Culture in vitro causes a slow, progressive hardening of the zona pellucida (ZP) of fully grown dictyate oocytes isolated from the mouse ovary. Hardening cannot be prevented by inhibitors of peroxidase or by a tyrosine analogue. Culture in anaerobic conditions is very effective in preventing ZP hardening. If the oocyte is cultured surrounded by its own follicle cells or in contact with cumuli oophori obtained from superovulated females, hardening is much reduced. The results suggest that the “spontaneous” hardening in cultured ovarian oocytes is not due to a cortical reaction, and that a diffusible factor is produced by follicle cells that protect the ZP from hardening.
{"title":"Spontaneous hardening of the zona pellucida of mouse oocytes during in vitro culture","authors":"M. De Felici, G. Siracusa","doi":"10.1002/MRD.1120060203","DOIUrl":"https://doi.org/10.1002/MRD.1120060203","url":null,"abstract":"Culture in vitro causes a slow, progressive hardening of the zona pellucida (ZP) of fully grown dictyate oocytes isolated from the mouse ovary. Hardening cannot be prevented by inhibitors of peroxidase or by a tyrosine analogue. Culture in anaerobic conditions is very effective in preventing ZP hardening. If the oocyte is cultured surrounded by its own follicle cells or in contact with cumuli oophori obtained from superovulated females, hardening is much reduced. The results suggest that the “spontaneous” hardening in cultured ovarian oocytes is not due to a cortical reaction, and that a diffusible factor is produced by follicle cells that protect the ZP from hardening.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"9 1","pages":"107-113"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87030065","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 effects of aphidicolin and α-amanitin on DNA synthesis by preimplantation mouse embryos were studied. It was found that both blastocyst and 8-cell embryos showed marked inhibition of 3H-thymidine incorporation into DNA by aphidicolin at concentrations of 20–50 μg/ml. However, aphidicolin did not inhibit the conversion of morula embryos to blastocyst embryos, although aphidicolin-treated blastocysts lost their blastocoel and collapsed into a compact form after prolonged exposure to the drug. Both 8-cell and blastocyst embryos were found to be susceptible to inhibition of DNA synthesis by α-amanitin.
{"title":"The effects of aphidicolin and α-amanitin on DNA synthesis in preimplantation mouse embryos","authors":"K. M. Cozad, C. Warner","doi":"10.1002/MRD.1120060209","DOIUrl":"https://doi.org/10.1002/MRD.1120060209","url":null,"abstract":"The effects of aphidicolin and α-amanitin on DNA synthesis by preimplantation mouse embryos were studied. It was found that both blastocyst and 8-cell embryos showed marked inhibition of 3H-thymidine incorporation into DNA by aphidicolin at concentrations of 20–50 μg/ml. However, aphidicolin did not inhibit the conversion of morula embryos to blastocyst embryos, although aphidicolin-treated blastocysts lost their blastocoel and collapsed into a compact form after prolonged exposure to the drug. Both 8-cell and blastocyst embryos were found to be susceptible to inhibition of DNA synthesis by α-amanitin.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"17 1","pages":"155-160"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78730317","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 problem of how the successive nucleoproteins enter and leave the nucleus of mammalian spermatids is studied with electron microscopy in thick and thin sections of testis, stained en bloc with the procedure of Thiery and Rambourg [1976], which is able to immobilize small molecules. Staining at different pH values reveals that the stain could demonstrate some spermatidspecific nucleoproteins in elongating nuclei and the spermatozoa-specific protamines in elongated nuclei of the boar, the ram, and the stallion. The stained substances enter or leave the nuclei at precise steps in spermiogenesis. They follow several ways inside a special apparatus made rigid with the manchette. The apparatus is composed of the endoplasmic reticulum, continuous with the nuclear envelope, and of the nuclear pockets continuous with the nuclear pores. In the stallion and the boar, cytoplasmic granules, surrounded by a double wall of membranes, fuse with the nuclear envelope at the time the protamines enter the nucleus.
