Ovarian follicles in horses are structures within the ovaries that contain the developing oocyte, or egg, and are essential for reproduction. These follicles undergo a series of growth stages, from primordial to preovulatory, influenced by hormonal changes within the mare's reproductive cycle. The development and maturation of ovarian follicles are regulated by hormones such as follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Monitoring follicular development is integral to understanding reproductive physiology and managing breeding in equine practice. This page compiles peer-reviewed research studies and scholarly articles that explore the physiology, development, and clinical management of ovarian follicles in horses.
Carnevale EM.Methods for the collection and transfer of equine oocytes have been developed, and uses of these techniques have resulted in new clinical and research possibilities. Because oocyte transfer avoids reproductive problems associated with the oviduct, uterus, and cervix, pregnancies can be produced from many mares that cannot carry a pregnancy or produce embryos. Oocytes for clinical transfers are usually collected from preovulatory follicles and cultured for a short interval or transferred directly into a recipient's oviduct. For oocyte transfer, the recipient is inseminated within the uterus. A ...
Stich KL, Wendt KM, Blanchard TL, Brinsko SP.Mares treated with subcutaneous deslorelin implants on the first postpartum estrus early in the breeding season had significant reductions in the number of large follicles at early pregnancy examinations and delayed return to estrus (in mares that failed to become pregnant); these adverse effects were attributed to a prolonged release of the drug from the implant. In 2003, an injectable short-term release (<24 h) deslorelin product became available. The objective of this study was to determine if this product would hasten ovulation in early foaling first postpartum estrus mares without reducin...
Pedersen HG, Watson ED, Telfer EE.The effects of ovary holding time and temperature on granulosa cell apoptosis, oocyte chromatin configuration and cumulus morphology were investigated through a series of experiments. Three experiments were performed to determine the effect of ovary holding time and temperature on granulosa cell apoptosis. Ovaries were held (1) at 20, 30 or 35-37 degrees C for up to 2h, (2) at 30 degrees C for 0-1, 1-2, 2-3, 3-4, 4-6 or 6-10h, and (3) granulosa cells were held for 0, 1, 2, 3, 5, 12 or 24h in M199 with Hank's salts at room temperature (suboptimal incubation). Granulosa cell DNA was analysed by ...
Ginther OJ, Gastal EL, Gastal MO, Bergfelt DR, Baerwald AR, Pierson RA.Deviation in growth rates of the follicles of the ovulatory wave begins at the end of a common growth phase and is characterized by continued growth of the developing dominant follicle (F1) and regression of the largest subordinate follicle (F2). Follicle diameters during an interovulatory interval were compared between 30 mares and 30 women, using similar methods for collecting and analyzing data. Follicles were tracked and measured daily by ultrasonography. Diameter at follicle emergence (mares, 13 mm; women, 6 mm) and the required minimal attained diameter for assessment of follicles (mares...
Acosta TJ, Beg MA, Ginther OJ.Color Doppler transrectal ultrasound was used to evaluate blood flow area in the wall of dominant anovulatory follicles versus ovulatory follicles in mares during the transition between anovulatory and ovulatory seasons. Daily examinations were done in 11 control mares toward the end of the anovulatory season. In 13 separate mares, follicular fluid was collected from 30-mm follicles, and blood flow areas from control mares were used as a basis for designating the sampled follicle as either anovulatory or ovulatory. Blood flow area in the controls ranged from 0.18 to 0.35 cm(2) in six mares on ...
Acosta TJ, Gastal EL, Gastal MO, Beg MA, Ginther OJ.Diameter deviation during a follicular wave is characterized by the continued growth of the developing dominant follicle and reduced growth and regression of the subordinate follicles. This study considered the hypothesis that reduced blood flow in the future largest subordinate follicle precedes the beginning of diameter deviation. The hypothesis was tested by quantifying the daily changes in blood-flow velocities and blood-flow area within the wall of follicles before and during diameter deviation in mares (n = 7). The blood-flow end points were quantified daily by transrectal color Doppler ...
