The effects of storage of hamster sperm in vitro at temperatures above freezing on fertilization and early development were studied. Initially, female hamsters were inseminated artificially with epididymal sperm stored up to five days at 5°, 16°, or 23–25°C. Eggs were examined at the pronuclear stage for evidence of fertilization. The best conditions for the storage of hamster sperm were found to be 16°C in a 5% CO2 in air gas phase. Pre- and postimplantation development were then assessed following insemination of females with sperm stored at 16°C. An increased frequency of eggs with three pronuclei, a decrease in the number of pregnant females, and an increase in the number of resorption sites were found with increasing sperm storage time. Chromosome mosaicism and triploidy were encountered in 9.5-day embryos resulting from eggs fertilized by stored sperm. No chromosome abnormalities were found in a control group of embryos. Thus, storage of hamster epididymal sperm resulted in decreased fertility and an increased frequency of abnormal development.
{"title":"The effect of in vitro storage of hamster sperm on fertilization and early development","authors":"E. Shaver, R. Yanagimachi","doi":"10.1002/MRD.1120010303","DOIUrl":"https://doi.org/10.1002/MRD.1120010303","url":null,"abstract":"The effects of storage of hamster sperm in vitro at temperatures above freezing on fertilization and early development were studied. Initially, female hamsters were inseminated artificially with epididymal sperm stored up to five days at 5°, 16°, or 23–25°C. Eggs were examined at the pronuclear stage for evidence of fertilization. The best conditions for the storage of hamster sperm were found to be 16°C in a 5% CO2 in air gas phase. Pre- and postimplantation development were then assessed following insemination of females with sperm stored at 16°C. An increased frequency of eggs with three pronuclei, a decrease in the number of pregnant females, and an increase in the number of resorption sites were found with increasing sperm storage time. Chromosome mosaicism and triploidy were encountered in 9.5-day embryos resulting from eggs fertilized by stored sperm. No chromosome abnormalities were found in a control group of embryos. Thus, storage of hamster epididymal sperm resulted in decreased fertility and an increased frequency of abnormal development.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"81 1","pages":"235-245"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81629113","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 simple method for the collection of a large number of ova from pig ovaries is described. The method consists of disrupting the tissue in an ordinary meat grinder, then separating the oocytes by sieving the disrupted tissue through two pore sizes of stainless steel mesh.
{"title":"A simple method for the isolation of a large number of ova from pig ovaries","authors":"T. Oikawa","doi":"10.1002/MRD.1120010306","DOIUrl":"https://doi.org/10.1002/MRD.1120010306","url":null,"abstract":"A simple method for the collection of a large number of ova from pig ovaries is described. The method consists of disrupting the tissue in an ordinary meat grinder, then separating the oocytes by sieving the disrupted tissue through two pore sizes of stainless steel mesh.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"7 1","pages":"265-267"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87272685","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 165,000g pellet fraction, isolated from lysed rooster spermatozoa, was shown to contain a deoxynucleotide polymerizing activity which banded in sucrose at a buoyant density of 1.15gm/ml. The solubilized enzyme, partially purified by gel filtration, was estimated to have a molecular weight of 33,000 and utilized dT10 or dG10 as exogenous primers in template-independent reactions sensitive to either sodium pyrophosphate or ethylenediaminetetra- cetic acid. Pyrimidine nucleotides (dTTP) were preferentially polymerized in the presence of Co++, while purine nucleotides (dGTP) were the preferred substrate when Mn++ was the divalent cation. These properties are all consistent with those of terminal deoxynucleotidyl transferase. An enzyme with similar properties was also obtained by subjecting a rooster sperm lysate to the procedure utilized to isolate terminal deoxynucleotidyl transferase from calf thymus. It is hypothesized that the enzyme may be involved in the regulation of gene expression during embryogenesis.
{"title":"Terminal deoxynucleotidyl transferase in rooster sperm","authors":"S. Witkin, B. Reiner, Cynthia A. Brown","doi":"10.1002/MRD.1120010302","DOIUrl":"https://doi.org/10.1002/MRD.1120010302","url":null,"abstract":"A 165,000g pellet fraction, isolated from lysed rooster spermatozoa, was shown to contain a deoxynucleotide polymerizing activity which banded in sucrose at a buoyant density of 1.15gm/ml. The solubilized enzyme, partially purified by gel filtration, was estimated to have a molecular weight of 33,000 and utilized dT10 or dG10 as exogenous primers in template-independent reactions sensitive to either sodium pyrophosphate or ethylenediaminetetra- cetic acid. Pyrimidine nucleotides (dTTP) were preferentially polymerized in the presence of Co++, while purine nucleotides (dGTP) were the preferred substrate when Mn++ was the divalent cation. These properties are all consistent with those of terminal deoxynucleotidyl transferase. An enzyme with similar properties was also obtained by subjecting a rooster sperm lysate to the procedure utilized to isolate terminal deoxynucleotidyl transferase from calf thymus. It is hypothesized that the enzyme may be involved in the regulation of gene expression during embryogenesis.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"53 1","pages":"227-233"},"PeriodicalIF":0.0,"publicationDate":"1978-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83167561","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}
Incorporation of [32P]-phosphate into proteins was enhanced when Sabellaria oocytes were stimulated with specific protease to continue from prophase I block to metaphase I block. The rate of incorporation was increased 50 fold between onset of treatment and germinal vesicle breakdown (GVB). The same result was obtained when release from prophase block involved fertilization, or activation with ionophore A 23187. In all cases, meiosis was associated with phosphorylation of an 18,000 dalton protein, which is perhaps not labeled in prophase-blocked oocytes. Phosphorylation of a 38,000–40,000 dalton doublet of membrane proteins, which are among the main phosphorylated proteins in intact oocytes, was also strongly enhanced in vitro in homogenates prepared from oocytes following release from prophase block.
{"title":"Stimulation of endogenous protein phosphorylation in oocytes of Sabellaria alveolata (polychaete annelid) at meiosis reinitiation induced by protease, fertilization, or ionophore A 23187","authors":"G. Peaucellier, M. Dorée, J. Demaille","doi":"10.1002/MRD.1120050202","DOIUrl":"https://doi.org/10.1002/MRD.1120050202","url":null,"abstract":"Incorporation of [32P]-phosphate into proteins was enhanced when Sabellaria oocytes were stimulated with specific protease to continue from prophase I block to metaphase I block. The rate of incorporation was increased 50 fold between onset of treatment and germinal vesicle breakdown (GVB). The same result was obtained when release from prophase block involved fertilization, or activation with ionophore A 23187. In all cases, meiosis was associated with phosphorylation of an 18,000 dalton protein, which is perhaps not labeled in prophase-blocked oocytes. Phosphorylation of a 38,000–40,000 dalton doublet of membrane proteins, which are among the main phosphorylated proteins in intact oocytes, was also strongly enhanced in vitro in homogenates prepared from oocytes following release from prophase block.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"35 1","pages":"115-123"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88172652","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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