Journal of reproduction and fertility. Supplement.
Discontinued
Publisher:
Blackwell Scientific Publications for Journals of Reproduction and Fertility,. Cambridge, UK : Journals of Reproduction and Fertility, Ltd.
Frequency: Irregular
Country: England
Language: English
Author(s):
Society for the Study of Fertility.
Start Year:1966 - 2001
Identifiers
| ISSN: | 0449-3087 (Print) 0449-3087 (Linking) |
| NLM ID: | 0225652 |
| (DNLM): | J37020000(s) |
| (OCoLC): | 01639439 |
| Coden: | JRFSAR |
| LCCN: | sn 82004517 |
| Classification: | W1 JO868KA |
Biophysical and 1H magnetic resonance spectroscopy characteristics of fractionated stallion ejaculates. The composition of seminal plasma must be determined to assess the possible roles of sex gland secretions in survival of stallion spermatozoa. In the present study, an automated semen collection device and 1H magnetic resonance spectroscopy were used to analyse and compare the composition of seminal plasma from fractionated and nonfractionated stallion ejaculates. The contribution of each semen component to the ejaculate (sequence of production of component and concentration) was evaluated and its relationship to biophysical parameters was determined. 1H magnetic resonance spectroscopy was use...
Equine viral arteritis: further characterization of the carrier state in stallions. Further characterization of the carrier state in stallions infected with equine arteritis virus revealed that there is considerable variation in the frequency of its occurrence among breeds. The frequency ranged from 12.5% (Holsteiner stallions) to 72.7% (Dutch Warmblood stallions), with a mean occurrence of 40.8% in the seropositive stallions (n=561) examined. More than 70% of the virus shedders were Standardbred stallions. The carrier state was not confirmed in any of the stallions that had been vaccinated against equine viral arteritis nor was there any evidence of intermittent virus sheddi...
Effect of cholesterol on the motility and plasma membrane integrity of frozen equine spermatozoa after thawing. The aim of the present study was to investigate the cryoprotectant properties of cholesterol after incorporation into the plasma membranes of equine spermatozoa. A cholesterol-methyl-beta-cyclodextrin complex was used to alter sperm plasma membrane cholesterol content. Ejaculates from six stallions were centrifuged in a non-fat skimmed milk glucose-sucrose extender (MK) or a modified Tyrode's medium (TALP). The sperm pellets were resuspended in the appropriate extender with or without added cholesterol (0.125 mmol cholesterol-methyl-beta-cyclodextrin complex l(-1)) and incubated at 24 degrees ...
Pregnancies produced from fertile and infertile stallions by intracytoplasmic sperm injection (ICSI) of single frozen-thawed spermatozoa into in vivo matured mare oocytes. The use of intracytoplasmic sperm injection (ICSI) for in vitro fertilization of equine oocytes and the developmental potential of these oocytes after transfer to the Fallopian tubes of synchronized mares were examined. Oocytes were aspirated from mature follicles 39 h after injection of a GnRH analogue and transported 190 km at 39 degrees C. Semen from a fertile and an infertile stallion was frozen and prepared for injection. Successfully injected oocytes were transferred surgically into the ampulla of the Fallopian tube either: (i) 4-8 h after semen injection; or (ii) after 24-48 h culture b...
Zona pellucida-sperm binding assay for equine oocytes. The binding of a spermatozoon to the zona pellucida is the first step in fertilization. The number of spermatozoa bound to a zona pellucida may reflect the functional status of both the oocyte and spermatozoa. The aim of the present study was to determine whether the stage of maturation of the equine oocyte affects the capacity of the zona pellucida to bind with spermatozoa. Cumulus-oocyte complexes (COCs) were collected from the ovaries of mares from abattoirs or were obtained in vivo by ultrasound-guided follicular aspiration. Oocytes were inseminated directly after collection or after 30 h ...
Freezing of stallion semen: interactions among cooling treatments, semen extenders and stallions. In the present study, the interactions among stallions, semen extenders and cooling treatments before stallion semen samples were frozen were studied. In Expt 1, the effects of four cooling treatments and three semen extenders were investigated (11 stallions x four split ejaculates), whereas in Expt 2, the effects of two semen extenders, two egg yolk concentrations and two glycerol concentrations were investigated (six stallions x five split ejaculates). Sperm motility after thawing was evaluated. In Expt 1, the extender x cooling treatment interaction was significant. Centrifugation and addit...
Ultrastructure, protein phosphorylation and mRNA status of equine oocytes matured in vivo and in vitro. Equine oocytes were collected by follicle aspiration in vivo or by dissection of material obtained from an abattoir, and the ultrastructure, protein phosphorylation and mRNA status of the oocytes were evaluated. Electron microscopy studies indicated that the nucleus had a smooth membrane in oocytes with a compact cumulus, whereas the nuclear membrane was undulated in all other groups. Oocytes with compact cumuli had only a few microvilli, whereas those with expanded cumuli had more microvilli. There were only small numbers of cortical granules close to the oolemma in oocytes with compact cumul...
