Reproduction in horses encompasses the biological processes and mechanisms involved in the breeding and development of equine offspring. This includes the study of reproductive anatomy, physiology, and endocrinology in both mares and stallions. Key areas of interest include the estrous cycle, ovulation, conception, gestation, and parturition. Researchers also examine factors influencing fertility, reproductive technologies such as artificial insemination and embryo transfer, and management practices that impact reproductive success. This page compiles peer-reviewed research studies and scholarly articles that explore the physiological, genetic, and environmental aspects of equine reproduction.
Griggers S, Paccamonti DL, Thompson RA, Eilts BE.Urospermia has been reported as a cause of infertility in numerous species. The detrimental effects of urine on spermatozoa are due, at least in part, to changes in pH and osmolarity. Semen was collected and subjected to conditions of varying pH (Experiment 1), of varying osmolarity (Experiment 2), and various quantities and concentrations of urine (Experiment 3) and effects on motility were recorded. Finally, semen was contaminated with urine and then either of 2 semen extenders was added, with or without centrifugation, in an attempt to alleviate the detrimental effect of urine on motility (...
Thway TM, Clay CM, Maher JK, Reed DK, McDowell KJ, Antczak DF, Eckert RL, Nilson JH, Wolfe MW.Immortalized cell lines have many potential experimental applications including the analysis of molecular mechanisms underlying cell-specific gene expression. We have utilized a recombinant retrovirus encoding the simian virus-40 (SV-40) large T antigen to construct several immortalized cell lines of equine chorionic girdle cell lineage - the progenitor cells that differentiate into the equine chorionic gonadotropin (eCG) producing endometrial cups. Morphologically, the immortalized cell lines appear similar to normal chorionic girdle cells. Derivation of the immortalized cell lines from a cho...
Lima SB, Verreschi IT, Ribeiro Neto LM.Equine unsaturated estrogens are the main components of brand formulations indicated for hormonal replacement therapy in both hypogonadic and postmenopausal women. These hormones are produced by the fetoplacental unit during equine gestation. A method is described for the quantitative determination of equilenin (EL), equilin (EQ), 17alpha-dihydroequilin (17dEQ), and estrone (El) in the plasma of a pregnant mare. Blood samples are obtained weekly during pregnancy by jugular venipuncture using sodium ethylenediaminetetracetic as the anticoagulant. For the quantitation of these estrogens, plasma ...
Acker DA, Curran S, Bersu ET, Ginther OJ.To describe the gross and histologic changes that develop in the equine embryo proper (ie, the portion of the embryo that becomes the fetus) from days 17 to 40 after ovulation and to compare the external features of equine embryos with those of porcine, ovine, and human embryos. Methods: 34 embryos collected from mixed-breed pony mares. Methods: External features for each embryo proper, including length, number of branchial arches, growth of appendages, face and head features, and body features, were examined, using a dissecting microscope, for embryos collected on days 17 to 40. Internal feat...
Allen WR.Four separate components combine to produce the progesterone and biologically active 5 alpha-reduced pregnanes needed to maintain pregnancy in the mare. The primary corpus luteum (CL) is prolonged beyond its cyclical lifespan by the down-regulation of endometrial oxytocin receptors to prevent activation of the luteolytic pathway and its waning progesterone production is supplemented from day 40 of gestation by the formation of a series of accessory CL which develop in the maternal ovaries as a result of the gonadotrophic actions of pituitary FSH and the equine chorionic gonadotrophin (eCG). Fr...
Tremoleda JL, Schoevers EJ, Stout TA, Colenbrander B, Bevers MM.Meiotic maturation of mammalian oocytes is a complex process during which microfilaments and microtubules provide the framework for chromosomal reorganisation and cell division. The aim of this study was to use fluorescence and confocal laser scanning microscopy to examine changes in the distribution of these important cytoskeletal elements and their relationship to chromatin configuration during the maturation of horse oocytes in vitro. Oocytes were cultured in M199 supplemented with pFSH and eLH and, at 0, 12, 24, and 36 hr after the onset of culture, they were fixed for immunocytochemistry ...
Dell'Aquila ME, Masterson M, Maritato F, Hinrichs K.There is a great variability in the success of horse oocyte maturation and fertilization among laboratories. This study was conducted to determine if the meiotic and developmental competence of horse oocytes could be dependent on the method of oocyte collection, i.e., aspiration of follicular fluid with a vacuum apparatus, or opening follicles and scraping the granulosa layer. Horse oocytes were recovered from abattoir ovaries by aspiration or scraping and classified as having compact (Cp), expanded (Ex), or partial (P) cumuli. In Experiment 1 (Part A in May and Part B in October), oocytes wer...
Ryan PL, Bennett-Wimbush K, Vaala WE, Bagnell CA.Tall fescue is one of the most widely grown forage grasses for horses in the United States. However, it is frequently infected with the endophyte Neotyphodium coenophialum which produces ergot alkaloids that cause severe adverse effects in the pregnant mare. The objectives of this study were to determine the effects of fescue toxicosis and fluphenazine on circulating relaxin in pregnant pony mares and evaluate the usefulness of relaxin as a monitor of treatment efficacy. Twelve mares were maintained on endophyte-infected tall fescue pasture. Group TRT (n = 6), received 25 mg of fluphenazine de...
Rigby SL, Barhoumi R, Burghardt RC, Colleran P, Thompson JA, Varner DD, Blanchard TL, Brinsko SP, Taylor T, Wilkerson MK, Delp MD.Persistent, postmating endometritis affects approximately 15% of mares and results in reduced fertility and sizable economic losses to the horse-breeding industry. Mares that are susceptible to postmating endometritis have delayed uterine clearance associated with reduced uterine contractility. Unfortunately, the mechanism for reduced uterine contractility remains an enigma. The present study examined the hypothesis that mares with delayed uterine clearance have an intrinsic contractile defect of the myometrium. Myometrial contractility was evaluated in vitro by measuring isometric tension gen...
Carneiro G, Lorenzo P, Pimentel C, Pegoraro L, Bertolini M, Ball B, Anderson G, Liu I.The effects of insulin-like growth factor-I (IGF-I) and its interaction with gonadotropins, estradiol, and fetal calf serum (FCS) on in vitro maturation (IVM) of equine oocytes were investigated in this study. We also examined the role of IGF-I in the presence or absence of gonadotropins, estradiol, and FCS in parthenogenic cleavage after oocyte activation with calcium ionophore combined with 6-dimethylaminopurine (6-DMAP), using cleavage rate as a measure of cytoplasmic maturation. Only equine cumulus-oocyte complexes with compact cumulus and homogenous ooplasm (n = 817) were used. In experim...
Katila T.The article reviews methods used for in vitro evaluation of sperm, with particular emphasis on frozen-thawed stallion sperm. The techniques, limitations of the methods and correlations with fertility results are discussed. Very few studies have tried to find correlation between fertility of frozen stallion semen and laboratory tests. It is difficult and expensive to inseminate an adequate number of mares to achieve statistically significant differences. Significant, but low correlations have been demonstrated between the foaling rate and subjective motility of sperm incubated for 2 h and 4 h a...
Nie GJ, Goodin AN, Braden TD, Wenzel JG.To determine whether administration of a microdose of prostaglandin at the BAI HUI acupuncture point offers any advantage over IM injections for luteolysis, ovulatory interval, or systemic response in mares. Methods: 17 mature cycling mares, 3 to 20 years of age and weighing 400 to 500 kg. Methods: Conventional and microdoses of the prostaglandin dinoprost tromethamine (PGF2alpha), the analogue cloprostenol, or sterile water (control) were administered to mares in 7 treatment groups. Treatments were assigned by dose, administration site (semimembranosus, semitendinosus, or lumbosacral region),...
Choi YH, Chung YG, Seidel GE, Squires EL.The objective was to compare culture media for in vitro maturation of equine oocytes and for in vitro culture of zygotes produced from IVF of partially zona-removed oocytes. Cumulus-oocyte complexes from slaughterhouse-derived ovaries were washed in m-Dulbecco's PBS and cultured in TCM-199, F10-DMEM or c-F10-DMEM (50% F10-DMEM + 50% F10-DMEM conditioned medium from culture of an equine trophoblast monolayer for 3 or 4 days). All media included FSH, LH, E2, and 10% FCS. After 28 to 30 h maturation, cumulus expansion was scored from 0 (no expansion) to 4 (fully expanded). Oocytes with a 1st pola...
Lear TL.Telomeres are a class of repetitive DNA sequences that are located at chromosome termini and that act to stabilize the chromosome ends. The rapid karyotypic evolution of the genus Equus has given rise to ten taxa, all with different diploid chromosome numbers. Using fluorescence in situ hybridization (FISH) we localized the mammalian telomere sequence, (TTAGGG)(n), to the chromosomes of nine equid taxa. TTAGGG signal was located at chromosome termini in all species, however additional signal was seen at interstitial sites on some chromosomes in the Burchell's zebra, Equus quagga burchelli, the...
Sartori Blanc N, Senn A, Leforestier A, Livolant F, Dubochet J.In human and other mammal sperm nuclei, DNA is packed in a highly condensed state, the structure of which remains unsolved. Cryoelectron microscopy of vitrified sections provides a first direct view of the local arrangement of the nucleoprotamine filament. DNA aligns in parallel in layers and its orientation rotates along a single-twist direction as in a cholesteric liquid crystal. The structure contains numerous defects, which introduce locally double-twist configurations. Destruction of the SS bonds with dithiotrehitol relaxes the twist and favors the extension of the hexagonal close packing...
Galosi CM, Vila Roza MV, Oliva GA, Pecoraro MR, EcheverrÃa MG, Corva S, Etcheverrigaray ME.Equine herpesvirus 1 (EHV-1) is the causative agent of abortion, perinatal foal mortality, neurological and acute respiratory diseases in horses. Conventional laboratory diagnosis involving viral isolation from aborted foetuses is laborious and lengthy and requires processing of samples within 24 h of collection, which is problematic for samples that come from long distances. The aim of this study was to develop a polymerase chain reaction (PCR) assay useful in Argentina to detect DNA sequences of EHV-1 in different tissues from aborted equine foetuses with variable quality of preservation and...
Watson ED, Barbacini S, Berrocal B, Sheerin O, Marchi V, Zavaglia G, Necchi D.Ninety five mares were inseminated with frozen semen either within 12 h before ovulation or within 8 h after ovulation. The effect of preovulatory versus postovulatory insemination (AI) on the subsequent detection of uterine fluid was studied. The overall pregnancy rate was 43% and this was not significantly influenced by preovulatory or postovulatory insemination. When mares were first examined 12 h after AI, 18 of 52 mares (35%) had accumulated uterine fluid. However, when mares were first examined 18 to 24 h after AI, only 6 of 43 mares (14%) had uterine fluid. Presence of intrauterine flui...
Baltsen M, Bøgh IB, Byskov AG.Meiosis activating sterols (MAS) are pre-cholesterol sterols that can be isolated from follicular fluid (FF-MAS) or testes (T-MAS). Meiosis activating sterols trigger the resumption of meiosis in cultured meiotically competent oocytes. In the present work MAS, cholesterol and progesterone were assayed by HPLC in follicular fluids collected from pony mares at fixed days after the last ovulation. Follicles were divided into two groups according to whether they were aspirated before or after Day 17 after the last ovulation. The latter group was further divided according to whether the follicle di...
