Biology of reproduction.
Publisher:
Society for the Study of Reproduction. New York : Oxford University Press (2017)
Frequency: Monthly
Country: United States
Language: English
Author(s):
Society for the Study of Reproduction.
Start Year:1969 -
ISSN:
0006-3363 (Print)
1529-7268 (Electronic)
0006-3363 (Linking)
1529-7268 (Electronic)
0006-3363 (Linking)
Impact Factor
3.6
2022
| NLM ID: | 0207224 |
| (DNLM): | B17900000(s) |
| (OCoLC): | 01536448 |
| Coden: | BIREBV |
| Classification: | W1 BI852N |
Partial complementary deoxyribonucleic acid cloning of equine relaxin messenger ribonucleic acid, and its localization within the equine placenta. To determine the site of relaxin gene expression in equine placentae, a set of degenerate oligonucleotide primers was made according to the published amino acid sequence of the A- and B-chain of equine relaxin (eRXN). Total cellular RNA (tcRNA) from equine placentae at about 120 and 300 days of pregnancy was subjected to reverse transcriptase-polymerase chain reaction (RT-PCR) with use of these primers. A single amplification product of approximately 430 bp was detected in each case by agarose gel electrophoresis. The PCR product was ligated into Bluescript plasmid and sequenced to confirm the...
Local initiation of spermatogenesis in the horse. Gross observation of testicular parenchyma of 1.5- to 2-yr-old horses reveals both light and dark regions. If this gross, differential shading reflects quantitative differences in the development of spermatogenesis and interstitial cell populations, the horse may prove to be a useful model for study of the paracrine relationships associated with initiation of spermatogenesis. The objective of this study was to characterize seminiferous tubules and interstitium of testes with gross, differential shading. Testes with both light and dark regions of parenchyma were obtained from horses 1.5-2 yr ol...
Horse trophoblasts produce tumor necrosis factor alpha but not interleukin 2, interleukin 4, or interferon gamma. The distribution of four cytokines was analyzed in the endometrium and trophoblast of the horse between Days 30 and 55 of gestation. Endometrial tissues, invasive trophoblast (chorionic girdle), and noninvasive trophoblast (chorion and allantochorion) were examined separately. Cytokine expression was determined by amplification of specific mRNA via the reverse transcriptase polymerase chain reaction (RT-PCR). Messenger RNA for interleukin 2 (IL-2), interleukin 4 (IL-4), and interferon gamma (IFN gamma) was detected in endometrial tissues, unstimulated peripheral blood lymphocytes, and control ...
Changes in equine endometrial retinol-binding protein RNA during the estrous cycle and early pregnancy and with exogenous steroids. A cDNA library was constructed from poly(A) RNA obtained from Day 14 nonbred equine endometrium. A cDNA probe for porcine retinol-binding protein (RBP) was used to screen the library, and a complete cDNA sequence (1133 bp, excluding the poly(A) tail) was obtained. Endometrial biopsies were obtained from cycling, nonbred mares at Days 0, 1, 4, 8, 10, 11, 13, and 15 and from pregnant mares at Days 11, 13, 15, and 17 after ovulation (n = 2 mares each day). Endometrial biopsies were also taken from 18 noncycling anestrous mares after the following treatments: C (vehicle control for 1 day, n = 3), ...
Interaction of equine spermatozoa with oviduct epithelial cell explants is affected by estrous cycle and anatomic origin of explant. Regulation of attachment of equine spermatozoa to homologous oviduct epithelium was investigated by co-culture of spermatozoa with oviductal epithelial cell explants. Stallion spermatozoa were incubated with explants derived from the isthmus and ampulla of follicular, postovulatory, and diestrous mares. Steroid treatments (estradiol, progesterone, or control) were applied across all explant groups. Estimates of motility and total numbers of attached spermatozoa were made 0.5, 24, and 48 h after initiation of co-culture. Equine spermatozoa attached by their rostral acrosomal region to both cili...