{"title":"Some nucleocytoplasmic exchanges during spermiogenesis of boar, ram, and stallion","authors":"J. Courtens","doi":"10.1002/MRD.1120050204","DOIUrl":"https://doi.org/10.1002/MRD.1120050204","url":null,"abstract":"The problem of how the successive nucleoproteins enter and leave the nucleus of mammalian spermatids is studied with electron microscopy in thick and thin sections of testis, stained en bloc with the procedure of Thiery and Rambourg [1976], which is able to immobilize small molecules. Staining at different pH values reveals that the stain could demonstrate some spermatidspecific nucleoproteins in elongating nuclei and the spermatozoa-specific protamines in elongated nuclei of the boar, the ram, and the stallion. The stained substances enter or leave the nuclei at precise steps in spermiogenesis. They follow several ways inside a special apparatus made rigid with the manchette. The apparatus is composed of the endoplasmic reticulum, continuous with the nuclear envelope, and of the nuclear pockets continuous with the nuclear pores. In the stallion and the boar, cytoplasmic granules, surrounded by a double wall of membranes, fuse with the nuclear envelope at the time the protamines enter the nucleus.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"53 1","pages":"137-152"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78893056","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}
G. Olson, J. Jonas‐Davies, L. H. Hoffman, M. Orgebin‐Crist
An enriched suspension of rat epididymal epithelial cells was prepared by sequential enzymatic removal of connective tissue and peritubular cells from the epididymal tubule. The viability, structural characteristics, and pattern of polypeptides synthesized by the isolated cells were determined. Electron microscopic analysis revealed that the isolated principal cells were intact and retained their polarized morphology. Several light microscopic protocols were employed to evaluate the percentage of epithelial cells in the suspensions. These included (1) the visualization of the pattern of FITC-lectin binding in which the principal cells could be identified by their polarized fluorescence; (2) the visualization of prominent autofluorescent granules in the cell cytoplasm which appeared to be characteristic of only epithelial cells; and (3) immunochemical staining with an antikeratin antibody which was reactive only with cells of epithelial origin. These structural probes indicated that between 80% and 90% of the isolated cells were epithelial in nature. Two-dimensional gel electrophoresis revealed a complex pattern of polypeptides synthesized by the epithelial cells; these results are compared to those of earlier studies utilizing minced whole epididymis.
{"title":"Structural characterization of isolated rat epididymal epithelial cells","authors":"G. Olson, J. Jonas‐Davies, L. H. Hoffman, M. Orgebin‐Crist","doi":"10.1002/MRD.1120060210","DOIUrl":"https://doi.org/10.1002/MRD.1120060210","url":null,"abstract":"An enriched suspension of rat epididymal epithelial cells was prepared by sequential enzymatic removal of connective tissue and peritubular cells from the epididymal tubule. The viability, structural characteristics, and pattern of polypeptides synthesized by the isolated cells were determined. Electron microscopic analysis revealed that the isolated principal cells were intact and retained their polarized morphology. Several light microscopic protocols were employed to evaluate the percentage of epithelial cells in the suspensions. These included (1) the visualization of the pattern of FITC-lectin binding in which the principal cells could be identified by their polarized fluorescence; (2) the visualization of prominent autofluorescent granules in the cell cytoplasm which appeared to be characteristic of only epithelial cells; and (3) immunochemical staining with an antikeratin antibody which was reactive only with cells of epithelial origin. These structural probes indicated that between 80% and 90% of the isolated cells were epithelial in nature. Two-dimensional gel electrophoresis revealed a complex pattern of polypeptides synthesized by the epithelial cells; these results are compared to those of earlier studies utilizing minced whole epididymis.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"79 1","pages":"161-178"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91225710","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}
Using a semi-chemically defined medium, the requirement of extracellular Ca2+ for survival, capacitation, and acrosome reaction of spermatozoa as well as various stages of fertilization in the hamster was studied. A Ca2+-deficient environment is unfavorable for long-term survival of spermatozoa. Sperm capacitation may occur in Ca2+-deficient media, but not as efficiently as in normal media. The acrosome reaction definitely requires extracellular Ca2+. Other processes or phenomena that require extracellular Ca2+ are initiation and maintenance of hyperactivated motility of spermatozoa, penetration of acrosome-reacted spermatozoa into the zona pellucida, fusion of the spermatozoa with eggs, and the development of pronuclear eggs into two-cell embryos. Extracellular Ca2+ is apparently unnecessary for the attachment of spermatozoa to the zona and egg surfaces, decondensation of the sperm nucleus, and the development of sperm and egg pronuclei within the egg. These results were compared with data obtained in other species such as the sea urchin, mouse, rat and guinea pig.
