Gamete Research最新文献_第2页

Lower temperature of the cauda epididymidis facilitates the storage of sperm by enhancing oxygen availability 附睾尾部较低的温度通过提高氧气的可用性来促进精子的储存

Gamete Research Pub Date : 1986-11-01 DOI: 10.1002/MRD.1120150305

D. Djakiew, R. Cardullo

引用次数: 27

Examination of living and fixed gametes and early embryos stained with supravital fluorochromes (Hoechst 33342 and 3,3′-dihexyloxacarbocyanine iodide) 活配子和固定配子及早期胚胎的荧光染色检查(Hoechst 33342和3,3 ' -二己基碳氰碘)

Gamete Research Pub Date : 1986-11-01 DOI: 10.1002/MRD.1120150308

S. Luttmer, F. Longo

{"title":"Examination of living and fixed gametes and early embryos stained with supravital fluorochromes (Hoechst 33342 and 3,3′-dihexyloxacarbocyanine iodide)","authors":"S. Luttmer, F. Longo","doi":"10.1002/MRD.1120150308","DOIUrl":"https://doi.org/10.1002/MRD.1120150308","url":null,"abstract":"We have examined living and fixed gametes and early embryos of surf clams, sea urchins, and hamsters stained with the supravital dyes Hoechst 33342 for DNA and 3,3′-dihexyloxacarbocyanine iodide (DIOC6) for mitochondria and endoplasmic reticulum. Hoechst staining (10 μM) was confined exclusively to egg and sperm chromatin and, in living marine specimens, did not interfere with sperm motility, fertilization, or nuclear activity during meiosis or early embryogenesis. Although Hoechst staining did not appear to affect the motility of hamster sperm, only zonae-free eggs inseminated. Because chromatin retained Hoechst 33342 stain during fertilization, the paternally and maternally derived chromosomes of living and fixed preparations fluoresced and their number, organization, and location within the zygote cytoplasm could be determined. Hence, polyspermy and other nuclear abnormalities were amenable to examination in these stained preparations. DIOC6 staining (8.7 μM) was restricted primarily to the mitochondria of spermatozoa. Eggs stained with DIOC6 (0.87 to 8.7 μM) were brightly fluorescent because of their size and the presence of large numbers of mitochondria and other DIOC6-positive organelles. Sea urchin and surf clam sperm stained with DIOC6 fertilized unstained eggs and the location of the incorporated sperm mitochondrion up to first cleavage was followed. Although hamster sperm stained with DIOC6 were less motile than unstained sperm, they were capable of inseminating only zonae-free eggs. These observations demonstrate that staining with supravital fluorochromes provides a rapid and useful method to analyze macromolecular and organelle changes in a variety of living and fixed gametes and embryos.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"69 1","pages":"267-283"},"PeriodicalIF":0.0,"publicationDate":"1986-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74127776","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}

引用次数: 44

Immunochemical localization of epididymal protein DE on rat spermatozoa: Its fate after induced acrosome reaction 附睾蛋白DE在大鼠精子上的免疫化学定位:诱导顶体反应后的命运

Gamete Research Pub Date : 1986-11-01 DOI: 10.1002/MRD.1120150306

M. Cameo, Fernanda González Echeverría, J. Blaquier, M. Burgos

引用次数: 30

Sperm-oocyte membrane fusion in the human during monospermic fertilization 人单精子受精过程中精子-卵母细胞膜融合

Gamete Research Pub Date : 1986-10-01 DOI: 10.1002/MRD.1120150208

A. Sathananthan, Christopher Chen

{"title":"Sperm-oocyte membrane fusion in the human during monospermic fertilization","authors":"A. Sathananthan, Christopher Chen","doi":"10.1002/MRD.1120150208","DOIUrl":"https://doi.org/10.1002/MRD.1120150208","url":null,"abstract":"Sperm-oocyte membrane fusion has been observed during monospermic fertilization of a human oocyte in vitro. \u0000 \u0000 \u0000 \u0000Women were stimulated with both clomiphene citrate and human menopausal gonadotropin and were given human chorionic gonadotropin before a LH-surge. Twelve oocytes, collected at laparoscopy from six women who became pregnant by IVF, were allowed to mature for 7–14 hours in vitro and inseminated with preincubated sperm, fixed between 1–3 hours after insemination, and examined by transmission electron microscopy. \u0000 \u0000 \u0000 \u0000Membrane fusion had occurred in one ovum 2 hours after insemination, and the oocyte had resumed maturation and was at anaphase II of meiosis. Cortical granules had been exocytosed, and some of their contents were visible at the surface close to the oolemma all around the oocyte. The sperm that fused with this oocyte was acrosome-reacted and had been partly incorporated into the ooplasm, while the anterior two-thirds of its head was phagocytosed by a tongue of cortical ooplasm. Membrane fusion had occurred between the oolemma and the plasma membrane overlying the postacrosomal segment of the sperm head, posterior to the equatorial vestige. Sperm chromatin had not decondensed, and serial sections revealed a midpiece attached to the basal plate and a tail located deeper in the ooplasm, all devoid of plasma membrane. Supplementary sperm penetrating the inner zona, approaching the perivitelline space, had undergone the acrosome reaction but had a persistent vestige of the equatorial segment of the acrosome with intact plasma membrane. Evidence of sperm chromatin decondensation was seen in other oocytes, 3 hours after insemination, which were at telophase II of meiosis. Eight oocytes penetrated by sperm were monospermic, while four were unfertilized. \u0000 \u0000 \u0000 \u0000The general pattern of sperm fusion and incorporation appears to conform to that seen in most other mammals. The study also reveals that sperm have to complete the acrosome reaction before fusing with the egg.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"56 1","pages":"177-186"},"PeriodicalIF":0.0,"publicationDate":"1986-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79832019","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}