A subpopulation of morphologically normal, motile spermatozoa attach to equine oviductal epithelial cell monolayers. Attachment of spermatozoa to oviductal epithelial cells (OEC) may be a prefertilization event in some species. We tested the hypothesis that spermatozoa that attach to equine OEC monolayers are a selected subpopulation of the initial inseminate, containing a higher proportion of morphologically normal, motile cells than the inseminate. Washed stallion spermatozoa were cocultured with monolayers of OEC or monolayers of Vero cells, and controls were incubated in wells coated with basement membrane extract (Matrigel [Mgel]) or in plastic (uncoated) wells. Unattached spermatozoa were removed by ri...
Initial characterization of equine inhibin. Inhibin has been characterized from a number of mammals; however, it has not been extensively studied in horses. Western blot analysis was used to examine the size heterogeneity of equine inhibin alpha- and beta-subunits. The distribution of equine inhibin activity from the initial sizing column (S-200, 25 x 94 cm) indicated that the majority of equine inhibin activity was present as larger-molecular-size forms. When the large forms were analyzed by Western blot in nonreducing conditions, alpha-subunit bands were detected at 40,000 M(r), 56,000 M(r), 80,000 M(r), and 90,000 M(r); beta a reacti...
Identification of phosphoseryl residues in protamines from mature mammalian spermatozoa. Protamines isolated from ejaculated human, stallion, bull, boar, and ram spermatozoa were subjected to phosphoserine conversion reaction and protein sequencing. Phosphoserines were detected as S-ethylcysteines. Endogenously phosphorylated protamines have previously been found only in ejaculated human sperm. In this study, we demonstrate that ejaculated sperm from other species also contain protamines phosphorylated at serine residues. In P1-protamines, the endogenously phosphorylated serines were located at the N-terminal region in all species studied, whereas in major forms of human and stall...
Platelet-activating factor acetylhydrolase activity in seminal plasma from the bull, stallion, rabbit, and rooster. Platelet-activating factor (PAF) acetylhydrolase, which inactivates PAF, has been detected in human and bovine seminal plasma and may represent a mechanism for regulating sperm-derived PAF. This study was designed to characterize further PAF acetylhydrolase in seminal plasma from domestic animal species. Sperm-free seminal plasma from the bull, stallion, rabbit, and rooster was assayed for acetylhydrolase activity based on the release of [3H]acetate from PAF. As reported previously for bull seminal plasma, activity in stallion, rabbit, and rooster seminal plasma was linear with both time and p...
Ovarian follicular growth and development in mammals. Evidence from several species indicates that the initial stages of follicular growth proceed very slowly. In contrast, the stages after antrum formation are much more rapid. Atresia seems to be most prevalent as follicles approach the size at which they could be recruited for potential ovulation. Although most follicles become atretic around that stage, a few are recruited into a cohort or wave of follicles that continue to grow beyond the stage at which atresia normally occurs. Next, a species-specific number of follicles is selected for dominance. In some species (e.g. rats, primates, pigs),...
Function of uterine and blood-derived polymorphonuclear neutrophils in mares susceptible and resistant to chronic uterine infection: phagocytosis and chemotaxis. In vitro phagocytosis and chemotaxis of uterine and blood-derived polymorphonuclear neutrophils (PMNs) were compared in mares with different resistance to chronic uterine infection (CUI). Both the primary in vitro function of PMNs and the role of uterine environmental factors on PMN function were investigated. The uteri of mares susceptible to (n = 6) and resistant to CUI (n = 5) were inoculated with 5 x 10(6) Streptococcus zooepidemicus when the mares were in estrus. Uterine secretions in addition to uterine and blood-derived PMNs were sampled at 5 and 24 h later. During a subsequent estrus, ...
Immunoglobulin (IgG and IgA) and complement (C3) concentrations in uterine secretion following an intrauterine challenge of Streptococcus zooepidemicus in mares susceptible to versus resistant to chronic uterine infection. The validity of measuring concentrations of immunoglobulins in undiluted uterine secretions was established. The concentrations of IgG, IgA, and cleavage factor C3 of the complement system in uterine secretions were compared in mares with different resistance to chronic uterine infection (CUI). The uteri of mares susceptible (n = 6) and resistant to CUI (n = 5) were inoculated with 5 x 10(6) Streptococcus zooepidemicus when the mares were in estrus. Uterine secretions were sampled, and sampling was immediately followed by a uterine lavage at 5 and 24 h after bacterial inoculation. During a sub...
