Search PubMed⌕ Search

Biomedical subjects

H D Moore

Publications and source records attributed to H D Moore.

At least 73 records · Page 4Linked to original sources

Administration of a gonadotropin releasing hormone analogue in oligozoospermic infertile males.

This study investigated the stimulatory potential of a superactive gonadotropin releasing hormone analogue in idiopathic oligozoospermia. In a double-blind trial, 19 men were randomized to receive buserelin (D-Ser(TBU)6-GnRH ethylamide) in one of two dosages or saline, twice weekly, for twelve weeks. Treatment did not lead to a significant increase in serum concentrations of the pituitary gonadotropins or in sperm concentrations. However, in the dosages and schedules investigated, there was no inhibitory effect of the analogue. Further assessment of this approach is suggested.

Buserelin↗

Hormonal and behavioural detection of oestrus in blackbuck, Antilope cervicapra, and successful artificial insemination with fresh and frozen semen.

Hormonal detection of urinary pregnanediol-3 alpha-glucuronide proved an effective method of monitoring the progress of oestrous cycles in the blackbuck; observation of sexual behaviour in a vasectomized male was, however, a more practical procedure. Good correlation was observed between the occurrence of minimal pregnanediol concentrations in females and the maximal behavioural response by the male. On the basis of intervals between periods of behaviourally detected oestrus, a mean cycle length of 16.9 days (+/- 0.62, s.e.m.) was derived from 12 cycles (4 animals). Eleven females were inseminated in this study, 8 with freshly collected semen and 3 with frozen semen; 6 calves were obtained, 1 after the use of frozen semen. Pregnancy was monitored by measurements of pregnanediol-3 alpha-glucuronide excretion and by ultrasound scanning. The mean interval between insemination and parturition was 183.3 days inclusive, ranging from 182 to 186 days.

Animals↗

Visualization and characterization of the acrosome reaction of human spermatozoa by immunolocalization with monoclonal antibody.

A monoclonal antibody generated against hamster epididymal spermatozoa and recognizing an antigen within the acrosome was used in conjunction with FITC-antimouse immunoglobulin as a marker of the human acrosome during sperm development, capacitation, and the acrosome reaction. The specificity of binding of the monoclonal antibody was assessed using immunolocalization by epi-fluorescence and electron microscopy. Immunofluorescence revealed that antibody bound over the entire anterior acrosome in hamster and human spermatozoa. Ultrastructural localization indicated that antigen was predominantly present on the inner face of the outer acrosomal membrane and within the acrosomal content. Qualitative specificity was studied using a highly purified preparation of hamster acrosomes in an enzyme-linked immunosorbent assay. Since the antibody rapidly visualized human acrosomes, it was used to detect abnormal acrosome morphology of mature spermatozoa and to mark spermatids present in the ejaculate. During incubation in capacitating medium, changes in the immunofluorescence of live or methanol fixed spermatozoa were correlated with incubation interval and the ability of spermatozoa to fuse with zona-free hamster oocytes. Spermatozoa bound to zona-free hamster oocytes displayed no fluorescence, confirming that acrosome loss occurred before spermatozoa attached to the vitellus.

Acrosome↗

The ultrastructure of early implantation in the marmoset monkey (Callithrix jacchus).

The ultrastructural morphology of the initial stages of implantation in the marmoset monkey (Callithrix jacchus) was studied in pregnant monkeys at known time intervals after ovulation. The earliest samples, obtained 13 days after ovulation, displayed both cytotrophoblast and syncytiotrophoblast. The cytotrophoblast was restricted to the blastocoel, whilst syncytiotrophoblast intruded to the endometrial basal lamina. At later stages, days 16 and 19 after ovulation, both cytotrophoblast and syncytiotrophoblast had extended laterally around the uterus, and the syncytiotrophoblast also extended deeper into the maternal tissues. The mesoderm layer was first discernible at 19 days after ovulation. At 23 days after ovulation the syncytiotrophoblast surrounded the maternal blood vessels entirely. In this study syncytiotrophoblast was not observed to breach the maternal blood vessels, even at 31 days after ovulation. Early cytotrophoblast columns could be seen at 31 days after ovulation. The endothelial cells lining the maternal blood vessels displayed hypertrophy from the earliest stages (day 13) onwards, although a true decidual response was only observed in samples of 23 and 31 days after ovulation.

Animals↗

Monoclonal antibody against a sperm antigen Mr 95,000 inhibits attachment of human spermatozoa to the zona pellucida.

A murine monoclonal antibody raised against hamster spermatozoa was found to cross-react with human spermatozoa. By immunofluorescence, the antigen was visualized over the equatorial segment of human sperm heads. In the presence of antibody, sperm binding to the zona pellucida of salt-stored human oocytes was significantly inhibited (P less than or equal to 0.005) compared with other antibodies or control preparations. Using SDS-PAGE of whole spermatozoa and membrane preparations followed by Western blot analysis, the antigen was identified as a determinant with a relative molecular weight of 95,000.

