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.
Zhang Y, Kobayashi K, Sasagawa K, Imai K, Kobayashi M.The physiological significance of the position and shape of the oxygen equilibrium curve (OEC) of horse hemoglobin (Hb) is considered from the viewpoint of oxygen (O2) transport efficiency and the effectiveness of the Bohr effect. In horse fetal and maternal bloods, their physiological O2 affinities are nearly optimized with respect to the effectiveness of the Bohr shift occurring at the O2 release site, when it is measured by the change in O2 saturation per unit change in P50. With relatively low cooperativity (n=2.69) of horse Hb under physiological conditions, the effectiveness of the Bohr ...
Ménard M, Nauc V, Lazure C, Vaillancourt D, Manjunath P.A family of bull seminal plasma (BSP) phospholipid-binding proteins (BSP proteins), potentiate heparin- and HDL-induced capacitation. The homologous proteins have been purified from stallion and boar seminal plasma, and detected in low concentrations in other mammalian seminal plasma. In this study, we developed a new isolation method for mammalian seminal plasma choline phospholipid-binding proteins wherein they are present in low concentrations. The method is based on the interaction of this family of proteins with egg yolk low-density lipoprotein fraction (LDF). In order to demonstrate the ...
Wieczorek M, Switoński M, Yang F.Fluorescence in situ hybridization with the use of the equine X whole chromosome painting probe was carried out on chromosome spreads originating from three mares with poor reproductive performance (infertility, miscarriage or stillbirth). The numbers of analysed spreads were high (105, 300 and 480) and in all three mares a low frequency of mosaicism was identified. The mares had the following karyotypes: 64,XX/63,X/65,XXX (93.6%/5.7%/0.7%), 64,XX/63,X (98.9%/1.1%) and 64,XX/63,X (94.3%/5.7%). The incidence and importance of the low percentage X chromosome mosaicism are discussed.
Boerboom D, Brown KA, Vaillancourt D, Poitras P, Goff AK, Watanabe K, Doré M, Sirois J.Luteolysis in domestic species is mediated by the release of luteolytic pulses of prostaglandin (PG) F(2alpha) by the uterus at the end of diestrus, which must be suppressed by the conceptus to permit maternal recognition of pregnancy. In many species, including the horse, both the conceptus and the endometrium also synthesize PGE(2), which may antagonize PGF(2alpha) by playing a luteotropic and/or antiluteolytic role. While the release of PGE(2) and PGF(2alpha) by the equine endometrium in late diestrus and early pregnancy has been previously studied, the underlying prostaglandin synthase gen...
Mansour GD, Henry M, Ferreira AM.This study was designed to identify the distribution of type IV collagen, laminin, and fibronectin with an avidin-biotin method in sections of equine endometrial samples, fixed in Bouin's solution and embedded in paraffin wax. Thirty endometrial biopsies were collected at three different stages of the oestrous cycle. The basement membrane of luminal epithelium reacted positively with antibody against type IV collagen. Both type IV collagen and laminin were found in the basement membranes of endometrial glands, and fibronectin occurred diffusely in the interstitial tissue. Blood vessels express...
Carluccio A, Zedda MT, Schiaffino GM, Pirino S, Pau S.Testicular biopsy is one of the supplementary examinations performed in the course
of andrological testing. In veterinary medicine, it is seldom used in low-fertility or
sterility testing, or in the investigation of suspected cancer. Biopsies are discouraged
in horses in particular, since they may result in testicular sclerosis and atrophy
(Galina, 1971; Smith, 1974; Marusi and Corradi, 1989; Varner, 1991; Del Vento et al.,
1992; Threlfall and Lopate, 1993; Roser, 2000). In humans (Foresta and Varotto,
1992; Foresta et al., 1992; Kessaris et al., 1995; Harrington et al., 1996) good resul...
Welland LM.A 6-year-old, primiparous standardbred mare was presented with a history of intestinal protrusion from the rectum shortly after parturition. A subacute grade IV rectal tear was diagnosed. The unusual nature of the tear led to speculation of a possible owner palpation accident. A grave prognosis was given and the mare was euthanized.