{"title":"Requirement of extracellular calcium ions for various stages of fertilization and fertilization-related phenomena in the hamster","authors":"R. Yanagimachi","doi":"10.1002/MRD.1120050404","DOIUrl":"https://doi.org/10.1002/MRD.1120050404","url":null,"abstract":"Using a semi-chemically defined medium, the requirement of extracellular Ca2+ for survival, capacitation, and acrosome reaction of spermatozoa as well as various stages of fertilization in the hamster was studied. A Ca2+-deficient environment is unfavorable for long-term survival of spermatozoa. Sperm capacitation may occur in Ca2+-deficient media, but not as efficiently as in normal media. The acrosome reaction definitely requires extracellular Ca2+. Other processes or phenomena that require extracellular Ca2+ are initiation and maintenance of hyperactivated motility of spermatozoa, penetration of acrosome-reacted spermatozoa into the zona pellucida, fusion of the spermatozoa with eggs, and the development of pronuclear eggs into two-cell embryos. Extracellular Ca2+ is apparently unnecessary for the attachment of spermatozoa to the zona and egg surfaces, decondensation of the sperm nucleus, and the development of sperm and egg pronuclei within the egg. These results were compared with data obtained in other species such as the sea urchin, mouse, rat and guinea pig.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"55 1","pages":"323-344"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83814124","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 spermatozoa of the annelid polychaete Ophryotrocha have been examined under the electron microscope. They show some primitive features (scarcely condensed nucleus, pores in the nuclear envelope, unmodified mitochondria) and are aflagellate. This aberrant structure is related to a very specialized type of external fertilization in which the spermatozoa are ejaculated close to the eggs.
{"title":"The aflagellate spermatozoon of ophryotrocha: A line of evolution of fertilization among polychaetes","authors":"G. Berruti, M. Ferraguti, C. L. Donin","doi":"10.1002/MRD.1120010309","DOIUrl":"https://doi.org/10.1002/MRD.1120010309","url":null,"abstract":"The spermatozoa of the annelid polychaete Ophryotrocha have been examined under the electron microscope. They show some primitive features (scarcely condensed nucleus, pores in the nuclear envelope, unmodified mitochondria) and are aflagellate. This aberrant structure is related to a very specialized type of external fertilization in which the spermatozoa are ejaculated close to the eggs.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"2010 1","pages":"287-292"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86286328","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 defined medium for growing defolliculated Xenopus laevis oocytes was developed by monitoring vitellogenin incorporation. Optimum conditions were acheived by use of 50% Liebovitz L-15 medium supplemented with 1 mM L-glutamine, 15 mM Hepes, 5 mg/ml vitellogenin, and 1 ..mu..g/ml insulin (final pH = 7.8). Stage IV/V oocytes remained morphologically normal in this medium for at least two weeks and grew at an average rate of 0.25 mm/sup 3/ per week. This is the first time defolliculated Xenopus laevis oocytes have been grown in vitro.
通过监测卵黄原蛋白的掺入，开发了一种用于卵泡去除的非洲爪蟾卵母细胞生长的培养基。在50% Liebovitz L-15培养基中，添加1 mM l -谷氨酰胺、15 mM Hepes、5 mg/ml卵黄原蛋白和1…g/ml胰岛素(终pH = 7.8)。IV/V期卵母细胞在该培养基中保持形态正常至少两周，并以平均0.25 mm/sup /周的速度生长。这是第一次在体外培养去卵泡非洲爪蟾卵母细胞。
{"title":"Development of a culture medium for growing xenopus laevis oocytes","authors":"R. Wallace, Z. Misulovin, D. Jared, H. Wiley","doi":"10.1002/MRD.1120010307","DOIUrl":"https://doi.org/10.1002/MRD.1120010307","url":null,"abstract":"A defined medium for growing defolliculated Xenopus laevis oocytes was developed by monitoring vitellogenin incorporation. Optimum conditions were acheived by use of 50% Liebovitz L-15 medium supplemented with 1 mM L-glutamine, 15 mM Hepes, 5 mg/ml vitellogenin, and 1 ..mu..g/ml insulin (final pH = 7.8). Stage IV/V oocytes remained morphologically normal in this medium for at least two weeks and grew at an average rate of 0.25 mm/sup 3/ per week. This is the first time defolliculated Xenopus laevis oocytes have been grown in vitro.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"37 1","pages":"269-280"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86095397","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 this study was to develop a modification of the medium of Toyoda and Chang which would permit high rates of fertilization in vitro in rats of our colony. Elevation of the calcium concentration from 1.71 to 3.42 mM increased the fertilization rate by 87%. Greater additions of calcium gave no further improvement. This effect was not due to reduction in phosphate activity, nor was it due to antagonism of heavy-metal contamination. Although calcium is known to enhance the fertilizing capacity of spermatozoa, our results suggest that it may also affect the oocyte.
