of Molecular Cell Biology, IBCB, SIBS, CAS. All rights reserved.

  15. Relationships between egg-recognition and egg-ejection in a grasp-ejector species.

    Directory of Open Access Journals (Sweden)

    Manuel Soler

    Full Text Available Brood parasitism frequently leads to a total loss of host fitness, which selects for the evolution of defensive traits in host species. Experimental studies have demonstrated that recognition and rejection of the parasite egg is the most common and efficient defence used by host species. Egg-recognition experiments have advanced our knowledge of the evolutionary and coevolutionary implications of egg recognition and rejection. However, our understanding of the proximate mechanisms underlying both processes remains poor. Egg rejection is a complex behavioural process consisting of three stages: egg recognition, the decision whether or not to reject the putative parasitic egg and the act of ejection itself. We have used the blackbird (Turdus merula as a model species to explore the relationship between egg recognition and the act of egg ejection. We have manipulated the two main characteristics of parasitic eggs affecting egg ejection in this grasp-ejector species: the degree of colour mimicry (mimetic and non-mimetic, which mainly affects the egg-recognition stage of the egg-rejection process and egg size (small, medium and large, which affects the decision to eject, while maintaining a control group of non-parasitized nests. The behaviour of the female when confronted with an experimental egg was filmed using a video camera. Our results show that egg touching is an indication of egg recognition and demonstrate that blackbirds recognized (i.e., touched non-mimetic experimental eggs significantly more than mimetic eggs. However, twenty per cent of the experimental eggs were touched but not subsequently ejected, which confirms that egg recognition does not necessarily mean egg ejection and that accepting parasitic eggs, at least sometimes, is the consequence of acceptance decisions. Regarding proximate mechanisms, our results show that the delay in egg ejection is not only due to recognition problems as usually suggested, given that experimental

  16. Epididymosomes: transfer of fertility-modulating proteins to the sperm surface

    OpenAIRE

    Patricia A Martin-DeLeon

    2015-01-01

    A variety of glycosylphosphatidylinositol (GPI)-linked proteins are acquired on spermatozoa from epididymal luminal fluids (ELF) during sperm maturation. These proteins serve roles in immunoprotection and in key steps of fertilization such as capacitation, acrosomal exocytosis and sperm-egg interactions. Their acquisition on sperm cells is mediated both by membrane vesicles (epididymosomes, EP) which were first reported to dock on the sperm surface, and by lipid carriers which facilitate the ...

  17. Egg laying sequence influences egg mercury concentrations and egg size in three bird species: Implications for contaminant monitoring programs

    Science.gov (United States)

    Ackerman, Joshua T.; Eagles-Smith, Collin A.; Herzog, Mark P.; Yee, Julie L.; Hartman, C. Alex

    2016-01-01

    Bird eggs are commonly used in contaminant monitoring programs and toxicological risk assessments, but intra-clutch variation and sampling methodology could influence interpretability. We examined the influence of egg laying sequence on egg mercury concentrations and burdens in American avocets, black-necked stilts, and Forster's terns. The average decline in mercury concentrations between the first and last egg laid was 33% for stilts, 22% for terns, and 11% for avocets, and most of this decline occurred between the first and second eggs laid (24% for stilts, 18% for terns, and 9% for avocets). Trends in egg size with egg laying order were inconsistent among species and overall differences in egg volume, mass, length, and width were mercury concentrations generally declined by 16% between the first and second eggs laid. Despite the strong effect of egg laying sequence, most of the variance in egg mercury concentrations still occurred among clutches (75%-91%) rather than within clutches (9%-25%). Using simulations, we determined that to accurately estimate a population's mean egg mercury concentration using only a single random egg from a subset of nests, it would require sampling >60 nests to represent a large population (10% accuracy) or ≥14 nests to represent a small colony that contained <100 nests (20% accuracy).

  18. [Female age - related fertility decline: Far from the myth of the "selfish working-girl" and the "right to have a child"].

    Science.gov (United States)

    Vialle, M; Perrin, J; Amar-Hoffet, A; Boyer, P; Courbiere, B

    2016-04-01

    To study the social dimension of age-related female infertility through an analysis of three key themes: the personal life histories of infertile women over 40 years of age; representations of age and the desire to become pregnant after age 40; opinions of French legislations framing Assisted Reproductive Technologies, age limits, egg donation, and egg freezing for non-medical reasons. This qualitative sociological study was based on semi-structured interviews with infertile women over age 40 going through fertility treatments. The interviews contained three parts: personal and relationship histories; experiences related to age; opinions related to French legislation. Twenty-three interviews were conducted; each lasting between 90 to 120minutes. Far from having similar life histories, the women interviewed had very different backgrounds leading to their desire for a pregnancy after 40 years of age. From the beginning of their fertility treatments, they perceived a "race against the clock". This feeling of urgency accompanied their experiences and was related to the desire to not be too old for their future child. The women interviewed were mainly in favor of loosening French bioethical laws in order to avoid the need to travel abroad to pursue fertility treatments. The profiles studied attest to a growing gap between biological and biographical temporalities, as well as an inability of women to reduce their desire for a child. Faced with this disparity, egg donation and egg freezing were seen as practical solutions. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  19. The Significance of Protein Components in Heterogeneous Eggs for Embrionic Development and Larvas Persistence of Cyprinidae

    Directory of Open Access Journals (Sweden)

    Zalepukhin Valeriy Vladimirovich

    2014-12-01

    Full Text Available Endogenous heterogeneity of females and their eggs is one of the most important factors of annual generation’s formation in nature-spawn and incubative centers. Its assessment is significant for comprehending the notion of biochemical changes in proximate prespawn period. Between the shares of egg’s biochemical composition of Cyprinidae species only protein and b-lipoproteids levels are incovering constant and authentic correlations with embrionic development and larval survival in the conditions of artificial reproduction. The promotion of protein’s level to 20 – 24 % (in wet weigth in percolating eggs positively affects the fertilization and larvae vitality. This trend is fair for the domesticated and nature-spawn fish. The same significance is important for the b-lipoproteids level in ovular eggs of grass carp (Ctenopharyngodon idella Val.. The optimal concentration is 100-200 mg % in wet weight.

  20. The role of egg-nest contrast in the rejection of brood parasitic eggs.

    Science.gov (United States)

    Aidala, Zachary; Croston, Rebecca; Schwartz, Jessica; Tong, Lainga; Hauber, Mark E

    2015-04-15

    Hosts of avian brood parasites can avoid the reproductive costs of raising genetically unrelated offspring by rejecting parasitic eggs. The perceptual cues and controls mediating parasitic egg discrimination and ejection are well studied: hosts are thought to use differences in egg color, brightness, maculation, size and shape to discriminate between their own and foreign eggs. Most theories of brood parasitism implicitly assume that the primary criteria to which hosts attend when discriminating eggs are differences between the eggs themselves. However, this assumption is confounded by the degree to which chromatic and achromatic characteristics of the nest lining co-vary with egg coloration, so that egg-nest contrast per se might be the recognition cue driving parasitic egg detection. Here, we systematically tested whether and how egg-nest contrast itself contributes to foreign egg discrimination. In an artificial parasitism experiment, we independently manipulated egg color and nest lining color of the egg-ejector American robin (Turdus migratorius), a host of the obligate brood parasitic brown-headed cowbird (Molothrus ater). We hypothesized that the degree of contrast between foreign eggs and the nest background would affect host egg rejection behavior. We predicted that experimentally decreasing egg-nest chromatic and achromatic contrast (i.e. rendering parasitic eggs more cryptic against the nest lining) would decrease rejection rates, while increasing egg-nest contrast would increase rejection rates. In contrast to our predictions, egg-nest contrast was not a significant predictor of egg ejection patterns. Instead, egg color significantly predicted responses to parasitism. We conclude that egg-egg differences are the primary drivers of egg rejection in this system. Future studies should test for the effects of egg-nest contrast per se in predicting parasitic egg recognition in other host-parasite systems, including those hosts building enclosed nests and

  1. Broiler performance, hatching egg, and age relationships of progeny from standard and dwarf broiler dams.

    Science.gov (United States)

    Tahir, M; Cervantes, H; Farmer, C W; Shim, M Y; Pesti, G M

    2011-06-01

    The relationship of egg and chick weights to the performance of broiler chickens from two 42-wk-old flocks (standard and dwarf dams) having male parents from the same genetic stock was investigated in this study. Fertility (91.7 vs. 94.7%) and hatchability (95.2 vs. 96.3%) were not significantly (P > 0.10) different for eggs from standard and dwarf dams, respectively. Egg weight contributed significantly to the variation in BW [BW = β(0) + β(i) (egg weight) + β(i) (dam) + β(i) (sex)]. Body weight as a function of chick weight was not significant. However, chick weight was significant when included in a model with egg weight, suggesting that significant differences in BW at 50 d could be attributed to both egg and chick weights. The negative coefficient for chick weight indicated that between the 2 broilers of the same egg weight, the one with the greater chick weight would have the smaller 50-d BW. Chick weight was a linear function of egg weight. Similarly, the effect of egg or chick weight on broiler BW at 35 or 50 d was best represented by a single linear function. Dam genotype did not contribute significantly to variation in 50-d BW after variation attributable to egg weight was removed from the model. Differences in BW attributable to egg weight increased with broiler age. The coefficients of egg weight and chick weight showed that the differences in BW per gram of egg were 1.43, 3.06, 6.24, and 7.61 g and those per gram of chick were 1.87, 3.99, 8.14, and 9.93 g, respectively, at 7, 21, 35, and 50 d. Body weight increased by 0.1563 times egg weight (and 0.2092 times chick weight) with each additional day of age for both sexes and genotypes. Clearly, both egg and chick weights are important for modeling or predicting market-age broiler BW and economic returns. The relatively small relationship between BW and egg weight demonstrates that genetic selection over the past 3 decades has decreased the influence of egg weight on broiler growth. The present dwarf

  2. Ostrich eggs geometry

    Directory of Open Access Journals (Sweden)

    Šárka Nedomová

    2013-01-01

    Full Text Available Precise quantification of the profile of egg can provide a powerful tool for the analysis of egg shape for various biological problems. A new approach to the geometry of a Ostrich’s egg profile is presented here using an analysing the egg’s digital photo by edge detection techniques. The obtained points on the eggshell counter are fitted by the Fourier series. The obtained equations describing an egg profile have been used to calculate radii of curvature. The radii of the curvature at the important point of the egg profile (sharp end, blunt end and maximum thickness are independent on the egg shape index. The exact values of the egg surface and the egg volume have been obtained. These quantities are also independent on the egg shape index. These quantities can be successively estimated on the basis of simplified equations which are expressed in terms of the egg length, L¸ and its width, B. The surface area of the eggshells also exhibits good correlation with the egg long circumference length. Some limitations of the most used procedures have been also shown.

  3. Adaptive immunity and histopathology in frog virus 3-infected Xenopus

    International Nuclear Information System (INIS)

    Robert, Jacques; Morales, Heidi; Buck, Wayne; Cohen, Nicholas; Marr, Shauna; Gantress, Jennifer

    2005-01-01

    Xenopus has been used as an experimental model to evaluate the contribution of adaptive cellular immunity in amphibian host susceptibility to the emerging ranavirus FV3. Conventional histology and immunohistochemistry reveal that FV3 has a strong tropism for the proximal tubular epithelium of the kidney and is rarely disseminated elsewhere in Xenopus hosts unless their immune defenses are impaired or developmentally immature as in larvae. In such cases, virus is found widespread in most tissues. Adults, immunocompromised by depletion of CD8 + T cells or by sub-lethal γ-irradiation, show increased susceptibility to FV3 infection. Larvae and irradiated (but not normal) adults can be cross-infected through water by infected adult conspecifics (irradiated or not). The natural MHC class I deficiency and the absence of effect of anti-CD8 treatment on both larval CD8 + T cells and larval susceptibility to FV3 are consistent with an inefficient CD8 + T cell effector function during this developmental period

  4. [The effect of palm oil and safflower oil in the feed of parent fattening hens on fertility, hatchability and growth of progeny].

    Science.gov (United States)

    Halle, I

    1999-01-01

    The aim of two experiments with broiler breeder hens was to evaluate the effect of diets containing palm butter or safflower oil (25 g and 50 g/kg feed, resp.) on fertility, hatchability and growth of progeny. Especially the incorporation of oleic and linoleic acid in egg yolk reflected the dietary fatty acid source. Eggs were collected and stored in the incubator at a hen age of 31, 40, 50, and 60 weeks. Hatched chicks were reared over 5 weeks. The number of fertile eggs (Experiment 1 and 2, 75 and 88%, resp.) differed between the experiments (P < or = 0.05). Neither embryonic mortality nor hatchability (Experiment 1 and 2, 76 and 78%, resp.) were significantly affected by fatty acid composition of yolk. No clear maternal dietary effect was recorded on chicken weight at hatching (Experiment 1 and 2, 43.3 g and 43.7 g, resp.) and at 35 days of age (Experimental 1 and 2, 1676 g and 1764 g, resp.) The fatty acid composition in the analysed egg yolk sac of chicks showed a different fatty level but corresponded to fatty acid composition of breeding eggs before incubation. According to a decreased level of docosahexaenoic acid in egg yolk due to increased incorporation of linoleic acid, the content of this fatty acid was also diminished in phospholipids of the brain of chicken on days 1 and 5 after hatching.

  5. Early development of Ensatina eschscholtzii: an amphibian with a large, yolky egg

    Directory of Open Access Journals (Sweden)

    Collazo Andres

    2010-08-01

    Full Text Available Abstract Background Comparative analyses between amphibians, concentrating on the cellular mechanisms of morphogenesis, reveal a large variability in the early developmental processes that were thought to be conserved during evolution. Increased egg size is one factor that could have a strong effect on early developmental processes such as cleavage pattern and gastrulation. Salamanders of the family Plethodontidae are particularly appropriate for such comparative studies because the species have eggs of varying size, including very large yolky eggs. Results In this paper, we describe for the first time the early development (from fertilization through neurulation of the plethodontid salamander Ensatina eschscholtzii. This species has one of the largest eggs known for an amphibian, with a mean ± SD diameter of 6 ± 0.43 mm (range 5.3-6.9; n = 17 eggs. Cleavage is meroblastic until approximately the 16-cell stage (fourth or fifth cleavage. At the beginning of gastrulation, the blastocoel roof is one cell thick, and the dorsal lip of the blastopore forms below the equator of the embryo. The ventral lip of the blastopore forms closer to the vegetal pole, and relatively little involution occurs during gastrulation. Cell migration is visible through the transparent blastocoel roof of the gastrula. At the end of gastrulation, a small archenteron spreading dorsally from the blastopore represents the relatively small and superficial area of the egg where early embryonic axis formation occurs. The resulting pattern is similar to the embryonic disk described for one species of anuran. Conclusions Comparisons with the early development of other species of amphibians suggest that an evolutionary increase in egg size can result in predictable changes in the patterns and rate of early development, but mainly within an evolutionary lineage.

  6. Becoming a morther by non-anonymous egg donation: secrecy and the relationship between egg recipient, egg donor and egg donation child

    NARCIS (Netherlands)

    Berkel, D.A.M. van; Candido, A.; Pijffers, W.H.

    The object of the study was to investigate secrecy in non-anonymous egg donation, to explore some characteristics of this kind of egg donation arrangement and the relationship of the recipient with her non-genetic child. Forty-four egg recipients and 62 IVF patients with a child conceived through

  7. 9 CFR 590.410 - Shell eggs and egg products required to be labeled.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Shell eggs and egg products required..., DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION INSPECTION OF EGGS AND EGG PRODUCTS (EGG PRODUCTS INSPECTION ACT) Identifying and Marking Product § 590.410 Shell eggs and egg products required to be labeled...

  8. Light-induced, GTP-binding protein mediated membrane currents of Xenopus oocytes injected with rhodopsin of cephalopods.

    Science.gov (United States)

    Ando, H; Seidou, M; Kito, Y

    1991-01-01

    Xenopus oocytes that were injected with rhabdomeric membranes of squid and octopus photoreceptors acquired light sensitivity. The injected oocytes showed a light-induced current having characteristics similar to other G-protein-mediated Cl- currents induced by the activation of other membrane receptors. Pretreatment of the oocytes with pertussis toxin before the injection suppressed the generation of the light-induced current, indicating an ability of cephalopod rhodopsin to cross-react with an endogenous G-protein of Xenopus oocytes.

  9. rab3 mediates cortical granule exocytosis in the sea urchin egg.

    Science.gov (United States)

    Conner, S; Wessel, G M

    1998-11-15

    Egg activation at fertilization in the sea urchin results in the exocytosis of approximately 15,000 cortical granules that are docked at the plasma membrane. Previously, we reported that several integral membrane proteins modeled in the SNARE hypothesis, synaptotagmin, VAMP, and syntaxin, in addition to a small GTPase of the ras superfamily, rab3, were present on cortical granules (Conner, S., Leaf, D., and Wessel, G., Mol. Reprod. Dev. 48, 1-13, 1997). Here we report that rab3 is associated with cortical granules throughout oogenesis, during cortical granule translocation, and while docked at the egg plasma membrane. Following cortical granule exocytosis, however, rab3 reassociates with a different population of vesicles, at least some of which are of endocytic origin. Because of its selective association with cortical granules in eggs and oocytes, we hypothesize that rab3 functions in cortical granule exocytosis. To test this hypothesis, we used a strategy of interfering with rab3 function by peptide competition with its effector domain, a conserved region within specific rab types. We first identified the effector domain sequence in Lytechinus variegatus eggs and find the sequence 94% identical to the effector domain of rab3 in Stronglocentrotus purpuratus. Then, with synthetic peptides to different regions of the rab3 protein, we find that cortical granule exocytosis is inhibited in eggs injected with effector domain peptides, but not with peptides from the hypervariable region or with a scrambled effector peptide. Additionally, effector-peptide-injected eggs injected with IP3 are blocked in their ability to exocytose cortical granules, suggesting that the inhibition is directly on the membrane fusion event and not the result of interference with the signal transduction mechanism leading to calcium release. We interpret these results to mean that rab3 functions in the regulation of cortical granule exocytosis following vesicle docking. Copyright 1998 Academic

  10. No detectable fertility benefit from a single additional mating in wild stalk-eyed flies.

    Directory of Open Access Journals (Sweden)

    Elisabeth Harley

    2010-12-01

    Full Text Available Multiple mating by female insects is widespread, and the explanation(s for repeated mating by females has been the subject of much discussion. Females may profit from mating multiply through direct material benefits that increase their own reproductive output, or indirect genetic benefits that increase offspring fitness. One particular direct benefit that has attracted significant attention is that of fertility assurance, as females often need to mate multiply to achieve high fertility. This hypothesis has never been tested in a wild insect population.Female Malaysian stalk-eyed flies (Teleopsis dalmanni mate repeatedly during their lifetime, and have been shown to be sperm limited under both laboratory and field conditions. Here we ask whether receiving an additional mating alleviates sperm limitation in wild females. In our experiment one group of females received a single additional mating, while a control group received an interrupted, and therefore unsuccessful, mating. Females that received an additional mating did not lay more fertilised eggs in total, nor did they lay proportionately more fertilised eggs. Female fertility declined significantly through time, demonstrating that females were sperm limited. However, receipt of an additional mating did not significantly alter the rate of this decline.Our data suggest that the fertility consequences of a single additional mating were small. We discuss this effect (or lack thereof, and suggest that it is likely to be attributed to small ejaculate size, a high proportion of failed copulations, and the presence of X-linked meiotic drive in this species.

