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.
Claes A, Ball BA, Scoggin KE, Roser JF, Woodward EM, Davolli GM, Squires EL, Troedsson MHT.Reproductive aging must be well understood to optimize the reproductive management of older mares and to predict their reproductive life-span. The objectives of this study were to: 1) examine age-related differences in follicular dynamics, endocrine profiles, and primordial follicle counts, 2) evaluate the influence of antral follicle count (AFC) on age-related changes in follicular parameters, and 3) determine the influence of diestrous ovulations on the estrous cycle. Young (3-8yr; n = 10), middle-aged (9-18 yr; n = 16), and old (>18 yr; n = 19) light horse mares were examined wit...
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 GD, Alves BG, Alves KA, Souza ME, Vieira AD, Varela AS, Figueiredo JR, Feugang JM, Lucia T, Gastal EL.The method of transportation and the conditions imposed on the ovarian tissue are pivotal aspects for the success of ovarian tissue cryopreservation (OTC). The aim of this study was to evaluate the effect of the size of the ovarian tissue (e.g. whole ovary, biopsy size and transplant size) during different times of storage (0, 6, 12 and 24 h) on the structural integrity of equine ovarian tissue transported at 4°C. Eighteen pairs of ovaries from young mares (<10 years old) were harvested in a slaughterhouse and processed to simulate the fragment sizes (biopsy and transplant size groups) o...
Piccinno M, Rizzo A, Cariello G, Sciorsci RL.In this study, the Authors investigated the modulatory effect of three single doses (10, 10, and 10 M) of neostigmine on the spontaneous contractility of equine pre-ovulatory follicles in an isolated organ bath, to establish the relationship between this acetylcholinesterase inhibitor and ovulation, in the mare. The results indicate that neostigmine increases pre-ovulatory contractility in equine follicles at each dose, but in a different manner. Indeed, the rise in contractility induced by neostigmine at 10 M and 10 M was phasic, while at 10 M it was tonic. The data obtained indicate poss...
Brito LF, Baldrighi JM, Wolf CA, Ginther OJ.The objective of the present study was to investigate the effect of reproductive hormones (GnRH, hCG, LH and progesterone) on the regulation of corpus luteum (CL) and ovarian blood flow. Diestrous mares received a single treatment of saline, 100μg gonadorelin (GnRH), or 1500IU hCG 10days after ovulation. Plasma LH and progesterone concentrations, resistance index (RI) for ovarian artery blood-flow, and percentage of corpus luteum (CL) with color-Doppler signals of blood flow were determined immediately before treatment (hour 0) and at hours 0.25, 0.5, 1, 1.5, 2, 3, 4, 5, and 6. In the GnRH gr...
Polasek TCM, Kozicki LE, Pedrosa VB, Weiss RR, Bertol MAF, Camargo CE, Talini R.This study aimed to correlate the inflammatory reaction (IR) caused by a progesterone-releasing intravaginal device (P4) with ovarian activity and pregnancy rate (PR) in embryo-recipient anestrus mares (to decrease the spring transitional period). 50 animals were assigned to three groups: GP4 (P4 group; n = 16), GP4OH (P4 + oxytetracycline hydrochloride and hydrocortisone sprayed onto the device; n = 14), and GNP4 (no intravaginal P4; n = 20). The administration protocol for GP4 was: Day 0, 750 mg P4 + ovarian examination by ultrasonography (US) + vaginal sample collection; Day 8, US;...
Scarlet D, Ille N, Ertl R, Alves BG, Gastal GDA, Paiva SO, Gastal MO, Gastal EL, Aurich C.The objective of this study was to determine whether (1) systemic and intrafollicular cortisol concentrations in horses are directly related and (2) supraphysiological levels of glucocorticoids affect in vitro maturation (IVM) rates of oocytes. Specifically, we studied the (1) changes in the intrafollicular cortisol and progesterone in context with granulosa cell gene expression during maturation of equine follicles (from 5-9 mm, 10-14 mm, 15-19 mm, 20-24 mm, and ≥25 mm in diameter) and (2) effects of cortisol supplementation on IVM rates and gene expression of equine cumulus-oocyte comple...
