G. R. Poirier, R. Robinson, R. Richardson, K. Hinds, D. Clayton
Murine cauda epididymal sperm preincubated in either a modified Krebs-Ringer or M 199 solution bind to cumulus-free, zona pellucida-intact eggs. Pretreatment of such eggs with an affinity purified preparation of the seminal inhibitor binding component (acceptor), isolated from epididymal sperm, reduces in a concentration dependent manner, the number of sperm that bind. Treatment of cauda sperm, preincubated in either of the above two media, with the seminal inhibitor, also reduces the number of sperm able to bind. Incubation of cauda sperm in the Krebs-Ringer solution for up to 4 h does not affect their ability to bind the seminal inhibitor. Omission of bovine serum albumin from the preincubation medium results in a significant reduction in sperm binding. These data are interpreted to mean that the seminal inhibitor acceptor sites on the sperm surface of incubated sperm function in the in vitro binding to the zona pellucida.
{"title":"Evidence for a binding site on the sperm plasma membrane which recognizes the murine zona pellucida: A binding site on the sperm plasma membrane","authors":"G. R. Poirier, R. Robinson, R. Richardson, K. Hinds, D. Clayton","doi":"10.1002/MRD.1120140307","DOIUrl":"https://doi.org/10.1002/MRD.1120140307","url":null,"abstract":"Murine cauda epididymal sperm preincubated in either a modified Krebs-Ringer or M 199 solution bind to cumulus-free, zona pellucida-intact eggs. Pretreatment of such eggs with an affinity purified preparation of the seminal inhibitor binding component (acceptor), isolated from epididymal sperm, reduces in a concentration dependent manner, the number of sperm that bind. Treatment of cauda sperm, preincubated in either of the above two media, with the seminal inhibitor, also reduces the number of sperm able to bind. Incubation of cauda sperm in the Krebs-Ringer solution for up to 4 h does not affect their ability to bind the seminal inhibitor. Omission of bovine serum albumin from the preincubation medium results in a significant reduction in sperm binding. These data are interpreted to mean that the seminal inhibitor acceptor sites on the sperm surface of incubated sperm function in the in vitro binding to the zona pellucida.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"51 1","pages":"235-243"},"PeriodicalIF":0.0,"publicationDate":"1986-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79807838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Eggs were isolated from the oviducts or ovaries of LT/Sv strain mice in order to investigate the pathways taken by them following spontaneous or induced parthenogenetic activation. The chromosome preparations from the ovarian oocytes that matured in vitro to metaphase I were all morphologically normal. Of 42 recently ovulated eggs that failed to activate parthenogenetically in culture, 57% on nuclear densitometric analysis were found to have the normal 2C amount of DNA, and 1N (haploid) number of chromosomes present, and were arrested at metaphase II. Somewhat unexpectedly, 43% had a 4C amount of DNA, and 2N (diploid) number of chromosomes present, had been arrested at metaphase I, and were evidently ovulated as primary oocytes. Following parthenogenetic activation, the majority of oocytes extruded a polar body and developed a single pronucleus. The activated eggs could be divided into two sub-populations according to the diameter (and therefore volume) of the pronucleus—in one group this was about one-third greater than in the other. The chromosome constitution of the two groups was determined separately at the first cleavage mitosis. Those with a normal-sized pronucleus were invariably haploid, while those with an enlarged pronuclear volume were invariably found to be diploid. The chromosomes in the diploid spreads often appeared to be associated in homologous pairs. We conclude that almost uniquely in LT/Sv strain females eggs may be activated parthenogenetically at either stage of meiotic maturation giving rise to diploid or haploid embryos, respectively.