  11. Commercially laid eggs vs. discarded hatching eggs: contamination by Salmonella spp.

    Science.gov (United States)

    Kottwitz, Luciana B M; Leão, Joice Aparecida; Back, Alberto; Rodrigues, Dalia dos P; Magnani, Marciane; de Oliveira, Tereza C R M

    2013-01-01

    Salmonella enterica is frequently associated with outbreaks of human salmonellosis, and products of avian origin, such as eggs and chicken meat, are the main vehicles of its transmission. The present study describes the occurrence of different serovars of Salmonella enterica and phagotypes of S. enterica serovar Enteritidis in eggs destined for human consumption. Four thousand eggs obtained from commercial egg laying farms and one thousand discarded hatching eggs from broiler farms, which were acquired at farmers' markets and informal shops, were analyzed. Salmonella spp. was isolated from 52.0% of the discarded hatching eggs, in which the predominant serovar was Enteritidis (84.6%), and the predominant Salmonella Enteritidis phagotype (PT) was PT7 (26.9%). Salmonella spp. was not isolated from eggs obtained from commercial egg laying farms. The antimicrobial resistance profile showed that 23.1% (n = 6) of the SE strains were resistant to nalidixic acid. The results suggest that the consumption of discarded hatching eggs represents an important source of Salmonella transmission to humans.

  12. The establishment of polarized membrane traffic in Xenopus laevis embryos.

    Science.gov (United States)

    Roberts, S J; Leaf, D S; Moore, H P; Gerhart, J C

    1992-09-01

    Delineation of apical and basolateral membrane domains is a critical step in the epithelialization of the outer layer of cells in the embryo. We have examined the initiation of polarized membrane traffic in Xenopus and show that membrane traffic is not polarized in oocytes but polarized membrane domains appear at first cleavage. The following proteins encoded by injected RNA transcripts were used as markers to monitor membrane traffic: (a) VSV G, a transmembrane glycoprotein preferentially inserted into the basolateral surface of polarized epithelial cells; (b) GThy-1, a fusion protein of VSV G and Thy-1 that is localized to the apical domains of polarized epithelial cells; and (c) prolactin, a peptide hormone that is not polarly secreted. In immature oocytes, there is no polarity in the expression of VSV G or GThy-1, as shown by the constitutive expression of both proteins at the surface in the animal and vegetal hemispheres. At meiotic maturation, membrane traffic to the surface is blocked; the plasma membrane no longer accepts the vesicles synthesized by the oocyte (Leaf, D. L., S. J. Roberts, J. C. Gerhart, and H.-P. Moore. 1990. Dev. Biol. 141:1-12). When RNA transcripts are injected after fertilization, VSV G is expressed only in the internal cleavage membranes (basolateral orientation) and is excluded from the outer surface (apical orientation, original oocyte membrane). In contrast, GThy-1 and prolactin, when expressed in embryos, are inserted or released at both the outer membrane derived from the oocyte and the inner cleavage membranes. Furthermore, not all of the cleavage membrane comes from an embryonic pool of vesicles--some of the cleavage membrane comes from vesicles synthesized during oogenesis. Using prolactin as a marker, we found that a subset of vesicles synthesized during oogenesis was only released after fertilization. However, while embryonic prolactin was secreted from both apical and basolateral surfaces, the secretion of oogenic prolactin

  13. Mother and offspring fitness in an insect with maternal care: phenotypic trade-offs between egg number, egg mass and egg care

    Science.gov (United States)

    2014-01-01

    Background Oviparous females have three main options to increase their reproductive success: investing into egg number, egg mass and/or egg care. Although allocating resources to either of these three components is known to shape offspring number and size, potential trade-offs among them may have key impacts on maternal and offspring fitness. Here, we tested the occurrence of phenotypic trade-offs between egg number, egg mass and maternal expenditure on egg care in the European earwig, Forficula auricularia, an insect with pre- and post-hatching forms of maternal care. In particular, we used a series of laboratory observations and experiments to investigate whether these three components non-additively influenced offspring weight and number at hatching, and whether they were associated with potential costs to females in terms of future reproduction. Results We found negative associations between egg number and mass as well as between egg number and maternal expenditure on egg care. However, these trade-offs could only be detected after statistically correcting for female weight at egg laying. Hatchling number was not determined by single or additive effects among the three life-history traits, but instead by pairwise interactions among them. In particular, offspring number was positively associated with the number of eggs only in clutches receiving high maternal care or consisting of heavy eggs, and negatively associated with mean egg mass in clutches receiving low care. In contrast, offspring weight was positively associated with egg mass only. Finally, maternal expenditure on egg care reduced their future reproduction, but this effect was only detected when mothers were experimentally isolated from their offspring at egg hatching. Conclusions Overall, our study reveals simultaneous trade-offs between the number, mass and care of eggs. It also demonstrates that these factors interact in their impact on offspring production, and that maternal expenditure on egg

  14. Visualisation of cerebrospinal fluid flow patterns in albino Xenopus larvae in vivo

    Directory of Open Access Journals (Sweden)

    Mogi Kazue

    2012-04-01

    Full Text Available Abstract Background It has long been known that cerebrospinal fluid (CSF, its composition and flow, play an important part in normal brain development, and ependymal cell ciliary beating as a possible driver of CSF flow has previously been studied in mammalian fetuses in vitro. Lower vertebrate animals are potential models for analysis of CSF flow during development because they are oviparous. Albino Xenopus laevis larvae are nearly transparent and have a straight, translucent brain that facilitates the observation of fluid flow within the ventricles. The aim of these experiments was to study CSF flow and circulation in vivo in the developing brain of living embryos, larvae and tadpoles of Xenopus laevis using a microinjection technique. Methods The development of Xenopus larval brain ventricles and the patterns of CSF flow were visualised after injection of quantum dot nanocrystals and polystyrene beads (3.1 or 5.8 μm in diameter into the fourth cerebral ventricle at embryonic/larval stages 30-53. Results The fluorescent nanocrystals showed the normal development of the cerebral ventricles from embryonic/larval stages 38 to 53. The polystyrene beads injected into stage 47-49 larvae revealed three CSF flow patterns, left-handed, right-handed and non-biased, in movement of the beads into the third ventricle from the cerebral aqueduct (aqueduct of Sylvius. In the lateral ventricles, anterior to the third ventricle, CSF flow moved anteriorly along the outer wall of the ventricle to the inner wall and then posteriorly, creating a semicircle. In the cerebral aqueduct, connecting the third and fourth cerebral ventricles, CSF flow moved rostrally in the dorsal region and caudally in the ventral region. Also in the fourth ventricle, clear dorso-ventral differences in fluid flow pattern were observed. Conclusions This is the first visualisation of the orchestrated CSF flow pattern in developing vertebrates using a live animal imaging approach. CSF flow

  15. NAA Comparison of Nutriens in Egg Yolk and Egg White

    International Nuclear Information System (INIS)

    Ruangdit, S.; Maijan, P.; Channuie, J.; Picha, R.

    2014-01-01

    Food we eat has significant effects on our wellbeing. Eggs are among food products widely consumed and contain many essential nutrients. Yet eggs often are involved in dietary controversy regarding benefit versus risk. The fear of serum cholesterol has driven a large number of people away from consuming egg yolks. In this study, we assessed the nutritional importance of eggs using instrumental neutron activation analysis (INAA) technique at Thai Research Reactor, operating at 1.2 MW. We investigated whether consumers miss any health benefits by choosing to eat only egg whites or yolks rather than whole natural eggs. Essential minerals such as sodium, potassium, manganese, magnesium, copper, selenium, iron and zinc are studied using three NAA loading methods: short-term individual pneumatic transfer system loading using inner-core tubes, medium-term CA3 loading and long-term Lazy Susan (LS) loading for 10 s, 7 h and 3 d, respectively. Two Standard Reference Materials (SRMs) were used to find the nutrient concentrations and validate the method. From the study, we found significant differences in nutritional contents between egg yolks and egg whites. Along with literature review conducted as part of this study, we evaluated the consumption choice when it comes to eggs.

  16. Dinosaur origin of egg color: oviraptors laid blue-green eggs.

    Science.gov (United States)

    Wiemann, Jasmina; Yang, Tzu-Ruei; Sander, Philipp N; Schneider, Marion; Engeser, Marianne; Kath-Schorr, Stephanie; Müller, Christa E; Sander, P Martin

    2017-01-01

    Protoporphyrin (PP) and biliverdin (BV) give rise to the enormous diversity in avian egg coloration. Egg color serves several ecological purposes, including post-mating signaling and camouflage. Egg camouflage represents a major character of open-nesting birds which accomplish protection of their unhatched offspring against visually oriented predators by cryptic egg coloration. Cryptic coloration evolved to match the predominant shades of color found in the nesting environment. Such a selection pressure for the evolution of colored or cryptic eggs should be present in all open nesting birds and relatives. Many birds are open-nesting, but protect their eggs by continuous brooding, and thus exhibit no or minimal eggshell pigmentation. Their closest extant relatives, crocodiles, protect their eggs by burial and have unpigmented eggs. This phylogenetic pattern led to the assumption that colored eggs evolved within crown birds. The mosaic evolution of supposedly avian traits in non-avian theropod dinosaurs, however, such as the supposed evolution of partially open nesting behavior in oviraptorids, argues against this long-established theory. Using a double-checking liquid chromatography ESI-Q-TOF mass spectrometry routine, we traced the origin of colored eggs to their non-avian dinosaur ancestors by providing the first record of the avian eggshell pigments protoporphyrin and biliverdin in the eggshells of Late Cretaceous oviraptorid dinosaurs. The eggshell parataxon Macroolithus yaotunensis can be assigned to the oviraptor Heyuannia huangi based on exceptionally preserved, late developmental stage embryo remains. The analyzed eggshells are from three Late Cretaceous fluvial deposits ranging from eastern to southernmost China. Reevaluation of these taphonomic settings, and a consideration of patterns in the porosity of completely preserved eggs support an at least partially open nesting behavior for oviraptorosaurs. Such a nest arrangement corresponds with our

  17. Dinosaur origin of egg color: oviraptors laid blue-green eggs

    Directory of Open Access Journals (Sweden)

    Jasmina Wiemann

    2017-08-01

    Full Text Available Protoporphyrin (PP and biliverdin (BV give rise to the enormous diversity in avian egg coloration. Egg color serves several ecological purposes, including post-mating signaling and camouflage. Egg camouflage represents a major character of open-nesting birds which accomplish protection of their unhatched offspring against visually oriented predators by cryptic egg coloration. Cryptic coloration evolved to match the predominant shades of color found in the nesting environment. Such a selection pressure for the evolution of colored or cryptic eggs should be present in all open nesting birds and relatives. Many birds are open-nesting, but protect their eggs by continuous brooding, and thus exhibit no or minimal eggshell pigmentation. Their closest extant relatives, crocodiles, protect their eggs by burial and have unpigmented eggs. This phylogenetic pattern led to the assumption that colored eggs evolved within crown birds. The mosaic evolution of supposedly avian traits in non-avian theropod dinosaurs, however, such as the supposed evolution of partially open nesting behavior in oviraptorids, argues against this long-established theory. Using a double-checking liquid chromatography ESI-Q-TOF mass spectrometry routine, we traced the origin of colored eggs to their non-avian dinosaur ancestors by providing the first record of the avian eggshell pigments protoporphyrin and biliverdin in the eggshells of Late Cretaceous oviraptorid dinosaurs. The eggshell parataxon Macroolithus yaotunensis can be assigned to the oviraptor Heyuannia huangi based on exceptionally preserved, late developmental stage embryo remains. The analyzed eggshells are from three Late Cretaceous fluvial deposits ranging from eastern to southernmost China. Reevaluation of these taphonomic settings, and a consideration of patterns in the porosity of completely preserved eggs support an at least partially open nesting behavior for oviraptorosaurs. Such a nest arrangement corresponds

  18. Transgenic Xenopus laevis Line for In Vivo Labeling of Nephrons within the Kidney

    Directory of Open Access Journals (Sweden)

    Mark E. Corkins

    2018-04-01

    Full Text Available Xenopus laevis embryos are an established model for studying kidney development. The nephron structure and genetic pathways that regulate nephrogenesis are conserved between Xenopus and humans, allowing for the study of human disease-causing genes. Xenopus embryos are also amenable to large-scale screening, but studies of kidney disease-related genes have been impeded because assessment of kidney development has largely been limited to examining fixed embryos. To overcome this problem, we have generated a transgenic line that labels the kidney. We characterize this cdh17:eGFP line, showing green fluorescent protein (GFP expression in the pronephric and mesonephric kidneys and colocalization with known kidney markers. We also demonstrate the feasibility of live imaging of embryonic kidney development and the use of cdh17:eGFP as a kidney marker for secretion assays. Additionally, we develop a new methodology to isolate and identify kidney cells for primary culture. We also use morpholino knockdown of essential kidney development genes to establish that GFP expression enables observation of phenotypes, previously only described in fixed embryos. Taken together, this transgenic line will enable primary kidney cell culture and live imaging of pronephric and mesonephric kidney development. It will also provide a simple means for high-throughput screening of putative human kidney disease-causing genes.

  19. Anosmin-1 is essential for neural crest and cranial placodes formation in Xenopus.

    Science.gov (United States)

    Bae, Chang-Joon; Hong, Chang-Soo; Saint-Jeannet, Jean-Pierre

    2018-01-15

    During embryogenesis vertebrates develop a complex craniofacial skeleton associated with sensory organs. These structures are primarily derived from two embryonic cell populations the neural crest and cranial placodes, respectively. Neural crest cells and cranial placodes are specified through the integrated action of several families of signaling molecules, and the subsequent activation of a complex network of transcription factors. Here we describe the expression and function of Anosmin-1 (Anos1), an extracellular matrix protein, during neural crest and cranial placodes development in Xenopus laevis. Anos1 was identified as a target of Pax3 and Zic1, two transcription factors necessary and sufficient to generate neural crest and cranial placodes. Anos1 is expressed in cranial neural crest progenitors at early neurula stage and in cranial placode derivatives later in development. We show that Anos1 function is required for neural crest and sensory organs development in Xenopus, consistent with the defects observed in Kallmann syndrome patients carrying a mutation in ANOS1. These findings indicate that anos1 has a conserved function in the development of craniofacial structures, and indicate that anos1-depleted Xenopus embryos represent a useful model to analyze the pathogenesis of Kallmann syndrome. Copyright © 2017. Published by Elsevier Inc.

  20. Sperm competition, but not major histocompatibility divergence, drives differential fertilization success between alternative reproductive tactics in Chinook salmon.

    Science.gov (United States)

    Lehnert, S J; Helou, L; Pitcher, T E; Heath, J W; Heath, D D

    2018-01-01

    Post-copulatory sexual selection processes, including sperm competition and cryptic female choice (CFC), can operate based on major histocompatibility (MH) genes. We investigated sperm competition between male alternative reproductive tactics [jack (sneaker) and hooknose (guard)] of Chinook salmon (Oncorhynchus tshawytscha). Using a full factorial design, we examined in vitro competitive fertilization success of paired jack and hooknose males at three time points after sperm activation (0, 15 and 60 s) to test for male competition, CFC and time effects on male fertilization success. We also examined egg-mediated CFC at two MH genes by examining both the relationship between competitive fertilization success and MH divergence as well as inheritance patterns of MH alleles in resulting offspring. We found that jacks sired more offspring than hooknose males at 0 s post-activation; however, jack fertilization success declined over time post-activation, suggesting a trade-off between sperm speed and longevity. Enhanced fertilization success of jacks (presumably via higher sperm quality) may serve to increase sneaker tactic competitiveness relative to dominant hooknose males. We also found evidence of egg-mediated CFC (i.e. female × male interaction) influencing competitive fertilization success; however, CFC was not acting on the MH genes as we found no relationship between fertilization success and MH II β 1 or MH I α 1 divergence and we found no deviations from Mendelian inheritance of MH alleles in the offspring. Our study provides insight into evolutionary mechanisms influencing variation in male mating success within alternative reproductive tactics, thus underscoring different strategies that males can adopt to attain success. © 2017 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2017 European Society For Evolutionary Biology.

  1. Sequence-specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98.

    Science.gov (United States)

    Takahashi, Y; Bigler, D; Ito, Y; White, J M

    2001-04-01

    ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion. Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs. Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain. Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop. Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface. Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop. Bead binding was also inhibited by the function-blocking anti-alpha6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-alpha6 mAb, or by mAbs against either the alphav or beta3 integrin subunits. We also present evidence that in addition to the tetraspanin CD9, two other beta1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs. Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain. Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg beta1 integrin-associated proteins in sperm-egg binding and fusion. We propose that an egg surface "tetraspan web" facilitates fertilization and that it may do so by fostering ADAM-integrin interactions.