Bae SE, Corcoran BM, Watson ED.Little is known about neurogenic regulation of uterine contractility in mares. The present study investigated the distribution of adrenergic and peptidergic nerves in the mare uterus. Samples from the uterine horn, body and cervix were collected from 18 cyclic mares for immunohistochemistry. The uterus was well supplied with adrenergic nerves. A large number of tyrosine hydroxylase- and dopamine beta-hydroxylase-immunoreactive nerve bundles and fibres were present in the myometrium and endometrium in all regions of the uterus and cervix. These adrenergic nerve bundles and fibres travelled para...
Rathi R, Colenbrander B, Bevers MM, Gadella BM.The primary aim of this study was to establish a flow cytometric technique for determining the capacitation status of stallion spermatozoa. To this end, a flow cytometric technique that demonstrates changes in plasma membrane fluidity; namely, merocyanine 540 staining, was compared with the more conventional Ca(2+)-dependent fluorescence microscopic technique, chlortetracycline (CTC) staining, for assessing capacitation status. In addition, the effect of bicarbonate/CO(2) on the progress of capacitation and the acrosome reaction (AR) and on temporal changes in sperm motility, with particular r...
Bergfelt DR, Gastal EL, Ginther OJ.The increase in LH concentrations at the time of the decrease in FSH concentrations during follicle deviation in mares was studied to determine the role of LH in the production of estradiol and immunoreactive inhibin (ir-inhibin). Ten days after ovulation, all follicles > or =6 mm were ablated, prostaglandin F(2 alpha) was given, and either 0 mg (control group, n = 15) or 100 mg of progesterone in safflower oil (treated group, n = 16) was given daily for 14 days, encompassing the day of diameter deviation. The follicular and hormonal data were normalized to the expected day of the beginning...
MacLeay JM.In 1992, a multiparous 13-year-old Thoroughbred mare and her 48-hour-old colt were examined because of possible neonatal isoerythrolysis (NI). Supportive treatment was administered, and the foal recovered without requiring a transfusion. According to the owners, the mare had delivered foals without incident during 1987 and 1991. The mare was barren during 1993, but in 1994, delivered a filly that developed severe NI. The foal was given 3 transfusions and eventually recovered without complications. Blood typing analysis of the mare and its foals indicated that all 4 foals were positive for the ...
Stout TA, Allen WR.Between at least day 9 and day 16 after ovulation the spherical equine conceptus migrates continuously throughout the uterine lumen, propelled by peristaltic myometrial contractions. This unusually long period of intrauterine movement ensures that the conceptus delivers its anti-luteolytic signal to the entire endometrium to achieve luteostasis. The present experiment tested the hypothesis that prostaglandins stimulate the myometrial contractions that result in the migration of the conceptus. Serial ultrasonographic examinations of the uteri of eight mares performed during 2 h periods between ...
Choi YH, Love CC, Varner DD, Thompson JA, Hinrichs K.Two different culture media (TCM-199 and follicular fluid), two activation treatments (10 and 50 micromol calcium ionophore l(-1)) and three culture periods with cycloheximide were evaluated to find effective culture conditions for activation of cumulus-free equine oocytes. Oocytes were collected by scraping the follicle walls of ovaries obtained from an abattoir. Oocytes with expanded cumuli were matured at 38.2 degrees C in a humidified atmosphere of 5% CO(2) in air, in either TCM-199 with 10% fetal bovine serum (FBS) and 5 microU FSH ml(-1), or in 100% follicular fluid derived from a preovu...
Thway TM, Wolfe MW.Primates and equids are the only species known to express the placental glycoprotein hormone, chorionic gonadotropin (CG), a heterodimeric glycoprotein composed of an alpha subunit linked to a hormone-specific beta subunit. The regulatory mechanisms involved in the induction of equine glycoprotein alpha subunit gene expression have not been identified. Epidermal growth factor (EGF) receptor is known to transduce signals that alter a number of different cellular functions (cell proliferation, differentiation, hormone secretion, and gene regulation). In the present study, we investigated the reg...