Antibodies, Monoclonal↗

Characterization of oestrus and timed collection of oocytes in the grey short-tailed opossum, Monodelphis domestica.

A system of test-pairing was used to detect reproductive behaviour in the grey short-tailed opossum. This enabled timing and characterization of the development of pro-oestrous and oestrous behaviour, and facilitated collection of ovulated and unovulated oocytes. Oestrus was induced 8.5 days (n = 80, 95% confidence limits 7.56-9.21) after the introduction of a male. Timed examination of the ovaries by laparotomy indicated that ovulation occurred 14-16 h after the first onset of oestrous behaviour. The development of follicles was linked to pro-oestrous behaviour, and ovulation occurred in the absence of copulation. Vaginal exfoliative cytology indicated that pro-oestrous behaviour was associated with an increasing number of keratinized epithelial cells, and at the time of maximum receptivity to males, a heavy infiltration of polymorphonuclear leucocytes was seen. Oocytes were typically marsupial: large (approximately 250 microns in diameter), with a yolky vitellus and thin zona pellucida. An average of 6 oocytes were ovulated per ovary.

Animals↗

Semen collection and spermatozoa characteristics in budgerigars (Melopsittacus undulatus).

Semen samples were obtained from budgerigars by applying gentle pressure to both sides of the cloaca. The semen appeared to be stored in the seminal glomera, two structures on either side of the proctodeum. These structures have been described before in Passeriformes. The spermatozoon in budgerigars is similar to the spermatozoon of the domestic fowl, but differs from the spermatozoon of Passeriformes.

Animals↗

Avian spermatozoa penetrate zona-free hamster oocytes in vitro.

After collection from the cloaca of adult males, budgerigar spermatozoa were incubated in modified BWW medium at pH 8.3. By light microscopy, many spermatozoa (greater than 100 per oocyte) were seen to be bound to the oolemma, and a small proportion (1-2 spermatozoa) were judged to have undergone fusion and penetration of the vitellus, as indicated by complete decondensation of their chromatin. The latter finding was confirmed at the electron microscope level by the presence of sperm nucleus, midpiece and tail in egg cytoplasm. This finding is discussed in relation to events at fertilization in the different Phyla.

Animals↗

In-vitro culture of hamster epididymal epithelium and induction of sperm motility.

Epithelium from the proximal corpus region of the epididymis of adult hamsters was cultured in modified RPMI 1640 medium supplemented with growth factors and androgens at 37 degrees C in 5% CO2 in air. Prepared plaques of epithelium formed spheres of tissue with epithelial cells outermost. At the light and electron microscope level, these epithelial balls displayed morphology consistent with continued secretory and absorptive function. After 3-5 days, cultured cells either plated out over the bottom of plastic wells or formed vesicles which expanded as their interior became fluid filled. Spermatozoa recovered from the caput epididymidis were co-cultured with epithelium. After 8 and 24 h, a proportion of spermatozoa (30%) exhibited slow but persistent flagellum beats with slow progressive motility. Spermatozoa in control incubations were immotile. This change in motility pattern would suggest that some sperm maturation processes had occurred in vitro.

Animals↗

A determinant of Mr 34,000 expressed by hamster epididymal epithelium binds specifically to spermatozoa in co-culture.

A murine monoclonal antibody raised against hamster cauda epididymal spermatozoa was shown to recognize an Mr 34,000 component of epididymal epithelium. Antigen was localized by immunocytochemistry on the surface and in the apical cytoplasm of principal cells in the proximal corpus epididymidis but not in the caput or initial segment regions. Spermatozoa from the corpus epididymidis expressed antigen on their post-acrosomal plasma membrane and annulus. Epididymal principal cells from the proximal corpus region when cultured in vitro bound antibody on their apical surface for at least 5 days. Spermatozoa from the caput epididymidis co-cultured with epithelium expressed antigen after incubation for 8 and 24 h. These results suggest that a surface change to epididymal spermatozoa during maturation in vivo may also be elicited during in-vitro culture.

Animals↗

In-vitro development of the fertilizing ability of hamster epididymal spermatozoa after co-culture with epithelium from the proximal cauda epididymidis.

The epididymides of adult male hamsters were surgically ligated at the junction of the distal corpus and proximal cauda regions. After 3 days, spermatozoa recovered from the distal corpus displayed greater progressive motility and head to head agglutination in capacitating medium than did those from intact controls, but had low fertilizing ability (3% fertilization rate) in vitro or in vivo. When these spermatozoa were incubated for 6 h with epithelial cells from the proximal cauda epididymidis, previously cultured for 3 days, they maintained motility and exhibited a significant increase in fertilizing ability (30% and 29% in vitro and in vivo respectively). The fertilizing ability of distal corpus spermatozoa incubated with 3-day-old cultures without androgens, or 8-12-day-old epithelial cells with fibroblast overgrowth, or without epithelial cells, remained low (5%). Increase in sperm fertilizing ability was associated with increased sperm binding to the zona pellucida in vitro. These results demonstrate that, under suitable culture conditions, the final stages in the development of hamster sperm fertilizing ability can be achieved in vitro. Factors secreted by cultured epithelium from the proximal cauda epididymidis are implicated in this maturation process.