Claes AN, Ball BA.Anti-Müllerian hormone (AMH) plays a major role in sexual differentiation, Leydig cell differentiation, and folliculogenesis. In addition, AMH has clinical value in equine practice. In stallions, AMH can serve as an endocrine marker for equine cryptorchidism and as an immunohistochemical marker for Sertoli cell tumors. Considering that AMH is also an ovarian specific product, intact mares can be differentiated from ovariectomized mares. Peripheral AMH concentrations reflect the follicular population in mares, and therefore, are useful in the assessment of ovarian reserve and reproductive life...
Rebordão MR, Galvão A, Pinto-Bravo P, Pinheiro J, Gamboa S, Silva E, Mateus L, Ferreira-Dias G.Oxytocin (OXT) has been used to prolong the luteal phase in mares, but its mechanism of action is unknown. The aim of this study was to evaluate the effect of chronic exogenous OXT administration to mid-luteal phase mares on luteal maintenance. Also, endometrial expression of prostaglandin endoperoxide synthase 2 (PTGS2), prostaglandin Fα, E and I synthases (AKR1C3, PTGES, and PTGIS), oxytocin receptor (OXTR), progesterone receptor (PGR), and estrogen receptors 1 (ESR1) and 2 (ESR2) were assessed in mares experiencing luteal maintenance 2 weeks after chronic exogenous OXT administration. Con...
Finan SA, Lamkin EL, McKinnon AO.To investigate the comparative efficacy of BioRelease Deslorelin® (BRD) and Ovuplant® for induction of ovulation in cyclic mares in Australia. Methods: Ovarian follicular activity of 60 mares for a total of 95 cycles was monitored by ultrasonography until they developed a follicle ≥30 mm and a uterine oedema pattern of 3. Mares were then randomly allocated to one of three treatment groups: (1) treatment with 1.25 mg BRD, (2) a single Ovuplant pellet or (3) 1 mL compound sodium lactate control. Follicular activity was monitored with ultrasonography every 12 h until ovulation was detec...
Ellinger I.Active placental transport of maternal serum calcium (Ca(2+)) to the offspring is pivotal for proper development of the fetal skeleton as well as various organ systems. Moreover, extracellular Ca(2+) levels impact on distinct processes in mammalian reproduction. The calcium-sensing receptor (CaSR) translates changes in extracellular Ca(2+)-concentrations into cellular reactions. This review summarizes current knowledge on the expression of CaSR and its putative functions in reproductive organs. CaSR was detected in placental cells mediating materno-fetal Ca(2+)-transport such as the murine int...
McGrath BM, Scott CJ, Wynn PC, Loy J, Norman ST.Managing the return to regular cyclicity after the winter anestrous period in the mare has been a challenge for the equine breeding industry. Specifically, efforts have been made to shift or shorten the vernal transition period and to have it followed by a predictable first ovulation at the commencement of the breeding season. Intravenous administration of kisspeptin is known to stimulate an LH response in both reproductively active and inactive mares. This study examined the effects of a constant rate infusion (CRI) of kisspeptin on mares during vernal transition. Mares were given a 30 hours...
Klein C.Relaxin is a peptide hormone best known for its action during the latter half of pregnancy, in particular for its softening effect on pelvic ligaments that aids in preparation of the birth canal for the impending delivery of the fetus. The source of relaxin during early pregnancy varies across species, with the CL being the main source in a number of species. The main source of relaxin during late equine pregnancy is the placenta. In mares with impaired placental function, circulating relaxin levels decline before abortion. During early pregnancy, relaxin promotes endometrial angiogenesis thro...
Martínez-Boví R, Cuervo-Arango J.The objectives were to determine: (i) whether intrafollicular administration of PGE2 and PGF2α to mares would hasten follicle collapse and (ii) the differences in reproductive hormone characteristics in mares with spontaneous and prostaglandin-induced follicle collapses. Six mares were followed for two oestrous cycles each: when the mares reached a follicle diameter of 30-35 mm and showed mild-to-moderate endometrial oedema, mares were administered a single 0.5 ml dose containing 500 μg PGE2 and 125 μg PGF2α (treatment cycle) or a placebo (0.5 ml of water for injection; control cycle) into...
Joonè CJ, Bertschinger HJ, Gupta SK, Fosgate GT, Arukha AP, Minhas V, Dieterman E, Schulman ML.Few studies have investigated ovarian function in the mare undergoing porcine zona pellucida (pZP) immunocontraception despite reported ovarian dysfunction in other species. Objective: This study aimed to describe ovarian function and oestrous cyclicity in pony mares following treatment with either the conventional pZP vaccine or a novel recombinant form of the vaccine derived from porcine ZP3 and ZP4 (reZP). In addition, the contraceptive efficacy of pZP vs. reZP was assessed. Methods: Blinded, randomised, prospective clinical trial. Methods: Mares (n = 21) were randomised into 3 groups of 7:...