Aurich C, Gerlach T, Aurich JE, Hoppen HO, Lange J, Parvizi N.In the non-breeding season, LH release is reduced via dopaminergic systems in the ram. On the other hand, our previous studies demonstrated an opioidergic inhibition of LH release in stallions outside the breeding season. Thus, in the present study we investigated the dopaminergic regulation of LH and prolactin secretion in stallions, considering interactions between dopamine and opioids. To achieve this, stallions (n=8) were treated with the dopamine antagonist sulpiride (0.6 mg/kg), the opioid antagonist naloxone (0.5 mg/kg), sulpiride plus naloxone or saline in December, March and June. Two...
Ramsay JD, Leib SR, Orfe L, Call DR, Tallmadge RL, Fraser DG, Mealey RH.Development of an accurate and efficient molecular-based equine MHC class I typing method would facilitate the study of T lymphocyte immune responses in horses. Here, a DNA microarray was designed to detect expressed classical MHC class I genes comprising serologically defined equine leukocyte antigen (ELA)-A haplotypes represented in a closed Arabian horse breeding herd. Initially, cloning and sequencing of RT-PCR products were used to identify sequences associated with the ELA-A1, A4, and W11 haplotypes, and one undefined haplotype, in six horses. Subsequently, sequence-specific, conserved (...
van Niekerk FE, van Niekerk CH.In the main experiment the total daily protein intake and quality (essential amino-acids) was varied in 4 groups of mares. The incidence of oestrus in mares during the transitional period was unaffected by protein nutrition. Ovarian activity, as evaluated by follicular development and size of the ovaries, was affected. Mares that received low-quality protein (Groups 1 and 2) had a higher number of smaller follicles (< 10 mm) that developed during the transitional period compared to mares on a high-quality protein intake (Groups 3 and 4). The mares that received the high quality protein ovulate...
Bertelsmann H, Sieme H, Behne D, Kyriakopoulos A.In the sperm nuclei, of mammalian species selenium has been found only in the form of sperm nuclei glutathione peroxidase (snGPx) where it is most likely bound to the chromatin of spermatozoa. Over 80% of selenium in sperm is bound to the selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) in the midpiece of rat sperm. Zinc in sperm is mainly contained in the outer dense fiber (ODF) proteins of the flagella of mammalian spermatozoa. In the sperm nuclei, zinc is predominately located in the chromatin to the protamine proteins. In order to investigate if the insertion of zinc...
McClure JJ, Koch C, Traub-Dargatz J.A red cell antigen of donkeys and mules was identified using antibodies in serum from a mare which produced a mule foal affected with neonatal isoerythrolysis (NI). Subsequently antibodies with similar activity were identified in the sera of other mares which had produced mule foals and were produced by immunization of horses with blood from donkeys. The antigen detected by these antibodies does not correspond to any recognized horse red cell alloantigen. This may be a xenoantigen since all donkeys (and mules) tested have shared this antigen and all horses tested have lacked the antigen. The r...
Bollinger A, Eckert J, Rüttimann B, Becker F.Intermittent claudication (IC) due to arterial occlusive disease was first diagnosed by the French veterinary surgeon Jean-François Bouley jeune in a horse drawing a cabriolet in the streets of Paris as early as 1831. The animal was repeatedly exercised and always started to limp with the hind legs at similar work loads. Autopsy revealed partially thrombosed aneurysm of the abdominal aorta and occlusions of both femoral arteries which were correctly identified as the cause of IC. In 1858 the famous neurologist Jean-Martin Charcot working at the Salpêtrière in Paris first discovered the cond...
Watson ED, Bae SE, Thomassen R, Thomson SR, Woad K, Armstrong DG.The period between seasonal anoestrus and cyclicity is characterized in many mares by cyclical growth and regression of large dominant follicles. The insulin-like growth factor (IGF) system plays a key role in follicular growth and regression; therefore, we hypothesized that changes in the IGF system and its binding proteins would modulate onset of cyclicity in mares. Ovaries were obtained from pony mares on the day after detection of an actively growing 30 mm transitional anovulatory follicle, and also at the second or third oestrus of the breeding season on the day after the preovulatory fol...