{"title":"The ovulation and activation of primary and secondary oocytes in LT/Sv strain mice","authors":"M. Kaufman, S. Howlett","doi":"10.1002/MRD.1120140309","DOIUrl":"https://doi.org/10.1002/MRD.1120140309","url":null,"abstract":"Eggs were isolated from the oviducts or ovaries of LT/Sv strain mice in order to investigate the pathways taken by them following spontaneous or induced parthenogenetic activation. The chromosome preparations from the ovarian oocytes that matured in vitro to metaphase I were all morphologically normal. Of 42 recently ovulated eggs that failed to activate parthenogenetically in culture, 57% on nuclear densitometric analysis were found to have the normal 2C amount of DNA, and 1N (haploid) number of chromosomes present, and were arrested at metaphase II. Somewhat unexpectedly, 43% had a 4C amount of DNA, and 2N (diploid) number of chromosomes present, had been arrested at metaphase I, and were evidently ovulated as primary oocytes. Following parthenogenetic activation, the majority of oocytes extruded a polar body and developed a single pronucleus. The activated eggs could be divided into two sub-populations according to the diameter (and therefore volume) of the pronucleus—in one group this was about one-third greater than in the other. The chromosome constitution of the two groups was determined separately at the first cleavage mitosis. Those with a normal-sized pronucleus were invariably haploid, while those with an enlarged pronuclear volume were invariably found to be diploid. The chromosomes in the diploid spreads often appeared to be associated in homologous pairs. We conclude that almost uniquely in LT/Sv strain females eggs may be activated parthenogenetically at either stage of meiotic maturation giving rise to diploid or haploid embryos, respectively.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"1 1","pages":"255-264"},"PeriodicalIF":0.0,"publicationDate":"1986-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90291528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The fertilizing capacity was compared between testicular and vas deferens sperm in Cynops pyrrhogaster. The testicular sperm was not capable of fertilizing jelly eggs. In contrast, the vas deferens sperm was already capable of fertilizing the newt jelly eggs. There was no inhibitory factor for fertilizing jelly eggs in the testis. These results suggest that the testicular sperm is immature as to the fertilizing capacity. The testicular sperm gained the fertilizing capacity for the jelly eggs by treatment with Holtfreter's solution or 1/20 strength Holtfreter's solution. The treatment may promote the step of maturation to achieve the fertilizing capacity. The treated testicular sperm did not fertilize dejellied eggs, although vas deferens sperm fertilized dejellied eggs. Therefore, the maturation state of the treated testicular sperm is different from that of vas deferens sperm. Newt sperm may be matured within the vas deferens, as the newt does not have an organ like the mammalian epididymis.
{"title":"Difference of fertilizing capacity between testicular sperm and vas deferens sperm in Cynops pyrrhogaster","authors":"M. Matsuda","doi":"10.1002/MRD.1120140304","DOIUrl":"https://doi.org/10.1002/MRD.1120140304","url":null,"abstract":"The fertilizing capacity was compared between testicular and vas deferens sperm in Cynops pyrrhogaster. The testicular sperm was not capable of fertilizing jelly eggs. In contrast, the vas deferens sperm was already capable of fertilizing the newt jelly eggs. There was no inhibitory factor for fertilizing jelly eggs in the testis. These results suggest that the testicular sperm is immature as to the fertilizing capacity. The testicular sperm gained the fertilizing capacity for the jelly eggs by treatment with Holtfreter's solution or 1/20 strength Holtfreter's solution. The treatment may promote the step of maturation to achieve the fertilizing capacity. The treated testicular sperm did not fertilize dejellied eggs, although vas deferens sperm fertilized dejellied eggs. Therefore, the maturation state of the treated testicular sperm is different from that of vas deferens sperm. Newt sperm may be matured within the vas deferens, as the newt does not have an organ like the mammalian epididymis.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"115 1","pages":"209-214"},"PeriodicalIF":0.0,"publicationDate":"1986-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89807660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A combination of autoradiography and air-dried techniques was used to calculate the duration of the major meiotic stages in the first wave of spermatogenesis in the newborn mouse. The data indicated that the entry into meiosis occurred asynchronously over 2 days, and the time required for each stage and the total cycle was constant. These time intervals were nearly identical with those estimated for adult animals in the present study and by other authors.