  2. Sequence-Specific Interaction between the Disintegrin Domain of Mouse ADAM 3 and Murine Eggs: Role of β1 Integrin-associated Proteins CD9, CD81, and CD98

    Science.gov (United States)

    Takahashi, Yuji; Bigler, Dora; Ito, Yasuhiko; White, Judith M.

    2001-01-01

    ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion. Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs. Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain. Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop. Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface. Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop. Bead binding was also inhibited by the function-blocking anti-α6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-α6 mAb, or by mAbs against either the αv or β3 integrin subunits. We also present evidence that in addition to the tetraspanin CD9, two other β1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs. Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain. Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg β1 integrin-associated proteins in sperm-egg binding and fusion. We propose that an egg surface “tetraspan web” facilitates fertilization and that it may do so by fostering ADAM–integrin interactions. PMID:11294888

  3. Toxicity of pesticides associated with potato production, including soil fumigants, to snapping turtle eggs (Chelydra serpentina).

    Science.gov (United States)

    de Solla, Shane Raymond; Palonen, Kimberley Elizabeth; Martin, Pamela Anne

    2014-01-01

    Turtles frequently oviposit in soils associated with agriculture and, thus, may be exposed to pesticides or fertilizers. The toxicity of a pesticide regime that is used for potato production in Ontario on the survivorship of snapping turtle (Chelydra serpentina) eggs was evaluated. The following treatments were applied to clean soil: 1) a mixture of the pesticides chlorothalonil, S-metolachlor, metribuzin, and chlorpyrifos, and 2) the soil fumigant metam sodium. Turtle eggs were incubated in soil in outdoor plots in which these mixtures were applied at typical and higher field application rates, where the eggs were subject to ambient temperature and weather conditions. The pesticide mixture consisting of chlorothalonil, S-metolachlor, metribuzin, and chlorpyrifos did not affect survivorship, deformities, or body size at applications up to 10 times the typical field application rates. Hatching success ranged between 87% and 100% for these treatments. Metam sodium was applied at 0.1¯ times, 0.3¯ times, 1 times, and 3 times field application rates. Eggs exposed to any application of metam sodium had 100% mortality. At typical field application rates, the chemical regime associated with potato production does not appear to have any detrimental impacts on turtle egg development, except for the use of the soil fumigant metam sodium, which is highly toxic to turtle eggs at the lowest recommended application rate. © 2013 SETAC.

  4. Effect of Linear Alkylbenzene Sulfonate on Mortality, Hatching Rate of Eggs and Abnormality of Catfish (Pangasius hypophthalmus Sauvage Larvae

    Directory of Open Access Journals (Sweden)

    E. Supriyono

    2007-01-01

    Full Text Available Linear alkylbenzene sulfonate (LAS surfactant in the water can affecting fish in all developmental stages.  This study was aimed to observe the effect of LAS on mortality, hatching rate of eggs, and abnormality of patin catfish (Pangasius hypophthalmus Sauvage larvae.   Fertilized eggs were incubated in water containing LAS at the dosages of 0.0, 0.5, 1.5, 3.0, 9.0, and 18.0 mg/L.  Eggs mortality was observed every 6 hours until larvae hatched (24 hours.  The results of study showed that the exposure of 18.0 mg LAS per liter water could put to death all the fertilized eggs and larvae be abnormal.  The exposure of LAS at concentration of 9.0 mg/L could kill 98% of eggs and hatching rate was only 2%.  The abnormality in larvae was bending in the body and tails. Keywords: patin catfish, Pangasius, Linear Alkylbenzene Sulfonate, LAS, abnormality   ABSTRAK Surfaktan Linear Alkylbenzene Sulfonate (LAS yang masuk ke dalam perairan sangat berpengaruh terhadap ikan dari stadia awal hidup ikan sampai dewasa. Penelitian ini bertujuan untuk mengetahui pengaruh LAS terhadap mortalitas, daya tetas telur dan abnormalitas larva ikan patin (Pangasius hypophthalmus Sauvage.  Telur ikan patin yang telah dibuahi di rendam dalam air yang mengandung LAS dengan konsentrasi 0,0; 0,5; 1,5; 3,0; 9,0 dan 18,0 mg/L.  Mortalitas telur dicatat setiap 6 jam sampai menetas (24 jam.  Hasil penelitian menunjukkan bahwa konsentrasi LAS sebesar 18,0 mg/L dapat mengakibatkan mortalitas telur dan abnormalitas pada larva secara total.  Konsentrasi LAS sebesar 9,0 mg/L dapat mematikan telur hingga 98% dan hanya menghasilkan daya tetas sebanyak 2%. Abnormalitas pada larva berupa pembengkokan pada tubuh dan ekor. Kata kunci: ikan patin, Pangasius, Linear Alkylbenzene Sulfonate, LAS dan abnormalitas

  5. Egg-laying sequence influences egg mercury concentrations and egg size in three bird species: Implications for contaminant monitoring programs.

    Science.gov (United States)

    Ackerman, Joshua T; Eagles-Smith, Collin A; Herzog, Mark P; Yee, Julie L; Hartman, C Alex

    2016-06-01

    Bird eggs are commonly used in contaminant monitoring programs and toxicological risk assessments, but intraclutch variation and sampling methodology could influence interpretability. The authors examined the influence of egg-laying sequence on egg mercury concentrations and burdens in American avocets, black-necked stilts, and Forster's terns. The average decline in mercury concentrations between the first and last eggs laid was 33% for stilts, 22% for terns, and 11% for avocets, and most of this decline occurred between the first and second eggs laid (24% for stilts, 18% for terns, and 9% for avocets). Trends in egg size with egg-laying order were inconsistent among species, and overall differences in egg volume, mass, length, and width were eggs laid. Despite the strong effect of egg-laying sequence, most of the variance in egg mercury concentrations still occurred among clutches (75-91%) rather than within clutches (9%-25%). Using simulations, the authors determined that accurate estimation of a population's mean egg mercury concentration using only a single random egg from a subset of nests would require sampling >60 nests to represent a large population (10% accuracy) or ≥14 nests to represent a small colony that contained <100 nests (20% accuracy). Environ Toxicol Chem 2016;35:1458-1469. Published 2015 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America. Published 2015 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America.

  6. Xenopus laevis embryos and tadpoles as models for testing for ...

    African Journals Online (AJOL)

    The toxicity of bio available Zn, Cu, Pb, and Cd on the life stages of Xenopus laevis embryos and tadpoles was investigated. Cu and Cd were found to affect the hatching success of the embryos, with a strong negative relationship existing between the increase in Cu concentrations and the hatching of the embryos.

  7. Membrane fusion is induced by a distinct peptide sequence of the sea urchin fertilization protein bindin

    NARCIS (Netherlands)

    Ulrich, AS; Glabe, CG; Hoekstra, D

    1998-01-01

    Fertilization in the sea urchin is mediated by the membrane-associated acrosomal protein bindin, which plays a key role in the adhesion and fusion between sperm and egg. We have investigated the structure/function relationship of an 18-amino acid peptide fragment "B18," which represents the minimal

  8. Patent gastrointestinal nematode infections in organically and conventionally pastured dairy cows and their impact on individual milk and fertility parameters.

    Science.gov (United States)

    May, Katharina; Brügemann, Kerstin; König, Sven; Strube, Christina

    2017-10-15

    Infections with gastrointestinal nematodes (GIN) can lead to production losses and impacts on product quality in affected cows, which has mainly been demonstrated during deworming experiments or via herd-level measurements. Here, a field study was carried out to explore the association between GIN infection status and milk production as well as fertility parameters in individual dairy cows. Different selection lines of Black and White cows were included in the study, which were distributed among 17 small and medium-sized organic and conventional German grassland farms. Faecal samples of 1166 dairy cows were examined twice, in July and September 2015. Nematode eggs were found in the faeces of 473 (40.6%) cows. As expected, strongylid eggs (Trichostrongylidae or Oesophagostomum and Bunostomum spp., respectively) were the predominant morphotype, followed by Strongyloides papillosus and Capillaria spp. eggs. In July, cows kept under organic conditions had a significantly lower GIN prevalence in comparison to cows kept on conventional farms. Faecal egg counts were generally low, with the highest value in September and an arithmetic mean of 11.3 eggs per gram faeces (EPG) for all observations. The relationships between GIN infection status and milk yield (kg milk/cow/day), milk protein content (%) and milk fat content (%) for each first test-day record after parasitological assessment were estimated by using linear mixed models. Milk protein content was estimated 0.05% lower in GIN positive compared to GIN negative cows, whereas no significant effect on milk yield or milk fat content was observed. The impact of GIN infection status on success in first insemination (SFI) was estimated by using a threshold model. No significant association was demonstrated between GIN infection status and SFI. Unexpectedly, the fertility parameter days from calving-to-first-service (CTFS) showed a significantly shorter average interval in GIN positive cows. However, these data on

  9. Advanced reproductive age and fertility.

    Science.gov (United States)

    Liu, Kimberly; Case, Allison

    2011-11-01

    To improve awareness of the natural age-related decline in female and male fertility with respect to natural fertility and assisted reproductive technologies (ART) and provide recommendations for their management, and to review investigations in the assessment of ovarian aging. This guideline reviews options for the assessment of ovarian reserve and fertility treatments using ART with women of advanced reproductive age presenting with infertility. The outcomes measured are the predictive value of ovarian reserve testing and pregnancy rates with natural and assisted fertility. Published literature was retrieved through searches of PubMed or Medline, CINAHL, and The Cochrane Library in June 2010, using appropriate key words (ovarian aging, ovarian reserve, advanced maternal age, advanced paternal age, ART). Results were restricted to systematic reviews, randomized controlled trials/controlled clinical trials, and observational studies. There were no date or language restrictions. Searches were updated on a regular basis and incorporated into the guideline to December 2010. The quality of evidence was rated using the criteria described in the Report of the Canadian Task Force on Preventive Health Care. Recommendations for practice were ranked according to the method described in that report (Table). Primary and specialist health care providers and women will be better informed about ovarian aging and the age-related decline in natural fertility and about options for assisted reproductive technology. 1. Women in their 20s and 30s should be counselled about the age-related risk of infertility when other reproductive health issues, such as sexual health or contraception, are addressed as part of their primary well-woman care. Reproductive-age women should be aware that natural fertility and assisted reproductive technology success (except with egg donation) is significantly lower for women in their late 30s and 40s. (II-2A) 2. Because of the decline in fertility and the

  10. Different routes lead to apoptosis in unfertilized sea urchin eggs.

    Science.gov (United States)

    Philippe, Laetitia; Tosca, Lucie; Zhang, Wen Ling; Piquemal, Marion; Ciapa, Brigitte

    2014-03-01

    Results obtained in various species, from mammals to invertebrates, show that arrest in the cell cycle of mature oocytes is due to a high ERK activity. Apoptosis is stimulated in these oocytes if fertilization does not occur. Our previous data suggest that apoptosis of unfertilized sea urchin eggs is the consequence of an aberrant short attempt of development that occurs if ERK is inactivated. They contradict those obtained in starfish, another echinoderm, where inactivation of ERK delays apoptosis of aging mature oocytes that are nevertheless arrested at G1 of the cell cycle as in the sea urchin. This suggests that the cell death pathway that can be activated in unfertilized eggs is not the same in sea urchin and in starfish. In the present study, we find that protein synthesis is necessary for the survival of unfertilized sea urchin eggs, contrary to starfish. We also compare the effects induced by Emetine, an inhibitor of protein synthesis, with those triggered by Staurosporine, a non specific inhibitor of protein kinase that is widely used to induce apoptosis in many types of cells. Our results indicate that the unfertilized sea urchin egg contain different mechanisms capable of leading to apoptosis and that rely or not on changes in ERK activity, acidity of intracellular organelles or intracellular Ca and pH. We discuss the validity of some methods to investigate cell death such as measurements of caspase activation with the fluorescent caspase indicator FITC-VAD-fmk or acidification of intracellular organelles, methods that may lead to erroneous conclusions at least in the sea urchin model.

  11. Beware of memes in the interpretation of your results - lessons from gene-disrupted mice in fertilization research.

    Science.gov (United States)

    Okabe, Masaru

    2018-05-22

    For decades, researchers in the fertilization field reported various candidate factors involved in sperm-egg interaction through experiments using enzyme inhibitors and/or antibodies. However, almost all of these factors have been shown to be nonessential by gene disruption experiments. Recently, attention has focused on the low reproducibility of papers in many research fields. In this Review, I retrospectively revisit how fertilization factors were misinterpreted and led to wrong hypotheses in relation to the reportedly low reproducibility of scientific papers. © 2018 Federation of European Biochemical Societies.

  12. Effects of gravity on meiosis, fertilization and early embryogenesis in Caenorhabditis elegans

    Science.gov (United States)

    Sasagawa, Y.; Saito, Y.; Shimizu, M.; Ishioka, N.; Yamashita, M.; Takahashi, H.; Higashitani, A.

    The embryonic development of the nematode Caenorhabditis elegans was examined under different gravitational conditions. The first cleavage plane in the 1-cell embryo was slid to some extent by re-orientation of liquid culture vessel, but the pattern and timing of cleavages were not affected. Under 100G of hypergravity condition with swing-centrifuge, the number of eggs laid from an adult hermaphrodite decreased and their hatching rate was drastically reduced. On the other hand, the embryonic development after fertilization normally occurred and grew to adulthood at more than 100G of hypergravity. When the adult hermaphrodites cultured under 100G of hypergravity transferred to a ground condition (1G), the newly fertilized embryos normally developed and their hatching rate was fully recovered. These results indicated that the reproductive process except spermatogenesis, oogenesis and embryogenesis after fertilization is impaired under 100G of hypergravity condition, and the effect is transient. Namely, the fertilization process including meiotic divisions I and II is sensitive to hypergravity in the nematode C. elegans.

  13. Water-borne sperm trigger vitellogenic egg growth in two sessile marine invertebrates.

    Science.gov (United States)

    Bishop, J D; Manríquez, P H; Hughes, R N

    2000-06-22

    A diverse array of sessile marine invertebrates mate by passive dispersal of sperm which fertilize the brooded eggs of neighbours. In two such species, a sea-mat (phylum Bryozoa) and an ascidian (phylum Chordata), vitellogenic egg growth is absent in reproductively isolated specimens, but is triggered by a water-borne factor released by conspecifics. In both of these colonial, hermaphroditic species, the active factor can be removed from water by filtration. The effect involves self-/non-self-recognition: water conditioned by a separate subcolony of the same genetic individual does not prompt oocyte growth. In each species, allosperm move from the surrounding water to the ovary and are then stored in close association with the growing oocytes. We concluded that sperm themselves are the water-borne factor that triggers the major phase of female reproductive investment. This mechanism is, to our knowledge, previously undescribed in animals, but has parallels with the initiation of maternal investment in flowering plants following the receipt of compatible pollen. The species studied may be representative of many other aquatic invertebrates which mate in a similar way. The stimulation of egg growth by allosperm could lead to intersexual conflict during oogenesis.

  14. Dinosaur origin of egg color: oviraptors laid blue-green eggs

    OpenAIRE

    Wiemann, Jasmina; Yang, Tzu-Ruei; Sander, Philipp N.; Schneider, Marion; Engeser, Marianne; Kath-Schorr, Stephanie; M?ller, Christa E.; Sander, P. Martin

    2017-01-01

    Protoporphyrin (PP) and biliverdin (BV) give rise to the enormous diversity in avian egg coloration. Egg color serves several ecological purposes, including post-mating signaling and camouflage. Egg camouflage represents a major character of open-nesting birds which accomplish protection of their unhatched offspring against visually oriented predators by cryptic egg coloration. Cryptic coloration evolved to match the predominant shades of color found in the nesting environment. Such a selecti...

  15. Islet-1 is required for ventral neuron survival in Xenopus

    International Nuclear Information System (INIS)

    Shi, Yu; Zhao, Shuhua; Li, Jiejing; Mao, Bingyu

    2009-01-01

    Islet-1 is a LIM domain transcription factor involved in several processes of embryonic development. Xenopus Islet-1 (Xisl-1) has been shown to be crucial for proper heart development. Here we show that Xisl-1 and Xisl-2 are differentially expressed in the nervous system in Xenopus embryos. Knock-down of Xisl-1 by specific morpholino leads to severe developmental defects, including eye and heart failure. Staining with the neuronal markers N-tubulin and Xisl-1 itself reveals that the motor neurons and a group of ventral interneurons are lost in the Xisl-1 morphants. Terminal dUTP nick-end labeling (TUNEL) analysis shows that Xisl-1 morpholino injection induces extensive apoptosis in the ventral neural plate, which can be largely inhibited by the apoptosis inhibitor M50054. We also find that over-expression of Xisl-1 is able to promote cell proliferation and induce Xstat3 expression in the injected side, suggesting a potential role for Xisl-1 in the regulation of cell proliferation in co-operation with the Jak-Stat pathway.

  16. Diet shifts during egg laying: Implications for measuring contaminants in bird eggs

    Energy Technology Data Exchange (ETDEWEB)

    Morrissey, Christy A. [Catchment Research Group, School of Biosciences, Cardiff University, Cardiff CF10 3AX (United Kingdom); Elliott, John E. [Pacific Wildlife Research Centre, Environment Canada, 5421 Robertson Road, Delta, British Columbia V4K 3N2 (Canada); Ormerod, Stephen J., E-mail: ormerod@cf.ac.u [Catchment Research Group, School of Biosciences, Cardiff University, Cardiff CF10 3AX (United Kingdom)

    2010-02-15

    We combined stable isotope tracers of blood plasma, blood cells and egg contents with faecal analysis during pre-breeding and egg laying phases in two dipper species Cinclus cinclus and Cinclus mexicanus to determine the occurrence of dietary shifts during egg production and to assess consequences for egg contaminant loads. In both species, changes in delta{sup 13}C (C. cinclus) or delta{sup 15}N (C. mexicanus) in female plasma relative to red blood cells indicated a dietary shift during laying that was not observed in males. Eurasian dippers increased prey consumption as breeding approached, shifting from primarily trichopteran insect larvae to ephemeropterans and plecopterans. In American dippers, egg-laying females switched to feeding at a higher trophic level by consuming more fish. Eggs derived from higher trophic level diets contained more mercury (American dipper), polychlorinated biphenyls and some organochlorines, especially DDT metabolites. The results demonstrate how dietary changes during egg laying accompany the demands for egg production with consequences for contaminant deposition in avian eggs. - Changes in laying diet influences contaminant deposition in bird eggs.