Alves KA, Alves BG, Gastal GD, de Tarso SG, Gastal MO, Figueiredo JR, Gambarini ML, Gastal EL.Ovarian tissue collected by biopsy procedures allows the performance of many studies with clinical applications in the field of female fertility preservation. The aim of the present study was to investigate the influence of reproductive phase (anestrous vs. diestrous) and ovarian structures (antral follicles and corpus luteum) on the quality, class distribution, number, and density of preantral follicles, and stromal cell density. Ovarian fragments were harvested by biopsy pick-up procedures from mares and submitted to histological analysis. The mean preantral follicle and ovarian stromal cell...
Bashir ST, Ishak GM, Gastal MO, Roser JF, Gastal EL.Changes in intrafollicular growth factors and hormones were evaluated in vivo in postdeviation and impending ovulation follicles. Mares (n = 30) were randomly assigned to five experimental groups based on target diameters of 25, 30, 35, 40 mm, and impending signs of ovulation. Furthermore, data belonging to two or more proximal diameter groups that were not different were combined and regrouped for each factor separately. Follicular fluid-free insulin-like growth factor 1 was highest (P 0.05) among groups. Additionally, follicular fluid activin A tended (P < 0.06) to be higher in i...
Aguiar FL, Lunardi FO, Lima LF, Rocha RM, Bruno JB, Magalhães-Padilha DM, Cibin FW, Nunes-Pinheiro DC, Gastal MO, Rodrigues AP, Apgar GA, Gastal EL....This study investigated the effect of adding different concentrations of bovine recombinant follicle-stimulating hormone on the IVC of equine preantral follicles enclosed in ovarian tissue fragments. Randomized ovarian fragments were fixed immediately (fresh noncultured control) or cultured for 1 or 7 days in α-MEM(+) supplemented with 0, 10, 50, and 100 ng/mL FSH and subsequently analyzed by classical histology. Culture media collected on Day 1 or Day 7 and were analyzed for steroids (estradiol and progesterone) and reactive oxygen species (ROS). After Day 1 and Day 7 of culture, 50-ng/mL FS...
Aguiar FL, Lunardi FO, Lima LF, Rocha RM, Bruno JB, Magalhães-Padilha DM, Cibin FW, Rodrigues AP, Gastal MO, Gastal EL, Figueiredo JR.This study investigated the effect of insulin concentration on the in vitro culture of equine preantral follicles enclosed in ovarian tissue. Ovarian tissue samples were immediately fixed (noncultured control) or cultured for 1 or 7 days in α-MEM(+) supplemented with 0 ng/mL, 10 ng/mL, or 10 μg/mL insulin. Ovarian tissues were processed and analyzed by classical histology. Culture medium samples were collected after 1 and 7 days of culture for steroid and reactive oxygen species (ROS) analyses. The percentage of morphologically normal follicles was greater (P < 0.001) in insulin-treated grou...
Diel de Amorim M, Nairn D, Manning S, Dedden I, Ripley E, Nielsen K, Card C.Ultrasound-guided biopsies of corpora lutea have been previously used for research purposes in the mare and cow. However, the health effects and fertility after transvaginal luteal biopsies (TVLB) or transvaginal ovarian biopsies (TVOB) obtained for diagnostic purposes in cases of suspected ovarian tumors have not been previously evaluated in the horse. The aim of this study was to determine the effects on health and fertility of TVLB and TVOB in mares; 53 mares were included in the study (11 control non-biopsied mares, 37 TVLB mares biopsied on one or more of the following Days 8, 10, 12, 15,...
Claes A, Ball BA, Troedsson MH, Curry TE, Squires EL, Scoggin KE.The wide variation in circulating anti-Müllerian hormone (AMH) concentrations between mares is attributed to differences in antral follicle count (AFC) which may reflect follicular function. There are few data regarding variations in AFC and associated regulatory factors for AMH in the equine follicle during follicular development. Objective: To examine molecular and hormonal differences in the equine follicle in relation to variations in AFC and circulating AMH concentrations during follicular development and to identify genes co-expressed with AMH in the equine follicle. Methods: Observatio...