Colleoni S, Luciano AM, Gandolfi F.Horse is a seasonal breeder and information on oocyte quality outside the breeding season is very limited. Ovaries obtained at the slaughterhouse are a convenient but often limited source of oocytes in this species. As the low quantity of ovaries leads to an intensive use of all available material, it would be useful to know whether ovaries collected during the non-breeding season are suitable for in vitro maturation (IVM). In an attempt to characterize the effect of season on oocyte quality, we investigated the permeability of the gap junctions (GJ) present between cumulus cells and oocytes b...
Catone G, Marino G, Mancuso R, Zanghì A.The present study describes the clinicopathological features of a mature cystic ovarian teratoma occurred in a 4 year old, cyclic, nulliparous Andalusian mare. The mass was discovered in the left ovary during a breeding soundness examination, and was hard in consistency and variously echogenic. Laparoscopy was performed to confirm the neoplastic change, followed by a unilateral laparotomic ovariectomy in the standing animal. On the cut surface the teratomatous ovary showed a large cyst filled with hair and sebaceous material and three small cysts containing mucous fluid, surrounded by solid ar...
Watson ED, Bae SE, Steele M, Thomassen R, Pedersen HG, Bramley T, Hogg CO, Armstrong DG.The period of spring transition, from the anovulatory to the ovulatory season, is characterized in many mares by cyclical growth and regression of large dominant follicles. These follicles produce only low concentrations of estradiol and it is thought that acquisition of steroidogenic competence by large follicles during spring transition is prerequisite in stimulating LH prior to first ovulation. In situ hybridization was used to localize and quantify expression of factors that play a key role in follicular steroidogenesis: StAR, P450scc (CYP11A1), P450c17 (CYP17), P450arom (CYP19), and LH re...
Preis KA, Carnevale EM, Coutinho da Silva MA, Caracciolo di Brienza V, Gomes GM, Maclellan LJ, Squires EL.Transportation of equine ovaries would allow shipment of oocytes for research purposes or transfer after the death of a valuable mare. The objective of this study was to compare two temperatures for maintaining ovaries during a transport interval of 18-24 h. The goal was to obtain pregnancies after transport of ovaries, maturation of oocytes in vitro, and transfer of oocytes. Each shipment was composed of ovaries four to seven mares collected from an abattoir. From each mare, one ovary was packaged at approximately 12 degrees C, and the other was packaged at approximately 22 degrees C. Upon ar...
Shirazi A, Gharagozloo F, Ghasemzadeh-Nava H.The Caspian breed of horses is believed to be the direct descendant of the earliest equine animals. Some special characteristics of Caspian horse differentiate this breed of horses from other breeds. In the current study the ultrasonically observed characteristics of a preovulatory dominant follicle and the lengths of estrus, diestrus as well as some related parameters were studied during 42 interovulatory intervals in 11 healthy Caspian mares. The preovulatory dominant follicle deviated from subordinate follicles and became the largest follicle in the ovaries at Day -8.7 +/- 0.53 (Day 0=ovula...
Bollwein H, Kolberg B, Stolla R.In recent years, a positive relationship between genital perfusion and fertility has been established; in species other than horses, uterine and ovarian perfusion was improved by exogenous estrogen but impaired by exogenous progestin. The goal of the present study was to investigate the effect of exogenous estrogen and progestin on uterine and ovarian blood flow in cycling mares. Five Trotter mares were examined daily during three estrous cycles. Mares were given no treatment, altrenogest (0.044 mg/kg BW) orally from Day 0 (ovulation) to Day 14 and estradiol benzoate (5mg i.m.) on Days 0, 5, a...
Riley SC, Thomassen R, Bae SE, Leask R, Pedersen HG, Watson ED.Profound hormonally controlled tissue remodelling occurs in the equine ovary for follicle growth and development, and also for the alteration in follicle shape directed towards the ovulation fossa, the site where ovulation occurs. The aim of this study was to examine the spatial and temporal regulation of matrix metalloproteinases (MMP)-2 and MMP-9, important enzymes in tissue remodelling, during follicle growth, and ovulation. Using gelatin substrate zymography, we measured these MMPs in follicular fluid of large anovulatory follicles collected during spring transition, early dominant follicl...
Shoemaker RW, Read EK, Duke T, Wilson DG.The feasibility of leaving the ovaries within the peritoneal cavity after laparoscopic coagulation and transection of the ovarian pedicle was assessed in the juvenile horse. Elective ovariectomy was performed on 10 quarter horses, aged 4 to 5 mo, with the fillies in a Trendelenburg position. The mesovarium was isolated, and multiple coagulation and transection cycles were performed until all ovarian attachments had been severed. The ovaries were dropped within the abdomen, and hemostasis of the transected mesovarium was evaluated before closure. The mean surgical time was 33 min (range, 23 to ...
Derar RI, Maeda Y, Hoque SM, Osawa T, Watanabe G, Taya K, Miyake Y.Two pony mares were immunized against recombinant porcine inhibin alpha subunit three times with 39 day intervals. Clinical findings and endocrinological changes before immunization were taken as the control. The first significant rise in the anti-inhibin titre (P<0.05) in the circulation was found 27 days after the first injection. Maximum binding activity was reached by the 12th day after the second booster dose. The number of small, medium and large sized follicles had increased significantly compared to before immunization (11.75 +/- 4.30, 2.75 +/- 0.69 and 2.51 +/- 0.63 vs 6.50 +/- 1.4...
Viñoles C, Quintans G, Paiva N, Cavestany D.The ovarian responses of anoestrus beef cows to a combined treatment with medroxy-progesterone acetate (MAP) sponges and oestradiol benzoate or equine chorionic gonadotrophin (eCG) were evaluated. Forty-five suckling Hereford cows were allocated to three equal groups. Group 1 received a MAP sponge for seven days plus an injection of 2 mg oestradiol benzoate when the sponge was inserted (day 0) and 1 mg when the sponge was withdrawn; group 2 received identical treatment until day 7, when a dose of 400 iu of eCG was administered, and group 3 were left untreated as control animals. From day 0 to ...
Brown KA, Boerboom D, Bouchard N, Doré M, Lussier JG, Sirois J.17Beta-hydroxysteroid dehydrogenase type 4 (17betaHSD4) has a unique multidomain structure, with one domain involved in 17beta-estradiol inactivation. The objective of the study was to investigate the regulation of 17betaHSD4 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine 17betaHSD4 cDNA was cloned and was shown to encode a 735-amino acid protein that is highly conserved (81-87% identity) compared with other mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of 17betaHSD4 transcripts in equine preovulatory follicles ...
Westermann CM, Parlevliet JM, Meertens NM, Sloet van Oldruitenborgh-Oosterbaan MM.Mares regularly have an enlarged ovary. The main causes are haematomas, anovulatory follicles, abscesses, and neoplasia. The granulosa-theca-cell tumour is by far the most common neoplasia of the ovary (about 97%) and accounts for 2.5% of all equine tumours. In this article the differential diagnosis of an enlarged ovary and the background of granulosa-theca cell tumours are reviewed. A case is described of a mare with a very large granulosa-theca cell tumour in the left ovary, which was discovered 1 month after delivery of a healthy foal. This case is special not only because the tumour was e...
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...
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...
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...
Pinto CR, Paccamonti DL, Eilts BE, Venugopal CS, Short CR, Gentry LR, Thompson DL, Godke RA.In the present study, follicular fluids of estrous mares treated with saline solution (Control) or nitric oxide synthase (NOS) inhibitors were analyzed for nitric oxide (NO), estradiol-17beta (E2) and progesterone (P4) concentrations before and 36h after administration of human chorionic gonadotropin (hCG). Follicular fluids obtained before (0h) hCG administration from control mares had lower concentrations of NO than those obtained 36h after administration of hCG (58.3+/-17.8 micromol versus 340.4+/-57.7 micromol; P<0.05). A similar pattern was also noted for intrafollicular P4 in control ...