Bedford SJ, Kurokawa M, Hinrichs K, Fissore RA.In oocytes from all mammalian species studied to date, fertilization by a spermatozoon induces intracellular calcium ([Ca(2+)](i)) oscillations that are crucial for appropriate oocyte activation and embryonic development. Such patterns are species-specific and have not yet been elucidated in horses; it is also not known whether equine oocytes respond with transient [Ca(2+)](i) oscillations when fertilized or treated with parthenogenetic agents. Therefore, the aims of this study were: (i) to characterize the activity of equine sperm extracts microinjected into mouse oocytes; (ii) to ascertain i...
Martoriati A, Caillaud M, Goudet G, Gérard N.Interleukin 1 beta (IL-1 beta) inhibits the LH-induced resumption of meiosis of equine oocytes in vitro. The present study was performed to clarify this inhibitory effect of IL-1 beta by testing increasing concentrations of IL-1 beta, and by measuring the effect of addition of IL-1 receptor antagonist (IL-1RA) to the culture medium. The effect of IL-1 beta on epidermal growth factor (EGF)-induced resumption of meiosis was also studied. Cumulus-oocyte complexes (COCs) were collected from subordinate follicles on ovaries obtained from an abattoir. In five distinct experiments, COCs were cultured...
Fumuso E, Giguère S, Wade J, Rogan D, Videla-Dorna I, Bowden RA.Endometrial mRNA expression of the pro-inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) was assessed in mares resistant (RM) or susceptible (SM) to persistent post-breeding endometritis (PPBE). Eight RM and eight SM, were selected based on reproductive records and functional tests out of a herd of 2,000 light cross-type mares. Three experiments were done to study transcription patterns in (i) basal conditions; (ii) after artificial insemination (AI); and (iii) after administration of an immunomodulator at time of artificial ...
Donadeu FX, Ginther OJ.The role of estradiol and inhibin in suppression of FSH and LH during the initiation of follicle deviation was examined in mares. In Experiment 1, the two largest follicles (F1, F2) were retained during a wave and the rest were ablated as they reached > or =10 mm. The largest follicle was left intact (control, n=12) or was ablated when it reached > or =20.0 mm (Day 0; expected beginning of deviation). The second largest follicle continued growing (n=9) or regressed (n=4) after F1 ablation. Circulating estradiol and total inhibin decreased after Day 0 in the F2-regressing group, whereas e...
Warnke C, Tuchscherer A, Alm H, Kanitz W, Blottner S, Torner H.The aim of the study was to compare different types of movement pattern and velocities of stallion spermatozoa depending on cryopreservation during breeding and non-breeding season. Ejaculates were collected from four stallions during May (n = 24) and December (n = 24). Parameters of sperm movement were evaluated by computer-aided sperm analysis (CASA) system, and included percentages of motile spermatozoa, different patterns of motility, the velocity, linearity (LIN), amplitude of lateral head displacement (ALH) and beat-cross frequency (BCF). In winter the average percentages of motility wer...
van Buiten A, Westers P, Colenbrander B.The "effect" of stallion, mare and management-related factors on the odds of pregnancy per cycle in the horse were identified and quantified from the breeding records of Dutch Warmblood (n=4491), Friesian (n=1467) and Shetland-pony mares (n=3267) mated either naturally or by artificial insemination to one of the 88 stallions between 1992 and 1996. A mare was considered to be pregnant when she did not return to oestrous within 28 days of the last insemination. For Dutch Warmblood horses, the percentage of mares that did not return for service within 28 days (NR28) varied between studfarms and r...
Bøgh IB, Brink P, Jensen HE, Lehn-Jensen H, Greve T.In the mare, ultrasound-guided transvaginal oocyte recovery and transfer might offer a way to circumvent the demanding procedures of in vitro embryo production. Before clinical application, the possible consequences for subsequent fertility have to be considered. Objective: To examine ovarian function and morphology in mares after repeated follicular punctures. Methods: A total of 14-26 follicular puncture sessions were conducted on each of 4 Norwegian pony mares over a period of 8 years. The ovaries of these mares were recovered by bilateral ovariectomy or at post mortem and subjected to macr...