Castro T, Oliveira FA, Siddiqui MA, Baldrighi JM, Wolf CA, Ginther OJ.A study was performed on the effect of a single dose per mare of 0 (n = 9), 100 (n = 8), or 300 (n = 9) of GnRH on Day 10 (Day 0 = ovulation) on concentrations of LH, FSH, and progesterone (P4) and blood flow to the CL ovary. Hormone concentration and blood flow measurements were performed at hours 0 (hour of treatment), 0.25, 0.5, 1, 2, 3, 4, and 6. Blood flow was assessed by spectral Doppler ultrasonography for resistance to blood flow in an ovarian artery before entry into the CL ovary. The percentage of the CL with color Doppler signals of blood flow was estimated from videotapes of real-t...
Lima AG, Costa LC, Alvarenga MA, Martins CB.The presence of anovulatory haemorrhagic follicles during the oestrous cycle of mares causes financial impacts, slowing conception and increasing the number of services per pregnancy. Non-steroidal anti-inflammatory drugs (NSAIDs) such as meloxicam and phenylbutazone are used in the treatment of several disorders in mares, and these drugs can impair the formation of prostaglandins (PGs) and consequently interfere with reproductive activity. This study aimed to evaluate the effects of treatment with NSAIDs on the development of pre-ovulatory follicles in mares. In total, 11 mares were studied o...
Ono M, Akuzawa H, Nambo Y, Hirano Y, Kimura J, Takemoto S, Nakamura S, Yokota H, Himeno R, Higuchi T, Ohtaki T, Tsumagari S.A three-dimensional internal structure microscopy (3D-ISM) can clarify the anatomical arrangement of internal structures of equine ovaries. In this study, morphological changes of the equine ovary over the first 12 months of life were investigated by 3D-ISM in 59 fillies and by histological analysis in 2 fillies. The weight and volume of the paired ovaries initially decreased from 0 to 1 months to 2 to 3 months of age and then significantly increased at 8 to 12 months of age. The ovulation fossa was first observed around the 3rd month and became evident after the 6th month. The number of folli...
Marth CD, Young ND, Glenton LY, Noden DM, Browning GF, Krekeler N.The physiological changes associated with the varying hormonal environment throughout the oestrous cycle are linked to the different functions the uterus needs to fulfil. The aim of the present study was to generate global gene expression profiles for the equine uterus during oestrus and Day 5 of dioestrus. To achieve this, samples were collected from five horses during oestrus (follicle >35 mm in diameter) and dioestrus (5 days after ovulation) and analysed using high-throughput RNA sequencing techniques (RNA-Seq). Differentially expressed genes between the two cycle stages were further inves...
Gomes RG, Lisboa LA, Silva CB, Max MC, Marino PC, Oliveira RL, González SM, Barreiros TR, Marinho LS, Seneda MM.The aim of this study was to evaluate the effects of different concentrations of ascorbic acid (25, 50, and 100 μg/mL) in supplemented minimum essential medium (MEM+) on the development of equine preantral follicles that were cultured in vitro for 2 or 6 days. The contralateral ovaries (n = 5) from five mares in seasonal anestrus were collected from a local abattoir. Nine ovarian tissue fragments of approximately 5 × 5 × 1 mm were obtained from each animal. One fragment was immediately fixed and subjected to histologic analysis (control group; Day 0), and the other eight were placed in PBS ...
da Silveira JC, Winger QA, Bouma GJ, Carnevale EM.Age-related decline in fertility is a consequence of low oocyte number and/or low oocyte competence resulting in pregnancy failure. Transforming growth factor (TGF)-β signalling is a well-studied pathway involved in follicular development and ovulation. Recently, small non-coding RNAs, namely microRNAs (miRNAs), have been demonstrated to regulate several members of this pathway; miRNAs are secreted inside small cell-secreted vesicles called exosomes. The overall goal of the present study was to determine whether altered exosome miRNA content in follicular fluid from old mares is associated wi...
Campos-Chillon F, Farmerie TA, Bouma GJ, Clay CM, Carnevale EM.We hypothesised that advanced mare age is associated with follicle and oocyte gene alterations. The aims of the study were to examine quantitative and temporal differences in mRNA for LH receptor (LHR), amphiregulin (AREG) and epiregulin (EREG) in granulosa cells, phosphodiesterase (PDE) 4D in cumulus cells and PDE3A, G-protein-coupled receptor 3 (GPR3), growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15) and mitochondrial (mt) DNA in oocytes. Samples were collected from dominant follicles of Young (3-12 years) and Old (≥20 years) mares at 0, 6, 9 and 12h after admi...