{"title":"Effect of elevated calcium concentration on fertilization of rat oocytes in vitro","authors":"R. Kaplan, P. Kraicer","doi":"10.1002/MRD.1120010308","DOIUrl":"https://doi.org/10.1002/MRD.1120010308","url":null,"abstract":"The purpose of this study was to develop a modification of the medium of Toyoda and Chang which would permit high rates of fertilization in vitro in rats of our colony. Elevation of the calcium concentration from 1.71 to 3.42 mM increased the fertilization rate by 87%. Greater additions of calcium gave no further improvement. This effect was not due to reduction in phosphate activity, nor was it due to antagonism of heavy-metal contamination. Although calcium is known to enhance the fertilizing capacity of spermatozoa, our results suggest that it may also affect the oocyte.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"37 1","pages":"281-285"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76820881","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}
Epididymal spermatozoa from two species of vole demonstrate cytoskeletal structures closely associated with the plasma membrane. Microtubule-like elements (2O–24 nm in diameter) form a parallel sheath radiating from the posterior ring to the inferior acrosomal margin just beneath the plasma membrane. Fine filaments (5–8 nm in diameter) are associated with the cell surface, especially in the region of the concave curvature of the acrosomal hook. Both of these elements may aid in stabilizing these respective areas and it is suggested that microtubule-like and filamentous structures may serve important cytoskeletal roles in the mature sperm head.
{"title":"Observations on the fine structure of vole spermatozoa with particular reference to cytoskeletal elements in the mature sperm head","authors":"J. Koehler","doi":"10.1002/MRD.1120010304","DOIUrl":"https://doi.org/10.1002/MRD.1120010304","url":null,"abstract":"Epididymal spermatozoa from two species of vole demonstrate cytoskeletal structures closely associated with the plasma membrane. Microtubule-like elements (2O–24 nm in diameter) form a parallel sheath radiating from the posterior ring to the inferior acrosomal margin just beneath the plasma membrane. Fine filaments (5–8 nm in diameter) are associated with the cell surface, especially in the region of the concave curvature of the acrosomal hook. Both of these elements may aid in stabilizing these respective areas and it is suggested that microtubule-like and filamentous structures may serve important cytoskeletal roles in the mature sperm head.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"1 1","pages":"247-257"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86993502","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}
Hamster and bovine zonae pellucidae were dissolved by heating them in buffered saline for 35 min at 65°C and 80°C, respectively. Pretreatment of noncapacitated sperm with these solutions did not affect their binding to eggs, but the binding of cumulus-capacitated hamster sperm and of bovine sperm that had undergone a capacitation-like surface change in response to hypertonic-treatment was severely curtailed. These experiments demonstrate that such zona solutions exhibit receptor activity, but only for the capacitated sperm.
{"title":"Bovine and hamster zona solutions exhibit receptor activity for capacitated but not for noncapacitated sperm","authors":"R. Gwatkin, D. T. Williams","doi":"10.1002/MRD.1120010305","DOIUrl":"https://doi.org/10.1002/MRD.1120010305","url":null,"abstract":"Hamster and bovine zonae pellucidae were dissolved by heating them in buffered saline for 35 min at 65°C and 80°C, respectively. Pretreatment of noncapacitated sperm with these solutions did not affect their binding to eggs, but the binding of cumulus-capacitated hamster sperm and of bovine sperm that had undergone a capacitation-like surface change in response to hypertonic-treatment was severely curtailed. These experiments demonstrate that such zona solutions exhibit receptor activity, but only for the capacitated sperm.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"35 1","pages":"259-263"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75151035","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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