摘要:

AbstractThe presence of actin filaments around mitochondria during vertebrate spermiogenesis was demonstrated by immunofluorescence and immuno‐electron microscopy and by heavy meromyosin decoration. The presence of actin is supposed to be related to mitochondrial rearrangements occurring in the spermatid stag

ISSN:0148-7280    

DOI:10.1002/mrd.1120030302

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

年代:1980

数据来源: WILEY

摘要:

AbstractBecause the acrosome of human sperm is too small to be directly visualized by phase‐contrast microscopy, acrosome reactions (that is loss of the acrosome) are generally not evaluated in studies of human sperm capacitation and fertilization. Nevertheless, it would be useful in such studies to have a technique for easily identifying and quantitating acrosome‐reacted sperm. In this paper, we describe a method for labeling the human sperm acrosome with fluorescein‐conjugated Ricinus communis agglutinin‐60 (FITC‐RCA); we show that in sperm without acrosomal caps, FITC‐RCA labeling occurs either not at all or only in the equatorial segment of the acrosome. To determine if the absence of FITC‐RCA labeling in the acrosomal cap region gives a reliable estimate of acrosome reactions, washed sperm or sperm incubated in a capacitating medium (BWW) were divided into two groups, which were then fixed for FITC‐RCA labeling or transmission electron microscopy. Counts of acrosome reactions made by each method were similar, and we observed an increase in the percentage of reactions following incubation in BWW. We conclude that the FITC‐TCA labeling technique is a reliable method for accurately scoring the percentage of acrosome‐

ISSN:0148-7280    

DOI:10.1002/mrd.1120030303

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

年代:1980

数据来源: WILEY

摘要:

AbstractA procedure is described for isolating milligram quantities of bovine and porcine zonae pellucidae, uncontaminated by follicle cells or their processes. On SDS‐polyacrylamide gel electrophoresis the isolated bovine zona material formed one major glycoprotein band with an estimated molecular weight of approximately 100,000 daltons and two minor lower molecular weight components. The isolated pig zonae formed only one glycoprotein band with a molecular weight of approximately 62,000 daltons. Rabbit antisera raised against the isolated zonae were zona‐specific and formed only a single precipitin line against the heat‐solubilized zonae on immunoelectrophoresis. An adjuvant was not required for high‐antibody titers. High titers were also obtained by injection of the dog and rhesus monkey. Anti‐zona antibody was detected by immunofluorescence, zona‐coating, double‐immunodiffusion, and the inhibition of spermbinding to eggs, including those of human origin. Antigenic and sperm receptor properties were stable at 100°C for five minutes, but some activity was lost after longer exposure. The serum antibody produced by rabbits immunized with pig zonae was predominantly IgG and cross‐reacted with the zonae of a variety of other species, including primates. Pregnancy was inhibited in female rabbits immunized with pig z

ISSN:0148-7280    

DOI:10.1002/mrd.1120030304

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

年代:1980

数据来源: WILEY

摘要:

AbstractThe relationship between cellular dispersion and steroidogenesis was studied in culture using oocyte‐cumulus complexes harvested from porcine follicles. The cells were cultured in modified TC 199 containing pig serum for one to two days. When oocyte‐cumulus complexes were cultured in the absence of hormone the oocytes resumed meiosis, the cumulus cells grew out forming monolayers, and progesterone accumulation was low. Addition of ovine LH, purified human LH, or purified human FSH stimulated expansion of the cumulus mass as well as enhanced progesterone accumulation. Oocyte maturation was not affected by the hormones. In absence of hormone, oocyte‐cumulus complexes obtained from large (6–12 mm) follicles showed increased cellular dispersion and higher progesterone accumulation as compared to complexes obtained from medium‐sized (3–5 mm

ISSN:0148-7280    

DOI:10.1002/mrd.1120030305

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

年代:1980

数据来源: WILEY

摘要:

AbstractAnti‐rabbit proacrosin IgG was prepared from goat serum following immunization with a homogeneous preparation of rabbit testis proacrosin. The “auto‐activation” products of purified testis proacrosin were separated into 68,000 and 34,000 molecular weight (mol wt) acrosins by Sephadex G‐100 column chromatography. Immunodiffusion analysis of testis and epididymal sperm proacrosins and acrosins on agarose gel against goat anti‐rabbit testis proacrosin showed immunological identity between rabbit testis and sperm proacrosins and the initial testis acrosin (mol wt 68,000). However, the 34,000 mol wt form of testis acrosin showed weaker reaction with the antibody and only partial identity with the proacrosin and the 68,000 mol wt form of acrosin. These results suggest that there is no major structural difference between testis and sperm proacrosins and between proacrosin and the 68,000 mol wt acrosin, but such a structual change occurs when the 34,000 mol wt acrosi

ISSN:0148-7280    

DOI:10.1002/mrd.1120030306

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

年代:1980

数据来源: WILEY

摘要:

AbstractThe motility of salmonid spermatozoa initiated by dilution of the milt with ovarian fluid or isotonic saline is brief duration; it was believed that it can be activated only once in the life of the spermatozoon. Dilution of the milt with an equal volume of isotonic saline (0.12 M‐NaCl) containing 5 mM‐3‐isobutyl‐1‐methylxanthine (MIX) prolonged and intensified sperm motiliy. When motility had stopped after initial mobilization with saline or ovarian fluid, it could be reactivated by addition of MIX; reactivated spermatozoa fertilized eggs. Dilution with saline containing K+(24 mEq/liter) did not initiate sperm motility even in the presence of MIX. The spermatozoa were mobilized by subsequent with 0.12 M‐NaCl. The concentration of adenosine triphosphate (ATP) in sperm suspensions dropped on dilution with saline and rose as motility ceased, but declined without subsequent recovery following dilution with MIX‐saline. The concentration of cyclic adenosine monophosphate (cAMP) rose and fell sharply on initiation of motility and rose again after motility had declined. While salmonid spermatozoa can be mobilized by dilition with saline alone, the effectiveness of MIX in reactivating “spent” spermatozoa supports the assumption that cAMP plays a role in the initiation

ISSN:0148-7280    

DOI:10.1002/mrd.1120030307

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

年代:1980

数据来源: WILEY

摘要:

AbstractNuclear vacuoles resembling the “crater defect” described in bull spermatozoa were observed in 14 boars. Both the incidence of the defect and semen quality were monitored with phase contrast microscopy over a three‐month period. The percentages of cratered spermatozoa varied widely both among boars and in ejaculates from the same boar taken on different days. The presence of cratered spermatozoa at a level of 5% or more appeared to be associated with low semen quality. The defect was studied with scanning and transmission electron microscopy and was found to consist of nuclear invaginations, about 0.5 μm in diameter, containing some scanty amorphous electron‐dense material. In boars showing a high incidence of spermatozoa with crater defects, abnormalities of the acrosome and perforatorium wer

ISSN:0148-7280    

DOI:10.1002/mrd.1120030308

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

年代:1980

数据来源: WILEY

摘要:

AbstractA mammalian ovoperoxiadase activity has been detected in ionophore activated mouse eggs. The peroxidase activity was demonstrated at the electron microscopic level using the 3,3′‐diaminobenzidine (DAB) histochemical method. A positive DAB reaction was detected in a portion of the intact cortical granules of untreated or DMSO treated control eggs. In the ionophore activated eggs, the DAB reaction product was routinely detected by electron microscopy, predominantly on the cell surface, that is on the zona pellucida, in the perivitelline space, and in association with the cortical granule exudates. Furthermore, the peroxidase inhibitors phenylhydrazine and sodium sulfite prevented DAB staining in ionophore activated oocytes. These results indicate the presence of an ovoperoxidase, possibly of cortical granule origin, on the surface of activated mammalian eggs, detectable by histochemical me

ISSN:0148-7280    

DOI:10.1002/mrd.1120030309

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

年代:1980

数据来源: WILEY

摘要:

AbstractOne consequence of fertilization or parthenogenetic activation of mammalian eggs is an altaration in the solubility proprieties of the zona pellucida, known as zona hardening. Several lines of evidence indicate that an ovoperoxidase, which is activated and/or secreted from mouse eggs. Following parthenogenetic activation, corss‐links tyrosine residues in the zona pellucida and results in hardening of the zona. First, zona hardening, as determined by decreased solubility of the zona in pronase, is inhibited by several compounds known to inhibit peroxidases. Inhibitors of hardening include phenylhydrazine, sodium sulfite, sodium azide, and glycine ethyl ester. Second, tyrosine analogs inhibit zona hardening, unless the phenolic hydroxyl group or ortho position is blocked. That is, O‐methyltyrosine (methyl substitution of phenolic hydroxyl) does not inhibit hardening; o‐methyltyrosine (methyl substitution of one ortho position) partially inhibits, whereas tyramine and N‐acetyltyrosine (free hydroxyl and ortho positions) effectively block hardening. Finally, exogenous horseradish peroxidasepromotes limited hardening of the zona in unactivated eggs. These results are consistent with a peroxidase catalyzed cross‐linking of tyrosines in the zona that results in hardening of the zona

ISSN:0148-7280    

DOI:10.1002/mrd.1120030310

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

年代:1980

数据来源: WILEY

摘要:

AbstractNuclei of pteridium sperm have been dispersed by turbulence in natural or slightly alkaline buffer after stripping off the cytoplasm with nonionic detergent. The nuclei tended to break up into fragments arranged in a linear order. These fragments fluoresced brightly with acridine orange as did intact nuclei. Grounds are given for identifying the smaller fragments with chromosomes. It is proposed that the sperm nucleus of British Pteridium, possibly an autotetrapolid, consists of a sequence of paired homologues.

ISSN:0148-7280    

DOI:10.1002/mrd.1120030311

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

年代:1980

数据来源: WILEY