Hejmej A, Wiszniewska B, Kosiniak-Kamysz K, Sadowska J, Bilińska B.Distribution of androgen receptors (ARs) in the epididymal duct and prostate of three entire stallions and one bilaterally cryptorchid horse was studied immunohistochemically using a polyclonal rabbit antiserum against the ARs. In both the healthy stallions and the cryptorchid, the epithelial cells of the epididymides showed nuclear staining for ARs. The intensity of AR-staining in the principal cells of the epididymis was stronger than that of the basal cells. In the prostate, the glandular secretory cells were moderately stained whereas the basal cells expressed weak AR-staining. Immunostain...
Tunón A-M , Katila T, Magnusson U, Nummijärvi A, Rodriguez-Martinez H.The T-cell response after the introduction of semen into the uterine cavity in the mare was studied by examining, immunohistochemically, the distribution of helper T-cells (CD4+) and cytotoxic T-cells (CD8+) in endometrial biopsy specimens. Endometrial tissue samples were obtained from twenty-five gynecologically healthy mares during estrus before and 6 or 48 h after deposition of a single dose of stallion semen. An increase (P=0.04) in the number of helper T-cells (CD4+) compared to pre-insemination values was observed in the uterine body in both groups, 6 and 48 h, after insemination. No sig...
Gallardo Bolaños JM, Balao da Silva C, MartÃn Muñoz P, Plaza Dávila M, Ezquerra J, Aparicio IM, Tapia JA, Ortega Ferrusola C, Peña FJ.To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h ...
Kraemer DC.The procedures that are collectively referred to as embryo transfer (ET) have many uses. They were first used as research tools to study fetal-maternal physiology. Since the first successful mammalian embryo transfer in 1890, ET has been utilized for enhancement of genetic selection; diagnosis and treatment of infertility; control of infectious disease transmission; screening for genetic defects; propagation of rare and endangered species; and the study of developmental biology. Most of the embryo transfers have been intraspecific. A listing of the species includes rabbit, rat, sheep, mouse, g...
Herrera C, Morikawa MI, Castex CB, Pinto MR, Ortega N, Fanti T, Garaguso R, Franco MJ, Castañares M, Castañeira C, Losinno L, Miragaya MH, Mutto AA.Normal mammalian early embryonic development involves apoptosis of blastomeres as a remodeling process during differentiation, starting at the blastocyst stage. Genomic DNA has been recently detected in the blastocele fluid of human embryos and has been amplified by real-time polymerase chain reaction (PCR) to diagnose the sex of in vitro-produced human embryos. This new approach varies from conventional preimplantation genetic diagnosis in that no cells are extracted from the embryo and only the blastocele fluid is aspirated and used as a DNA sample for diagnosis. In the present work, we inv...
Drumm NJ, Embertson RM, Woodie JB, Ruggles AJ, Hopper SA, Fimmers R, Handler J.Clinical experience has suggested that foaling rates following colic surgery in the pregnant mare are influenced by days of gestation. This premise has not been supported in previous studies. We also aimed to determine the effects of other potential influencing factors. Objective: To determine the influence of age of gestation on foaling rates in pregnant mares following colic surgery and evaluate the relationships between other factors and foaling rates. Methods: Medical records of Thoroughbred mares, which had colic surgery performed from 1993 to 2007 at the Rood and Riddle Equine Hospital, ...
Castro T, Jacob JC, Domingues RR, Ginther OJ.Prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) are involved in equine embryo mobility throughout the uterus on Days 11-15 (ovulation = Day 0). On a day (Day 12) of maximal embryo mobility in pregnant mares (n = 13) and before luteolysis in nonbred mares (n = 10), gene expressions were compared between the uterine horns that did and did not contain the mobile embryo and between pregnant and nonbred mares. A cytobrush was used to collect an endometrial sample from the middle of each uterine horn. In nonbred mares, there was no difference for any of the considered gene expressions ...