Dopamine antagonist-induced reproductive function in anoestrous mares: gonadotrophin secretion and the effects of environmental cues. The effect of the dopamine antagonist sulpiride on FSH secretion and onset of reproductive activity in anoestrous mares under different environmental conditions was investigated. In Expt 1, sulpiride (0.5 mg (-)-sulpiride kg(-1) twice a day) had no affect on FSH pulse frequency, mean FSH concentration, basal FSH concentration or FSH pulse amplitude in anoestrous mares. These data do not support the hypothesis that dopamine inhibits reproductive activity by suppressing GnRH secretion, as it does in other species. In Expt 2, the interval to first ovulation (14.8 +/- 1.9 days; range 12-22 days) i...
Testicular secretion of inhibin in stallions. In this study, clear seasonal changes in the circulating concentrations of immunoreactive inhibin were observed in stallions throughout the year. The highest plasma concentrations of immunoreactive inhibin were observed during the breeding season and the lowest concentrations were observed during the nonbreeding season. There was positive correlation between the circulating concentrations of immunoreactive inhibin, steroid hormones and gonadotrophins. Inhibin alpha subunit positive staining was observed in Sertoli cells and more clearly in Leydig cells. Both cell types also stained positively ...
Effect of timing of follicle aspiration on pregnancy rate after oocyte transfer in mares. Mares with preovulatory follicles >33 mm in diameter were administered hCG and were randomly assigned for aspiration of the dominant follicle at 24 h or 35 h after hCG administration. Oocytes recovered at 24 h were cultured for 12 h before transfer and oocytes recovered at 35 h were cultured for 1 h. Oocytes were transferred by flank laparotomy to the oviduct of the same mare, or to the oviduct of another oocyte donor. Recipient mares were inseminated before and after transfer. The oocyte recovery rates at 24 h and 35 h after hCG administration were not significantly different (10/15 (66%) and...
Influence of the dopamine antagonist domperidone on the vernal transition in seasonally anoestrous mares. The aim of the present study was to determine the effect of prolonged administration of the dopamine antagonist domperidone on follicular development, ovulation and endocrine profiles in anoestrous mares. Anoestrous mares (n=16) were maintained under natural photoperiod and ambient temperature. Eight of the mares were treated with domperidone each day from 15 January until the first ovulation of the year. The mean number and size of follicles > or =20 mm in diameter were significantly greater in domperidone-treated mares than in control mares by day 14 of treatment. The day of first ovulati...
Testicular biopsy in stallions: diagnostic potential and effects on prospective fertility. Testicular biopsy was performed as a standing procedure on fertile, mature stallions (n=7) under sedation and local anaesthesia. The 'Biopty' instrument was used to collect three samples from the left testis of each stallion. The oestradiol, testosterone and inhibin content in each testis were assessed by validated radioimmunoassay (RIA); protein concentrations were determined and the testes were prepared histologically for haematoxylin and eosin staining. The fertility status of each stallion was assessed before and after testicular biopsy by semen evaluation, changes in total scrotal width, ...
Immunolocalization of oxytocin and neurophysin in the mare uterus. The presence of oxytocin in the mare uterus was determined by radioimmunoassay of tissue extracts and uterine lavage fluid, and by immunohistochemistry. Samples were obtained from mares during the oestrous cycle and on day 14 of pregnancy. Immunoreactive oxytocin was detected in extracts of endometrium and myometrium, and in uterine lavage fluid. In tissue sections, the luminal epithelium, the epithelium of the superficial, but not deep, glands, blood vessel walls and myometrium all stained positively for oxytocin, whereas only the luminal epithelium and epithelium of the superficial glands st...
Exposure of progesterone receptors on the plasma membranes of stallion spermatozoa as a parameter for prediction of fertility. Subfertility in stallions is attributed to the inability of spermatozoa to undergo the acrosome reaction in response to progesterone. In the present study, it was assessed whether there is a correlation between stallion fertility, defined on the basis of first cycle foaling rate and first cycle 'non-return rate', and the proportion of spermatozoa with exposed progesterone receptors on their plasma membranes. Semen from Dutch Warmblood (n=10) and Friesian (n=4) stallions was analysed. Progesterone 3-(o-carboxymethyl) oxime-BSA coupled with fluorescein isothiocyanate was used as a progesterone r...