{"title":"The kinetics of the first wave of spermatogenesis in the newborn male mouse","authors":"W. Sung, M. Komatsu, G. Jagiello","doi":"10.1002/MRD.1120140308","DOIUrl":"https://doi.org/10.1002/MRD.1120140308","url":null,"abstract":"A combination of autoradiography and air-dried techniques was used to calculate the duration of the major meiotic stages in the first wave of spermatogenesis in the newborn mouse. The data indicated that the entry into meiosis occurred asynchronously over 2 days, and the time required for each stage and the total cycle was constant. These time intervals were nearly identical with those estimated for adult animals in the present study and by other authors.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"54 1","pages":"245-254"},"PeriodicalIF":0.0,"publicationDate":"1986-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78683920","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The effect of the calmodulin antagonist W-7 on the capacitation and the acrosome reaction of guinea pig spermatozoa was examined. The characteristic features of the acrosome reaction induced by W-7 were the dependence on the composition and pH of the medium and on the presence of sodium bicarbonate. The most effective concentration of W-7 for inducing the acrosome reaction was approximately 5 μM, which is far less than the Kd for calmodulin. Moreover, W-7 enhanced the ability of spermatozoa to acquire capacitation in a Ca2+-free medium. The spermatozoa induced to undergo the acrosome reaction by W-7 were capable of penetrating the zona-free hamster eggs. W-5, which has a lower affinity for calmodulin than W-7, also induced the acrosome reaction in the same manner as W-7. These results suggest that the naphthalenesulfonamide derivatives W-7 and W-5 can induce the acrosome reaction in guinea pig spermatozoa via capacitation in a pH-dependent, Ca2+-calmodulin-independent manner.
{"title":"Induction of the acrosome reaction in guinea pig spermatozoa by calmodulin antagonist W-7","authors":"T. Nagae, P. N. Srivastava","doi":"10.1002/MRD.1120140303","DOIUrl":"https://doi.org/10.1002/MRD.1120140303","url":null,"abstract":"The effect of the calmodulin antagonist W-7 on the capacitation and the acrosome reaction of guinea pig spermatozoa was examined. The characteristic features of the acrosome reaction induced by W-7 were the dependence on the composition and pH of the medium and on the presence of sodium bicarbonate. The most effective concentration of W-7 for inducing the acrosome reaction was approximately 5 μM, which is far less than the Kd for calmodulin. Moreover, W-7 enhanced the ability of spermatozoa to acquire capacitation in a Ca2+-free medium. The spermatozoa induced to undergo the acrosome reaction by W-7 were capable of penetrating the zona-free hamster eggs. W-5, which has a lower affinity for calmodulin than W-7, also induced the acrosome reaction in the same manner as W-7. These results suggest that the naphthalenesulfonamide derivatives W-7 and W-5 can induce the acrosome reaction in guinea pig spermatozoa via capacitation in a pH-dependent, Ca2+-calmodulin-independent manner.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"20 1","pages":"197-208"},"PeriodicalIF":0.0,"publicationDate":"1986-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81254917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
We have developed a procedure for the long-term storage of lobster (Homarus) sperm. Sperm are collected from males by electrically stimulating the area around the gonopore. They are transferred with a bamboo stick to a plastic test tube containing paraffin oil and are stored for variable periods of time at 4–7°C. Sperm stored up to 289 days appear morphologically normal (equivalent to unstored sperm) when examined by phase contrast microscopy and electron microscopy, and morphologically normal sperm are able to undergo acrosome reactions. After longer periods of storage, degenerative changes begin to develop in sperm. These include loss of the nuclear spikes, condensation of the subacrosomal material, and distortion of the acrosome. Sperm stored better in spermatophores that had thick walls than in those with thin walls. In some spermatophores, bacteria were found in the sperm mass after storage; in general, sperm in these spermatophores were morphologically abnormal. This technique provides a means for saving lobster sperm for subsequent use in experiments or for artificial insemination of female lobsters. It may be adaptable to other invertebrate species that produce spermatophores.