  17. Diet shifts during egg laying: Implications for measuring contaminants in bird eggs

    International Nuclear Information System (INIS)

    Morrissey, Christy A.; Elliott, John E.; Ormerod, Stephen J.

    2010-01-01

    We combined stable isotope tracers of blood plasma, blood cells and egg contents with faecal analysis during pre-breeding and egg laying phases in two dipper species Cinclus cinclus and Cinclus mexicanus to determine the occurrence of dietary shifts during egg production and to assess consequences for egg contaminant loads. In both species, changes in δ 13 C (C. cinclus) or δ 15 N (C. mexicanus) in female plasma relative to red blood cells indicated a dietary shift during laying that was not observed in males. Eurasian dippers increased prey consumption as breeding approached, shifting from primarily trichopteran insect larvae to ephemeropterans and plecopterans. In American dippers, egg-laying females switched to feeding at a higher trophic level by consuming more fish. Eggs derived from higher trophic level diets contained more mercury (American dipper), polychlorinated biphenyls and some organochlorines, especially DDT metabolites. The results demonstrate how dietary changes during egg laying accompany the demands for egg production with consequences for contaminant deposition in avian eggs. - Changes in laying diet influences contaminant deposition in bird eggs.

  18. Assessing the impact of egg sweating on Salmonella Enteritidis penetration into shell eggs.

    Science.gov (United States)

    Gradl, Janet A; Curtis, Patricia A; Jones, Deana R; Anderson, Kenneth E

    2017-07-01

    Salmonella Enteritidis (SE) prevalence in eggs is a major concern to the egg industry. Some research has shown that egg sweating can increase Salmonella penetration into egg contents when refrigerated eggs are moved to a warmer temperature. This occurs when eggs are tempered before wash, to minimize thermal cracks. The effect of egg sweating on SE penetration into shell eggs over a 6 week storage period at 4°C was assessed. A 2 × 2 factorial of SE inoculation and egg sweating was utilized. Treatments included (SES) nalidixic acid (NA)-resistant SE inoculated and sweated, (SENS) NA-resistant SE inoculated and not sweated, (NSES) buffered peptone water (BPW) inoculated and sweated, and (NSENS) BPW inoculated and not sweated. Eggs were inoculated with 108 SE. Eggs formed condensation for approximately 17 min in a 32°C incubator. Shell rinse, shell emulsion, and egg contents were sampled then enumerated and assessed for prevalence of SE over a 6 wk storage period at 4°C. After wk 1, the SENS shell rinse had higher SE counts (0.32 log10 CFU/mL) than the other 3 treatments, where no SE was enumerated. A significant week by treatment interaction was found for the shell rinse SE detection (P egg shell rinse, shell emulsion, or egg contents. The SENS shell rinses had significantly higher SE prevalence than the SES rinses in weeks 1 (100% vs. 34.3%), 2 (57.6% vs. 22.2%), and 3 (38.2% vs. 11.1%) (P Egg sweating did not increase SE penetration into the shell emulsion across treatment or week (P egg sweating occurring under common US egg handling practices is not harmful to egg safety. © 2017 Poultry Science Association Inc.

  19. The influence of a light and dark cycle on the egg laying activity of Aedes aegypti (Linnaeus, 1762 (Diptera: Culicidae

    Directory of Open Access Journals (Sweden)

    Luana Cristina Farnesi

    2018-02-01

    Full Text Available BACKGROUND The epidemiological importance of the mosquito Aedes aegypti as a vector of multiple human pathogens has generated a growing number of studies on the physiology and behaviour of its blood-feeding females. The activity of oviposition is one of the critical elements contributing to the expansion of Ae. aegypti's populations. Although there is a vast literature about oviposition behaviour, significant specific knowledge about egg viability and female fertility under light and dark conditions is still lacking. OBJECTIVES We studied, in controlled laboratory conditions, the effect that light and dark cycles have on the efficiency of oviposition by Ae. aegypti females. METHODS Physiological assays were performed using synchronised eggs obtained from forced egg laying. The number and viability of eggs was analysed under three different light/dark regimes: LD12:12 (12 h of light and 12 h of dark, DD (constant darkness and LL (constant light. FINDINGS and CONCLUSIONS Our results show that females prefer to lay their eggs in dark conditions, but maximising the number and viability of eggs requires the occurrence of a light/dark cycle. Ongoing research on this theme has the potential of contributing to the proposition of new strategies for control based on the failure of egg laying and hatching.

  20. Balancing Eggs

    Science.gov (United States)

    Mills, Allan

    2014-01-01

    Theory predicts that an egg-shaped body should rest in stable equilibrium when on its side, balance vertically in metastable equilibrium on its broad end and be completely unstable on its narrow end. A homogeneous solid egg made from wood, clay or plastic behaves in this way, but a real egg will not stand on either end. It is shown that this…

  1. XenDB: Full length cDNA prediction and cross species mapping in Xenopus laevis

    Directory of Open Access Journals (Sweden)

    Giegerich Robert

    2005-09-01

    Full Text Available Abstract Background Research using the model system Xenopus laevis has provided critical insights into the mechanisms of early vertebrate development and cell biology. Large scale sequencing efforts have provided an increasingly important resource for researchers. To provide full advantage of the available sequence, we have analyzed 350,468 Xenopus laevis Expressed Sequence Tags (ESTs both to identify full length protein encoding sequences and to develop a unique database system to support comparative approaches between X. laevis and other model systems. Description Using a suffix array based clustering approach, we have identified 25,971 clusters and 40,877 singleton sequences. Generation of a consensus sequence for each cluster resulted in 31,353 tentative contig and 4,801 singleton sequences. Using both BLASTX and FASTY comparison to five model organisms and the NR protein database, more than 15,000 sequences are predicted to encode full length proteins and these have been matched to publicly available IMAGE clones when available. Each sequence has been compared to the KOG database and ~67% of the sequences have been assigned a putative functional category. Based on sequence homology to mouse and human, putative GO annotations have been determined. Conclusion The results of the analysis have been stored in a publicly available database XenDB http://bibiserv.techfak.uni-bielefeld.de/xendb/. A unique capability of the database is the ability to batch upload cross species queries to identify potential Xenopus homologues and their associated full length clones. Examples are provided including mapping of microarray results and application of 'in silico' analysis. The ability to quickly translate the results of various species into 'Xenopus-centric' information should greatly enhance comparative embryological approaches. Supplementary material can be found at http://bibiserv.techfak.uni-bielefeld.de/xendb/.

  2. Nuclear translocation of phospholipase C-zeta, an egg-activating factor, during early embryonic development

    International Nuclear Information System (INIS)

    Sone, Yoshie; Ito, Masahiko; Shirakawa, Hideki; Shikano, Tomohide; Takeuchi, Hiroyuki; Kinoshita, Katsuyuki; Miyazaki, Shunichi

    2005-01-01

    Phospholipase C-zeta (PLCζ), a strong candidate of the egg-activating sperm factor, causes intracellular Ca 2+ oscillations and egg activation, and is subsequently accumulated into the pronucleus (PN), when expressed in mouse eggs by injection of RNA encoding PLCζ. Changes in the localization of expressed PLCζ were investigated by tagging with a fluorescent protein. PLCζ began to translocate into the PN formed at 5-6 h after RNA injection and increased there. Observation in the same embryo revealed that PLCζ in the PN dispersed to the cytoplasm upon nuclear envelope breakdown and translocated again into the nucleus after cleavage. The dynamics was found in the second mitosis as well. When RNA was injected into fertilization-originated 1-cell embryos or blastomere(s) of 2-8-cell embryos, the nuclear localization of expressed PLCζ was recognized in every embryo up to blastocyst. Thus, PLCζ exhibited alternative cytoplasm/nucleus localization during development. This supports the view that the sperm factor could control cell cycle-dependent generation of Ca 2+ oscillations in early embryogenesis

  3. Structure and expression of the Xenopus retinoblastoma gene.

    Science.gov (United States)

    Destrée, O H; Lam, K T; Peterson-Maduro, L J; Eizema, K; Diller, L; Gryka, M A; Frebourg, T; Shibuya, E; Friend, S H

    1992-09-01

    We have cloned a Xenopus homology (XRb1) of the human retinoblastoma susceptibility gene. DNA sequence analysis shows that the XRb1 gene product is highly conserved in many regions. The leucine repeat motif and many of the potential cdc2 phosphorylation sites, as well as potential sites for other kinases, are retained. The region of the protein homologous to the SV40 T antigen binding site and the basic region directly C-terminal to the E1A binding site are all conserved. XRb1 gene expression at the RNA level was studied by Northern blot analysis. Transcripts of 4.2 and 10-kb are present as maternal RNA stores in the oocyte. While the 4.2-kb product is stable until at least the mid-blastula stage, the 10-kb transcript is selectively degraded. Between stages 11 and 13 the 10-kb transcript reappears and also a minor product of approximately 11 kb becomes apparent. Both the 4.2- and the 10-kb transcripts remain present until later stages of development and are also present in all adult tissues examined, although at differing levels. Antibodies raised against human p105Rb which recognize the protein product of the XRb1 gene, pXRb1, detect the Xenopus 99-kDa protein prior to the mid-blastula stage, but at lower levels than at later stages in development.

  4. Caffeine alleviates the deterioration of Ca2+ release mechanisms and fragmentation of in vitro aged mouse eggs

    Science.gov (United States)

    Zhang, Nan; Wakai, Takuya; Fissore, Rafael. A.

    2011-01-01

    The developmental competence of mammalian eggs is compromised by postovulatory aging. We and others found that in these eggs the intracellular calcium ([Ca2+]i) responses required for egg activation and initiation of development are altered. Nevertheless, the mechanism(s) underlying this defective Ca2+ release is not well known. Here, we investigated if the function of IP3R1, the major Ca2+ release channel at fertilization, was undermined in in vitro aged mouse eggs. We found that in aged eggs IP3R1 displayed reduced function, as many of the changes acquired during maturation that enhance IP3R1 Ca2+ conductivity such as phosphorylation, receptor reorganization and increased Ca2+ store content ([Ca2+]ER) were lost with increasing postovulatory time. IP3R1 fragmentation, possibly associated with the activation of caspase-3, was also observed in these eggs. Many of these changes were prevented when the postovulatory aging of eggs was carried out in the presence of caffeine, which minimized the decline in IP3R1 function and maintained [Ca2+]ER content. Caffeine also maintained mitochondrial membrane potential as measured by JC-1 fluorescence. We therefore conclude that [Ca2+]i responses in aged eggs are undermined by reduced IP3R1 sensitivity, decreased [Ca2+]ER and compromised mitochondrial function, and that addition of caffeine ameliorates most of these aging-associated changes. Understanding the molecular basis of the protective effects of caffeine will be useful in elucidating, and possibly reversing, the signaling pathway(s) compromised by in vitro culture of eggs. PMID:22095868

  5. Asymmetries in Cell Division, Cell Size, and Furrowing in the Xenopus laevis Embryo.

    Science.gov (United States)

    Tassan, Jean-Pierre; Wühr, Martin; Hatte, Guillaume; Kubiak, Jacek

    2017-01-01

    Asymmetric cell divisions produce two daughter cells with distinct fate. During embryogenesis, this mechanism is fundamental to build tissues and organs because it generates cell diversity. In adults, it remains crucial to maintain stem cells. The enthusiasm for asymmetric cell division is not only motivated by the beauty of the mechanism and the fundamental questions it raises, but has also very pragmatic reasons. Indeed, misregulation of asymmetric cell divisions is believed to have dramatic consequences potentially leading to pathogenesis such as cancers. In diverse model organisms, asymmetric cell divisions result in two daughter cells, which differ not only by their fate but also in size. This is the case for the early Xenopus laevis embryo, in which the two first embryonic divisions are perpendicular to each other and generate two pairs of blastomeres, which usually differ in size: one pair of blastomeres is smaller than the other. Small blastomeres will produce embryonic dorsal structures, whereas the larger pair will evolve into ventral structures. Here, we present a speculative model on the origin of the asymmetry of this cell division in the Xenopus embryo. We also discuss the apparently coincident asymmetric distribution of cell fate determinants and cell-size asymmetry of the 4-cell stage embryo. Finally, we discuss the asymmetric furrowing during epithelial cell cytokinesis occurring later during Xenopus laevis embryo development.

  6. Parental perceptions in egg allergy: does egg challenge make a difference?

    Science.gov (United States)

    Kemp, Andrew Stewart; Allen, Clare Wendy; Campbell, Dianne Elisabeth

    2009-11-01

    The aim of the study was to determine the effect of an oral egg challenge in egg sensitized children on parental perceptions relating to their child's allergy. A questionnaire was completed by parents for 167 children attending a tertiary paediatric clinic with egg sensitization. The questionnaires included 10 questions concerning parental perceptions of their child's egg allergy. Parental perceptions of those children who had not had an egg challenge (n = 83) were compared with those whose children had a positive (n = 27) and those with a negative (n = 57) egg challenge. A significant difference (p = parents in the CN group expected little or no future inconvenience for the child. The responses of parents whose child had undergone an egg challenge differed significantly (p = expectation of little or no future discomfort for the child and whether others treated the child differently. The performance of an egg challenge was associated with reduced adverse parental concerns. For 6/10 parameters, expectations concerning egg allergy in children who had been challenged were significantly better than those who had never been challenged irrespective of the challenge outcome. The greater certainty provided by the performance of a food challenge may be a positive outcome in both CP and CN children.

  7. cis- and trans-acting elements of the estrogen-regulated vitellogenin gene B1 of Xenopus laevis.

    Science.gov (United States)

    Wahli, W; Martinez, E; Corthésy, B; Cardinaux, J R

    1989-01-01

    Vitellogenin genes are expressed under strict estrogen control in the liver of female oviparous vertebrates. Gene transfer experiments using estrogen-responsive cells have shown that the 13 bp perfect palindromic element GGTCACTGTGACC found upstream of the Xenopus laevis vitellogenin gene A2 promoter mediates hormonal stimulation and thus, was called the estrogen-responsive element (ERE). In the Xenopus vitellogenin genes B1 and B2 there are two closely adjacent EREs with one or more base substitutions when compared to the consensus ERE GGTCANNNTGACC. On their own, these degenerated elements have only a low or no regulatory capacity at all but act together synergistically to form an estrogen-responsive unit (ERU) with the same strength as the perfect palindromic 13 bp element. Analysis of estrogen receptor binding to the gene B1 ERU revealed a cooperative interaction of receptor dimers to the two adjacent imperfect EREs which most likely explains the synergistic stimulation observed in vivo. Furthermore, a promoter activator element located between positions --113 and --42 of the gene B1 and functional in the human MCF-7 and the Xenopus B3.2 cells has been identified and shown to be involved in the high level of induced transcription activity when the ERE is placed at a distance from the promoter. Finally, a hormone-controlled in vitro transcription system derived from Xenopus liver nuclear extracts was exploited to characterize two additional novel cis-acting elements within the vitellogenin gene B1 promoter. One of them, a negative regulatory element (NRE), is responsible for repression of promoter activity in the absence of hormone. The second is related to the NF-I binding site and is required, together with the ERE, to mediate hormonal induction. Moreover, we detected three trans-acting activities in Xenopus liver nuclear extracts that interact with these regions and demonstrated that they participate in the regulation of the expression of the vitellogenin

  8. The Xenopus oocyte: a model for studying the metabolic regulation of cancer cell death.

    Science.gov (United States)

    Nutt, Leta K

    2012-06-01

    Abnormal metabolism and the evasion of apoptosis are both considered hallmarks of cancer. A remarkable biochemical model system, the Xenopus laevis oocyte, exhibits altered metabolism coupled to its apoptotic machinery in a similar fashion to cancer cells. This review considers the theory that these two hallmarks of cancer are coupled in tumor cells and provides strong proof that the Xenopus laevis oocyte system is an appropriate model in which to dissect the biochemical events underlying the connection between the two hallmarks. By further elucidating the mechanisms through which metabolism suppresses apoptotic machinery, we may gain a better understanding about how normal cells transform into cancer cells. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. Effects of egg storage on hatchability, chick quality, performance and immunocompetence parameters of broiler chickens.

    Science.gov (United States)

    Goliomytis, Michael; Tsipouzian, Theofania; Hager-Theodorides, Ariadne L

    2015-09-01

    Pre-incubation egg storage is a necessity for the poultry industry. This study evaluated the effects of pre-incubation storage length of broiler eggs on hatchability, 1-day-old chick quality, subsequent performance, and immunocompetence. To this end, a total of 360 hatching eggs were stored for 4, 12, or 16 d prior to incubation. Hatchability and chick quality were assessed at hatch, and growth performance and immunocompetence parameters were assessed during a 35 d rearing period. Hatchability of set and fertile eggs, and embryonic mortality, were not affected by egg storage. On the contrary, 1-day-old chick BW and length were linearly negatively correlated with egg storage length (P-linearegg weight prior to setting was unaffected, and corrected chick length was positively affected by storage length. One-day-old chick Tona score, navel quality, and post-hatch growth performance (BW at 7 and 35 d, cumulative feed intake, and feed conversion ratio at 35 d) were unaffected by egg storage (P, P-linear>0.05). Lymphoid organ weights at 2 and 35 d, the titre of maternal anti-NDV antibodies, most of the thymocyte subpopulations defined by CD3, CD4, and CD8 cell surface expression in the thymus of 2-d-old chicks, cellular responses to the PHA skin test, humoral responses to primary SRBC, and NDV immunizations were also not influenced by length of storage (P, P-linear>0.05). On the contrary, the length of egg storage was found to negatively influence the abundance of CD3+CD4-CD8- thymocytes that represent the majority of γδ-T cells in the thymus of 2-day-old chicks, as well as the humoral response to booster NDV immunization of the birds. In brief, pre-incubation storage of broiler hatching eggs for up to 16 d did not affect most developmental and growth parameters investigated, except for BW and length at hatch. Egg storage was found to suppress some aspects of the immunocompetence of the birds, particularly aspects of acquired immunity. © 2015 Poultry Science

  10. Effects of S-adenosylmethionine decarboxylase, polyamines, amino acids, and weak bases (amines and ammonia) on development and ribosomal RNA synthesis in Xenopus embryos.

    Science.gov (United States)

    Shiokawa, Koichiro; Aso, Mai; Kondo, Takeshi; Takai, Jun-Ichi; Yoshida, Junki; Mishina, Takamichi; Fuchimukai, Kota; Ogasawara, Tsukasa; Kariya, Taro; Tashiro, Kosuke; Igarashi, Kazuei

    2010-02-01

    We have been studying control mechanisms of gene expression in early embryogenesis in a South African clawed toad Xenopus laevis, especially during the period of midblastula transition (MBT), or the transition from the phase of active cell division (cleavage stage) to the phase of extensive morphogenesis (post-blastular stages). We first found that ribosomal RNA synthesis is initiated shortly after MBT in Xenopus embryos and those weak bases, such as amines and ammonium ion, selectively inhibit the initiation and subsequent activation of rRNA synthesis. We then found that rapidly labeled heterogeneous mRNA-like RNA is synthesized in embryos at pre-MBT stage. We then performed cloning and expression studies of several genes, such as those for activin receptors, follistatin and aldolases, and then reached the studies of S-adenosylmethionine decarboxylase (SAMDC), a key enzyme in polyamine metabolism. Here, we cloned a Xenopus SAMDC cDNA and performed experiments to overexpress the in vitro-synthesized SAMDC mRNA in Xenopus early embryos, and found that the maternally preset program of apoptosis occurs in cleavage stage embryos, which is executed when embryos reach the stage of MBT. In the present article, we first summarize results on SAMDC and the maternal program of apoptosis, and then describe our studies on small-molecular-weight substances like polyamines, amino acids, and amines in Xenopus embryos. Finally, we summarize our studies on weak bases, especially on ammonium ion, as the specific inhibitor of ribosomal RNA synthesis in Xenopus embryonic cells.

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

    Science.gov (United States)

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

    2014-06-01

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

  12. 21 CFR 160.100 - Eggs.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Eggs. 160.100 Section 160.100 Food and Drugs FOOD... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.100 Eggs... identity for the food commonly known as eggs. ...

  13. Mirror-image duplication of the primary axis and heart in Xenopus embryos by the overexpression of Msx-1 gene.

    Science.gov (United States)

    Chen, Y; Solursh, M

    1995-10-01

    The Msx-1 gene (formerly known as Hox-7) is a member of a discrete subclass of homeobox-containing genes. Examination of the expression pattern of Msx-1 in murine and avian embryos suggests that this gene may be involved in the regionalization of the medio-lateral axis during earlier development. We have examined the possible functions of Xenopus Msx-1 during early Xenopus embryonic development by overexpression of the Msx-1 gene. Overexpression of Msx-1 causes a left-right mirror-image duplication of primary axial structures, including notochord, neural tube, somites, suckers, and foregut. The embryonic developing heart is also mirror-image duplicated, including looping directions and polarity. These results indicate that Msx-1 may be involved in the mesoderm formation as well as left-right patterning in the early Xenopus embryonic development.

  14. Experimental Shifts in Intraclutch Egg Color Variation Do Not Affect Egg Rejection in a Host of a Non-Egg-Mimetic Avian Brood Parasite

    Science.gov (United States)

    Croston, Rebecca; Hauber, Mark E.

    2015-01-01

    Avian brood parasites lay their eggs in the nests of other birds, and impose the costs associated with rearing parasitic young onto these hosts. Many hosts of brood parasites defend against parasitism by removing foreign eggs from the nest. In systems where parasitic eggs mimic host eggs in coloration and patterning, extensive intraclutch variation in egg appearances may impair the host’s ability to recognize and reject parasitic eggs, but experimental investigation of this effect has produced conflicting results. The cognitive mechanism by which hosts recognize parasitic eggs may vary across brood parasite hosts, and this may explain variation in experimental outcome across studies investigating egg rejection in hosts of egg-mimicking brood parasites. In contrast, for hosts of non-egg-mimetic parasites, intraclutch egg color variation is not predicted to co-vary with foreign egg rejection, irrespective of cognitive mechanism. Here we tested for effects of intraclutch egg color variation in a host of nonmimetic brood parasite by manipulating egg color in American robins (Turdus migratorius), hosts of brown-headed cowbirds (Molothrus ater). We recorded robins’ behavioral responses to simulated cowbird parasitism in nests where color variation was artificially enhanced or reduced. We also quantified egg color variation within and between unmanipulated robin clutches as perceived by robins themselves using spectrophotometric measures and avian visual modeling. In unmanipulated nests, egg color varied more between than within robin clutches. As predicted, however, manipulation of color variation did not affect rejection rates. Overall, our results best support the scenario wherein egg rejection is the outcome of selective pressure by a nonmimetic brood parasite, because robins are efficient rejecters of foreign eggs, irrespective of the color variation within their own clutch. PMID:25831051

  15. Experimental shifts in intraclutch egg color variation do not affect egg rejection in a host of a non-egg-mimetic avian brood parasite.

    Directory of Open Access Journals (Sweden)

    Rebecca Croston

    Full Text Available Avian brood parasites lay their eggs in the nests of other birds, and impose the costs associated with rearing parasitic young onto these hosts. Many hosts of brood parasites defend against parasitism by removing foreign eggs from the nest. In systems where parasitic eggs mimic host eggs in coloration and patterning, extensive intraclutch variation in egg appearances may impair the host's ability to recognize and reject parasitic eggs, but experimental investigation of this effect has produced conflicting results. The cognitive mechanism by which hosts recognize parasitic eggs may vary across brood parasite hosts, and this may explain variation in experimental outcome across studies investigating egg rejection in hosts of egg-mimicking brood parasites. In contrast, for hosts of non-egg-mimetic parasites, intraclutch egg color variation is not predicted to co-vary with foreign egg rejection, irrespective of cognitive mechanism. Here we tested for effects of intraclutch egg color variation in a host of nonmimetic brood parasite by manipulating egg color in American robins (Turdus migratorius, hosts of brown-headed cowbirds (Molothrus ater. We recorded robins' behavioral responses to simulated cowbird parasitism in nests where color variation was artificially enhanced or reduced. We also quantified egg color variation within and between unmanipulated robin clutches as perceived by robins themselves using spectrophotometric measures and avian visual modeling. In unmanipulated nests, egg color varied more between than within robin clutches. As predicted, however, manipulation of color variation did not affect rejection rates. Overall, our results best support the scenario wherein egg rejection is the outcome of selective pressure by a nonmimetic brood parasite, because robins are efficient rejecters of foreign eggs, irrespective of the color variation within their own clutch.

  16. Xenopus laevis Retinal Ganglion Cell Dendritic Arbors Develop Independently of Visual Stimulation

    Directory of Open Access Journals (Sweden)

    Barbara Lom

    2004-01-01

    Full Text Available Newly formed neurons must locate their appropriate target cells and then form synaptic connections with these targets in order to establish a functional nervous system. In the vertebrate retina, retinal ganglion cell (RGC dendrites extend from the cell body and form synapses with nearby amacrine and bipolar cells. RGC axons, however, exit the retina and synapse with the dendrites of midbrain neurons in the optic tectum. We examined how visual stimulation influenced Xenopus RGC dendritic arborization. Neuronal activity is known to be an important factor in shaping dendritic and axonal arborization. Thus, we reared tadpoles in dark and light environments then used rhodamine dextran retrograde labeling to identify RGCs in the retina. When we compared RGC dendritic arbors from tadpoles reared in dark and light environments, we found no morphological differences, suggesting that physiological visual activity did not contribute to the morphological development of Xenopus RGC dendritic arbors.

  17. The masquerade game: marine mimicry adaptation between egg-cowries and octocorals

    Directory of Open Access Journals (Sweden)

    Juan A. Sánchez

    2016-08-01

    Full Text Available Background. Background matching, as a camouflage strategy, is one of the most outstanding examples of adaptation, where little error or mismatch means high vulnerability to predation. It is assumed that the interplay of natural selection and adaptation are the main evolutionary forces shaping the great diversity of phenotypes observed in mimicry; however, there may be other significant processes that intervene in the development of mimicry such as phenotypic plasticity. Based on observations of background mismatching during reproduction events of egg-cowries, sea snails of the family Ovulidae that mimic the octocoral where they inhabit, we wondered if they match the host species diversity. Using observations in the field and molecular systematics, we set out to establish whether the different egg-cowrie color/shape polymorphisms correspond to distinct lineages restricted to specific octocoral species. Methods. Collection and observations of egg-cowries and their octocoral hosts were done using SCUBA diving between 2009 and 2012 at two localities in the Tropical Eastern Pacific (TEP, Malpelo Island and Cabo Corrientes (Colombia. Detailed host preference observations were done bi-annually at Malpelo Island. We analyzed the DNA sequence of the mitochondrial genes COIand 16S rDNA, extensively used in phylogenetic and DNA barcoding studies, to assess the evolutionary relationship among different egg-cowrie colorations and morphologies. Results. No genetic divergence among egg-cowries associated to different species of the same octocoral genus was observed based on the two mitochondrial genes analyzed. For instance, all egg-cowrie individuals from the two sampled localities observed on 8 different Pacifigorgia-Eugorgia species showed negligible mitochondrial divergence yet large morphologic divergence, which suggests that morphologies belonging to at least two sea snail species, Simnia avena(=S. aequalis and Simnialena rufa, can cross-fertilize

  18. Effects of water salinity on hatching of egg, growth and survival of larvae and fingerlings of snake head fish, Channa striatus

    Directory of Open Access Journals (Sweden)

    Thumronk Amornsakun

    2017-04-01

    Full Text Available A study on the effect of water salinity ranging from 0-30 ppt on hatching success of snake head fish, Channa striatus was conducted in a 15-liter glass aquarium (water volume 10 liters containing 500 eggs for various levels of water salinity. Fertilization rates at 0, 5, 10, 11, 12, 13 and 14 ppt were 69.33, 72.67, 71.33, 72.67, 82.00, 73.33 and 10.67%, respectively. The fertilization rate at 12-13 ppt salinity was significantly (P0.05 among 0, 5 and 10 ppt.

  19. The origin and evolution of queen and fertility signals in Corbiculate bees.

    Science.gov (United States)

    Caliari Oliveira, Ricardo; Oi, Cintia Akemi; do Nascimento, Mauricio Meirelles Castro; Vollet-Neto, Ayrton; Alves, Denise Araujo; Campos, Maria Claudia; Nascimento, Fabio; Wenseleers, Tom

    2015-11-16

    In social Hymenoptera (ants, bees and wasps), various chemical compounds present on the cuticle have been shown to act as fertility signals. In addition, specific queen-characteristic hydrocarbons have been implicated as sterility-inducing queen signals in ants, wasps and bumblebees. In Corbiculate bees, however, the chemical nature of queen-characteristic and fertility-linked compounds appears to be more diverse than in ants and wasps. Moreover, it remains unknown how queen signals evolved across this group and how they might have been co-opted from fertility signals in solitary ancestors. Here, we perform a phylogenetic analysis of fertility-linked compounds across 16 species of solitary and eusocial bee species, comprising both literature data as well as new primary data from a key solitary outgroup species, the oil-collecting bee Centris analis, and the highly eusocial stingless bee Scaptotrigona depilis. Our results demonstrate the presence of fertility-linked compounds belonging to 12 different chemical classes. In addition, we find that some classes of compounds (linear and branched alkanes, alkenes, esters and fatty acids) were already present as fertility-linked signals in the solitary ancestors of Corbiculate bees, while others appear to be specific to certain species. Overall, our results suggest that queen signals in Corbiculate bees are likely derived from ancestral fertility-linked compounds present in solitary bees that lacked reproductive castes. These original fertility-linked cues or signals could have been produced either as a by-product of ovarian activation or could have served other communicative purposes, such as in mate recognition or the regulation of egg-laying.

  20. Vision drives correlated activity without patterned spontaneous activity in developing Xenopus retina.

    Science.gov (United States)

    Demas, James A; Payne, Hannah; Cline, Hollis T

    2012-04-01

    Developing amphibians need vision to avoid predators and locate food before visual system circuits fully mature. Xenopus tadpoles can respond to visual stimuli as soon as retinal ganglion cells (RGCs) innervate the brain, however, in mammals, chicks and turtles, RGCs reach their central targets many days, or even weeks, before their retinas are capable of vision. In the absence of vision, activity-dependent refinement in these amniote species is mediated by waves of spontaneous activity that periodically spread across the retina, correlating the firing of action potentials in neighboring RGCs. Theory suggests that retinorecipient neurons in the brain use patterned RGC activity to sharpen the retinotopy first established by genetic cues. We find that in both wild type and albino Xenopus tadpoles, RGCs are spontaneously active at all stages of tadpole development studied, but their population activity never coalesces into waves. Even at the earliest stages recorded, visual stimulation dominates over spontaneous activity and can generate patterns of RGC activity similar to the locally correlated spontaneous activity observed in amniotes. In addition, we show that blocking AMPA and NMDA type glutamate receptors significantly decreases spontaneous activity in young Xenopus retina, but that blocking GABA(A) receptor blockers does not. Our findings indicate that vision drives correlated activity required for topographic map formation. They further suggest that developing retinal circuits in the two major subdivisions of tetrapods, amphibians and amniotes, evolved different strategies to supply appropriately patterned RGC activity to drive visual circuit refinement. Copyright © 2011 Wiley Periodicals, Inc.

  1. Migratory and adhesive properties of Xenopus laevis primordial germ cells in vitro

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    Aliaksandr Dzementsei

    2013-11-01

    The directional migration of primordial germ cells (PGCs to the site of gonad formation is an advantageous model system to study cell motility. The embryonic development of PGCs has been investigated in different animal species, including mice, zebrafish, Xenopus and Drosophila. In this study we focus on the physical properties of Xenopus laevis PGCs during their transition from the passive to the active migratory state. Pre-migratory PGCs from Xenopus laevis embryos at developmental stages 17–19 to be compared with migratory PGCs from stages 28–30 were isolated and characterized in respect to motility and adhesive properties. Using single-cell force spectroscopy, we observed a decline in adhesiveness of PGCs upon reaching the migratory state, as defined by decreased attachment to extracellular matrix components like fibronectin, and a reduced adhesion to somatic endodermal cells. Data obtained from qPCR analysis with isolated PGCs reveal that down-regulation of E-cadherin might contribute to this weakening of cell-cell adhesion. Interestingly, however, using an in vitro migration assay, we found that movement of X. laevis PGCs can also occur independently of specific interactions with their neighboring cells. The reduction of cellular adhesion during PGC development is accompanied by enhanced cellular motility, as reflected in increased formation of bleb-like protrusions and inferred from electric cell-substrate impedance sensing (ECIS as well as time-lapse image analysis. Temporal alterations in cell shape, including contraction and expansion of the cellular body, reveal a higher degree of cellular dynamics for the migratory PGCs in vitro.

  2. Subcellular localization of class I histone deacetylases in the developing Xenopus tectum

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    Xia eGuo

    2016-01-01

    Full Text Available Histone deacetylases (HDACs are thought to localize in the nucleus to regulate gene transcription and play pivotal roles in neurogenesis, apoptosis and plasticity. However, the subcellular distribution of class I HDACs in the developing brain remains unclear. Here, we show that HDAC1 and HDAC2 are located in both the mitochondria and the nucleus in the Xenopus laevis stage 34 tectum and are mainly restricted to the nucleus following further brain development. HDAC3 is widely present in the mitochondria, nucleus and cytoplasm during early tectal development and is mainly distributed in the nucleus in stage 45 tectum. In contrast, HDAC8 is broadly located in the mitochondria, nucleus and cytoplasm during tectal development. These data demonstrate that HDAC1, HDAC2 and HDAC3 are transiently localized in the mitochondria and that the subcellular distribution of class I HDACs in the Xenopus tectum is heterogeneous. Furthermore, we observed that spherical mitochondria accumulate in the cytoplasm at earlier stages, whereas elongated mitochondria are evenly distributed in the tectum at later stages. The activity of histone acetylation (H4K12 remains low in mitochondria during tectal development. Pharmacological blockades of HDACs using a broad spectrum HDAC inhibitor of Trichostatin A (TSA or specific class I HDAC inhibitors of MS-275 and MGCD0103 decrease the number of mitochondria in the tectum at stage 34. These findings highlight a link between the subcellular distribution of class I HDACs and mitochondrial dynamics in the developing optic tectum of Xenopus laevis.

  3. Modification of fecundity and fertility during oogenesis by γ radiation and/or ozone with a cytological analysis in the ectoparasitic wasp, Habrobracon juglandis (Ashmead)

    International Nuclear Information System (INIS)

    Ofuoku, E.E.

    1984-01-01

    In Experiment I, adult female wasps were exposed to ozone for 0, 2, 4, 6, 8, 10, 12, 16, 24, and 27 h. The results indicated that the 27 h of ozone exposure produced 100% lethality on the first day. Exposures below 27 h progressively decreased life span with increasing length of exposure. In Experiment II A, adult virgin Habrobracon females were exposed to ozone for 0, 2, 4, 6, 8, 10, 12, 16, and 24 h to determine the effects of ozone on fecundity (egg laying ability) and fertility (egg hatching ability). The results showed that ozone significantly decreased fecundity and fertility in all meiotic stages except metaphase I. In Experiment II B, adult virgin Habrobracon females were exposed to Co-60 γ radiation. All treated wasps showed significant progressive decreases in fecundity and fertility with increases in radiation dose. In Experiment II C, adult virgin Habrobracon females were exposed to Co-60 γ radiation, to ozone, or to combinations thereof to determine the effects of these insults on fecundity and fertility. Together or singly ozone and radiation reduced fecundity and fertility. In Experiment III, adult virgin Habrobracon females were exposed to the conditions of Experiment II C to correlate by cytological examination of the ovarioles the effects of ionizing radiation and/or ozone on the germ cells at specific meiotic stages. Results obtained from the cytological study explain the fecundity and fertility observations

  4. Dicty_cDB: Contig-U01127-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ( BJ076721 ) Xenopus laevis cDNA clone:XL058i17, 3' end, singl... 44 5.9 1 ( FG291907 ) 1108800220021 New World... Screwworm Egg 9261 ESTs C... 44 5.9 1 ( FG291539 ) 1108793348396 New World Sc...rewworm Egg 9261 ESTs C... 44 5.9 1 ( FG286660 ) 1108770723740 New World Screwworm Egg 9261 ESTs C... 44 5.9

  5. Fast egg collection method greatly improves randomness of egg sampling in Drosophila melanogaster

    DEFF Research Database (Denmark)

    Schou, Mads Fristrup

    2013-01-01

    When obtaining samples for population genetic studies, it is essential that the sampling is random. For Drosophila, one of the crucial steps in sampling experimental flies is the collection of eggs. Here an egg collection method is presented, which randomizes the eggs in a water column...... and diminishes environmental variance. This method was compared with a traditional egg collection method where eggs are collected directly from the medium. Within each method the observed and expected standard deviations of egg-to-adult viability were compared, whereby the difference in the randomness...... and to obtain a representative collection of genotypes, the method presented here is strongly recommended when collecting eggs from Drosophila....