Animals↗

Early implantation stages in the marmoset monkey (Callithrix jacchus).

The morphology of the initial stages of implantation in the marmoset monkey (Callithrix jacchus) was studied by obtaining embryos and associated endometrium at timed intervals after ovulation. Estrus cycles were detected by measuring daily levels of plasma progesterone. Following a short follicular phase, circulating levels of progesterone above 20 ng/ml were taken as representing day 1 after ovulation. On this basis, single, twin, and triplet embryos were recovered from six perfused-fixed females on days 13, 16, 19, 23, and 29 after ovulation and prepared in resin for light microscopy. Early implantation stages, 13 and 16 days after ovulation, were characterized by the intrusion of syncytial trophoblast between epithelial cells of the endometrium with minimal cellular damage. Some hyperplasia of epithelium at the margin of the implantation site was evident. The consolidation of the initial attachment was achieved by an increase in syncytial trophoblast underlying the inner cell mass of the embryo which rapidly surrounded and breached maternal capillaries. Although initially separate, the chorions of twin or triplet embryos started to fuse by day 19 after ovulation. This process was complete by day 29 such that embryos shared a common uterine exocoelom surrounded by continuous trophoblast. It was concluded that implantation in the marmoset monkey commenced on days 11-12.5 after ovulation and involved an intrusive mechanism. Although trophoblast penetration of endometrium was superficial, maternal capillaries were tapped at an early stage of implantation. The fusion of chorions of twins and triplets first occurred around day 19 after ovulation.

Animals↗

Maturation and function of the hamster spermatozoon probed with monoclonal antibodies.

To investigate the surface of the mammalian spermatozoon during its maturation in the epididymis, seven monoclonal antibodies were raised in mice against intact spermatozoa (and isolated sperm heads) recovered from the cauda epididymis of the golden hamster. These antibodies, which with one exception were species specific, recognized various membrane components over restricted regions of the sperm head and tail. During epididymal transit the localisation of four antigens bound by these antibodies was considerably altered. In two cases, antigens were first detected on spermatozoa in the distal corpus epididymis where sperm fertilising capacity is initially acquired. A monoclonal antibody which bound to the acrosome and midpiece specifically blocked fertilisation of intact oocytes of the hamster in vitro. Preliminary characterisation of the antigens was undertaken.

Animals↗

Computer-assisted measurement of sperm swimming speed in human semen: correlation of results with in vitro fertilization assays.

A semiautomatic computerized technique for the measurement of sperm swimming speed is presented. The equipment is easy to use and would be suitable for routine clinical laboratories. The value of the sperm speed measurements obtained from over 100 individuals in relation to fertility has been studied by the correlation of these results with human in vitro fertilization (IVF) data and sperm penetrating capability in the zona-free hamster egg assay. The results show that sperm speed measurements correlate very well with those of the IVF, both human and hamster, and can be used successfully, in conjunction with multivariate statistical methods, to predict the outcomes of such techniques with about 75% accuracy.

Animals↗

Clinical evaluation of the heterologous oocyte penetration (HOP) test.

The heterologous oocyte penetration (HOP) test, using zona-free hamster oocytes, was used to assess the fertilising capacity of human spermatozoa. There was good correlation between the ability of ejaculated spermatozoa to penetrate the zona-free hamster oocytes and intact human oocytes. Using epididymal spermatozoa, the HOP test results showed that the ability to penetrate oocytes was acquired during their passage through the epididymis. Applied clinically, the HOP test enables a group of infertile men to be identified with a functional defect of their spermatozoa; these men may not be identified at routine seminal fluid analysis. Men with two negative HOP test results were confirmed as being infertile since their wives, if normal, conceived rapidly when donor spermatozoa were artificially inseminated.

Animals↗

Expression of sperm antigens during spermatogenesis and maturation detected with monoclonal antibodies.

The expression of hamster sperm antigens was investigated during spermatogenesis and sperm maturation with the use of monoclonal antibodies generated in culture from mice immunized with hamster cauda epididymal spermatozoa or sperm heads. Antigens were localized by immunofluorescence and immunoperoxidase techniques which were first visualized on isolated spermatids over the developing acrosome. In one case, antibody inhibited fertilization in vitro although localization on testicular or epididymal spermatozoa was minimal compared with the early spermatid. Antibodies also recognized surface antigens first expressed in the epididymis whose localization on the spermatozoon altered during epididymal transit or incubation in capacitating medium. The results were discussed in relation to the expression and function of surface determinants on the haploid germ cell.

Animals↗