Baumber J, Sabeur K, Vo A, Ball BA.The objective of this study was to examine the influence of reactive oxygen species (ROS) on equine sperm capacitation. Motile equine spermatozoa were separated on a discontinuous Percoll gradient, resuspended at 10 x 10(6)ml in Tyrode's medium supplemented with BSA (0.5%) and polyvinyl alcohol (0.5%) and incubated at 39 degrees C for 2h with or without the xanthine (X; 0.1mM)-xanthine oxidase (XO; 0.01 U/ml) system or NADPH (0.25 mM). The importance of hydrogen peroxide or superoxide for capacitation was determined by the addition of catalase (CAT; 150 U/ml) or superoxide dismutase (SOD; 150 ...
Castillo-Olivares J, Tearle JP, Montesso F, Westcott D, Kydd JH, Davis-Poynter NJ, Hannant D.Equine arteritis virus (EAV) causes a systemic infection in equids with variable outcome, ranging from subclinical infections to severe disease, and also has the capacity to induce abortion in pregnant mares and persistent infections in stallions. The serum virus-neutralizing antibody response that invariably develops in the infected animal lasts for many months or years and is believed to play an important role in virus clearance. However, very little is known about cellular immunity against EAV because of a lack of methods for evaluating these immune responses. In the present study, we descr...
Schröder HJ.The growth of the fetus is determined by substrate supply mostly for mass accretion and energy gain, and by control systems. Experiments with whole animal models will face the following problems: (1) The fetus, like a three compartmental "Russian doll", is at the end of a long supply chain. There are interactions (e.g. hormones) and partitioning of substrates between the compartments. (2) The fetal organism is growing and differentiating at the same time and not in a steady-state. Experimental results thus depend on gestational age. (3) About 75% of animal experiments on fetal growth restricti...
Heidler B, Parvizi N, Sauerwein H, Bruckmaier RM, Heintges U, Aurich JE, Aurich C.In this study, growth hormone (GH), insulin-like growth factor 1 (IGF-1), leptin, luteinising hormone (LH) and prolactin were analyzed in mares from late pregnancy throughout lactation (group 1, n=46) and in non-lactating mares (group 2, n=11). Plasma GH concentrations in group 1 mares during gestation and lactation were lower than in mares of group 2 (P<0.05). Highest IGF-1 levels were found in lactating mares in the week of foaling. IGF-1 concentrations decreased continuously thereafter. Plasma leptin concentrations decreased after foaling and, for 4 weeks, were lower in lactating than in...
Veronesi MC, Battocchio M, Faustini M, Gandini M, Cairoli F.The aim of this study was to evaluate the possible relationship of pharmacological induction of estrous and/or ovulation with the occurrence of twin pregnancies in Thoroughbred mares. Out of 680 mares, 356 received one of the following treatments during the estrous cycle in which they became pregnant: injection of 0.5mg of cloprostenol at the ultrasonographic detection of a CL (n=86); injection of 5000 IU human chorionic gonadotropin (hCG) immediately before mating (n=221); injection of 0.5mg of cloprostenol at the ultrasonographic detection of a CL plus injection of 5000 IU hCG immediately be...
Ginther OJ, Bergfelt DR, Beg MA, Meira C, Kot K.In cattle and mares, free insulin-like growth factor 1 (IGF-1) is higher in the future dominant follicle (F1) than in the future largest subordinate follicle (F2) before deviation in diameter or selection is manifested between the two follicles. The effect of IGF-1 on other follicular-fluid factors and on the destiny of F2 were studied in two experiments in each species, using a total of 40 heifers and 42 mares. An injection of IGF-1 was made into F2 at the expected beginning of deviation (heifers, F1 >or= 8.5 mm; mares, F1 >or= 20.0 mm; Hour 0). In heifers, follicular fluid was taken fr...
Camillo F, Vannozzi I, Rota A, Di Luzio B, Romagnoli S, Aria G, Allen WR.Mules, hybrids resulting from the mating of a horse mare (Equus caballus, 2n = 64) to a Jack donkey (E. asinus, 2n = 62), are generally infertile. Five horse embryos were transferred non-surgically to two cyclic and one acyclic recipient mules. In the mares and cycling mules, oestrus and ovulation were induced with, respectively, d-cloprostenol and human chorionic gonadotrophin (hCG). The acyclic mule, on the other hand, received oestradiol benzoate when the embryo donor was showing oestrus and progesterone after the donor had ovulated and until pregnancy diagnosis. Non-surgical embryo collect...