Ultrasonographic measurement of stallion accessory sex glands and excurrent ducts during seminal emission and ejaculation. Accessory sex gland and excurrent duct activity of 10 stallions was visualized with transrectal ultrasonography during seminal emission and ejaculation. First and last penile urethral contractions were defined as the start (0 sec) and end of ejaculation, respectively. Duration of ejaculation (n = 63 ejaculations) averaged 6.15 +/- 2.98 sec. Penile urethral contractions occurred an average of 6.2 +/- 2.2 times per ejaculation. Right bulbourethral glands (n = 19 ejaculations) alternated repeatedly from ovoid to flattened, irregular shapes during ejaculation. Cross-sectional area of right bulbour...
Modulation of gonadotropin-releasing hormone-stimulated luteinizing hormone release in cultured male equine anterior pituitary cells by gonadal steroids. The objective of the present study was to determine whether the testicular steroids, i.e., testosterone (T), dihydrotestosterone (DHT), estradiol (E2), estrone (E1), and estrone sulfate (E1SO4), play a physiological role in regulating LH release in the male horse by direct actions at the anterior pituitary gland. Enzymatically dispersed anterior pituitary cells from stallions (n = 4) or geldings (n = 3) were cultured for 48 h in alpha-modified Eagle's medium containing 10% steroid-free horse medium. To determine the effects of the steroids on the LH response to GnRH, the cells were incubated f...
In vitro maturation of horse oocytes: characterization of chromatin configuration using fluorescence microscopy. The chromatin configuration of resting horse oocytes and the time course of in vitro oocyte maturation was characterized using a fluorescent, DNA-specific label. Oocytes were classified as having either compact (CP) or expanded (EX) cumuli at the time of collection. Centrifugation of oocytes was effective in allowing visualization of the germinal vesicle. Two main chromatin configurations were found in oocytes known to have a germinal vesicle: condensed chromatin (CC), in which the chromatin formed a dense mass surrounding the nucleolus; and fluorescing nucleus (FN), in which the entire nucleu...
Breed differences in circulating equine relaxin. Equine relaxin has been previously determined in a small number of pregnant Thoroughbred mares. To better define the normal pregnancy pattern of relaxin, the current study reports on a much larger number of mares. It also was designed to determine if all equids have the same gestational pattern of relaxin secretion. Plasma samples were collected weekly in 24 Standardbred mares, every 7-10 days in 10 pony mares, and daily in late pregnancy from 16 burros. Standardbreds had higher concentrations of relaxin than that reported for Thoroughbreds during most of gestation and did not exhibit the midp...
Prostaglandin E2 secretion by oviductal transport-stage equine embryos. This study was conducted to identify embryonic products whose secretion was temporally associated with the oviductal transport period of the mare. Chemicals secreted by oviductal-transport-stage equine embryos were identified by incubating Day 6 or Day 7 early uterine embryos with 35S-methionine/cysteine, 3H-progesterone, or 3H-arachidonic acid for 24 h, and subsequently identifying radioactively labeled proteins (SDS-PAGE; n = 3 embryos), steroids (HPLC; n = 3 embryos), or prostaglandins (HPLC; n = 3 embryos) in the culture medium. Early uterine embryos secreted 116.1 +/- 45.5 pg of prostagla...
Effects of carbohydrates on the pharmacokinetics and biological activity of equine chorionic gonadotropin in vivo. The sialylation of eCG was examined to determine its influence on the in vivo metabolism and biological activity of the molecule. Sialic acid was decrementally removed from eCG by incubation with agarose-linked neuraminidase for varying time periods. Pharmacokinetic parameters for the disappearance of 4,000 IU (267 micrograms) of three desialylated eCG preparations (20%, 53%, and 80% sialic acid removed) and control eCG were determined in sheep. The clearance rate of eCG increased (p less than 0.05) with each decrement of sialic acid. The removal of 53% sialic acid enhanced the distribution of...