{"title":"Technique for the long-term storage of lobster (Homarus) spermatophores","authors":"Tamako Ishida, P. Talbot, Marcia J. Kooda-Cisco","doi":"10.1002/MRD.1120140302","DOIUrl":"https://doi.org/10.1002/MRD.1120140302","url":null,"abstract":"We have developed a procedure for the long-term storage of lobster (Homarus) sperm. Sperm are collected from males by electrically stimulating the area around the gonopore. They are transferred with a bamboo stick to a plastic test tube containing paraffin oil and are stored for variable periods of time at 4–7°C. Sperm stored up to 289 days appear morphologically normal (equivalent to unstored sperm) when examined by phase contrast microscopy and electron microscopy, and morphologically normal sperm are able to undergo acrosome reactions. After longer periods of storage, degenerative changes begin to develop in sperm. These include loss of the nuclear spikes, condensation of the subacrosomal material, and distortion of the acrosome. Sperm stored better in spermatophores that had thick walls than in those with thin walls. In some spermatophores, bacteria were found in the sperm mass after storage; in general, sperm in these spermatophores were morphologically abnormal. This technique provides a means for saving lobster sperm for subsequent use in experiments or for artificial insemination of female lobsters. It may be adaptable to other invertebrate species that produce spermatophores.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"71 1","pages":"183-195"},"PeriodicalIF":0.0,"publicationDate":"1986-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86152418","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Glutathione (GSH), the major low-molecular-weight thiol in mammalian cells, is believed to be a necessary factor for the transformation of the disulfide-stabilized sperm nucleus into the male pronucleus after fertilization. Its concentration in mouse ova, isolated from the ampulla of the oviduct after hormone-induced superovulation of 3–4-week-old mice, has been determined by an enzymic cycling microassay. The level found was 1.80 pmol per ovum. Mean ovum diameter was estimated as 71–72 μm, indicating a GSH concentration of 9–10 mM in the mouse egg. Administration of L-buthionine S, R-sulfoximine, an inhibitor of GSH biosynthesis, during the 2 days preceding ovulation, reduced ovum GSH content below 0.20 pmol (<1.0 mM). The mean GSH concentration of the hormone-stimulated ovaries was reduced from 3.2 mM to 0.2 mM under these conditions. It has also been demonstrated that measurement and manipulation of ovum and ovarian levels of GSH can aid in studying its function in ovaries, ova, and early embryos. Hormone-induced superovulation was achieved in BSO-treated prepuberal C57B1/6 X SJL mice whose ovaries contained less than 10% of control levels of GSH. Over 50% of the isolated ova were fertilized in vitro. However, abnormal one-cell embryos resulted in which the maternally derived pronucleus coexisted with an unchanged sperm nucleus, thus confirming that adequate levels of GSH are necessary for initiating transformation of the fertilizing sperm nucleus.
谷胱甘肽(GSH)是哺乳动物细胞中主要的低分子量硫醇，被认为是受精后二硫化物稳定的精子核向雄性原核转化的必要因素。3 - 4周龄小鼠经激素诱导超排卵后，从输卵管壶腹分离出小鼠卵，用酶循环微量测定法测定了其在小鼠卵中的浓度。发现的浓度为每颗卵子1.80 pmol。卵的平均直径为71 ~ 72 μm，表明卵中谷胱甘肽的浓度为9 ~ 10 mM。在排卵前2天给予抑制GSH生物合成的l -丁硫氨酸S, r -亚砜胺，可使卵细胞GSH含量降至0.20 pmol (<1.0 mM)以下。在这些条件下，激素刺激卵巢的GSH平均浓度从3.2 mM降低到0.2 mM。研究还表明，测量和控制卵子和卵巢的谷胱甘肽水平有助于研究其在卵巢、卵子和早期胚胎中的功能。bso处理的青春期前C57B1/6 X SJL小鼠卵巢中GSH含量低于对照水平的10%，实现了激素诱导的超排卵。超过50%的分离卵子在体外受精。然而，异常的单细胞胚胎导致母源原核与不变的精子核共存，从而证实了足够水平的谷胱甘肽是启动受精精子核转化所必需的。
{"title":"ESTIMATION AND MANIPULATION OF GLUTATHIONE LEVELS IN PREPUBERAL MOUSE OVARIES AND OVA: RELEVANCE TO SPERM NUCLEUS TRANSFORMATION IN THE FERTILIZED EGG","authors":"H. Calvin, K. Grosshans, E. J. Blake","doi":"10.1002/MRD.1120140310","DOIUrl":"https://doi.org/10.1002/MRD.1120140310","url":null,"abstract":"Glutathione (GSH), the major low-molecular-weight thiol in mammalian cells, is believed to be a necessary factor for the transformation of the disulfide-stabilized sperm nucleus into the male pronucleus after fertilization. Its concentration in mouse ova, isolated from the ampulla of the oviduct after hormone-induced superovulation of 3–4-week-old mice, has been determined by an enzymic cycling microassay. The level found was 1.80 pmol per ovum. Mean ovum diameter was estimated as 71–72 μm, indicating a GSH concentration of 9–10 mM in the mouse egg. Administration of L-buthionine S, R-sulfoximine, an inhibitor of GSH biosynthesis, during the 2 days preceding ovulation, reduced ovum GSH content below 0.20 pmol (<1.0 mM). The mean GSH concentration of the hormone-stimulated ovaries was reduced from 3.2 mM to 0.2 mM under these conditions. \u0000 \u0000It has also been demonstrated that measurement and manipulation of ovum and ovarian levels of GSH can aid in studying its function in ovaries, ova, and early embryos. Hormone-induced superovulation was achieved in BSO-treated prepuberal C57B1/6 X SJL mice whose ovaries contained less than 10% of control levels of GSH. Over 50% of the isolated ova were fertilized in vitro. However, abnormal one-cell embryos resulted in which the maternally derived pronucleus coexisted with an unchanged sperm nucleus, thus confirming that adequate levels of GSH are necessary for initiating transformation of the fertilizing sperm nucleus.