引用次数: 44

Partial purification and characterization of dipeptidyl peptidase II (DPP II) from guinea pig testes 豚鼠睾丸二肽基肽酶II (DPP II)的部分纯化和鉴定

Gamete Research Pub Date : 1986-10-01 DOI: 10.1002/MRD.1120150207

G. Dicarlantonio, P. Talbot, E. Dudenhausen

{"title":"Partial purification and characterization of dipeptidyl peptidase II (DPP II) from guinea pig testes","authors":"G. Dicarlantonio, P. Talbot, E. Dudenhausen","doi":"10.1002/MRD.1120150207","DOIUrl":"https://doi.org/10.1002/MRD.1120150207","url":null,"abstract":"Depeptidyl peptidase (DPP II) was partially purified from guinea pig testes by (NH4)2SO4 precipitation, Con A-Sepharose 4B chromatography, and Sephadex G-200 chromatography to a specific activity of 27.4 μmol Ala3 hydrolyzed min−1 mg−1 protein. Chromatography on a calibrated G-200 column yielded a molecular weight of 135,000 daltons for the enzyme. Sodium dodecyl sulfate polyacrylamide electrophoresis showed an enrichment of a broad doublet at 64–66,000 daltons. The enzyme had optimal activity toward hydrolysis of L-alanyl-alanyl-alanine at pH 4.5 and showed sensitivity to cations of increasing size with Tris producing the most inhibition of those tested. The enzyme was moderately inhibited by serine proteinase inhibitors. Thin-layer chromatography revealed the dipeptidase nature of the enzyme's activity on tripeptides and dipeptidyl arylamides. A doublet of activity occurred when nitrocellulose electroblots of nondenaturing gel electrophoresis of the (NH4)2SO4 fraction were reacted with the specific DPP II substrate, lysyl-alanyl-4-methoxy-2-napthylamide. Analytical isoelectric focusing of the G-200 fraction followed by fluorescent enzyme activity detection that used cellulose triacetate overlay membranes impregnated with the specific DPP II substrate, lysyl-alanyl-7-amino-4-trifluoromethylcou-marin, revealed multiple isoforms focusing at pI = 4.8–5.6. Two prominent bands focused at pI = 4.9 and pI = 5.1. The properties of guinea pig testicular DPP II are compared and contrasted with similar dipeptidyl peptidases from other sources.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"15 1","pages":"161-175"},"PeriodicalIF":0.0,"publicationDate":"1986-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84141883","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}

引用次数: 6

Measurement of intracellular calcium in human spermatozoa 人精子细胞内钙的测定

Gamete Research Pub Date : 1986-09-01 DOI: 10.1002/MRD.1120150107

D. Irvine, R. Aitken

引用次数: 41

Comparison of some properties of oocytes segregating in F2 hybrids between KE and CBA/Kw inbred strains of mice KE与CBA/Kw自交系小鼠F2杂种卵母细胞分离特性的比较

Gamete Research Pub Date : 1986-09-01 DOI: 10.1002/MRD.1120150110

B. Wabik-Sliz

引用次数: 4

Vital staining and acrosomal evaluation of bovine sperm 牛精子的生命染色和顶体评价

Gamete Research Pub Date : 1986-09-01 DOI: 10.1002/MRD.1120150108

E. Aalseth, R. G. Saacke

{"title":"Vital staining and acrosomal evaluation of bovine sperm","authors":"E. Aalseth, R. G. Saacke","doi":"10.1002/MRD.1120150108","DOIUrl":"https://doi.org/10.1002/MRD.1120150108","url":null,"abstract":"Dried smears prepared from vitally stained sperm were evaluated as a method of simultaneously determining sperm viability and acrosomal morphology. A combination Fast Green FCF-Eosin B stain was used. The stained smears were examined at × 1, 250 using differential interference contrast microscopy (DIC). For comparison, the percentage of sperm with intact acrosomes was also determined from wet smears using DIC. Acrosomal morphology was not altered by the staining procedure, as the percentage of intact acrosomes was similar whether quantitated from wet or stained smears. Absence of eosinophilic staining in the acrosome was used as an indication of sperm viability. The percentage of sperm with unstained acrosomes was highly correlated with the percentage of intact acrosomes quantitated from stained smears. Thus, vital staining provided an indication of sperm viability comparable to acrosomal integrity, a highly reliable technique. The major advantages of using dried stained smears were more thorough examination of individual sperm without sperm activity interference, simultaneous evidence of sperm viability and morphology, and the opportunity to delay evaluation. In addition, diluting spermatozoa in complex or simple media with or without egg yolk or follicular fluid did not interfere with subsequent staining or acrosomal evaluation.","PeriodicalId":12668,"journal":{"name":"Gamete Research","volume":"40 1","pages":"73-81"},"PeriodicalIF":0.0,"publicationDate":"1986-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86281285","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}

引用次数: 46

Effect of centrifugation of mouse eggs on their development in vitro and in vivo 小鼠卵离心对体外和体内发育的影响

Gamete Research Pub Date : 1986-09-01 DOI: 10.1002/MRD.1120150109

K. Nakamura, Y. Tsunoda, T. Nagai, T. Sugie

引用次数: 10

Electron microscopic study of the in vivo zona pellucida binding ability of epididymal ram spermatozoa 附睾公羊精子体内透明带结合能力的电镜研究

Gamete Research Pub Date : 1986-07-01 DOI: 10.1002/MRD.1120140306

S. Fournier-delpech, N. Crozet, M. Courot

引用次数: 4