Sessions-Bresnahan DR, Carnevale EM.Age-related changes in follicle paracrine signalling are not defined, and follicular gene transcript abundance could predict oocyte viability. Granulosa cells from preovulatory follicles of mares considered Young (n=12; 4-14 years), Mid-aged (n=9; 15-19 years) and Old (n=14; 20-27 years) were evaluated for transcript abundance related to systemic and follicle-specific pathways. Gene transcript abundance for receptors of insulin, adiponectin and peroxisome proliferating factor-γ were higher or tended to be higher in Mid-aged or Old than Young mares. Transcript abundance for interleukin (IL)-6 ...
Scarlet D, Walter I, Hlavaty J, Aurich C.In mares, FSH and its receptor (FSHR) are essential for ovarian function. The objective of the present study was to analyse FSHR gene expression at the mRNA and protein levels in ovarian tissue from newborn and adult horses. Expression of mRNA was analysed by reverse transcription polymerase chain reaction, whereas FSHR protein was visualised by immunohistochemistry (IHC), immunofluorescence labelling (IF) and western blot. FSHR mRNA was detected in ovarian follicles and luteal tissue from adult mares, as well as in the ovaries of neonates. Follicular growth up to 4mm in diameter was already p...
Lefebvre R, Theoret C, Doré M, Girard C, Laverty S, Vaillancourt D.An 8-year-old Arabian mare was admitted for a large ovarian anovulatory follicle. A clinical diagnosis of ovarian tumor and endometritis was established. Histological examinations revealed an ovarian teratoma and a grade II endometritis. Three months after unilateral ovariectomy, the mare was confirmed pregnant and eventually gave birth uneventfully. Une jument Arabe de 8 ans fut présentée pour un gros ovaire. Un diagnostic de tumeur ovarienne et d’endométrite fut posé. L’examen histologique confirma un tératoma ovarien et une endométrite grade II. Trois mois après une ovariectomie...
McLennan MW, Kelly WR.A case of hypertrophic osteopathy is described in a mare with a vague history of abdominal discomfort over the preceding 12 months. Bony swellings were obvious in the pastern area of both forelegs. There was a dysgerminoma of the left ovary that had metastasised to several abdominal lymph nodes and had infiltrated the posterior vena cava. There was no gross thoracic involvement although there was histologic evidence of tumour metastasis in pulmonary arterioles.
Donadeu FX, Ginther OJ.The temporal relationships in the changes in concentrations of follicular fluid factors during follicle selection were characterized in mares. All follicles > or =5 mm were ablated 10 days after ovulation, followed by follicular fluid collection from the three largest follicles (F1, F2, and F3) when F1 of the new wave reached a diameter of 8.0-11.9, 12.0-15.9, 16.0-19.9, 20.0-23.9, 24.0-27.9, or 28.0-31.9 mm (n = 4-8 mares/range). Diameter deviation between F1 and F2 began during the 20.0- to 23.9-mm range, as indicated by a greater difference in diameter between the two follicles at the 24...
Collins A, Palmer E, Bézard J, Burke J, Duchamp G, Buckley T.Samples of blood and follicular fluid were recovered from 27 Welsh Pony mares at 4 distinct stages of follicular development. Eighteen biochemical parameters were measured in each sample, including sodium, potassium, chloride, glucose, urea, creatinine, calcium, inorganic phosphate, total bilirubin, total protein, albumin, magnesium, triglyceride, total cholesterol, nonesterified fatty acids, alkaline phosphatase, gamma-glutamyltransferase and aspartate aminotransferase. The concentrations of progesterone, 17beta oestradiol and testosterone, pH and osmolarity, were also measured in all the fol...
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 ...
Dawod A, Miro J, Elbaz HT, Fahmy H, Abdoon AS.This study was designed to examine the effect of the intrauterine infusion of platelet-rich plasma (PRP) or equine lyophilized growth factor (L-GFequina) on the follicular growth, endometrial thickness, estrus cycle length, and pregnancy rate in purebred Arabian mares. A total of 73 purebred Arabian mares who experienced repeat breeding for three successive cycles were randomly divided into the following three groups: control group, without treatment; second Group (PRP group), in which mares were intrauterine infused with 20 mL of fresh PRP on the second day after the end of physic estrus phas...