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"2011 1","pages":"265-275"},"PeriodicalIF":0.0,"publicationDate":"1986-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87952438","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sperm-egg interaction in Rhynchocinetes typus was studied with the phase-contrast and scanning electron microscopes. R typus spermatozoa present in the vas deferens have the shape of a round-headed nail. After contact with seawater it is possible to observe the unfolding of the rays or stellate arms, giving the spermatozoon the appearance of an inverted umbrella. From the center of the flat face of the umbrella emerges a spike with longitudinal striations. Ovarian eggs and spermatozoa were mixed in vitro by agitating them for two minutes in Millipore-filtered seawater. The first gamete contact was established by the spermatozoon through the tip of the spike, which exerted a lytic action on the egg envelopes. After the rigid spike was completely inside the egg, the rays became aligned parallel to each other and began to enter the eggs. Toward the final stages of ray entry, it was possible to observe fusion of the ray membranes with one another, and later the fusion process continued toward the tip of the radial spines. Concomitantly, the egg surface that surrounds the sperm swelled in a circular fashion and formed a fertilization cone. After the spermatozoon entry was complete, a scarlike mark appeared at the place on the egg surface through which penetration occurred. The whole penetration process was completed within 45-60 minutes.
{"title":"Sperm-egg interactions in the shrimp Rhynchocinetes typus","authors":"C. Barros, E. Dupré, L. Viveros","doi":"10.1002/MRD.1120140208","DOIUrl":"https://doi.org/10.1002/MRD.1120140208","url":null,"abstract":"Sperm-egg interaction in Rhynchocinetes typus was studied with the phase-contrast and scanning electron microscopes. R typus spermatozoa present in the vas deferens have the shape of a round-headed nail. After contact with seawater it is possible to observe the unfolding of the rays or stellate arms, giving the spermatozoon the appearance of an inverted umbrella. From the center of the flat face of the umbrella emerges a spike with longitudinal striations. Ovarian eggs and spermatozoa were mixed in vitro by agitating them for two minutes in Millipore-filtered seawater. The first gamete contact was established by the spermatozoon through the tip of the spike, which exerted a lytic action on the egg envelopes. After the rigid spike was completely inside the egg, the rays became aligned parallel to each other and began to enter the eggs. Toward the final stages of ray entry, it was possible to observe fusion of the ray membranes with one another, and later the fusion process continued toward the tip of the radial spines. Concomitantly, the egg surface that surrounds the sperm swelled in a circular fashion and formed a fertilization cone. After the spermatozoon entry was complete, a scarlike mark appeared at the place on the egg surface through which penetration occurred. The whole penetration process was completed within 45-60 minutes.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"40 1","pages":"171-180"},"PeriodicalIF":0.0,"publicationDate":"1986-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74462571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The purpose of the present work was to study the feasibility of using hamster oocytes stored at 4°C in M-2 culture medium for 24 and 48 hours in the evaluation of human sperm fertilizability. A total of 1,394 oocytes were stored for 24 hours and 1,234 were stored for 48 hours. After the storage period all the ooctyes were stained with fluorescein diacetate, proving the physical integrity of the egg plasma membrane. Twenty-five and 22 semen samples were used to compare their ability to penetrate freshly ovulated and 24-and 48-hour-stored hamster oocytes. Freshly ovulated and 24-hour-stored oocytes were penetrated at percentages that in more than 95% of the cases showed no significant differences. The same experiments carried out with oocytes stored for 48 hours showed that in 75% of the cases no significant differences were found. The use of oocytes preserved at 4°C when large numbers of semen samples are to be tested for fertilizability is recommended.