  6. 21 CFR 160.115 - Liquid eggs.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Liquid eggs. 160.115 Section 160.115 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.115 Liquid eggs. (a) Liquid eggs, mixed eggs, liquid whole eggs, mixed whole eggs are eggs of the domestic...

  7. Feeding management strategy for male broiler breeders and its effects on body weight, hatchability and fertility

    Directory of Open Access Journals (Sweden)

    MM Silveira

    2014-12-01

    Full Text Available The objective of the present study was to evaluate the supply of a specific-male diet on rooster body weight, hatchability, and fertility. Two diets were supplied during the egg production period: treatment 1, diet formulated for females and containing, in average, 2,830 kcal ME/kg and 15% CP; and treatment 2, diet specifically formulated for male broiler breeders, containing 2,750 kcal ME/kg and 13.5% CP. In the experiment, 26 flocks, with approximately 7,000 females and 840 males each, belonging to the same broiler breeder commercial strain, were distributed into 26 houses (1200m² on the same farm, and submitted to the same management practices. Body weight was evaluated in 13 flocks per treatment and for five weeks (45, 50, 55, 60, 65, and hatchability and fertility in four incubation lots per treatment and for the same weeks. Data were evaluated by the analysis of mixed models for repeated measurements, using the PROC MIXED of SAS 9.3 (SAS Institute Inc., Cary, NC, USA, and means were compared by the test of Tukey-Kramer at 5% significance level. The specific-male diet allowed body weight control during all analyzed weeks and promoted high hatchability and egg fertility. The increase in the number of chicks hatched demonstrates the economic viability of feeding a specific diet for male broiler breeders.

  8. In vitro fertilization in Japan — Early days of in vitro fertilization and embryo transfer and future prospects for assisted reproductive technology —

    Science.gov (United States)

    SUZUKI, Masakuni

    2014-01-01

    Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and embryo transfer (ET) has been essential in the treatment of infertility. The world’s first IVF-ET baby was born in 1978 based on the technique developed by Dr. Robert Edwards and Dr. Patrick Steptoe.1) In Japan, the first IVF-ET birth was reported in 1983 by Prof. Masakuni Suzuki at Tohoku University School of Medicine.2,3) IVF-ET is a procedure used to achieve pregnancy that consists of extracting oocytes from an infertile woman, fertilizing them in vitro, and transferring fertilized eggs into the patient’s uterine cavity (Fig. 1). Since the first report of successful IVF-ET, numerous techniques related to ART, such as cryopreservation of oocytes and embryos, gamete intrafallopian transfer (GIFT), and microinsemination, have been developed and refined (Table 1). Herein we describe the history of basic research in IVF-ET that led to human applications, how the birth of the first IVF-ET baby was achieved in Japan, the current status of ART in Japan, issues related to ART, and future prospects for ART. PMID:24814992

  9. Comparative performances of eggs and embryos of sea urchin (Paracentrotus lividus) in toxicity bioassays used for assessment of marine sediment quality.

    Science.gov (United States)

    Khosrovyan, A; Rodríguez-Romero, A; Salamanca, M J; Del Valls, T A; Riba, I; Serrano, F

    2013-05-15

    The potential toxicity of sediments from various ports was assessed by means of two different liquid-phase toxicity bioassays (acute and chronic) with embryos and eggs of sea urchin Paracentrotus lividus. Performances of embryos and eggs of P. lividus in these bioassays were compared for their interchangeable applicability in integrated sediment quality assessment. The obtained endpoints (percentages of normally developed plutei and fertilized eggs) were linked to physical and chemical properties of sediments and demonstrated dependence on sediment contamination. The endpoints in the two bioassays were strongly correlated and generally exhibited similar tendency throughout the samples. Therein, embryos demonstrated higher sensitivity to elutriate exposure, compared to eggs. It was concluded that these tests could be used interchangeably for testing toxicity of marine sediments. Preferential use of any of the bioassays can be determined by the discriminatory capacity of the test or vulnerability consideration of the test subject to the surrounding conditions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. A highly conserved Poc1 protein characterized in embryos of the hydrozoan Clytia hemisphaerica: localization and functional studies.

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    Cécile Fourrage

    Full Text Available Poc1 (Protein of Centriole 1 proteins are highly conserved WD40 domain-containing centriole components, well characterized in the alga Chlamydomonas, the ciliated protazoan Tetrahymena, the insect Drosophila and in vertebrate cells including Xenopus and zebrafish embryos. Functions and localizations related to the centriole and ciliary axoneme have been demonstrated for Poc1 in a range of species. The vertebrate Poc1 protein has also been reported to show an additional association with mitochondria, including enrichment in the specialized "germ plasm" region of Xenopus oocytes. We have identified and characterized a highly conserved Poc1 protein in the cnidarian Clytia hemisphaerica. Clytia Poc1 mRNA was found to be strongly expressed in eggs and early embryos, showing a punctate perinuclear localization in young oocytes. Fluorescence-tagged Poc1 proteins expressed in developing embryos showed strong localization to centrioles, including basal bodies. Anti-human Poc1 antibodies decorated mitochondria in Clytia, as reported in human cells, but failed to recognise endogenous or fluorescent-tagged Clytia Poc1. Injection of specific morpholino oligonucleotides into Clytia eggs prior to fertilization to repress Poc1 mRNA translation interfered with cell division from the blastula stage, likely corresponding to when neosynthesis normally takes over from maternally supplied protein. Cell cycle lengthening and arrest were observed, phenotypes consistent with an impaired centriolar biogenesis or function. The specificity of the defects could be demonstrated by injection of synthetic Poc1 mRNA, which restored normal development. We conclude that in Clytia embryos, Poc1 has an essentially centriolar localization and function.

  11. Biology and fertility life table of Agrotis ipsilon on artificial diet; Biologia e tabela de vida de fertilidade de Agrotis ipsilon em dieta artificial

    Energy Technology Data Exchange (ETDEWEB)

    Bento, Flavia de Moura Manoel; Fortes, Priscila; Zerio, Neide Graciano; Parra, Jose Roberto Postali [Universidade de Sao Paulo (USP), Piracicaba, SP (Brazil). Escola Superior de Agricultura Luiz de Queiroz (ESALQ). Dept. Entomologia, Fitopatologia e Zoologia Agricola]. E-mail: flaviam@esalq.usp.br, pfortes@esalq.usp.br, ngzerio@esalq.usp.br, jrpparra@esalq.usp.br; Magro, Sandra Regina [Universidade Camilo Castelo Branco, Fernandopolis, SP (Brazil)]. E-mail: sandra.magro@gmail.com

    2007-10-15

    The objective of this work was to develop an artificial diet to rear Agrotis ipsilon in laboratory using biological parameters and fertility life table. The artificial diet was prepared with bean, casein, soybean protein, yeast and wheat germ as protein sources. The biological aspects duration and viability of larval and pupal stages, pupal weight, sex ratio, life span of adults, preoviposition period, egg laying capacity and fertility life table were evaluated. Six larval instars were observed comprising larval duration of 28,4 days and 93% of viability mean pupal duration of 12,4 days and viability of 96%. The total viability of the life cycle was 72%. The pupae weight was 387 mg for males and 484 mg for females. The sex ratio was 0,46 and the preoviposition period lasted one day and egg laying was 1,806 eggs per female. The net reproductive rate per generation and increase finite rate were 616,9 and 1,14, respectively. Artificial diet is adequate for rearing A. ipsilon in laboratory. (author)

  12. Notochord-derived hedgehog is essential for tail regeneration in Xenopus tadpole.

    Science.gov (United States)

    Taniguchi, Yuka; Watanabe, Kenji; Mochii, Makoto

    2014-06-18

    Appendage regeneration in amphibians is regulated by the combinatorial actions of signaling molecules. The requirement of molecules secreted from specific tissues is reflected by the observation that the whole process of regeneration can be inhibited if a certain tissue is removed from the amputated stump. Interestingly, urodeles and anurans show different tissue dependencies during tail regeneration. The spinal cord is essential for tail regeneration in urodele but not in anuran larva, whereas the notochord but not the spinal cord is essential for tail regeneration in anuran tadpoles. Sonic hedgehog is one of the signaling molecules responsible for such phenomenon in axolotl, as hedgehog signaling is essential for overall tail regeneration and sonic hedgehog is exclusively expressed in the spinal cord. In order to know whether hedgehog signaling is involved in the molecular mechanism underlying the inconsistent tissue dependency for tail regeneration between anurans and urodeles, we investigated expression of hedgehog signal-related genes in the regenerating tail of Xenopus tadpole and examined the effect of the hedgehog signal inhibitor, cyclopamine, on the tail regeneration. In Xenopus, sonic hedgehog is expressed exclusively in the notochord but not in the spinal cord of the regenerate. Overall regeneration was severely impaired in cyclopamine-treated tadpoles. Notochord maturation in the regenerate, including cell alignment and vacuolation, and myofiber formation were inhibited. Proliferation of spinal cord cells in the neural ampulla and of mesenchymal cells was also impaired. As in the axolotl, hedgehog signaling is required for multiple steps in tail regeneration in the Xenopus tadpole, although the location of the Shh source is quite different between the two species. This difference in Shh localization is the likely basis for the differing tissue requirement for tail regeneration between urodeles and anurans.

  13. Growth-arrest-specific protein 2 inhibits cell division in Xenopus embryos.

    Directory of Open Access Journals (Sweden)

    Tong Zhang

    Full Text Available Growth-arrest-specific 2 gene was originally identified in murine fibroblasts under growth arrest conditions. Furthermore, serum stimulation of quiescent, non-dividing cells leads to the down-regulation of gas2 and results in re-entry into the cell cycle. Cytoskeleton rearrangements are critical for cell cycle progression and cell division and the Gas2 protein has been shown to co-localize with actin and microtubules in interphase mammalian cells. Despite these findings, direct evidence supporting a role for Gas2 in the mechanism of cell division has not been reported.To determine whether the Gas2 protein plays a role in cell division, we over-expressed the full-length Gas2 protein and Gas2 truncations containing either the actin-binding CH domain or the tubulin-binding Gas2 domain in Xenopus laevis embryos. We found that both the full-length Gas2 protein and the Gas2 domain, but not the CH domain, inhibited cell division and resulted in multinucleated cells. The observation that Gas2 domain alone can arrest cell division suggests that Gas2 function is mediated by microtubule binding. Gas2 co-localized with microtubules at the cell cortex of Gas2-injected Xenopus embryos using cryo-confocal microscopy and co-sedimented with microtubules in cytoskeleton co-sedimentation assays. To investigate the mechanism of Gas2-induced cell division arrest, we showed, using a wound-induced contractile array assay, that Gas2 stabilized microtubules. Finally, electron microscopy studies demonstrated that Gas2 bundled microtubules into higher-order structures.Our experiments show that Gas2 inhibits cell division in Xenopus embryos. We propose that Gas2 function is mediated by binding and bundling microtubules, leading to cell division arrest.

  14. Egg origin determination efforts

    International Nuclear Information System (INIS)

    Horvath, A.; Futo, I.; Vodila, G.; Palcsu, L.

    2012-01-01

    Complete text of publication follows. As a co-operation with the Poultry Product Board, egg and drinking water samples were received in order to investigate whether the country of origin of the egg can be determined based on its stable isotope composition with the aim of market protection of the Hungarian eggs against the mislabelled foreign ones. The scientific background is that drinking water of egg laying hens is assumed to reflect the composition of regional precipitation, and it is also an input data in the process of egg formation. In the first sampling, altogether 23 sets of egg and drinking water samples were received from different production sites covering the whole area of Hungary. The egg white samples were vacuum distilled and frozen out by liquid nitrogen at -196 deg C. The process was monitored by two vacuum gauges. Water frozen out together with the drinking water samples was measured were measured by a Thermo Finnigan Delta PLUS XP isotope ratio mass spectrometer using a GasBench II peripheral unit equipped with a GC-autosampler. As a second issue, additionally, elemental composition of egg shells were also performed for series of Hungarian, Czech and Polish egg samples by energy dispersive X-ray fluorescence. The drinking waters fit well to the Global Meteoric Water Line indicating their precipitation origin. It was experienced that the water in egg white gets enriched compared to the drinking water (Δ 18 O = -4.9 ± 1.0 per thousand and Δ D = -21.8 ± 6.4 per thousand), however, this shift is independent of the type of the hens, since the mean shifts in the eggs of Tetra and Hy-line hens are similar within error bar. For more depleted drinking water, the shift of the egg white was higher than for more enriched ones. This can be due to the contribution of the nutriment isotopic composition. The water isotope composition of the Hungarian eggs investigated was δ 18 O = -4.8 - -7.3 per thousand and δD = -46.0 - -70.7 per thousand, therefore egg

  15. Effect of BioSentry 904 and ethylenediaminetetraacetic acid-tris disinfecting during incubation of chicken eggs on microbial levels and productivity of poultry.

    Science.gov (United States)

    Walker, S E; Sander, J E

    2004-01-01

    Proper sanitation practices and the use of efficacious disinfectants in a hatchery have an effect on chick quality. Aerosol bacterial counts, egg moisture loss, hatchability, chick quality, and broiler productivity were evaluated when egg surfaces were contaminated by immersion of each egg into a broth medium containing a field isolate of Pseudomonas aeruginosa and incubated with exposure to one of three disinfectant treatments administered by fine spray: distilled water, BioSentry 904 (904), and a 1:1 ratio of 904 and ethylenediaminetetraacetic acid (EDTA)-Tris. The aerosol bacteria levels were statistically greater on day 21 of incubation in the group treated with distilled water than in those receiving disinfectants. Overall hatch of fertile eggs and egg moisture loss were comparable among all three treatments. At 1 day of age, the chicks incubated with 904 had a statistically lower yolk sac contamination rate than those incubated with 904+EDTA-Tris or distilled water. The 2-wk mortality rates, body weights, feed conversion ratios, yolk sac weights, and yolk sac contamination rates were all similar among the three treatments.

  16. Influence of spawning procedure on gametes fertilization success in Salminus hilarii Valenciennes, 1850 (Teleostei: Characidae: Implications for the conservation of this species

    Directory of Open Access Journals (Sweden)

    Renato M. Honji

    Full Text Available Artificial reproduction and gamete fertilization were evaluated in Salminus hilarii wild and domesticated broodstocks. Wild and domesticated broodstocks were artificially induced to reproduction using a carp pituitary treatment. Four groups were considered: Group 1 (G1, fish caught in the wild maintained for three years in the same conditions as the domesticated broodstocks and spawned naturally; Group 2 (G2, broodstock born and raised in captivity and spawned naturally; Group 3 (G3, wild broodstocks, which were manually stripped for gamete collection and dry fertilization; and Group 4 (G4, domesticated males and females, also manually stripped. Oocytes, eggs, and larvae were sampled at different time intervals throughout embryonic development. Yolk sac absorption occurred approximately 24-29 h after hatching. Twenty-six h after hatching, the larvae mouths opened. Cannibalism was identified just 28-30 h after hatching. There was no morphological difference in embryonic development among all groups. The number of released eggs per gram of female was: G1: 83.3 ± 24.5 and G2: 103.8 ± 37.4; however, the fertilization success was lower in G2 (42.0 ± 6.37 % compared with G1 (54.7 ± 3.02% (P = 0.011. Hand-stripping of oocytes was not successful and the fertilization rate was zero. The reproduction of this species in captivity is viable, but it is necessary to improve broodstock management to enhance fertilization rates and obtain better fingerling production for restocking programs.

  17. Radionuclides in Canada goose eggs

    International Nuclear Information System (INIS)

    Rickard, W.H.; Sweany, H.A.

    1975-01-01

    Low levels of radionuclides were measured in Canada goose eggs taken from deserted nests from Columbia River islands on the Energy Research and Development Administration's Hanford Reservation. Potassium-40, a naturally occurring radionuclide, was the most abundant radionuclide measured in egg contents and egg shell. Strontium-90 was incorporated into egg shells and cesium-137 into inner egg contents. Manganese-54, cobalt-60, and zinc-65 were more abundant in inner egg contents than in egg shell. Cerium-144 was detected in egg shell but not in inner shell

  18. Using 3D printed eggs to examine the egg-rejection behaviour of wild birds

    Directory of Open Access Journals (Sweden)

    Branislav Igic

    2015-05-01

    Full Text Available The coevolutionary relationships between brood parasites and their hosts are often studied by examining the egg rejection behaviour of host species using artificial eggs. However, the traditional methods for producing artificial eggs out of plasticine, plastic, wood, or plaster-of-Paris are laborious, imprecise, and prone to human error. As an alternative, 3D printing may reduce human error, enable more precise manipulation of egg size and shape, and provide a more accurate and replicable protocol for generating artificial stimuli than traditional methods. However, the usefulness of 3D printing technology for egg rejection research remains to be tested. Here, we applied 3D printing technology to the extensively studied egg rejection behaviour of American robins, Turdus migratorius. Eggs of the robin’s brood parasites, brown-headed cowbirds, Molothrus ater, vary greatly in size and shape, but it is unknown whether host egg rejection decisions differ across this gradient of natural variation. We printed artificial eggs that encompass the natural range of shapes and sizes of cowbird eggs, painted them to resemble either robin or cowbird egg colour, and used them to artificially parasitize nests of breeding wild robins. In line with previous studies, we show that robins accept mimetically coloured and reject non-mimetically coloured artificial eggs. Although we found no evidence that subtle differences in parasitic egg size or shape affect robins’ rejection decisions, 3D printing will provide an opportunity for more extensive experimentation on the potential biological or evolutionary significance of size and shape variation of foreign eggs in rejection decisions. We provide a detailed protocol for generating 3D printed eggs using either personal 3D printers or commercial printing services, and highlight additional potential future applications for this technology in the study of egg rejection.