Effect of L-glutamine for freezing equine embryos: evaluation by DAPI staining and transfer of multiple embryos to recipient mares. Day 6.5 equine embryos (n=30) were frozen in a medium containing glycerol (2.5-10.0%) supplemented with 0, 20 or 100 mmol L-glutamine 1(-1). After thawing, the embryos were tested individually, using 4',6'-diamidino-2-phenylindole (DAPI) staining to evaluate cell death. Three embryos (one frozen at each L-glutamine concentration) were transferred together into individual recipient mares. Pregnancy diagnosis was performed at day 12 (age of embryo). Embryos were collected at day 14 (age of embryo) and were identified by PCR amplified microsatellite analysis. Nine of ten recipient mares that rece...
Inhibin localization in equine granulosa-theca cell tumours and inhibin forms in tumour fluid. The aim of this study was to examine inhibin production in granulosa-theca cell tumours (GTCT). The experimental aims were: (i) to determine GTCT cell types that produce inhibin alpha- and betaA-subunits; (ii) to determine whether alpha- and betaA-subunit forms differ in GTCT fluid and normal equine follicular fluid (eFF); and (iii) to determine whether dimeric inhibin (alpha betaA) is present in GTCT plasma and tumour fluid. Plasma, tumour fluid and tumour tissue were collected from mares (n=6) with GTCT. Plasma and eFF were collected during the follicular phase from mares (n=4) undergoing no...
Evaluation of equine fetal growth from day 100 of gestation to parturition by ultrasonography. Transrectal and transabdominal ultrasonography were performed on normal pregnant mares (n=10) at 2 week intervals from day 100 of gestation to parturition to evaluate fetal growth. Several fetal anatomical regions (head, eye, aorta, abdomen, rib, gonad, kidney and femur) were imaged and measured using standardized scan plans. The results of these analyses indicate that all of the biometric parameters correlate strongly with the day of gestation. Growth charts were developed, which demonstrate that the following variables have linear relationships with the day of gestation on which they were me...
Relationship between sperm nuclear protamine free -SH status and susceptibility to DNA denaturation. Data from the sperm chromatin structure assay (SCSA), a flow cytometric measurement of susceptibility of sperm nuclear DNA to denaturation, show strong correlation with the fertility potential of bulls, boars, men and stallions. Previous studies showed a strong relationship between stallion spermatozoa with denatured DNA and the presence of DNA strand breaks. In the present study, the relationship between stallion sperm DNA denaturation and the redox status of -SH groups on the cysteine residues of sperm nuclear protamines that are thought to stabilize chromatin was investigated. Semen samples...
Effect of oestradiol on LH secretion and pituitary responsiveness to GnRH in ovariectomized mares. Long-term ovariectomized Pony mares were treated with oestradiol (0.2-5.0 mg; i.m.) at 12 h intervals for 10 days. Blood samples were collected by jugular venepuncture three times a day throughout the experiment and additional blood samples were collected at 15 min intervals for 12 h on days 0 and 10 (sampling periods 1 and 2, respectively). There were significant effects of oestradiol treatment (P < 0.05) and oestradiol treatment x day (P < 0.0001) on the mean LH concentrations each day. Regression analysis of LH time trends each day indicates that there is heterogeneity (P < 0.001) ...
Immunolocalization of a novel protein (P19) in the endometrium of fertile and subfertile mares. One of the major progesterone-dependent endometrial proteins in the mare is a novel 19 kDa lipocalin (P19). This protein is secreted by the endometrial glands and is readily detectable in uterine secretions during the luteal phase of the oestrous cycle and early pregnancy. The function of P19 is unknown, but since most lipocalins act as carriers of small hydrophobic molecules, it probably transports a maternal factor to the conceptus during pregnancy. In this study, a high titre antiserum raised against recombinant-derived P19 was used to detect by immunohistochemistry the protein in endometri...
Development of equine endometrial glands from fetal life to ovarian cyclicity. Branched tubular endometrial glands are key features of the uterine lining in mares. The epithelium in the neck regions of the glands is similar to that lining the lumen of the uterus, whereas the epithelium in the deeper, coiled portions of the glands secretes protein rich histotrophe or 'uterine milk' during dioestrus and pregnancy. The present study was performed to characterize endometrial histology in perinatal and peripubertal fillies and to investigate the influences of specific steroid treatments on endometrial gland development in the prepubertal period. Uteri obtained from equine fet...
Which insemination results in fertilization when several are performed before ovulation? The aim of the present study was to determine which artificial insemination results in fertilization when mares are inseminated several times before ovulation. Mares in oestrus were inseminated over 62 cycles with fresh semen at 48 h intervals from when a follicle > or =30 mm in diameter was detected until ovulation. The number of inseminations was limited to three. Three fertile stallions were used and a different stallion was used for each artificial insemination. The order of the three stallions was changed for each cycle. Embryos were collected between day 10 and day 12 after ovulation and...