Prostaglandin E2 hastens oviductal transport of equine embryos. The hypothesis that treatment of pregnant mares with prostaglandin E2 (PGE2) hastens the oviductal transport of equine embryos was tested by treating bred mares with PGE2 on Day 3 after ovulation and subsequently measuring the rate of hastened oviductal transport (estimated by the uterine embryo recovery rate on Day 4 after ovulation). In a preliminary, noncontrolled experiment, oviductal transport was apparently not hastened after intramuscular, intrauterine, or intraperitoneal PGE2 administration to bred mares (0/6, 0/3, and 0/3 mares, respectively). Oviductal transport appeared to be hasten...
Season but not age affects Sertoli cell number in adult stallions. To evaluate the effect of age and season on Sertoli cell number per paired testes, ratio of germ cells per Sertoli cell, and daily sperm production, testes were obtained from 184 adult (4-20 yr) stallions at slaughter throughout one year. Numbers of Sertoli cells or germ cells were derived from nuclear volume density, volume of individual nuclei, and parenchymal volume. Germ cell to Sertoli cell ratios were calculated from cell numbers. Regression analysis was used to detect age-related differences in the breeding season (May-Jul) or throughout the year. A two-way analysis of variance was used...
Biological and immunoactive substances resembling chorionic gonadotropin are present in full-term horse and zebra placentas. This study describes the presence of immunoactive and bioactive eCG-like material in full-term placentas of both domestic horses and zebras. Term placental extracts were immunoreactive in an LH monoclonal antibody RIA, and methods successfully used previously for the purification of eCG and eLH were employed to further concentrate the immunoreactive materials to the point where additional characterization studies could be performed. Sufficient equine material was obtained to perform a final fractionation on a concanavalin A Sepharose column yielding an unadsorbed fraction, e17A, and an adsorbe...
Effects of steroid administration on pituitary luteinizing hormone and follicle-stimulating hormone in ovariectomized pony mares in the early spring: pituitary responsiveness to gonadotropin-releasing hormone and pituitary gonadotropin content. These experiments tested the hypothesis that administration of steroid hormones to ovariectomized (OVX) mares during the vernal transition to the breeding season would influence LH and FSH secretion. Circulating gonadotropin concentrations, response to exogenous GnRH, and pituitary gonadotropin content were monitored. Experiments 1 and 2 were conducted, beginning 10 March, and 3 February, respectively, utilizing a total of 30 long-term OVX pony mares. In experiment 1, mares were administered vehicle (n = 5) or estradiol-17 beta (E2, n = 5, 5 mg/3 ml sesame oil), twice daily for 16 days. Blood ...
Zebra chorionic gonadotropin: partial purification and characterization. Six samples of pregnant zebra (z) serum from the first and second trimesters of pregnancy were analyzed by RIA and shown to have chorionic gonadotropin levels comparable to that of the mare (0.9-5.3 micrograms/ml); first trimester levels in most cases were higher than second trimester levels. A pool of the sera (10 ml) was fractionated by methods previously employed for the purification of equine (e) and donkey (d) chorionic gonadotropin to achieve a concentration of the zebra chorionic gonadotropin (zCG). A yield of 1.0 mg of glycoprotein was obtained. HPLC analysis of the material indicated ...
Follicle-stimulating hormone pulse amplitude decreases with the onset of the breeding season in the mare. The relationship between daily mean FSH concentrations in serum and the pattern of FSH detected by frequent sampling for 12-h periods (samples every 15 min) was examined in five mares during the transition into the breeding season. The five mature anestrous mares were exposed to a natural increase in daylength. Blood samples were collected daily from February 1 until the first ovulation of the breeding season (April 14 +/- 3.7 days, Mean +/- SEM). Periods of frequent blood collection were performed every two weeks. Blood samples were obtained daily by jugular venipuncture or jugular cannula (f...