Shore MD, Macpherson ML, Combes GB, Varner DD, Blanchard TL.It has become a common practice in the equine breeding industry to send 2 insemination doses for breeding with transported cooled semen, one to be used for the initial insemination upon arrival, and the other to be held a second insemination the next day. One fertile stallion and 36 fertile mares were used to determine if breeding once with 1 dose of semen cooled for 24 h would improve fertility compared with breeding twice, 1 d apart, with half the dose of semen cooled for 24 h on the first day of breeding and half cooled for 48 h on the second day of breeding. Mares were given two intramuscu...
Witt MC, Bollwein H, Probst J, Baackmann C, Squires EL, Sieme H.The aim of the present study was to investigate the effects of a gonadotropin treatment to induce superovulation on ovarian and uterine blood flow and its relationship with steroid hormone levels and ovarian response in mares, using color Doppler sonography. Each of six mares were examined sonographically in five cycles for 3 d (t1 to t3) during the follicular development phase (FDP) beginning at a follicle size of ≥ 22 mm, and for 4 d (D-4 to D-1; D0 = Ovulation) in the preovulatory phase (POP). After each examination, total estrogens (E(tot)) and progesterone (P(4)) levels were determined ...
Jones I, Madej A.Microtitre plates were coated with antiserum against oestradiol-17 beta-6-(O-carboxymethyl)-oxime bovine serum albumin raised in sheep. The plasma samples (0.2-1.0 ml) were extracted with peroxide-free diethyl ether prepared daily by treatment with Al2O3. The enzyme conjugate was prepared by coupling oestradiol-17 beta-6-(O-carboxymethyl)-oxime to horse-radish peroxidase. The conjugate was chromatographed on a Sephadex G-25 column. The standard curve ranged from 0.37 to 18.40 fmol/well of oestradiol-17 beta. The amount of oestradiol-17 beta causing a 50% reduction of maximum binding was 4.4 fm...
Ababneh M, Ababneh H, Shidaifat F.Phospholipase A2 (PLA2) is a key enzyme for biosynthesis of PGF2α. Real-time RT-PCR and immunohistochemistry were used to determine transcription and cellular distribution of cytosolic PLA2 (cPLA2) in the equine endometrium during the oestrous cycle and early pregnancy. Endometrial biopsy and blood samples were collected from cycling mares on days (d) 8, 11, 15 and 18 (oestrus) (Day 0 = Day of ovulation; n = 5 for each day) and from pregnant mares (n = 4) on d15. Except for mares on d18 and some cyclic mares (n = 2) on d15 with low progesterone (P₄) concentrations (< 3.18 nm), P₄ conce...
Bousfield GR, Liu WK, Ward DN.LH hybrids were prepared by combining eLH alpha and eLH beta with the corresponding subunits of oLH, pLH and hCG. Recombinants were isolated by gel filtration and assessed by SDS-polyacrylamide gel electrophoresis under both dissociating and non-dissociating conditions. All combinations of subunits produced hybrid LH molecules. Hybrids prepared by combining eLH beta with oLH alpha, pLH alpha or hCG alpha were very inactive in rat radioligand and Leydig cell in vitro bioassays. Hybrids prepared with eLH alpha were very active in both assays. The greatest potentiating activity was observed when ...
Richard F, Martinat N, Remy JJ, Salesse R, Combarnous Y.Among all mammalian FSH receptors (FSH-R; including donkey (dk) FSH-R), only horse (hs) FSH-R does not bind hsLH/chorionic gonadotrophin (CG). In order to delineate the structural origin of hsFSH-R specificity precisely, we have cloned dkFSH-R cDNA from donkey testis mRNA by RT-PCR. Transiently expressed dkFSH-R endowed COS-7 cells with both hsLH/CG- and FSH-binding activity, as well as FSH-induced cAMP production. The deduced dkFSH-R amino acid sequence shares 96% identity with the hsFSH-R: notably, in the hormone-binding domain, the specificity of hsFSH-R may be ascribed to only four diverge...