Guillaume D, Chavatte-Palmer P, Combarnous Y, Duchamp G, Martinat N, Nagy P, Daels PF.The aim of this study was to compare the effects of treatment with repeated injections of sulpiride (a dopamine D2 antagonist) on prolactin secretion and induced lactation in ovariectomized and intact adult mares and to verify if this induction was possible at the beginning and at the end of the birth season. Two experiments were carried out in September [experiment (expt) 1], and in March (expt 2), in France (48 degrees N). In expt 1, three groups of five mares were tested: intact-control, intact-treated and ovariectomized-treated mares. In expt 2, mares previously subjected to artificial pho...
Hoque S, Senba H, Tsunoda N, Derar RI, Watanabe G, Taya K, Osawa T, Miyake Y.To clarify the endocrinological characteristics of the mares with granulosa theca cell tumor (GTCT), peripheral plasma samples from the 6 mares affected with GTCT were collected before and after the surgical removal of the affected ovary. Concentrations of testosterone (T), follicle stimulating hormone (FSH), luteinizing hormone (LH), immunoreactive-inhibin (ir-INH), progesterone (P) and estradiol-17beta (E(2)) in the plasma samples were measured by radioimmunoassay. Before removal of GTCT in all cases, the concentrations of T were significantly higher than those of normal mares at the breedin...
Gurfield N, Benirschke K.The placenta from a 300-day-gestational age, female, Arabian equine fetus was examined. Multifocal to coalescing, 0.5- to 4-cm-diameter, white, smooth nodules covered 50% of the placenta. Microscopic evaluation of the nodules revealed undifferentiated germ cells and a haphazard arrangement of immature, mesenchymal stroma, cartilage, squamous cornifying epithelium, scattered ducts and secretory acini lined by cuboidal to columnar epithelium, and mineralized foci. No umbilicus, arrangement about an axial skeleton, or organized polarity of structures was present. The lesion was diagnosed as a pla...
Wilsher S, Gower S, Allen WR.Previous reports documenting progesterone receptors (PR) and oestrogen receptors (ER) in the endometrium of early pregnant mares included specimens only up to Day 20 post ovulation. This study aimed to localise PR and ERα on equine feto-maternal tissues between Days 20 and 68 to encompass the period around fixation of the conceptus, development of the endometrial cups and attachment and initial interdigitation of the allantochorion. During early pregnancy mares had the same pattern of PR in the endometrium as that reported for other mammals; namely, a loss of PR from the endometrial epithelia...
Krakowski L, Krawczyk CH, Wrona Z, Dabrowski R, Jarosz Ł.The objective of the present study was to analyse the immune status of mares in the early stages of pregnancy and mares which experienced early embryonic death (EED). Forty-eight fertile purebred Arabian mares were used for the study. The mares were divided into two groups. Group I was the control group composed of 32 mares in the early stages of a normal pregnancy. Group II was composed of 12 mares which had experienced EED. In both groups, subpopulations of T lymphocytes containing CD2(+), CD4(+) and CD8(+) were characterized using flow cytometry. The percentages of CD3(+) lymphocytes (addin...
Antczak DF, Allen WR.There is little evidence for maternal immunological recognition of pregnancy in most species with the striking exception of the members of the genus Equus. Almost all mares make strong cytotoxic antibody responses to paternally inherited fetal antigens by Day 60 of gestation. Most of these responses are directed against antigens of the Major Histocompatibility Complex (MHC), which constitutes the primary immunogenetic barrier to successful organ transplantation. The source of fetal MHC antigens in the pregnant mare appears to be the specialized trophoblast cells of the chorionic girdle region ...