{"title":"Hamster oocyte fertilizability after 4°C storage","authors":"C. Barros, E. Herrera, I. Fuenzalida, B. Argüello","doi":"10.1002/MRD.1120140206","DOIUrl":"https://doi.org/10.1002/MRD.1120140206","url":null,"abstract":"The purpose of the present work was to study the feasibility of using hamster oocytes stored at 4°C in M-2 culture medium for 24 and 48 hours in the evaluation of human sperm fertilizability. A total of 1,394 oocytes were stored for 24 hours and 1,234 were stored for 48 hours. After the storage period all the ooctyes were stained with fluorescein diacetate, proving the physical integrity of the egg plasma membrane. Twenty-five and 22 semen samples were used to compare their ability to penetrate freshly ovulated and 24-and 48-hour-stored hamster oocytes. Freshly ovulated and 24-hour-stored oocytes were penetrated at percentages that in more than 95% of the cases showed no significant differences. The same experiments carried out with oocytes stored for 48 hours showed that in 75% of the cases no significant differences were found. The use of oocytes preserved at 4°C when large numbers of semen samples are to be tested for fertilizability is recommended.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"22 1","pages":"149-157"},"PeriodicalIF":0.0,"publicationDate":"1986-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86036782","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nicotinamide at a concentration of 20 mM prevented progesterone-induced germinal vesicle breakdown (GVBD) of Xenopus laevis oocytes in vitro in a reversible manner, whereas nicotinic acid did not. The vitamin was inhibitory even when applied 30 min after the addition of progesterone and blocked the induction of GVBD by microinjected maturation-promoting factor (MPF). Nicotinamide also prevented the burst of protein phos-phorylation associated with MPF expression and increased the oocyte cAMP level, whereas intracellular NAD content and protein synthesis were not affected. The present results suggest that nicotinamide acts by increasing the intracellular level of cAMP and by blocking MPF action.
{"title":"Nicotinamide inhibition of Xenopus laevis oocyte maturation","authors":"F. Godeau, M. K. Sahni, P. Boquet, S. Koide","doi":"10.1002/MRD.1120140207","DOIUrl":"https://doi.org/10.1002/MRD.1120140207","url":null,"abstract":"Nicotinamide at a concentration of 20 mM prevented progesterone-induced germinal vesicle breakdown (GVBD) of Xenopus laevis oocytes in vitro in a reversible manner, whereas nicotinic acid did not. The vitamin was inhibitory even when applied 30 min after the addition of progesterone and blocked the induction of GVBD by microinjected maturation-promoting factor (MPF). Nicotinamide also prevented the burst of protein phos-phorylation associated with MPF expression and increased the oocyte cAMP level, whereas intracellular NAD content and protein synthesis were not affected. The present results suggest that nicotinamide acts by increasing the intracellular level of cAMP and by blocking MPF action.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"71 1","pages":"159-170"},"PeriodicalIF":0.0,"publicationDate":"1986-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75657277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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