  19. Irradiation of shell egg on the physicochemical and functional properties of liquid egg white.

    Science.gov (United States)

    Min, B; Nam, K C; Jo, C; Ahn, D U

    2012-10-01

    This study was aimed at determining the effect of irradiation of shell eggs on the physiochemical and functional properties of liquid egg white during storage. Color and textural parameters of irradiated liquid egg white after cooking were also determined. Shell eggs were irradiated at 0, 2.5, 5, or 10 kGy using a linear accelerator. Egg white was separated from yolk and stored in at 4°C up to 14 d. Viscosity, pH, turbidity, foaming properties, color, and volatile profile of liquid egg white, and color and texture properties of cooked egg white were determined at 0, 7, and 14 d of storage. Irradiation increased the turbidity but decreased viscosity of liquid egg white. Foaming capacity and foam stability were not affected by irradiation at lower dose (2.5 kGy), but were deteriorated at higher doses (≥5.0 kGy) of irradiation. Sulfur-containing volatiles were generated by irradiation and their amounts increased as the irradiation dose increased. However, the sulfur volatiles disappeared during storage under aerobic conditions. Lightness (L* value) and yellowness (b* value) decreased, but greenness (-a* value) increased in cooked egg white in irradiation dose-dependent manners. All textural parameters (hardness, adhesiveness, cohesiveness, chewiness, and resilience) of cooked egg white increased as the irradiation dose increased, but those changes were marginal. Our results indicated that irradiation of shell egg at lower doses (up to 2.5 kGy) had little negative impact on the physiochemical and functional properties of liquid egg white, but can improve the efficiency of egg processing due to its viscosity-lowering effect. Therefore, irradiation of shell eggs at the lower doses has high potential to be used by the egg processing industry to improve the safety of liquid egg without compromising its quality.

  20. Using 3D printed eggs to examine the egg-rejection behaviour of wild birds

    Science.gov (United States)

    Nunez, Valerie; Voss, Henning U.; Croston, Rebecca; Aidala, Zachary; López, Analía V.; Van Tatenhove, Aimee; Holford, Mandë E.; Shawkey, Matthew D.; Hauber, Mark E.

    2015-01-01

    The coevolutionary relationships between brood parasites and their hosts are often studied by examining the egg rejection behaviour of host species using artificial eggs. However, the traditional methods for producing artificial eggs out of plasticine, plastic, wood, or plaster-of-Paris are laborious, imprecise, and prone to human error. As an alternative, 3D printing may reduce human error, enable more precise manipulation of egg size and shape, and provide a more accurate and replicable protocol for generating artificial stimuli than traditional methods. However, the usefulness of 3D printing technology for egg rejection research remains to be tested. Here, we applied 3D printing technology to the extensively studied egg rejection behaviour of American robins, Turdus migratorius. Eggs of the robin’s brood parasites, brown-headed cowbirds, Molothrus ater, vary greatly in size and shape, but it is unknown whether host egg rejection decisions differ across this gradient of natural variation. We printed artificial eggs that encompass the natural range of shapes and sizes of cowbird eggs, painted them to resemble either robin or cowbird egg colour, and used them to artificially parasitize nests of breeding wild robins. In line with previous studies, we show that robins accept mimetically coloured and reject non-mimetically coloured artificial eggs. Although we found no evidence that subtle differences in parasitic egg size or shape affect robins’ rejection decisions, 3D printing will provide an opportunity for more extensive experimentation on the potential biological or evolutionary significance of size and shape variation of foreign eggs in rejection decisions. We provide a detailed protocol for generating 3D printed eggs using either personal 3D printers or commercial printing services, and highlight additional potential future applications for this technology in the study of egg rejection. PMID:26038720

  1. Studies on the effect of gamma rays on egg plant

    Energy Technology Data Exchange (ETDEWEB)

    Ramaswamy, N [Agricultural Coll. and Research Inst., Madurai (India); Sayed, S [Tamil Nadu Agricultural Univ., Madurai (India)

    1977-01-01

    The dry seeds of the cultivar 'Annamalai' of egg plant (Solanum melongena L.) were treated with 10 to 50 krad gamma radiation in steps of 10 krad. The irradiated seeds were sown and M1 plants were selfed. Progeny of each M1 plant was raised for M2 generation. Germination percentage, pollen fertility and survival of seedlings decreased with increased radiation dose. 40 krad and beyond, complete lethality was observed. Flowering period, fruit size and fruit weight decreased with increase in radiation dose upto 20 krad while there was an increase at 30 krad. Mutants isolated in M2 were dwarf and short fruited.

  2. Egg introduction: differential allergic responses

    OpenAIRE

    Dosanjh, Amrita

    2017-01-01

    Amrita Dosanjh Medical Center, Rady Childrens Hospital, San Diego, CA, USA Abstract: The use of egg protein preparations in clinical trials to reduce the incidence of egg allergy among infants includes a number of preparations of egg. These include whole egg, egg white protein, and egg yolk preparations. The study of the differential immune responses to these allergenic proteins in comparison is suggested as a future research area of investigation. Keywords: food allergy, egg allergy, clinica...

  3. Maternal effects of egg size on emu Dromaius novaehollandiae egg composition and hatchling phenotype.

    Science.gov (United States)

    Dzialowski, Edward M; Sotherland, Paul R

    2004-02-01

    Parental investment in eggs and, consequently, in offspring can profoundly influence the phenotype, survival and ultimately evolutionary fitness of an organism. Avian eggs are excellent model systems to examine maternal allocation of energy translated through egg size variation. We used the natural range in emu Dromaius novaehollandiae egg size, from 400 g to >700 g, to examine the influence of maternal investment in eggs on the morphology and physiology of hatchlings. Female emus provisioned larger eggs with a greater absolute amount of energy, nutrients and water in the yolk and albumen. Variation in maternal investment was reflected in differences in hatchling size, which increased isometrically with egg size. Egg size also influenced the physiology of developing emu embryos, such that late-term embryonic metabolic rate was positively correlated with egg size and embryos developing in larger eggs consumed more yolk during development. Large eggs produced hatchlings that were both heavier (yolk-free wet and dry mass) and structurally larger (tibiotarsus and culmen lengths) than hatchlings emerging from smaller eggs. As with many other precocial birds, larger hatchlings also contained more water, which was reflected in a greater blood volume. However, blood osmolality, hemoglobin content and hematocrit did not vary with hatchling mass. Emu maternal investment in offspring, measured by egg size and composition, is significantly correlated with the morphology and physiology of hatchlings and, in turn, may influence the success of these organisms during the first days of the juvenile stage.

  4. 21 CFR 160.180 - Egg yolks.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Egg yolks. 160.180 Section 160.180 Food and Drugs... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.180 Egg yolks. (a) Egg yolks, liquid egg yolks, yolks, liquid yolks are yolks of eggs of the domestic hen so...

  5. Estimating the Number of Eggs in Blow Fly (Diptera: Calliphoridae) Egg Masses Using Photographic Analysis.

    Science.gov (United States)

    Rosati, J Y; Pacheco, V A; Vankosky, M A; Vanlaerhoven, S L

    2015-07-01

    Little work has been done to quantify the number of eggs oviposited by blow flies (Diptera: Calliphoridae) in studies examining colonization behavior. Egg counting methods currently available are time-consuming and destructive. This study used ImageJ software and analysis of covariance to relate the volume of egg masses to the number of eggs laid by three different blow fly species: Lucilia sericata (Meigen), Phormia regina (Meigen), and Chrysomya rufifacies (Macquart). Egg mass volume, species, and the interaction of species and egg mass volume all affected the number of blow fly eggs deposited in egg masses. Both species identity and egg mass volume are important when predicting egg number, as such a single regression equation cannot be used to estimate egg number for these three species. Therefore, simple linear regression equations were determined for each species. The volume of individual eggs was incorporated into the model, yet differences between species were observed, suggesting that the orientation of the eggs oviposited by multiple conspecific females within egg masses influences egg estimates. Based on our results, we expect that imaging software can be used for other blow fly species, as well as other insect species; however, equations specific to each species must be developed. This study describes an important tool for quantifying egg deposition in a nondestructive manner, which is important in studying the colonization behavior and life history of insects of ecological and forensic importance. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  6. Mechanisms underlying the noradrenergic modulation of longitudinal coordination during swimming in Xenopus laevis tadpoles

    DEFF Research Database (Denmark)

    Merrywest, Simon D; McDearmid, Jonathan R; Kjaerulff, Ole

    2003-01-01

    Noradrenaline (NA) is a potent modulator of locomotion in many vertebrate nervous systems. When Xenopus tadpoles swim, waves of motor neuron activity alternate across the body and propagate along it with a brief rostro-caudal delay (RC-delay) between segments. We have now investigated the mechani......Noradrenaline (NA) is a potent modulator of locomotion in many vertebrate nervous systems. When Xenopus tadpoles swim, waves of motor neuron activity alternate across the body and propagate along it with a brief rostro-caudal delay (RC-delay) between segments. We have now investigated...... might promote postinhibitory rebound firing. The synaptic inputs during swimming were simulated using a sustained positive current, superimposed upon which were brief negative currents. When these conditions were held constant NA enhanced the probability of rebound firing--indicating a direct effect...

  7. Calcium waves.

    Science.gov (United States)

    Jaffe, Lionel F

    2008-04-12

    Waves through living systems are best characterized by their speeds at 20 degrees C. These speeds vary from those of calcium action potentials to those of ultraslow ones which move at 1-10 and/or 10-20 nm s(-1). All such waves are known or inferred to be calcium waves. The two classes of calcium waves which include ones with important morphogenetic effects are slow waves that move at 0.2-2 microm s(-1) and ultraslow ones. Both may be propagated by cycles in which the entry of calcium through the plasma membrane induces subsurface contraction. This contraction opens nearby stretch-sensitive calcium channels. Calcium entry through these channels propagates the calcium wave. Many slow waves are seen as waves of indentation. Some are considered to act via cellular peristalsis; for example, those which seem to drive the germ plasm to the vegetal pole of the Xenopus egg. Other good examples of morphogenetic slow waves are ones through fertilizing maize eggs, through developing barnacle eggs and through axolotl embryos during neural induction. Good examples of ultraslow morphogenetic waves are ones during inversion in developing Volvox embryos and across developing Drosophila eye discs. Morphogenetic waves may be best pursued by imaging their calcium with aequorins.

  8. The effect of age on sperm stock and egg laying in the parasitoid wasp, Dinarmus basalis

    Directory of Open Access Journals (Sweden)

    D. Damiens

    2003-07-01

    Full Text Available Sperm quantity and quality during storage may be constraints acting on female fecundity and hence fitness. In Hymenoptera, the importance of sperm quality has rarely been considered, despite its central role in reproductive strategies and especially in sex ratio control. In these insects, fertilized eggs develop into females and unfertilized eggs into males. Experiments were conducted on the female wasp, Dinarmus basalis, in the laboratory with and without egg-laying resources (hosts. The first point was to test if sperm age influenced sperm storage by measuring sperm count and viability using a sperm viability test (SYBR-14 : propidium iodide. The second point was the influence of prolonged storage in the female genital tract on the quantity, sex ratio and fitness of offspring produced. Results show that sperm viability in the spermatheca does not change significantly with maternal age, and that the sperm stock is not affected when females are deprived of hosts. Egg-laying is gradually restored after 21 days of host deprivation but remains at a low level after 115 days. The fitness of mated D. basalis females is therefore not constrained by sperm quantity or quality and seems to depend on host availability and female age.

  9. Enumeration of Salmonellae in Table Eggs, Pasteurized Egg Products, and Egg-Containing Dishes by Using Quantitative Real-Time PCR

    DEFF Research Database (Denmark)

    Jakočiūnė, Džiuginta; Pasquali, Frédérique; da Silva, Cristiana Soares

    2014-01-01

    PCR) was employed for enumeration of salmonellae in different matrices: table eggs, pasteurized egg products, and egg-containing dishes. Salmonella enterica serovar Enteritidis and S. enterica serovar Tennessee were used to artificially contaminate these matrices. The results showed a linear regression between...... the numbers of salmonellae and the quantification cycle (Cq) values for all matrices used, with the exception of pasteurized egg white. Standard curves were constructed by using both stationary-phase cells and heat-stressed cells, with similar results. Finally, this method was used to evaluate the fate...

  10. Studies on improving ostrich egg hatch ability and its relation with some factors affecting embryonic development during artificial incubation

    International Nuclear Information System (INIS)

    Amer, N.S.I.

    2012-01-01

    The present study was carried out in co-operation between the Ostrich Production Farm, Nuclear Research Center, Atomic Energy Authority, Cairo, Egypt and the Faculty of Agriculture, AL-Azhar University, Nasr City, Cairo, Egypt. Ostrich eggs were obtained from Resk Company for Ostrich Production and set for incubation at ElShfie Farm, Belbas, Sharkia, Egypt. The objectives are:1- To follow up changes in some vital physiological parameters and blood components associated with ostrich embryonic development during incubation and to provide reference blood biochemical baseline values for future studies of avian species and to document novel information on some normal changes associated with growth of the developing ostrich embryo during the incubation, as no similar and complete data could be found on this aspect in the literature. 2- In an effort to improve the hatch ability and hatching performance of ostrich eggs by testing the effect of in ovo injection of several nutrients. Two trials were carried out: 1-First trial To follow up changes in some vital physiological parameters and blood components associated with ostrich embryonic development during incubation. A total number of 60 ostrich eggs weighed between 1300 and 1500 g were obtained from from Resk Company for Ostrich Production. Eggs were collected weekly in patches of 25 eggs and Egg incubation was performed in ElShfie Farm, Belbas, Egypt. Egg weight and egg weight loss during incubation were determined on each eggs.2- Second Trial In vivo injection In an effort to improve the hatch ability and hatching performance of ostrich eggs by testing the effect of in ovo injection of several nutrients. A total of 100 fertile ostrich eggs weighed between 1300 and 1500 g were obtained from from Resk Company for Ostrich Production. Eggs were collected weekly in patches of 25 eggs and egg incubation was performed in ElShfie Farm, Belbas, Egypt. Eggs were injected at the 7 th day of incubation to deposit test material in

  11. Egg number-egg size: an important trade-off in parasite life history strategies.

    Science.gov (United States)

    Cavaleiro, Francisca I; Santos, Maria J

    2014-03-01

    Parasites produce from just a few to many eggs of variable size, but our understanding of the factors driving variation in these two life history traits at the intraspecific level is still very fragmentary. This study evaluates the importance of performing multilevel analyses on egg number and egg size, while characterising parasite life history strategies. A total of 120 ovigerous females of Octopicola superba (Copepoda: Octopicolidae) (one sample (n=30) per season) were characterised with respect to different body dimensions (total length; genital somite length) and measures of reproductive effort (fecundity; mean egg diameter; total reproductive effort; mean egg sac length). While endoparasites are suggested to follow both an r- and K-strategy simultaneously, the evidence found in this and other studies suggests that environmental conditions force ectoparasites into one of the two alternatives. The positive and negative skewness of the distributions of fecundity and mean egg diameter, respectively, suggest that O. superba is mainly a K-strategist (i.e. produces a relatively small number of large, well provisioned eggs). Significant sample differences were recorded concomitantly for all body dimensions and measures of reproductive effort, while a general linear model detected a significant influence of season*parasite total length in both egg number and size. This evidence suggests adaptive phenotypic plasticity in body dimensions and size-mediated changes in egg production. Seasonal changes in partitioning of resources between egg number and size resulted in significant differences in egg sac length but not in total reproductive effort. Evidence for a trade-off between egg number and size was found while controlling for a potential confounding effect of parasite total length. However, this trade-off became apparent only at high fecundity levels, suggesting a state of physiological exhaustion. Copyright © 2014 Australian Society for Parasitology Inc. Published

  12. Intraspecific variation in egg size and egg composition in birds: effects on offspring fitness.

    Science.gov (United States)

    Williams, T D

    1994-02-01

    1. There is little unequivocal evidence to date in support of a positive relationship between egg size and offspring fitness in birds. Although 40 studies (of 34 species) have considered the effect of variation in egg size on chick growth and/or survival up to fledgling only 12 studies have controlled for other characters potentially correlated both with egg size and offspring fitness. Of these only two have reported a significant residual effect of egg size on chick growth (in the roseate tern and European blackbird) and three a residual effect on chick survival (all in seabirds: common tern, lesser black-backed gull and kittiwake). 2. More consistent evidence exists, though from fewer studies, for a positive relationship between egg size and offspring fitness early in the chick-rearing period; chick growth and chick survival being dependent on egg size in 8 of 10 studies and 4 of 5 studies respectively. It is suggested that the most important effect of variation in egg size might be in determining the probability of offspring survival in the first few days after hatching. 3. Egg size explains on average 66% of the variation in chick mass at hatching (n = 35 studies) but only 30% of the variation in chick body size (n = 18). When effects of hatching body size are controlled for chick mass remains significantly correlated with egg size, though the reverse is not true. This supports the hypothesis that large eggs give rise to heavier chicks at hatching, i.e., chicks with more nutrient (yolk) reserves, rather than structurally larger chicks. 4. Egg composition increased isometrically with increasing egg size in about half the studies so far reported (n equals approximately 20). However, in seabirds, and some passerines, larger eggs contain disproportionately more albumen, whilst in some waterfowl percentage yolk content increases with increasing egg size. Changes in albumen content largely reflect variation in the water content of eggs, but changes in yolk content

  13. Pentachlorophenol (PCP) bioaccumulation and effect on heat production on salmon eggs at different stages of development

    Energy Technology Data Exchange (ETDEWEB)

    Maeenpaeae, Kimmo A.; Penttinen, Olli-Pekka; Kukkonen, Jussi V.K

    2004-05-28

    In this study, pentachlorophenol (PCP) bioaccumulation and its effect on heat dissipation was studied in eggs of the lake salmon (Salmo salar m. sebago). In bioaccumulation studies, the eggs were exposed to low concentrations (0.051-0.056 {mu}mol/l, 13.583-14.915) of waterborne [{sup 14}C]-labeled PCP at two developmental stages: (1) 3 weeks after fertilization, and (2) just before hatching. The effect of PCP on egg heat dissipation was measured by a microcalorimeter after exposing the eggs to gradual concentrations (0-0.992 {mu}mol/l) of PCP for 48 h. After both the bioaccumulation and heat dissipation experiments, the eggs were dissected and the concentrations of PCP in tissue were determined separately for eggshell, yolk and embryo. The bioaccumulation studies showed that PCP accumulates more in the eggs at the late developmental stage. Bioconcentration factors (BCF) for different tissues were 3-42 times higher for the eggs at the late developmental stage compared with the eggs that were incubated only for 3 weeks. In early developmental stage, the eggshell adsorbs a large portion of the chemical. In late developmental stage, the actual embryo accumulated both proportionately and totally more than other dissected tissues in the beginning of the exposure, but eventually the yolk accumulated highest total amount of the chemical. A probable reason for the higher PCP body burden in the late developmental stage is that the respiration rate and metabolic activity of the embryo increases as it grows. The salmon eggs responded to an exposure to PCP with an elevated rate of heat dissipation. The threshold concentration above which the embryo heat dissipation was amplified was 29.64 {mu}mol/kg embryo wet weight (ww) or 0.28 {mu}mol/l. The highest embryo heat production was measured at the exposure concentration of 0.992 {mu}mol/l. At higher exposure concentrations the heat dissipation decreased. The basic findings of the study are that PCP accumulates in growing embryonic

  14. 9 CFR 590.45 - Prohibition on eggs and egg products not intended for use as human food.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Prohibition on eggs and egg products... INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION INSPECTION OF EGGS AND EGG PRODUCTS (EGG PRODUCTS INSPECTION ACT) Eggs and Egg Products Not Intended for Human Food § 590.45 Prohibition on...