Seasonal differences in equine spermatocytogenesis. Spermatocytogenesis plays a pivotal role in regulation of spermatogenesis; however, its details remain relatively obscure in nonrodent species. The equine testis contains approximately 100% more spermatogonia in summer than in winter and appears to be a good model to identify the flexible components of spermatocytogenesis that cause seasonal changes in daily sperm production. Testes were taken from horses in the winter (n = 47) and in summer (n = 43). Tissues were fixed by glutaraldehyde-perfusion and submission in osmium, embedded in Epon or methacrylate, sectioned at 0.5 micron or 5 microns,...
Effect of aspiration of the preovulatory follicle on luteinization, corpus luteum function, and peripheral plasma gonadotropin concentrations in the mare. Follicular fluid from small- to medium-sized follicles has been shown to have an inhibiting effect on luteinization of granulosa cells in vitro. This study was conducted to investigate the effect of in vivo removal of follicular fluid on luteinization, peripheral gonadotropin concentrations, and ovulation of secondary follicles in the mare. Follicular fluid was aspirated from the preovulatory follicles of mares when the diameter of the follicle was 30-34 mm (Group A), 35-39 mm (Group B), or 40-44 mm (Group C). Mares in Group D served as controls and the preovulatory follicle was not aspirated....
Different combinations of regulatory elements may explain why placenta-specific expression of the glycoprotein hormone alpha-subunit gene occurs only in primates and horses. Expression of the glycoprotein hormone alpha-subunit gene occurs in the pituitary of all mammals but in placenta of only primates and horses. In humans, two different elements, termed upstream regulatory element (URE) and cAMP response element (CRE), are required for placenta-specific expression of the alpha-subunit gene. The URE binds a protein unique to placenta whereas the CRE binds a ubiquitous protein. Comparative analysis of the promoter-regulatory region of the alpha-subunit gene from a number of mammals indicates that a functional URE has been retained and suggests the potential for pl...
Partial purification and characterization of rhinoceros gonadotropins, growth hormone, and prolactin: comparison with the horse and sheep. The rhinoceros is an endangered species related to the horse family. Little is known of its reproductive endocrinology. The objectives of this study were to partially purify rhinoceros pituitary hormones, determine which assays could be used for their assessment, and to ascertain whether rhinoceros LH possesses the intrinsic FSH activity of equine LH. A single pituitary each from a White (1.3 g) and a Black (1.2 g) Rhinoceros was homogenized and extracted (pH 9.5), then subjected to pH and salt fractionation, and ion-exchange chromatography (DEAE and Sephadex SP-C50) to yield partially purifie...
Equine granulosa-theca cell tumors express inhibin alpha- and beta A-subunit messenger ribonucleic acids and proteins. The association of equine granulosa-theca cell tumors with atrophied contralateral ovaries and abnormal estrous cycles suggests that these tumors produce hormones that affect pituitary gonadotropin production. Because inhibin, a heterodimer protein secreted by granulosa cells, decreases FSH production, we examined the presence of inhibin alpha- and beta A-subunits and their mRNAs in ovarian tumors obtained from three mares. These tumors contained neoplastic cords and nodules, multiple fluid-filled cysts, and a predominance of neoplastic granulosa cells. Reduced proteins from tumor-conditioned ...
Suppression of lymphocyte proliferation by a greater than 30,000 molecular weight factor in horse conceptus-conditioned medium. In this experiment we have identified and partially characterized the immunosuppressive activity of preimplantation horse conceptus-conditioned medium (HCCM). Horse conceptuses were nonsurgically flushed from mares at Days 9-10 (n = 6), 15-16 (n = 3), and 25-26 (n = 3). After incubating the conceptuses for 24 h in RPMI-1640 supplemented with 15% fetal calf serum (FCS) and 1% penicillin/streptomycin, HCCM was obtained from cultures and tested for immunosuppressive activity in lymphocyte proliferation assays. Peripheral blood lymphocytes obtained from randomly selected mares were stimulated with...