摘要:

AbstractNormal adult human testis has been separated using a combination of mechanical and enzymatic procedures to yield a suspension of viable single cells. The predominant cell types comprising this suspension are as follows: primary pachytene spermatocytes (7% of total cells), round spermatids (17%), residual bodies and condensing spermatids (31%), and Leydig cells (15%). Separated human germ cells viewed by Nomarski differential interference microscopy closely resemble mouse spermatogenic cells in relative size and appearance. Isolation of an enriched population of human pachytene spermatocytes has been achieved using unit gravity sedimentation (STA‐PUT) according to protocols originally developed for murine tissue. Pachytene cells are enriched to 75% and are contaminated only with Leydig cells and binucleated spermatid symplasts. Ultrastructural examination of isolated human pachytene spermatocytes indicates that these cells, as well as isolated round spermatids, exhibit a normal in situ morphology. Spermatocytes, for example, show numerous synaptonemal complexes, nuclear pores, annulate lamellae, and dictyosome‐like saccules. Round spermatids after isolation exhibit peripheral mitochondria, annulate lamellae, developing acrosomes, and other morphological features characteristic of early spermiogenesis. Therefore, enriched populations of human spermatogenic cells seem suitable for analysis using immunofluorescent, autoradiographic, or serological methods. In particular, isolated human spermatocytes should be useful for the analysis of molecular events involved in meiosis and should facilitate investigations concerning the pathophysiology of certain human infertility conditi

ISSN:0148-7280    

DOI:10.1002/mrd.1120040602

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractMammalian spermatozoa contain nonmuscle actin and many of the components of regulatory systems thought to be involved in nonmuscle actin‐myosin function. An actin‐stimulated adenosine triphosphate hydrolase (ATPase) activity has been fractionated from bovine ejaculated spermatozoa by immobilized‐actin affinity chromatography. The actinstimulated ATPase activity has a specific activity (0.04 ± 0.02 mM phosphate released/min/mg protein) similar to nonmuscle myosins from other mammalian cells and tissues, but it does not have appreciable K+‐EDTA ATPase activity. The sperm actin‐myosin may function in sperm morphogenesis in the seminiferous tubule, in capacitated spermatozoa undergoing an acrosome reaction, or in decondensation of the sperm nucleus after fer

ISSN:0148-7280    

DOI:10.1002/mrd.1120040603

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractOpossum and rabbit sperm sonicates dispersed the mouse cumulus oophorus in vitro at the same rate on a per sperm basis, despite much higher activities of the glycosidic acrosomal enzymes N‐acetylhexosaminidase (350 ×) and arylsulphatase (40 ×) in the opossum preparation. Activities of another glycosidase, hyaluronidase, and the protease acrosin where similar in sperm extracts from both species. However, specific inhibitors of N‐acetylhexosaminidase (iodoacetate) and arylsulphatase (SO42−, PO43−) markedly reduced the rate of cumulus dispersal by rabbit sperm sonicates. These findings suggest that, while hyaluronidase action probably represents the rate‐limiting (slow) step, several glycosidases act together to disperse the cumulus and in the passage of the fertilizing spermatozoon through it in eutherian mammals. The likely role of these acrosomal enzymes in the marsupial, where the ovulated oocytes lack a cumulus oophorus, is mor

ISSN:0148-7280    

DOI:10.1002/mrd.1120040604

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractStudies were made of the effects of D(+)‐glucose, L‐lactate and pyruvate on in vitro capacitation and acrosome reactions (AR) of hamster sperm using a more “defined” medium that that used in previous similar studies. In the absence of glucose or lactate, sperm underwent very few AR and activation (whiplash‐like motility characteristic of capacitated hamster sperm) was reduced compared to those events in sperm preincubated in the presence of glucose plus lactate plus pyruvate. Glucose and pyruvate supported more AR than glucose alone, but less than glucose, lactate, and pyruvate. The glycolytic inhibitor α‐chlorohydrin (10 μm) inhibited AR by 50% and reduced activation by less. When glucose was added to sperm incubated 2 hr with pyruvate and lactate, the number of AR observed after 4 hr was the same as that obtained when glucose was present throughout the incubation. When glucose was added after 3.5 hr, AR were delayed for 1 hr and lower numbers of sperm underwent AR. In the presence of lactate and pyruvate, 0.38 mM glucose was able to support activation and AR as well as 3.24 mM glucose. These results indicate that exogenous glucose and lactate are necessary for in vitro capacitation and AR of hamster sperm; only low levels of exogenous glucose are required; exogenous glucose is not required during the first 2 hr of capacitation; and glycolytic activity is necessary for capacitati