Butt TD, Wilson DG.An 11-year-old Trakehner gelding required 2 ventral midline celiotomies for correction of a large colon volvulus and a large colon displacement, respectively. Laparoscopic colopexy was performed 50 days following the 2nd celiotomy. Delayed laparoscopic colopexy is minimally invasive and does not disrupt the ventral midline incision following abdominal exploration.
Curtin DJ.An aggressive mare, presented for prebreeding examination, was found to have a significantly enlarged ovary (soccer ball size). A granulosa thecal cell tumor was initially suspected. Following laboratory and repeated clinical examination, the mare was diagnosed with an ovarian hematoma, which regressed with treatment.
Hoque S, Derar RI, Senba H, Osawa T, Kano K, Taya K, Miyake Y.To clarify the morphological and immunohistochemical characteristics in mares with granulosa theca cell tumor (GTCT), the localization of inhibin subunits (alpha, betaA, betaB) and aromatase in the granulosa cell layers and theca layers in the ovarian follicles were determined by immunohistochemical staining. The follicles were obtained from the ovaries of 6 mares with GTCT and 4 normal mares as controls. Immunohistochemically, inhibin alpha-subunit was localized in the granulosa cells of all follicles showing different sizes in all GTCT cases and betaA- subunit was localized in two GTCT cases...
Bollwein H, Sowade C, Stolla R.Transrectal color Doppler sonography was used to evaluate the effect of intrauterine infusion of skim milk semen extender, seminal plasma and raw semen on the endometrium and blood flow in the uterine and ovarian arteries in mares. Six Trotter mares (mean age: 12 years) were examined during estrus in three cycles. Each mare received an intrauterine infusion of 20 ml of skim milk semen extender, seminal plasma or raw semen during estrus in one of three cycles. Blood flow measurements in both uterine and ovarian arteries and the determination of intrauterine fluid via sonography were performed b...
Ginther OJ, Beg MA, Donadeu FX, Bergfelt DR.Diameter deviation is a distinctive change in growth rates among the follicles of a wave, heralding the formation of a dominant follicle and subordinate follicles. When the follicles are about 5mm in cattle and 13 mm in horses, the wave-stimulating FSH surge reaches peak concentrations. Follicle and FSH manipulation studies in both species have shown that the declining portion of the surge before the beginning of deviation is a function of multiple growing follicles that require the decreasing FSH. During this time, all follicles of the wave have the potential for future dominance. Deviation b...
Düsterdieck KF, Pleasant RS, Lanz OI, Saunders G, Howard RD.To evaluate use of the Harmonic Scalpel (Ethicon Endo-Surgery Inc., Cincinnati, OH) for performing laparoscopic bilateral ovariectomy in standing horses. Methods: Experimental study. Methods: Eight mares aged 2-20 years and weighing 410-540 kg. Methods: Standing laparoscopic bilateral ovariectomy was performed in 8 mares with normal reproductive tract anatomy. The Harmonic Scalpel (an ultrasonically activated instrument) was used to simultaneously transect and obtain hemostasis of the ovarian pedicle. Necropsy was performed on 4 mares 3 days after surgery and 4 mares 30 days after surgery. Gro...
Johnson CA, Thompson DL, Cartmill JA.Three experiments were performed to test the following hypotheses: 1) stallions and/or progesterone-estradiol-treated geldings could serve as models for the effects of a single implant of the GnRH analog, deslorelin acetate, on LH and FSH secretion by mares; and 2) multiple implants of deslorelin acetate could be used as a means of inducing ovarian atrophy in mares for future study of the mechanisms involved in the atrophy observed in some mares after a single implant. In Exp. 1, nine light horse stallions received either a single deslorelin implant (n = 5) or a sham injection (n = 4) on d 0. ...
Pastorello M, Gastal MO, Godoi DB, Gastal EL.Among female livestock, the mare has the shortest interval from parturition to first ovulation. Due to the scarcity of research on postpartum mares, little progress has been made on the characterization of the resumption of ovarian cyclicity after parturition. This study compared follicular and gonadotropin dynamics during follicle emergence and deviation in postpartum lactating (PP Lactating) versus non-postpartum cycling (N-PP Cycling) mares. On the day of parturition, every PP Lactating mare was paired with a N-PP Cycling mare. Comparisons were made by considering the partum-ovulation inter...
Watson ED, Buckingham J, Björkstén TS.Oxytocin has been identified in the ovaries of many species
including ruminants (Wathes and Swann 1982; Rodgers et al.
1983), pigs (Jarry etal. 1990), sub-human primates (Khan-Dawood
1998), rats (Ho and Lee 1992) and women (Schaeffer et al. 1984).
In these species ovarian oxytocin has several roles, including
control of luteal regression (Fht and Sheldrick 1983) and possibly
luteinisation (Aladin Chandrase.kher and Fortune 1990; Eispanier et
al. 1997), and involvement in ovulation (viggiano et al. 1989).
In the mare, circulating oxytocin is reported to be high in
oestrus and early dio...
Song L, Porter DG, Coomber BL.Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) play very important roles in extracellular matrix (ECM) remodeling in ovarian follicle growth and ovulation. Equine follicles are embedded in cortex that is at the center of the ovary, and they must expand/emigrate to the fossa, the only site in the ovary for ovulation. Therefore, equine ovarian stromal cells (EOSC) are probably involved in ECM remodeling during follicle growth. This study examined whether cultured EOSC synthesize gelatinases and TIMPs, molecules essential for ECM remodeling in other systems. Results showed...
Tharasanit T, Colenbrander B, Bevers MM, Stout TA.The only gonadotrophin preparation shown to stimulate commercially useful multiple ovulation in mares is equine pituitary extract (EPE); even then, the low and inconsistent ovulatory response has been ascribed to the variable, but high, LH content. This study investigated the effects of an LH-free FSH preparation, recombinant human follicle stimulating hormone (rhFSH), on follicle development, ovulation and embryo production in mares. Five mares were treated twice-daily with 450 i.u. rhFSH starting on day 6 after ovulation, coincident with PGF(2alpha) analogue administration; five control mare...
Dowsett KF, Tshewang U, Knott LM, Jackson AE, Trigg TE.A series of experiments using an ovalbumin conjugated gonadotrophin releasing hormone was used to stimulate antibody production, suppress testosterone secretion and depress testicular function in yearling and 2 year old colts and fillies. In the preliminary experiment, an injectable oil-based formulation was administered to yearling colts. Testicular development and testosterone secretion were retarded for a period of approximately 28-32 weeks while antibody titres were greater than 1:1000. An implant and water-soluble vaccine (200 and 400 mg) is presently being tested in 2 year old colts. Tes...
McCue PM, LeBlanc MM, Squires EL.Equine follicle stimulating hormone (eFSH) has been used to induce follicular development in transitional mares and problem acyclic mares, as well as superovulate cycling mares. The most efficacious protocol is to administer 12.5 mg eFSH, intramuscularly, twice daily beginning 5 to 7 days after ovulation when the diameter of the largest follicle is 20 to 25 mm. Prostaglandins are to be administered on the second day of eFSH therapy. Treatment with eFSH is continued for 3 to 5 days until follicle(s) are >or=35 mm in diameter. The mare is subsequently allowed to 'coast' for 36 h, after which hum...
Brück I, Greve T, Hyttel P.The present study characterised the oocyte-follicular connection (i.e., oocyte fixation site) in Graafian follicles of the mare morphologically. Antral follicles were dissected in toto from ovaries obtained from oestrous, dioestrous and transitional mares after slaughter. The location of the cumulus oophorus complex in relation to the ovulation fossa, the width and density of the blood vessels surrounding the cumulus oophorus complex, the relative dimensions and histological aspects of the cumulus oophorus were investigated. For ultrastructural analysis of the junctional regions, cumulus-oocyt...
Chopin JB, Chopin LK, Knott LM, de Kretser DM, Dowsett KF.An 8-year-old mare, with a foal at foot, was inseminated on foal heat with frozen semen, with the resultant pregnancy lost between days 34 and 41. The right ovary developed a large anovulatory follicle that was non-responsive to multiple doses of ovulating agents. The follicle eventually appeared to luteinise, although plasma progesterone concentrations did not reflect this. Another follicle developed, responded to GnRH and resulted in a pregnancy from frozen semen that went to term with a healthy foal. When the mare was examined after foaling, the structure on the right ovary appeared to be a...
Widders PR, Stokes CR, David JS, Bourne FJ.The effect of cycle stage on immunoglobulin and albumin levels in serum, follicular fluid, oviductal, uterine and vaginal secretions was measured. There was no variation in serum immunoglobulin levels during the oestrous cycle, although IgM levels were elevated in cyclic mares compared to non-cyclic (immature and anoestrous) animals. Similarly, there was no cyclical variation in follicular or oviductal protein concentrations. In the uterus, IgG and IgA levels relative to total protein were higher in oestrogenic than in progestagenic secretions, while the trend in relative IgM concentrations wa...
Del Campo MR, Del Campo CH, Mapletoft RJ, Ginther OJ.Morphology and location of the attached cumulus-oocyte complex (COC) were studied in slaughter-house ovaries in horses (49 follicles, 9 to 44 mm), cattle (68 follicles, 6 to 18 mm), and llamas (38 follicles, 3 to 14 mm). The expected point of ovulation was marked, using the ovulation fossa in mares and the center of the projecting follicular surface in cattle and llamas. A follicle was dissected from an ovary, and tissue was removed from the follicle until the COC became visible by transillumination. However, most llama follicles protruded prominently from the ovarian surface so that dissectio...
Goudet G, Belin F, Młodawska W, Bézard J.The effect of epidermal growth factor (EGF) on the in vitro maturation rate of equine oocytes was examined. Oocytes were collected from an abattoir (Expt 1) or using ultrasound-guided follicular puncture in vivo (Expt 2). All oocytes with a compact or expanded cumulus at recovery were cultured for 30 h in: medium 1 (TCM199 + fetal calf serum (FCS) + crude equine gonadotrophin (CEG) + oestradiol + antibiotics); medium 2 (TCM199 + EGF); medium 3 (medium 1 without FCS + EGF); or medium 4 (medium 1 without CEG + EGF). In Expt 1, 84% (37/44) and 87% (40/46) cumulus expansion (P > 0.05), and 39% (22...
Górecka A, Jezierski TA, Słoniewski K.Weak or equivocal expression of oestrous behaviour, related to different level of mares' reactivity, may cause problems in oestrus detection and thus influence the reproductive efficiency. The aim of the study was to test whether a breed characterised by low pregnancy rate and high emotional reactivity (Thoroughbreds) differs in oestrous behaviour from a primitive breed with higher reproductive efficiency (Koniks). Additionally, the follicle size was examined to determine how it influences the intensity of oestrus and uterine ultrasonic image and whether the pregnancy rate differs in relation ...
Pader K, Lescun TB, Freeman LJ.To develop and assess the feasibility of standing transvaginal Natural Orifice Transluminal Endoscopic Surgery (NOTES®) ovariectomy in the mare. Methods: Descriptive study. Methods: Mares (n=10). Methods: The technique was developed in 6 mares and then evaluated in a short-term study in 4 mares. Abdominal access was developed using controlled access vaginal cannula placement under endoscopic guidance. With viewing provided by a flexible endoscope, hemostasis, and transection of ovarian pedicles was performed using a customized bipolar vessel-sealing device. The ovaries were retrieved and the ...
Pereira BC, Ortiz I, Dorado J, Consuegra C, Diaz-Jimenez M, Demyda-Peyras S, Gosalvez J, Hidalgo M.DNA fragmentation of granulosa cells might be related to developmental competence of the equine oocyte. Granulosa cells are commonly stored before DNA fragmentation assessment, but the effect of preservation methods on this parameter remains unexplored. The aim of this study was to evaluate whether or not cryopreservation of granulosa cells affects the DNA damage. Equine oocytes were recovered from postmortem ovaries of five mares. Granulosa cells were washed by centrifugation and then analyzed (control) or stored in cryovials following four different protocols: P1 = directly plunged in liqui...
Alexander SL, Irvine CH.Mean plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels and their pulse frequencies and the size of the largest ovarian follicle increased during spring in 12 mares studied twice monthly from deep anoestrus to the occurrence of the first ovulation of the breeding season. Mean FSH levels were reduced significantly in deep anoestrus and when a pre-ovulatory follicle was present, whereas mean LH levels were highest close to ovulation. Five of these 12 research mares and 20 of 40 maiden or barren Standardbred mares at a commercial studfarm were given daily intramuscular ...
Aguilar JJ, Woods GL, Miragaya MH, Olsen LM, Vanderwall DK.The objective of this study was to test the hypothesis that incubating equine cumulus-oocyte complexes (COCs) in medium containing 50% or 100% homologous preovulatory follicular fluid would improve cumulus expansion and nuclear maturation. Oocytes were incubated in one of three media: 1) supplemented TCM-199 (control), 2) 50% (v/v) follicular fluid in control medium or 3) 100% follicular fluid. Cumulus expansion was evaluated subjectively, and nuclear maturation was evaluated by staining oocytes with Hoechst 33258. The hypothesis that incubating COCs in medium containing follicular fluid would...
Sayasith K, Bouchard N, Doré M, Sirois J.The objectives of the study were to clone the primary structure of the prostaglandin E2 receptor subtype 2 (PTGER2) cDNA and to characterize its regulation in equine follicles during gonadotropin-induced ovulation. Results from DNA isolation indicated that the equine PTGER2 cDNA encodes a predicted 353-amino acid protein, which is highly similar (76-85%) to known mammalian homologues. The regulation of PTGER2 was studied by semi-quantitative RT-PCR/Southern blot using preparations of theca interna and mural granulosa cells isolated from equine follicles 0-39 hr post-treatment with human chorio...
Lemes KM, Silva LA, Alonso MA, Celeghini ECC, Pugliesi G, Carvalho HF, Affonso FJ, Silva DF, Leite TG, de Arruda RP.Follicular development and deviation processes during the postpartum period are not completely known in horses. Thus, we aimed to study the characteristics of follicular dynamics and ovarian vascular perfusion during the postpartum period in mares that demonstrated estrous behavior and had early ( 0.1) was detected between the groups when the data were normalized for the days preceding the first postpartum ovulation (from D-7 to D-1). However, when the data were normalized to days postpartum, the dominant follicle was larger (P 25 mm diameter was greater (P 0.1) was detected between grou...
Gastal GDA, Aguiar FLN, Ishak GM, Cavinder CA, Willard ST, Ryan PL, Feugang JM, Gastal EL.Preservation of cellular integrity and its mechanisms after ovarian tissue cryopreservation (OTC) and in vitro culture (IVC) procedures are crucial aspects for the success of preservation and recovery of female fertility. This study aimed to evaluate the effects of two cryopreservation methods (slow-freezing, SF, and vitrification, VIT) on the equine ovarian tissue after 1, 3, and 7 days of IVC by assessing: (i) preantral follicle morphology and distribution of follicle classes; (ii) protein expression of markers of cell proliferation for EGFR and Ki-67; (iii) markers of apoptosis for Bax and...
Gentry PA, Zareie M, Liptrap RM.The amounts of total protein, albumin, fibronectin, alpha 2-macroglobulin (alpha 2-M), immunoglobulin G, ceruloplasmin and antithrombin were determined in fluids collected from 53 preovulatory equine follicles and compared with the contents of estradiol-17 beta, progesterone and androstenedione, with follicle size and the amounts of the equivalent proteins in normal equine plasma. The concentration of fibronectin and the fibronectin/albumin ratios increased significantly with follicle size and with follicular estradiol levels. The alpha 2-M levels and alpha 2-M/albumin ratios correlated with f...
Donadeu FX, Ginther OJ.The role of the number of follicles and circulating immunoreactive inhibin in the decrease in plasma FSH concentrations that occurs during development of a follicular wave was studied in mares. All follicles > or = 6 mm in diameter were ablated by ultrasound-guided transvaginal aspiration of follicular fluid on day 10 after ovulation. During the subsequent wave, all follicles, the three largest follicles (three follicle group), the largest follicle (single follicle group) or no follicles were retained and the remaining follicles were ablated before they reached > 10 mm in diameter (n = 1...
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...
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...
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...
King SS, Jones KL, Mullenix BA, Heath DT.Dopamine (DA) blockade during anestrus or early spring transition can facilitate ovarian recrudescence and advance the timing of the first ovulation of the season. Some laboratories have reported variable results using DA antagonists to stimulate follicular growth during the mid-portion of the anestrual period. Differences in DA antagonist efficacy may be due to the FSH secretory status of the anestrous mare and the presence or absence of functional ovarian FSH receptors. We hypothesize that direct ovarian dopaminergic input can affect follicular growth through regulation of FSH receptor (FSHr...
Ginther OJ.The mechanism for selection of the future dominant or ovulatory follicle in mares involves a relatively abrupt separation in growth rates between the future dominant follicle and several subordinate follicles and is termed diameter deviation. The event is used to illustrate that a coordinated complex of many follicular, hormonal, and biochemical factors interact and interbalance during a single physiological mechanism. For example, a positive effect of follicle stimulating hormone (FSH) on development of all follicles during the growing phase can later involve a positive effect of luteinizing ...
Pinto CR, Paccamonti DL, Eilts BE, Short CR, Godke RA.Recent studies suggest that nitric oxide (NO) may have a role in regulating ovarian physiology. To investigate the role of NO during ovulation in mares, inhibitors of the nitric oxide synthase (NOS) were administered to estrous mares. Forty cycling mares (20 horses and 20 pony mares) were allotted to one of the three treatment groups. Once a follicle was at least 27 mm in diameter, but smaller than 35 mm, mares were given one of the following treatments: saline solution 0.9% (n = 20, w/v, i.v., every 12 h), Nomega-nitro-L-arginine methyl ester hydrochloride (L-NAME; n = 10, 148 micromol/kg, i....
Waller CA, Thompson DL, Cartmill JA, Storer WA, Huff NK.Previous results from our laboratory indicated that a majority of mares with high body condition scores (BCS) displayed estrous cycles or had considerable follicular activity during the winter. Among these high BCS mares, about 35% of them exhibited a persistent hyperleptinemia and hyperinsulinemia. The current experiment was designed to compare the reproductive characteristics of high BCS mares with hyperleptinemia to those with normal (low) plasma concentrations of leptin during the winter and the first estrous cycle (or the first full cycle encountered for those already cycling). Light hors...
Souza SS, Aguiar FLN, Alves BG, Alves KA, Brandão FAS, Brito DCC, Raposo RDS, Gastal MO, Rodrigues APR, Figueiredo JR, Teixeira DÍA, Gastal EL.Ovarian tissue transplantation methods using cooled and cryopreserved samples have been attractive options for fertility preservation in animal models and humans. The aim of this study was to evaluate the impact of previous exposure to cooling, cryopreservation, and VEGF on the overall efficiency of equine ovarian tissue after heterotopic xenotransplantation in mice. The end points evaluated were follicular morphology and development, follicular and stromal cell densities, angiogenesis (i.e. the density of new and mature blood vessels), collagen types I and III fiber densities, and total fibro...
Watson ED, Thomson SR.Aromatase P-450 (P-450arom) is a crucial regulatory enzyme that is necessary for conversion of androgens to oestrogens. Corpora lutea and follicles were obtained from the ovaries of cyclic mares and from mares at day 20 and days 40-70 of pregnancy. The presence of P-450arom within specific cell types was investigated by immunostaining to determine potential sites of oestrogen synthesis. Immunoreactivity for P-450arom was confined to the granulosa layer of non-atretic follicles > 5 mm in diameter and to corpora lutea at all stages of the oestrous cycle and during pregnancy. These findings co...