Localization and cellular distribution of a unique hyaluronidase in stallion spermatozoa during epididymidal transit. Three protein bands with hyaluronidase activity and molecular masses of 87, 48 and 43 kDa were isolated from purified equine sperm plasma membranes. Indirect immunofluorescence was used to assess sperm labelling patterns using a polyclonal antibody to sperm hyaluronidase. In ejaculated spermatozoa, surface-associated hyaluronidase was localized to the posterior head region of 98 +/- 2% of spermatozoa (n=10). Epididymides were isolated from mature stallions (n=5) and divided into caput, corpus and cauda epididymides in separate Petri dishes. The epididymidal tubules were dissected and washed us...
Motility, morphology and triple stain analysis of fresh, cooled and frozen-thawed stallion spermatozoa. The aim of the present study was to determine whether there are characteristics of fresh, cooled and frozen-thawed semen samples that can be used to predict the suitability of stallion semen for preservation by cooling or freezing. Each of three ejaculates obtained from 12 stallions was divided into aliquots to be analysed for sperm motility, morphology and membrane integrity as fresh, cooled and frozen-thawed samples. The percentage of morphologically normal spermatozoa was similar in fresh and cooled samples and both were greater than in the frozen samples. There were no strong linear relati...
Maternal influence on pre- and postnatal growth of foals born after embryo transfer. In the present study the growth and development of three pairs of matching gender foals from birth to maturity were compared. One Polish Pony embryo of each pair was transferred from a donor mare (mass 387-400 kg) to a much larger heavy type recipient mare (mass 561-780 kg). The other embryo of each pair underwent normal gestation (control). The transferred and control horses were examined at 9 and 13 years of age, and X-ray pictures were taken of their front legs to measure the length of the bones. On the basis of the results of this study, it is concluded that the processes of growth and dev...
The role of endogenous opioids in the ovulatory LH surge in mares. Removal of opioid inhibition of GnRH neurones is thought to be a critical event in generating the ovulatory surge in some species. In the present study, a nonsurgical technique was used to collect pituitary venous blood samples from eight mares every 0.5-1.0 min for 1 h before and after administration of the opioid receptor antagonist naloxone (0.2 mg kg(-1), i.v.), to investigate whether opioid inhibition is also important in mares. Jugular blood samples were taken at 10-15 min intervals. Mares were studied 0, 1 or 2 days before ovulation. Naloxone administration increased mean rates of GnRH ...
Partial characterization of an equine conceptus prostaglandin inhibitory factor. Equine conceptuses are thought to produce antiluteolytic factors that inhibit endometrial PGF2alpha and, thus, prevent luteolysis in pregnant mares. The aim of the present study was to characterize partially the chemical nature of the prostaglandin inhibitory factor (PIF) produced by equine conceptuses in vitro. Embryos were collected from pregnant mares 13 +/- 0.5 days after ovulation and were cultured for 24 h. Harvested equine conceptus conditioned media (CCM) were assayed for antiluteolytic activity by determining the inhibition of endometrial PGF2alpha synthesis in vitro. Significant anti...
Relationship between thyroid function and seasonal reproductive activity in mares. The relationship between thyroid function and seasonal reproductive activity in mares was investigated by comparing plasma triiodothyronine and thyroxine concentrations in anoestrous mares and in mares with cyclic ovarian activity during the anovulatory season. In study 1, the diurnal changes in plasma triiodothyronine and thyroxine concentrations on a single day were compared in anoestrous and cyclic mares. The mean thyroxine concentrations were significantly higher (P < 0.001) in the cyclic mares than in the anoestrous mares, whereas mean triiodothyronine concentrations were similar in bo...
Apoptosis in equine granulosa cells and its relationship to cumulus expansion and oocyte chromatin configuration in ovarian follicles. During the oestrous cycle follicles grow and either ovulate or regress. Regressing follicles undergo atresia and in many species apoptosis has been identified as the underlying mechanism in this process. The aims of this study were to establish whether equine granulosa cells degenerate via an apoptotic mechanism and whether the presence of apoptotic cell death in granulosa cells is correlated with oocyte quality. Ovaries from mares at unknown stages of the oestrous cycle were obtained from an abattoir. In Expt 1, follicles (n=352) from 37 mares were processed. DNA was extracted from granulosa ...
Ovulation synchrony after follicle ablation in mares. Two experiments were performed to determine the efficacy of ultrasound-guided transvaginal follicle ablation for synchronizing ovarian function in mares. The experiments were initiated at random stages of the oestrous cycle in control (nonablated) and follicle-ablated mares. On day 0, all follicles > or =10 mm in diameter were punctured, aspirated and curettaged in ablated mares, and, on day 4, two doses of PGF2alpha were administered with a 12 h interval between the doses to both ablated and nonablated (control) mares. In Expt 1, hCG was administered to the ablated mares on the first or se...