  15. Genomic analysis of Xenopus organizer function

    Directory of Open Access Journals (Sweden)

    Suhai Sándor

    2006-06-01

    Full Text Available Abstract Background Studies of the Xenopus organizer have laid the foundation for our understanding of the conserved signaling pathways that pattern vertebrate embryos during gastrulation. The two primary activities of the organizer, BMP and Wnt inhibition, can regulate a spectrum of genes that pattern essentially all aspects of the embryo during gastrulation. As our knowledge of organizer signaling grows, it is imperative that we begin knitting together our gene-level knowledge into genome-level signaling models. The goal of this paper was to identify complete lists of genes regulated by different aspects of organizer signaling, thereby providing a deeper understanding of the genomic mechanisms that underlie these complex and fundamental signaling events. Results To this end, we ectopically overexpress Noggin and Dkk-1, inhibitors of the BMP and Wnt pathways, respectively, within ventral tissues. After isolating embryonic ventral halves at early and late gastrulation, we analyze the transcriptional response to these molecules within the generated ectopic organizers using oligonucleotide microarrays. An efficient statistical analysis scheme, combined with a new Gene Ontology biological process annotation of the Xenopus genome, allows reliable and faithful clustering of molecules based upon their roles during gastrulation. From this data, we identify new organizer-related expression patterns for 19 genes. Moreover, our data sub-divides organizer genes into separate head and trunk organizing groups, which each show distinct responses to Noggin and Dkk-1 activity during gastrulation. Conclusion Our data provides a genomic view of the cohorts of genes that respond to Noggin and Dkk-1 activity, allowing us to separate the role of each in organizer function. These patterns demonstrate a model where BMP inhibition plays a largely inductive role during early developmental stages, thereby initiating the suites of genes needed to pattern dorsal tissues

  16. Fertility desires and fertility outcomes.

    Science.gov (United States)

    Bracher, M; Santow, G

    1991-05-01

    An Australian 1-in-1000 national probability sample conducted in 1986 yielded 2547 women aged 20-59 who provided detailed life histories on marital unions, childbearing, and contraception. Age specific fertility rates, desired family size, differentials in desired family size, desired fertility and achieved fertility, and sequential family building are examined. The results indicate that the desired family size at 1st marriage has declined only slightly over the past 30 years. 3 children are generally desired, and ver few desire 2. The constance of fertility desires in contrasted with the fertilitydecline to below replacement levels. Several reasons are suggested for the desired family size: the desire is for a family size within the family tradition and modified by the desire to have 1 of each sex, the desire reflects less on intentions but more on normative pressure to become a parent. Marrying is self selecting on the desire for a traditional family of at least 2 children. There is a rising age at marriage as well as a decline in marriages. Desired family size exceeds completed fertility. Period factors and personal circumstances affect fertility intentions. Future inquires should explore the multiple factors relating to fertility, rather than in comparing fertility desires and actual fertility. The data collected on age specific fertility were comparable to official estimates. The fertility decline was evidenced in all groups except teenagers. The decline was nearly 50% for those 20-24 years between the 1050's-80's, 33% for ages 25-29. Marriage patterns explain this decline in part. Between 1971-76, women aged 20-25 were married 37 months out of 60 months in 1971-76 versus 25 out of 60 months in 1981-86. Within the 25 year age group, marital fertility has declined and unmarried fertility, which is low, has risen, Women in a marital union of any kind has remained stable. Fertility within de facto unions, which is lower than within marriage, is higher than

  17. 9 CFR 590.920 - Importer to make application for inspection of imported eggs and egg products.

    Science.gov (United States)

    2010-01-01

    ... inspection of imported eggs and egg products. 590.920 Section 590.920 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION INSPECTION OF EGGS AND EGG PRODUCTS (EGG PRODUCTS INSPECTION ACT) Imports § 590.920 Importer to make application for inspection of...

  18. Revisiting "The fertilization fairytale:" an analysis of gendered language used to describe fertilization in science textbooks from middle school to medical school

    Science.gov (United States)

    Campo-Engelstein, Lisa; Johnson, Nadia L.

    2014-03-01

    Emily Martin's (Signs J Women Cult Soc 16(31):485-501, 1991) article, "The Egg and the Sperm: How Science Has Constructed a Romance Based on Stereotypical Male-Female Roles," was published in Signs over 20 years ago. In this groundbreaking article, she discusses how gender roles are often projected onto reproductive biology, leading to the portrayal of eggs as passive and sperm as active. We were interested in seeing if many of her findings are still relevant today. We analyzed science textbooks from the middle school to the medical school level to determine if fertilization in human reproduction is described in gender-biased language regarding the sentence structure, amount of information provided for female and male processes/parts, and neutrality in describing female and male processes/parts. Although there has been much improvement, there is still a long way to go. Sexist language in scientific textbooks is troubling because it negatively affects both female and male students and undermines teachers' ability to teach in an accurate and gender-neutral way.

  19. Effect of Albendazole at different concentrations on fertilization and early cleavage in Tetrapygus niger “sea urchin”

    Directory of Open Access Journals (Sweden)

    Gina Zavaleta Espejo

    2013-09-01

    Full Text Available In this study we evaluated the effect of Albendazole at different concentrations and exposure times on the process of fertilization and early cleavage in Tetrapygus niger "sea urchin". Each experimental group consisted of 200 mL, of previously filtered seawater at pH 7.3 and temperature of 20 ± 2 °C, plus five drops of eggs and two drops of spermatozoa exposed to different concentrations of Albendazole 400 ppm, 800 ppm and 1200 ppm. Determining the effect of Albendazole was conducted by counting the number of cones of fertilization, as well as the number of cleaving embryos with normal and abnormal. The ANOVA and multiple comparison test Tukey averages showed significant differences between the treatments, namely that increasing the concentration of Albendazole fertilization rate decreases and increases the percentage of embryos with abnormal cleavage, so therefore concluded that the Albendazole at different concentrations and exposure times affects the process of fertilization and early cleavage in T, niger "sea urchin"

  20. Effects of preincubation heating of broiler hatching eggs during storage, flock age, and length of storage period on hatchability.

    Science.gov (United States)

    Gucbilmez, M; Ozlü, S; Shiranjang, R; Elibol, O; Brake, J

    2013-12-01

    The effects of heating of eggs during storage, broiler breeder age, and length of egg storage on hatchability of fertile eggs were examined in this study. Eggs were collected from Ross 344 male × Ross 308 broiler breeders on paper flats, held overnight (1 d) at 18°C and 75% RH, and then transferred to plastic trays. In experiment 1, eggs were obtained at 28, 38, and 53 wk of flock age. During a further 10 d of storage, eggs either remained in the storage room (control) or were subjected to a heat treatment regimen of 26°C for 2 h, 37.8°C for 3 h, and 26°C for 2 h in a setter at d 5 of storage. In experiment 2, eggs from a flock at 28 wk of age were heated for 1 d of a 6-d storage period. Eggs from a 29-wk-old flock were either heated at d 1 or 5 of an 11-d storage period in experiment 3. In experiment 4, 27-wk-old flock eggs were heated twice at d 1 and 5 of an 11-d storage period. Control eggs stored for 6 or 11 d were coincubated as appropriate in each experiment. Heating eggs at d 5 of an 11-d storage period increased hatchability in experiment 1. Although no benefit of heating 28-wk-old flock eggs during 6 d of storage in experiment 2 was observed, heating eggs from a 29-wk-old flock at d 1 or 5 of an 11-d storage period increased hatchability in experiment 3. Further, heating eggs from a 27-wk-old flock twice during 11 d of storage increased hatchability in experiment 4. These effects were probably due to the fact that eggs from younger flocks had been reported to have many embryos at a stage of development where the hypoblast had not yet fully developed (less than EG-K12 to EG-K13), such that heating during extended storage advanced these embryos to a more resistant stage.

  1. 9 CFR 590.510 - Classifications of shell eggs used in the processing of egg products.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Classifications of shell eggs used in the processing of egg products. 590.510 Section 590.510 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG PRODUCTS INSPECTION INSPECTION OF EGGS AND EGG PRODUCTS...

  2. Trichuris trichiura egg (image)

    Science.gov (United States)

    ... is the classical appearance of the Trichuria (whipworm) egg. The eggs are highly infectious. After a person eats contaminated food, the worms hatch from the eggs and live in the intestine, causing vomiting and ...

  3. PHENOBARBITAL AFFECTS THYROID HISTOLOGY AND LARVAL DEVELOPMENT IN THE AFRICAN CLAWED FROG XENOPUS LAEVIS

    Science.gov (United States)

    The abstract highlights our recent study to explore endocrine disrupting effects of phenobarbital in the African clawed frog, Xenopus laevis. In mammals, this chemical is known to induce the biotransforming enzyme UDP-glucuronosyltransferase (UDPGT) resulting in increased thyroid...

  4. Geographical variation in egg mass and egg content in a passerine bird.

    Directory of Open Access Journals (Sweden)

    Suvi Ruuskanen

    Full Text Available Reproductive, phenotypic and life-history traits in many animal and plant taxa show geographic variation, indicating spatial variation in selection regimes. Maternal deposition to avian eggs, such as hormones, antibodies and antioxidants, critically affect development of the offspring, with long-lasting effects on the phenotype and fitness. Little is however known about large-scale geographical patterns of variation in maternal deposition to eggs. We studied geographical variation in egg components of a passerine bird, the pied flycatcher (Ficedula hypoleuca, by collecting samples from 16 populations and measuring egg and yolk mass, albumen lysozyme activity, yolk immunoglobulins, yolk androgens and yolk total carotenoids. We found significant variation among populations in most egg components, but ca. 90% of the variation was among individuals within populations. Population however explained 40% of the variation in carotenoid levels. In contrast to our hypothesis, we found geographical trends only in carotenoids, but not in any of the other egg components. Our results thus suggest high within-population variation and leave little scope for local adaptation and genetic differentiation in deposition of different egg components. The role of these maternally-derived resources in evolutionary change should be further investigated.

  5. Effects of dietary humic substances on egg production and egg shell ...

    African Journals Online (AJOL)

    Effects of dietary humic substances on egg production and egg shell quality of hens after peak laying period. ... Ninety Isa Brown layers from 51 to 61 weeks of age were allocated to three treatment groups, namely H0, H1 and H2. ... Egg production (% hen-day) in the H2 group was higher (P < 0.05) than control group.

  6. Gene expression analysis of the ovary of hybrid females of Xenopus laevis and X. muelleri

    Directory of Open Access Journals (Sweden)

    Malone John H

    2008-03-01

    Full Text Available Abstract Background Interspecific hybrids of frogs of the genus Xenopus result in sterile hybrid males and fertile hybrid females. Previous work has demonstrated a dramatic asymmetrical pattern of misexpression in hybrid males compared to the two parental species with relatively few genes misexpressed in comparisons of hybrids and the maternal species (X. laevis and dramatically more genes misexpressed in hybrids compared to the paternal species (X. muelleri. In this work, we examine the gene expression pattern in hybrid females of X. laevis × X. muelleri to determine if this asymmetrical pattern of expression also occurs in hybrid females. Results We find a similar pattern of asymmetry in expression compared to males in that there were more genes differentially expressed between hybrids and X. muelleri compared to hybrids and X. laevis. We also found a dramatic increase in the number of misexpressed genes with hybrid females having about 20 times more genes misexpressed in ovaries compared to testes of hybrid males and therefore the match between phenotype and expression pattern is not supported. Conclusion We discuss these intriguing findings in the context of reproductive isolation and suggest that divergence in female expression may be involved in sterility of hybrid males due to the inherent sensitivity of spermatogenesis as defined by the faster male evolution hypothesis for Haldane's rule.

  7. Effects of gamma radiation on codling moth (Cydia pomonella, Lepidoptera: Tortricidae) fertility and reproductive behaviour

    International Nuclear Information System (INIS)

    Mansour, M.

    2002-01-01

    Studies were conducted with codling moth, Cydia pomonella (L.), to examine the effects of gamma radiation on fertility and reproductive behaviour. Data accumulated during these studies showed that egg production and hatch decreased with increasing radiation dose. Females were more sensitive to radiation treatment than were males. A dose of 150 Gy caused 100% sterility in females and significantly reduced fecundity, and a dose of 350 Gy reduced male fertility to less than 1%. Radiation dosages up to 400 Gy had no adverse effect on male longevity or competitiveness in cages using laboratory reared moths. However, males exposed to a dose of 350 or 400 Gy mated fewer times than unirradiated males. (author)

  8. Heterotrimeric Kinesin II Is the Microtubule Motor Protein Responsible for Pigment Dispersion in Xenopus Melanophores

    Science.gov (United States)

    Tuma, M. Carolina; Zill, Andrew; Le Bot, Nathalie; Vernos, Isabelle; Gelfand, Vladimir

    1998-01-01

    Melanophores move pigment organelles (melanosomes) from the cell center to the periphery and vice-versa. These bidirectional movements require cytoplasmic microtubules and microfilaments and depend on the function of microtubule motors and a myosin. Earlier we found that melanosomes purified from Xenopus melanophores contain the plus end microtubule motor kinesin II, indicating that it may be involved in dispersion (Rogers, S.L., I.S. Tint, P.C. Fanapour, and V.I. Gelfand. 1997. Proc. Natl. Acad. Sci. USA. 94: 3720–3725). Here, we generated a dominant-negative construct encoding green fluorescent protein fused to the stalk-tail region of Xenopus kinesin-like protein 3 (Xklp3), the 95-kD motor subunit of Xenopus kinesin II, and introduced it into melanophores. Overexpression of the fusion protein inhibited pigment dispersion but had no effect on aggregation. To control for the specificity of this effect, we studied the kinesin-dependent movement of lysosomes. Neither dispersion of lysosomes in acidic conditions nor their clustering under alkaline conditions was affected by the mutant Xklp3. Furthermore, microinjection of melanophores with SUK4, a function-blocking kinesin antibody, inhibited dispersion of lysosomes but had no effect on melanosome transport. We conclude that melanosome dispersion is powered by kinesin II and not by conventional kinesin. This paper demonstrates that kinesin II moves membrane-bound organelles. PMID:9852150

  9. The repetitive portion of the Xenopus IgH Mu switch region mediates orientation-dependent class switch recombination.

    Science.gov (United States)

    Zhang, Zheng Z; Pannunzio, Nicholas R; Lu, Zhengfei; Hsu, Ellen; Yu, Kefei; Lieber, Michael R

    2015-10-01

    Vertebrates developed immunoglobulin heavy chain (IgH) class switch recombination (CSR) to express different IgH constant regions. Most double-strand breaks for Ig CSR occur within the repetitive portion of the switch regions located upstream of each set of constant domain exons for the Igγ, Igα or Igϵ heavy chain. Unlike mammalian switch regions, Xenopus switch regions do not have a high G-density on the non-template DNA strand. In previous studies, when Xenopus Sμ DNA was moved to the genome of mice, it is able to support substantial CSR when it is used to replace the murine Sγ1 region. Here, we tested both the 2kb repetitive portion and the 4.6 kb full-length portions of the Xenopus Sμ in both their natural (forward) orientation relative to the constant domain exons, as well as the opposite (reverse) orientation. Consistent with previous work, we find that the 4.6 kb full-length Sμ mediates similar levels of CSR in both the forward and reverse orientations. Whereas, the forward orientation of the 2kb portion can restore the majority of the CSR level of the 4.6 kb full-length Sμ, the reverse orientation poorly supports R-looping and no CSR. The forward orientation of the 2kb repetitive portion has more GG dinucleotides on the non-template strand than the reverse orientation. The correlation of R-loop formation with CSR efficiency, as demonstrated in the 2kb repetitive fragment of the Xenopus switch region, confirms a role played by R-looping in CSR that appears to be conserved through evolution. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Eggs: good or bad?

    Science.gov (United States)

    Griffin, Bruce A

    2016-08-01