Saint-Dizier M, Foulon-Gauze F, Lecompte F, Combarnous Y, Chopineau M.Pituitary equine luteinizing hormone (eLH) and fetal chorionic gonadotrophin (eCG) have identical polypeptidic chains, but different linked carbohydrates. In equine tissues, eCG and eLH bind only to the LH/CG receptor (eLH/CG-R) and have no FSH activity. However, radio-receptor assays on equine luteal or testicular tissues have shown that eCG binds to the eLH/CG-R with only 2-4% of the binding activity of eLH. In order to study the structure-function relationship of eLH and eCG in a homologous system, we undertook the cloning and functional expression of the eLH/CG-R. Based on sequence homolog...
Taira T, Fujinaga T, Tamura K, Izumi M, Itoh H, Tsunoda N, Yamashita K, Okumura M, Mizuno S.Equine alpha 1-acid glycoprotein (alpha 1AG) was isolated from equine serum by successive ammonium precipitation, anion- and cation-exchange chromatographies, and gel filtration. Purified equine alpha 1AG had a molecular weight of 46,000 +/- 1,000, and contained 31.4% carbohydrate. Gel isoelectric focusing revealed an isoelectric point range of 2.8 to 3.7. With immunoelectrophoresis, it was found that alpha 1AG migrated to the alpha 1-globulin region. Single radial immunodiffusion was used for quantitative measurement of alpha 1AG in equine serum. In clinically normal foals, serum alpha 1AG wa...
Parrilla Hernández S, Franck T, Munaut C, Feyereisen É, Piret J, Farnir F, Reigner F, Barrière P, Deleuze S.Myeloperoxidase (MPO), as a marker of neutrophil activation, has been associated with equine endometritis. However, in absence of inflammation, MPO is constantly detected in the uterine lumen of estrous mares. The aim of this study was to characterize MPO in the uterus of mares under physiological conditions as a first step to better understand the role of this enzyme in equine reproduction. Total and active MPO concentrations were determined, by ELISA and SIEFED assay, respectively, in low-volume lavages from mares in estrus ( = 26), diestrus ( = 18) and anestrus ( = 8) in absence of endometr...
Squires EL, McKinnon AO, Carnevale EM, Morris R, Nett TM.For embryos collected from mares 7 days after ovulation, embryo recovery for single-ovulating mares was 53% compared to 106% for double-ovulating mares. Pregnancy rates 50 days after surgical transfer were 68 and 129%, respectively. Concentrations of LH were similar during the periovulatory period for cycles which included single or double ovulations. Horse pituitary extract given for 5.5 days resulted in greater than or equal to 2 ovulations (mean 3.8) in 26 of 28 mares and 2.0 embryos were recovered per donor compared to 0.65 for controls. Non-surgical pregnancy rates for embryos collected f...
Cecio A, Vittoria A.Amines and/or peptide-producing cells, deserving to be called paraneurons, were demonstrated in the urethro-prostatic complex of the man, rabbit, buffalo and sheep and in the uterine horns of the pig, horse and mouse, by means of histochemical, immunohistochemical, immunofluorescent and double labeling immunofluorescent techniques. In particular, the urethro-prostatic complex of the sheep contains cells producing serotonin, chromogranin A and somatostatin. Often the amine and the "marker"-protein were colocalized in the same cells. Chromogranin A- and somatostatin-containing cells were found i...
Bracher V, Mathias S, Allen WR.Videoendoscopy of the reproductive tract was performed in 87 Thoroughbred mares with histories of reduced fertility. During hysteroscopy samples for cytological, microbiological and histological examinations were obtained under visual control. Common findings in these broodmares included: (a) endometrial degeneration, as assessed by an uneven distribution or atrophy of endometrial folds and/or a scarred appearance of the endometrium (49 mares, 56%); (b) endometrial cysts of various sizes and locations within the uterus with the most common location being at the base of the uterine horns (48 ma...