ISSN:0148-7280    

DOI:10.1002/mrd.1120040605

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractThe temperature dependence of capacitation in bat sperm (Myotis lucifugus lucifugus) was studied by monitoring fertilizations rates of zona‐free hamster ova at different temperatures. Spermatozoa were cultured in BWW medium at temperatures 4°C, 24°C, 32°C, 42°C, and 55°C from 0–24 hr. Activation of sperm could be determined visually due to the change in movement seen through light microscopy. Activation was later confirmed by higher rates of fertilization. Preincubation of the bat sperm was found to have a direct effect on the success of penetration of the zona‐free hamster ova. Holding bat spermatozoa at low temperature for long intervals allowed them to remain motile but unable to fertilize. Sperm are not irreversibly damaged, however, and activation, when the temperature is increased to 32°C, is faster than when sperm are intitially put at 32°C, resulting in good fertili

ISSN:0148-7280    

DOI:10.1002/mrd.1120040606

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractInitial changes in the fine morphology of apparently normal mouse zygotes and embryos were studied in serial sections of mouse oviducts that had been fixed in situ. These changes included extra sperm in the perivitelline space, nuclear budding, the presence of large nucleoli, perinuclear vesicles, the concentration of cytoplasmic organelles, and the extrusion of cytoplasmic ground substance. Initial changes in the nucleus and cytoplasm were undetectable when the zygotes and embryos were examined with the light microscope. It was concluded that serious abnormalities in zygotes and embryos, which may not be identified at the light microscope level, may be detected if they are examined in the electron microscope. Therefore, zygotes and embryos should be critically evaluated before being rated as normal.

ISSN:0148-7280    

DOI:10.1002/mrd.1120040607

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractThe sperm of Luidia clathrata are morphologically typical of asteroid sperm. The head is spherical and contains the nucleus and acrosomal complex. The nucleus has an anterior indentation in which rests the acrosomal complex. There is no evidence of a centriolar fossa along the posterior border of the nucleus. The acrosome is a cup‐shaped structure containing a less electron dense central region. The periacrosomal material is homogeneous in nature, and the subacrosomal specialization of the periacrosomal materials appear as bands of varying electron density. The middle piece is an annular band of mitochondria which surrounds the proximal and distal centrioles. The centrioles exhibit the typical nine triplet arrangement. Both the centrioles and the axoneme project to one side of the middle piece region. Associated with the distal centriole is an elaborate pericentriolar proces

ISSN:0148-7280    

DOI:10.1002/mrd.1120040608

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractLow molecular weight, acid‐stable proteinase inhibitors from epididymal and seminal vesicle homogenates were isolated and characterized. The isolation procedure consisted of gel filtration, trypsin affinity, and ion exchange chromatography. The inhibitor from seminal vesicle homogenates has a molecular weight of approximately 6,200, and that of the epididymal inhibitor was estimated at 4,000. Antiserum directed against the seminal vesicle inhibitor did not react with epididymal components. The epididymal inhibitor shows competitive, whereas the seminal vesicle inhibitor shows noncompetitive inhibition against trypsin on double reciprocal plots. Both inhibitors are effective against trypsin and acrosin but not against chymotrypsin, kallikrein, thrombin, or plasmin. To verify site of origin and to investigate androgen dependency of the epididymal inhibitor, mice were efferentiectomized, orchiectomized, or orchiectomized with androgen supplementation. Gel filtration profiles of acid‐treated epididymal homogenates from normal and efferentiectomized animals show inhibitor peaks in the same regions. The concentration of acid‐stable inhibitor from epididymal homogenates decreased with orchiectomy but returned to normal values when exogenous androgen was supplied. These observations suggest that the low molecular weight inhibitor in the epididymal homogenates is distinct from that in the seminal vesicles. Furthermore, the inhibitor associated with epididymal homogenates is androgen‐dependent, and the epididymis is the site of origin of this in

ISSN:0148-7280    

DOI:10.1002/mrd.1120040609

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

ISSN:0148-7280    

DOI:10.1002/mrd.1120040610

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

ISSN:0148-7280    

DOI:10.1002/mrd.1120040611

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY