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Topic:Spermatogenesis
Spermatogenesis in horses is the process by which spermatozoa are produced in the testes. This complex sequence of events involves the proliferation and differentiation of germ cells, supported by Sertoli cells and influenced by hormonal regulation. Key stages include spermatogonial mitosis, meiosis, and spermiogenesis, resulting in the formation of mature sperm cells. Hormones such as testosterone and follicle-stimulating hormone (FSH) are integral to regulating this process. This topic encompasses research studies and scholarly articles that explore the mechanisms, hormonal influences, and factors affecting spermatogenesis in horses, as well as its implications for equine reproduction and breeding management.
Immunohistochemical localization of the spermadhesin AWN-1 in the equine male genital tract. Spermadhesins are proteins with various functions in sperm capacitation and zona pellucida binding. In this study the cellular localization of the spermadhesin AWN-1 has been examined in the equine male genital tract. Results obtained by immunohistochemical methods reveal that in the horse AWN-1 is synthesized in spermatogonia, in the rete testis, the ductus epididymidis and the seminal vesicles. These findings indicate that the cellular origin of spermadhesins is species-specific.
Expression of inhibin alpha-subunit in horse testis. Inhibin is believed to play roles in the pituitary secretion of FSH and in the paracrine regulation of testicular function. Although it has been generally accepted that inhibin is produced in Sertoli cells, there was a recent evidence for the localization of inhibin in Leydig cells of primates, rat and sheep. However, there is no report on the expression of inhibin in the adult horse testis. Therefore, using immunohistochemistry, western blotting and in situ hybridization techniques, the present study examined inhibin alpha-subunit (Ih-alpha) expression in the adult horse testis. For the detec...
[The development of the horse testis]. The aim of the study was to answer the open questions concerning the development of the horse's testis. This study revealed that the seminiferous tubules originate from the sex cords of the coelomic epithelium and Leydig cells from the proximal part of mesonephric nephrons, whereas the rete and the ductuli efferentes derive from intermediate and distal parts of the mesonephric tubules. During the development the Leydig cells undergo an enormous proliferation due to the PMSG secretion in the mare. The proliferation of these cells prevent the deep penetration of the rete into the medulla and is ...
Prostasome-like particles in stallion semen. Human semen contains membranous vesicles called prostasomes. They are secreted by the prostate gland and contain large amounts of cholesterol, sphingomyelin, and Ca2+. Prostasomes enhance the motility of ejaculated spermatozoa and are involved in a number of additional biological functions. No prostasome-like vesicles have been described in horse semen up to now. We have demonstrated the presence of prostasome-like vesicles in the equine semen and characterized them as to size, morphology, and lipid composition; we have found that they are similar to human prostasomes in many respects. We prop...
Testicular inhibin in the stallion: cellular source and seasonal changes in its secretion. The cellular localization of inhibin alpha, betaA, and betaB subunits, 3beta-hydroxysteroid dehydrogenase (3beta-HSD), and cytochrome P450 aromatase (aromatase) in stallion testes was investigated. In addition, detailed seasonal changes in circulating immunoreactive (ir)-inhibin were investigated in correlation with testosterone, estradiol, LH, and FSH. Inhibin alpha subunit-positive staining was observed in Sertoli cells, and more clearly positive staining was noted in Leydig cells. Inhibin betaA and betaB subunits were also stained in both types of cells. Immunoreactivity of 3beta-HSD and ar...
Lectin-staining pattern in extratesticular rete testis and ductuli efferentes of prepubertal and adult horses. This study was undertaken to determine the lectin affinity of the extratesticular rete testis and ductuli efferentes epithelial cells in adult and prepubertal horses, using ten different lectin horseradish peroxidase conjugates: Con-A, LCA, WGA, GSA-II, SBA, PNA, RCA-I, DBA, UEA-I, and LTA. In some cases, treatments with sialidase and KOH preceded the lectin staining. In sexually mature and immature horses the results showed the presence of different kinds of sialoglycoconjugates with the terminal sialic acid linked to D-GalNAc and beta-D-Gal residues in the rete testis. In the apical surface ...
Cytoplasmic extrusion and the switch from creatine kinase B to M isoform are completed by the commencement of epididymal transport in human and stallion spermatozoa. Although in several species there is a relationship between epididymal sperm transport and fertility, in human in vitro fertilization (IVF), spermatozoa recovered from the caput epididymidis or even the rete testis are fertile. We studied two objective markers of sperm maturity in the sperm of men and stallions: creatine kinase (CK) concentrations, which are a measure of cytoplasmic retention in immature spermatozoa, and the ratio of CK-M and CK-B isoforms (% CK-M/[CK-M + CK-B]), which is proportional to the incidence of mature sperm. The CK markers and the fertilizing function are closely rel...
Gonadal and pituitary responsiveness of stallions is not down-regulated by prolonged pulsatile administration of GnRH. The objective of this study was to determine if prolonged pulsatile administration of homologous gonadotropin-releasing hormone (GnRH) at therapeutic or 5x therapeutic doses would cause down-regulation of the stallion's hypothalamic-pituitary-testicular axis. Fifteen stallions were randomly assigned to three treatment groups (n=5/group) and received a 0.5 ml subcutaneous dose of saline (group 1), 50 microg GnRH (group 2), or 250 microg GnRH (group 3) every 2 hours for 75 days. Weekly evaluations of follicle stimulating hormone, luteinizing hormone, and testosterone and monthly evaluations of d...
[Clinical aspects of fertility in stallions]. The studies described in this thesis investigated the factors that can affect the fertility of stallions. The introduction describes the male gamete and the processes that occur during maturation of sperm and fertilization. Methods to evaluate the quality of sperm and ova are then discussed. Fertility can be expressed in various ways and is also affected by many factors such as the stallion, the mare and management factors. The fertility of stallions is usually assessed a good year after they have served mares, because then the number of foals is known. However, it would be preferable to be ab...
Biochemical and conformational characterisation of HSP-3, a stallion seminal plasma protein of the cysteine-rich secretory protein (CRISP) family. HSP-3 is a member of the cysteine-rich secretory protein (CRISP) family from stallion seminal plasma. We report a large-scale purification protocol for native HSP-3. This protein is a non-glycosylated polypeptide chain with a pI of 8-9 and an isotope-averaged molecular mass of 24987 +/- 3 Da. The molecular mass of HSP-3, determined by equilibrium sedimentation, is 26 kDa, showing that the protein exists in solution as a monomer. The concentration of HSP-3 in the seminal plasma of different stallions ranged from 0.3 to 1.3 mg/ml. On average, 0.9-9 million HSP-3 molecules/cell coat the postacros...
Hydrolysis of extracellular adenine nucleotides by equine epidydimal spermatozoa. Ectoenzymic activities capable of hydrolyzing ATP sequentially to adenosine are present on equine epidydimal spermatozoa membranes. Kinetic parameters for ATPase, ADPase and 5'-nucleotidase were obtained by analysis of progress reactions curve when ATP, ADP and AMP were supplied as initial substrates. These values are not different from those found when the substrates were supplied from the preceding reactions. Feed-forward inhibition on 5'-nucleotidase by ATP/ADP was taken into account to fit simulated data to the experimental results. None of the substrates supplied by the preceding reaction...
Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa. Evidence is provided that the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase is covalently linked to the fibrous sheath. The fibrous sheath is a typical structure of mammalian spermatozoa surrounding the axoneme in the principal piece of the flagellum. More than 90% of boar sperm glyceraldehyde 3-phosphate dehydrogenase activity is sedimented after cell disintegration by centrifugation. Detergents, different salt concentrations or short term incubation with chymotrypsin do not solubilize the enzyme, whereas digestion with trypsin or elastase does. Short term incubation with trypsin...
Estrogen concentrations in semen of the stallion. Large amounts of estrogens are secreted by the tests of the mature stallion. In a recent study by Claus et al. [Claus, Dimmick, T., Gimenez, T., Hudson, L.W., 1992. Estrogens and prostaglandin F2 alpha in the semen and blood plasma of stallions. Theriogenology 38, 687-693.], it was stated that high levels of estrogens were also present in semen. As a preliminary step to study possible implications for fertility in the stallion, we have measured estrone sulphate (E1S), the principal estrogen in blood, in both seminal plasma and spermatozoa. Semen was collected from four Standardbred stallions a...
Regional differentiation of the blood-epididymis barrier in stallion (Equus caballus). The occurrence, localization and ultrastructural characteristics of a blood-tissue barrier throughout the stallion proximal seminal excurrent duct system were studied by the exclusion of electron-dense tracers and freeze-fracture techniques. Striking differences were observed in the distribution of lanthanum tracer and in the geometrical organization of the zonulae occludentes along the ductus efferentes, epididymides and vas deferens. The zonulae occludentes domain, the principal structural component of the blood-epididymis barrier, differed in permeability, width and strand numbers along the...
Isolation and characterization of heparin- and phosphorylcholine-binding proteins of boar and stallion seminal plasma. Primary structure of porcine pB1. In the bovine, seminal plasma heparin-binding proteins bind to sperm lipids containing the phosphorylcholine group and mediate the capacitating effects of heparin-like glycosaminoglycans during sperm residence in the female genital tract. We report the characterization of heparin- and phosphorylcholine-binding proteins of stallion and boar seminal plasma. Horse seminal plasma proteins HSP-1 and HSP-2, and boar protein pB1, belong to the same family as the bull heparin- and phosphorylcholine-binding proteins BSP-A1/2, BSP-A3, and BSP-30K. We have determined the amino acid sequence and posttrans...
Determination of daily sperm production in stallion testes by enumeration of germ cells in homogenates. Thirty adult stallion testes were selected with high (n = 15) and low (n = 15) Daily Sperm Production (DSP)/testis. Parenchymal samples were prepared for morphometric analysis, and the numbers of germ cells and Sertoli cells were determined. Testicular samples were homogenized, and germ cells and Sertoli cells were enumerated using phase contrast microscopy. Numbers of germ cells and Sertoli cells and potential DSP during spermatogenesis were determined. Significant correlations existed between morphometric and homogenate determinations of number per testis of preleptotene, leptotene plus zygo...
Effect of 19-norandrostenololylaurate on semen characteristics of colts. The long-term effects of the anabolic steroid 19-norandrostenololylaurate on semen characteristics of Finnhorse colts were studied in 3 experiments. Semen was collected initially at 24 months of age and then twice a year. In experiment I, 500 mg or 100 mg of steroid per animal was given every 3rd week from 12 or 16 months to 24 months of age. In colts treated with 500 mg of anabolic steroid every 3rd week, azoospermia was observed in 3 out of 5 colts in the first semen collections, immediately after the end of treatment. The other 2 colts had low sperm numbers and a high percentage of proximal...
Effect of 19-norandrostenololylaurate on testicular growth in colts. The effect of anabolic steroid on testicular growth was investigated in 3 experiments. In experiment I, 500 mg of the anabolic steroid was given to 4 colts and 100 mg to another 4 colts, every 3rd week, starting at age 16 months and ending at age 24 months. Six colts served as controls. Both treatments decreased total scrotal width (TSW) within 6 weeks. Seasonal testicular growth during spring partly overcame the effect of steroid treatment. Cessation of anabolic steroid treatment was followed by testicular growth at the same time as TSW in untreated colts was decreasing by virtue of the effec...
Primary structure of stallion seminal plasma protein HSP-7, a zona-pellucida-binding protein of the spermadhesin family. The primary-structure of HSP-7, a 14-kDa protein isolated from stallion seminal plasma, has been determined, HSP-7 belongs to the spermadhesin protein family, shares 98% sequence identity with the boar seminal plasma protein AWN, and, like its boar homolog, displays zona-pellucida-binding activity. Despite these conserved structural and functional features, the equine and porcine spermadhesins differ in their topography on spermatozoa.
Seasonal rhythm of semen characteristics of a Brazilian breed (“Mangalarga”) stallion. An attempt has been made to define semen seasonality in a horse in the Southern Hemisphere. Repeated measurements of three variables in the semen were made for 36 months (Jan/90-Dec/92) in a 21-year old "Mangalarga" stallion living under natural photoperiod and temperature conditions in a farm situated in São José do Rio Pardo, São Paulo, Brazil (latitude 21 degrees) 36'S; longitude 46 degrees 53' W). The horse fed on natural pasture and a nutritionally balanced feed twice a day (11:00 and 17:00 h). Water and mineral supplement were available ad libitum. Semen was collected almost daily by ...
[The effect of the season and sexual stress on the concentration of testosterone and estradiol-17beta in the seminal plasma of stallions]. Semen from three stallions was collected weekly for six months (December through May) to determine semen parameters and hormones (testosterone and estradiol-17 beta) in seminal plasma. Once a month three ejaculates were collected at intervals of one hour and examined accordingly. Testosterone and estradiol-17 beta were also determined in peripheral blood plasma (V. jugularis) collected twice a week. Semen parameters (volume, gel-free volume and sperm concentration) were clearly influenced by season. The testosterone concentration in peripheral blood plasma was lowest during December whereas in...
Seminal plasma affects membrane integrity and motility of equine spermatozoa after cryopreservation. Effects of seminal plasma on post-thaw motility and membrane integrity of cryopreserved horse spermatozoa were investigated. Carboxyfluorescein diacetate staining was used for the assessment of sperm membrane integrity. Adding 30% of seminal plasma from stallions with high post-thaw sperm motility to ejaculates from stallions with low post-thaw sperm motility increased progressive motility from 24.0 +/- 1.6 to 34.5 +/- 1.9% (P < 0.05) and membrane integrity from 27.0 +/- 2.1 to 34.3 +/- 2.3% membrane-intact spermatozoa (P < 0.05). Conversely, the addition of seminal plasma from stallions...
Prolactin involvement with the increase in seminal volume after sexual stimulation in stallions. To test the hypothesis that prolactin mediates the increase in seminal volumes induced by sexual stimulation in stallions, semen was collected from six stallions every other day for 26 d. The last eight collection days were treatment days. For each stallion, four treatments were randomly assigned to the first four of the eight treatment collection days, and then repeated in reverse order on the last four collection days; 1) CONTROL: semen collected per normal procedures; 2) Sexually stimulated: stallions were presented to mares in a chute for 10 min before collection; 3) Bromocriptine (dopamin...
Suppression of testicular function using two dose rates of a reversible water soluble gonadotrophin releasing hormone (GnRH) vaccine in colts. To investigate the effect of two dose rates (200 and 400 ng) of a gonadotrophin releasing hormone (GnRH) vaccine on testicular function. Methods: A vaccination dose rate experiment. Methods: Two injections were administered 4 weeks apart to six colts in each treatment group. To maintain immunosuppression until the end of the breeding season, a third injection was given if antibody titres fell below 1000. Results: Effective antibody titres were present for 12 to 27 weeks. Testosterone concentrations decreased from 2.22 to 0.31 nmol/L 6 weeks after primary vaccination. Androstenedione concentrat...
Characterisation of proteins in the seminal plasma of stallions, geldings and supplemented with testosterone. The major proteins in stallion seminal plasma were characterised by two-dimensional polyacrylamide gel electrophoresis, and compared with the patterns of proteins in normal geldings (castrated males) and geldings supplemented with testosterone. The major proteins or groups of proteins identified according to their approximate relative molecular weight in kilodaltons (kDa) and apparent isoelectric point (pl) were: 1) 60 kDa. pl 7; 2) 23 kDa, pl 4-5; 3) 25-30 kDa, pl 5.5-6; 4) 23 kDa, pl 7-8; and 5) 15-20 kDa, pl 6-7.5. Protein groups 1 and 2 were more prominent in the seminal plasma from the st...
In vitro induction of acrosome reactions in stallion spermatozoa by heparin and A23187. The ability of the glycosaminoglycan, heparin, and the calcium ionophore, A23187, to induce acrosome reaction in equine spermatozoa was assessed using semen from 3 warmblood stallions of known high fertility. After collection of semen, the spermatozoa were washed and incubated in vitro with heparin or A23187. Incubation periods were 0, 4, 6 or 8 h with 0, 1, 10 or 100 microg/ml heparin or 0, 10, 30 or 60 min with 0, 0.01, 0.1, 1 or 10 microM A23187, respectively. Acrosome reactions were determined by staining the spermatozoa with naphthol yellow S plus erythrosin B, and sperm viability was ass...
Methods for induction of capacitation and the acrosome reaction of stallion spermatozoa. Methodologies to capacitate bovine spermatozoa, induce the acrosome reaction, and fertilize bovine oocytes in vitro have been established. The capability to do the same with stallion spermatozoa, however, is not available. Several different methods have been used to capacitate stallion spermatozoa with variable results. More basic research needs to be done to establish in vitro conditions necessary to capacitate and induce an acrosome reaction in stallion spermatozoa. Although much progress can be expected in this area, it is unlikely that the general practitioner will use these technologies i...
Immunohistolocalization of the carbonic anhydrase isoenzymes (CA-I, CA-II, and CA-III) in the reproductive tract of male horses. To elucidate locations of cytosolic carbonic anhydrase isoenzyme (CA-I, CA-II, and CA-III)-positive epithelial cells in equine male reproductive organs. Methods: Descriptive and immunohistochemical study. Methods: 4 clinically normal male horses. Methods: The testis (seminiferous tubules, rete tubules), epididymis (initial, middle, and terminal segments), proximal and distal portions of the ductus deferens, ampulla ductus deferentis, seminal vesicle, prostate, and bulbourethral gland were excised from euthanatized horses after administration of an overdose of pentobarbital. The tissue specimen...