Ginther OJ.Mares are superb models for study of follicle selection owing to similarities between mares and women in relative follicle diameters at specific events during the follicular wave and follicle accessibility for experimental sampling and manipulation. Usually, only 1 major follicular wave with a dominant follicle (DF) greater than 30 mm develops during the 22 to 24 d of the equine estrous cycle and is termed the primary or ovulatory wave. A major secondary wave occasionally (25%) develops early in the cycle. Follicles of the primary wave emerge at 6 mm on day 10 or 11 (day 0 = ovulation). The...
Johnson CA, Thompson DL, Cartmill JA.The present experiment characterized the pituitary responsiveness to exogenous GnRH in the first 10 d after ovulation following commercially available deslorelin acetate implantation at the normal dosage for hastening ovulation in mares. Twelve mature, cyclic mares were assessed daily for estrus and three times weekly for ovarian activity starting May 1. Mares achieving a follicle at least 25 mm in diameter or showing signs of estrus were checked daily thereafter for ovarian characteristics. When a follicle >30 mm was detected, mares were administered either a single deslorelin acetate impl...
Magnusson U.The ongoing emergence of antibiotic resistance jeopardizes efficacy of antibiotics in curing animals from bacterial infections that threaten their health, welfare and productivity. This review gives an overview of antimicrobial use data in food-producing animals, a discussion on how antimicrobials are used for some infections in the reproductive system in cattle, the horse and pig in Europe where there are differences in treatment practices including options to refine the use of antibiotics, and finally, a presentation of the antibiotic resistance for some bacteria collected from the reproduct...
Choi YH, Chung YG, Walker SC, Westhusin ME, Hinrichs K.This study was conducted to evaluate the effects of insulin-like growth factor I (IGF-I) and other media factors during oocyte maturation, and the presence of different compositions of amino acids in embryo culture medium, on the development of equine embryos. Oocytes recovered from slaughterhouse-derived ovaries were matured in vitro for 24 h and those with a polar body were subjected to intracytoplasmic sperm injection (ICSI) or nuclear transfer with adult fibroblasts (NT). For ICSI embryos, there were no significant differences in rates of morphological cleavage, cleavage with normal nuclei...
Drudge JH, Lyons ET, Swerczek TW, Tolliver SC.Cambendazole (CBZ) treatments (20 mg/kg) given at 8-week intervals were used for parasite control in a breeding band of ponies (n = 33 to 43) during the period July 1974 to August 1978. Pre- and posttreatment worm egg counts on feces were used to evaluate efficacy of treatments after every 2nd treatment interval by monitoring changes in strongyle egg counts. Initially, effective reductions (expressed as 92% to 96% fewer worm eggs) were lessened to 70% by the 9th treatment (18 months) and to 28% by the 12th treatment, and thereafter, they fluctuated between 0% and 38%. Critical tests on animals...
Lydka M, Kotula-Balak M, Kopera-Sobota I, Tischner M, Bilińska B.Specific patterns of cytoskeletal filaments reflect a functional state of the cell. In testicular cells intermediate filaments (IFs) are of the vimentin type. Since it is known that Sertoli cells regulate the spermatogenic function in the male gonad, it became important to propose a system that could quantify the state of seminiferous tubular quality. To date, a Johnsen score system has never been used to equine testes. Objective: To demonstrate the expression pattern of vimentin in testes of mature Arabian stallions and correlate its distribution with grade of seminiferous tubule impairment a...
Peretti V, Satué K, Ciotola F, Cristarella S, De Majo M, Biondi V, D'Anza E, Albarella S, Quartuccio M.A 3-year-old Arabian mare underwent medical examinations due to the presence of abnormalities of the reproductive apparatus and stallion behavior (nervous temperament, aggressiveness, masculine attitude). During the clinical visit, an anovulvar distance shorter than normal was observed; moreover, vulvar lips were dorsally fused except for the lower neckline, showing a blind ending from which a penis-like structure protruded. The ultrasound examination revealed the presence of a cervix and corpus of a uterus, hypoplastic uterine horns, and small gonads with an echogenicity similar to a testis. ...
Baczynska A, Fedder J, Schougaard H, Christiansen G.The reproduction rate of horses is one of the lowest within domestic livestock despite advances the veterinary medicine. Infertility in horses may be due mainly to the lack of suitable selection criteria in the breeding of horses. However, acquired infertility due to genital, bacterial infections may occur. Mycoplasmas have been implicated in genital disorders and infertility of many species including humans and horses. However, their role as commensals or pathogens of the genital tract of horses is still not determined. Bacteriological examinations made on the fossa glandis, urethra, penis an...
Kumar CS, Sivaramakrishna D, Ravi SK, Swamy MJ.Seminal fibronectin type-II (Fn-II) proteins interact with choline phospholipids present on the sperm plasma membrane and play a crucial role in sperm capacitation. Crystal structure of phosphorylcholine (PrC) complex of PDC-109, the major bovine Fn-II protein, together with fluorescence spectroscopic studies has shown that tryptophan residues are crucial for its specific interaction with choline phospholipids. In the present study, the heterogeneity and microenvironment of tryptophan residues in HSP-1/2, a major protein of horse seminal plasma (which is homologous to PDC-109) were investigate...
Teubner A, Müller K, Bartmann CP, Sieme H, Klug E, Zingrebe B, Schoon HA.The aim of this study was to determine the effect of the anabolic steroid testosterone on the testicular vascularization, angiogenesis and expression of angiogenic factors, and their receptors in testes of peripubertal stallions. Seven peripubertal stallions were treated with Durateston and castrated either 4 (treatment group 1 [TG1]) or 12 weeks (TG2) after the last injection. The castration of seven untreated control stallions (control group [CG]) took place within the same time. In the testicular specimens, volume density (VD), numerical density (ND), and area of vessels were determined mor...
Retamal C, Urzúa J, Lorca C, López ML, Alves EW.The present paper reports modifications in the electrophoretic and cytochemical characteristics of mature and immature stallion spermatozoa. Some sperm surface glycoproteins (36, 32, 29, 21, 20, 18 kDa) detected in cauda epididymidis spermatozoa, were either absent or present in a very low relative concentration in immature sperm cells. A major 14 kDa protein band, observed in sperm extracts obtained from ductus efferentes, progressively decreased along the epididymal ductus. The nature and distribution of carbohydrate residues on the sperm membrane, during epididymal maturation, was also stud...
Blanchard TL, Elmore RG, Youngquist RS, Loch WE, Hardin DK, Bierschwal CJ, Ganjam VK, Balke JM, Ellersieck MR, Dawson LJ, Miner WS.Fifty mature pony stallions were randomly assigned to one of five treatment groups: Group 1- controls (no treatment), Group 2 - 0.55 mg/kg stanozolol weekly for 13 treatments, Group 3 - 1.1 mg/kg stanozolol every 3 weeks for 5 treatments, Group 4 - 1.1 mg/kg boldenone undecylenate every 3 weeks for 5 treatments, and Group 5 - 0.55 mg/kg boldenone undecylenate weekly for 13 treatments. Scrotal widths (SW), combined testis weights (CTW), and daily sperm productions (DSP) were not different between Groups 1 and 2. Ponies in Group 5 had smaller SW (P<0.01), smaller CTW and decreased DSP compare...
Wilson M, Williams J, Montrose VT, Williams J.Most stallions within breeding programmes are expected to breed and compete concurrently. The exercising of stallions with regards to training regimes during the breeding season is a controversial subject. Daily exercise at low intensities is important for the mental and reproductive well-being of the stallion, however higher intensities of exercise, as seen in competing stallions, may have detrimental effects on seminal quality. To calculate if competition does affect semen quality, this study investigated the effect that equestrian discipline and timing of competition had on a range of stall...
Xie T, Zhang S, Shen W, Zhang G, Guo R, Zhang W, Cao Y, Pan Q, Liu F, Sun Y, Liu S.Twinning trait in donkeys is an important manifestation of high fecundity, but few reports are available elucidating its genetic mechanism. To explore the genetic mechanism underlying the twin colt trait in Dezhou donkeys, DNA from 21 female Dezhou donkeys that had birthed single or twin colts were collected for whole-genome resequencing. FST, θπ and Tajima’s D were used to detect the selective sweeps between single and twin colt fecundity in the Dezhou donkey groups. Another set of 20 female Dezhou donkeys with single or multiple follicles during estrus were selected to compare concentrat...
Alvarenga MA, Landim-Alvarenga FC, Moreira RM, Cesarino MM.The present experiments aimed to examine the substitution of glycerol (G) by ethylene glycol (E) as a cryoprotective agent for stallion spermatozoa. Two different ethylene glycol concentrations (5% and 10%) and also the association of glycerol (2%) and ethylene glycol (3%) (E/G) were studied (Experiment 1). In Experiment 2, two packing systems (0.5 x 4.0 ml) were evaluated using both cryoprotectors. In both experiments, the sperm membrane integrity after freezing was evaluated using transmission electron microscopy. The mean post-thaw motility was 34.25, 36.5, 29.25 and 34.75% for G5%, E5%, E1...
Vanderwall DK, Woods GL, Freeman DA, Weber JA, Rock RW, Tester DF.The objectives of this study were: 1) to document age-related ovulation failure in mares and 2) to contrast the number of ovarian follicles, occurrence of ovulations, and postovulatory concentrations of progesterone in aged versus young mares. In Experiment 1, 4 of 10 aged (25- to 33-years-old) mares were anovulatory between July 1 and September 1, 1989. In Experiment 2, two of 25 aged (20- to 30-years-old) and none of 21 young (3- to 12-years-old) mares were anovulatory between February 1 and June 30, 1990. The average (+/- SEM) day of the first ovulation was later (P<0.05) for aged versus...
Rose BV, Cabrera-Sharp V, Firth MJ, Barrelet FE, Bate S, Cameron IJ, Crabtree JR, Crowhurst J, McGladdery AJ, Neal H, Pynn J, Pynn OD, Smith C....Early pregnancy loss occurs in 6-10% of equine pregnancies making it the main cause of reproductive wastage. Despite this, reasons for the losses are known in only 16% of cases. Lack of viable conceptus material has inhibited investigations of many potential genetic and pathological causes. We present a method for isolating and culturing placental cells from failed early equine pregnancies. Trophoblast cells from 18/30 (60%) failed equine pregnancies of gestational ages 14-65 days were successfully cultured in three different media, with the greatest growth achieved for cells cultured in Amnio...
Desantis S, Ventriglia G, Zizza S, Guaricci AC, Losurdo M, Zarrilli A, Albrizio M.The presence of the mu-opioid receptor (MOR) was investigated in the mare oviduct during oestrus and anoestrus, by means of immunoblotting and immunohistochemistry. Immunoblotting analysis showed that the MOR protein is expressed as 65, 50 and 30 kDa forms in the infundibulum and ampulla both in oestrus and anoestrus, while the 30 kDa form is absent in the isthmus. Moreover, different levels of expression were observed along the ampulla in the two periods examined. Immunohistochemistry revealed MOR in the mucosal epithelium, stromal cells, myocytes and blood vessels. Ciliated cells expressed M...
Gibb Z, Morris LH, Maxwell WM, Grupen CG.Cryopreserved, sex-sorted stallion sperm has been shown to have poor fertility. During this study, the effects of cryoprotectant (glycerol [GLY] and dimethyl formamide [DMF]), cryoprotectant equilibration time (0, 30, 60, 90, or 120 minutes), and cryoprotectant concentration (2%, 3%, or 4% vol/vol) on stored sex-sorted and stored nonsorted stallion sperm were evaluated. Total motility, viability, and DNA integrity (determined using sperm chromatin structure assay) of sperm were assessed after thawing. Equilibration for 90 minutes improved total motility (33.8%) compared with 0 (28.5%) or 120 m...
Rahmanian MS, Thompson DL, Melrose PA.Gonadotropin-specific primary antisera and gold-conjugated secondary antibodies were used to immunocytochemically localize gonadotropins in the anterior pituitary of intact pony mares. Electron microscopy was then used to characterize the ultrastructure and immunoreactive staining characteristics of equine gonadotropes. Cells containing LH were morphologically indistinguishable from those containing FSH. Gonadotropes were relatively large and commonly had eccentric nuclei. The rough endoplasmic reticulum was well developed and dilated. Secretory granules were present in two morphologically dis...
Ryan PL, Klonisch T, Yamashiro S, Renaud RL, Wasnidge C, Porter DG.Immunoreactive, chromatographic and molecular techniques were used to study the expression of relaxin in mare ovaries at different stages of the oestrous cycle. Relaxin in follicular fluid ranged from 1.6 to 2.5, from 1.4 to 5.2, from 1.2 to 6.7 and from 1.0 to 3.5 ng ml-1 in small ( 2 3 4 cm) follicles, respectively, and total content of fluid relaxin per follicle increased (P < 0.05) with follicular size. When subjected to reverse phase HPLC analysis, follicular fluid yielded absorbance profiles corresponding closely to those of purified relaxin, and immunoreactive peaks in follicular f...
Tachibana Y, Sakurai T, Bai H, Shiota K, Nambo Y, Nagaoka K, Imakawa K.Extensive studies have been conducted to characterize the unique phenomena of equine pregnancy. Most studies have focused on embryo transmigration when the embryo is covered with a mucin-like glycoprotein capsule and on the characterization of the chorionic girdle and chorionic gonadotropin (CG) secretion. However, the events preceding and following capsule disappearance have not been well studied. In this study, the mRNA expression in conceptus membranes at days 19, 21, and 25 (day 0 = day of ovulation) was analyzed by RNA-seq (SOLiD3), and transcript levels on these three days and day 13...
Fay JE, Douglas RH.Individual antral follicles from 11 horse mares were studied at three stages of the oestrous cycle to determine the characteristics of the presumptive ovulatory follicle. Mares were ovariectomized (ovex) during the late luteal phase on Day 14 after ovulation (Group 1) and on the 1st (Group 2) or 4th (Group 3) day of oestrus. Every follicle greater than 5 mm in diameter was dissected from each ovary; follicles greater than or equal to 15 mm in diameter were analysed separately while others were pooled by size for subsequent analyses. The presumptive ovulatory follicle possessed the following ch...
Choi YH, Penedo MC, Daftari P, Velez IC, Hinrichs K.Preimplantation genetic diagnosis has great potential in the horse, but information on evaluation of equine embryo biopsy samples is limited. Blastocysts were biopsied using a Piezo drill and methods for whole-genome amplification (WGA) investigated. Results for 33 genetic loci were then compared between biopsy samples from in vitro-produced (IVP) and in vivo-recovered (VIV) blastocysts. Under the experimental conditions described, WGA using the Qiagen Repli-g Midi kit was more accurate than that using the Illustra Genomiphi V2 kit (98.2% vs 25.8%, respectively). Using WGA with the Qiagen kit,...
Koskinen E, Huhtinen M, Katila T.Weekly blood samples were collected for serum progesterone determination from 68 mares during the period from September 1990 to June 1991. A total of 78% (53/68) of the mares became anoestrous: 88% (15/17) of mares with foals and 75% (38/51) of dry mares (p = 0.323). Six mares of the 15 which continued cycling showed persistent corpus luteum for 77 days on average (range 42-106 days). Anoestrus began between September and February, most frequently in December, and ended in January to June, most frequently in March. Mares with foal at foot became anoestrous in 2 separate groups, in September an...
Albrizio M, Lacalandra GM, Cinone M.In this study, we defined the composition of the culture medium that yield a significant percentage of alive and functional equine spermatozoa during enough time before artificial insemination. The effects of sodium bicarbonate were analyzed in fresh and frozen semen in respect to sperm viability, capacitation, spontaneous acrosome reaction and several kinetic parameters such as total motility, progressive motility, VCL, VSL, ALH, BCF, LIN. Moreover, employing Bayk-6844 and Nifedipine, the involvement of L-type voltage-gated calcium channels in the modulation of intracellular calcium concentra...
Rajabi-Toustani R, Watanabe H, Tsogtgerel M, Gao Y, Canbo L, Haneda S, Cheong SH, Nambo Y.The main difficulty of large equine embryo cryopreservation is the replacement of blastocoel fluid with cryoprotectant solution. The objective of this study was to improve the cryopreservation of large equine embryos with PMAP and/or LAP. Embryos were collected via the non-surgical transcervical procedure and divided into three groups based on their size (A ≤ 300 µm, 300 µm<B 300 µm). However, more research is required to find the best method for embryos ≥700 µm.