Dalin AM, Andresen O, Malmgren L.The aim of the present study was to investigate the effect of active immunization against GnRH in mature Standardbred mares (three experimental and one control mare) on antibody titres, ovarian function, hormonal levels and oestrous behaviour. The mares were individually teased with a stallion once each day. During the first part of the experiment (period I: late April until November), blood was sampled every third day during the first 3 months, thereafter once per week. In the second part of the experiment (period II: December until August), sampling was carried out every second week. Progest...
Ababneh MM, Troedsson MH.Real-time PCR was used to investigate the role of progesterone (P4) and oestradiol (E2) in regulation of endometrial cytosolic, secretory and calcium-independent phospholipase A2 (PLA2G4A, PLA2G2A and PLA2G6, respectively) gene expression. Ovariectomized mares underwent 6 days of E2 pre-treatment followed by 14 days of P4 supplementation. At the start of P4 treatment (Day 1), mares were assigned in a 2 × 2 factorial design to receive either E2 or vehicle starting on Day 11 and endometrial biopsy collection on either Day 14 when P4 concentrations remained high (>4 ng/ml) or Day 16 when P4 conc...
Martoriati A, Lalmanach AC, Goudet G, Gérard N.A growing body of evidence suggests that the ovary is a site of inflammatory reactions, and thus, ovarian cells could represent sources and targets of the interleukin-1 (IL-1) system. The aim of the present work was to investigate the expression of IL-1alpha, IL-1beta, IL-1ra, IL-1R1, and IL-1R2 genes in equine cumulus cells and oocytes. Moreover, the influence of IL-1beta on in vitro maturation of cumulus-oocytes complexes (COCs) was examined. COCs were collected using ultrasound-guided follicular puncture in vivo. Oocytes and cumulus cells were isolated from preovulatory and subordinate foll...
Młodawska W, Grzesiak M, Kochan J, Nowak A.Steroidogenic activity in the equine ovary from birth to puberty has been poorly investigated. This study aimed to examine the capability of the ovarian follicles of prepubertal and pubertal fillies to produce steroid hormones and to evaluate the expression and cellular localization of androgen receptor (AR) in their ovaries. The ovaries of 6-18 month-old fillies were divided into two groups: prepubertal (PrP) - without preovulatory follicle (pF) and corpus luteum (CL), and ovulating/postpubertal (Ov/pB) - with pF and/or CL in at least one of the gonads. Adult mares (Me) were used as a control...
Hinrichs K, Williams KA.Horse oocytes with expanded (EX) cumuli appear to have greater meiotic competence than do horse oocytes with compact (CP) cumuli but are thought to come from atretic follicles. We evaluated the relationships among cumulus expansion, follicle viability, initial chromatin configuration, and meiotic competence of horse oocytes. Follicle walls were sectioned for histological examination, and the follicles were scraped to obtain the oocytes. Half of the oocytes were evaluated immediately and half were matured for 24 h in vitro. Cumulus expansion was significantly associated with follicle atresia. I...
Grøndahl C, Hyttel P, Grøndahl ML, Eriksen T, Gotfredsen P, Greve T.The objectives were to describe the ultrastructure of equine oocytes aspirated from small and preovulatory follicles, and to relate the ultrastructural features to follicle size and follicular fluid steroid concentrations. Mares were examined every second day by transrectal ultrasonography, and follicles measuring > 30 mm were aspirated (in vivo) using a 20-cm-long 12-gauge needle through the flank. Following slaughter, both large and small follicles were aspirated (in vitro) from six mares. The oocytes were isolated under a stereomicroscope and processed for transmission electron microscop...
Deleuze S, Goudet G, Caillaud M, Lahuec C, Duchamp G.In vivo techniques, such as intraoviductal oocyte transfer (OT) and intrafollicular oocyte transfer (IFOT), can be considered as alternatives to bypass the lack of efficient superovulation treatments and the inadequacy of conventional in vitro fertilization techniques in the horse. We compared embryo production after transfer of in vivo recovered oocytes (1) into a recipient's oviduct or (2) into her preovulatory follicle either immediately after ovum pick-up or (3) after in vitro maturation (IVM). Recipients were inseminated with fresh semen of a stallion with a known normal fertility. Ten da...
Salazar-Ortiz J, Monget P, Guillaume D.Feed intake affects the GH-IGF system and may be a key factor in determining the ovarian follicular growth rate. In fat mares, the plasma IGF-1 concentration is high with low GH and a quick follicular growth rate, in contrast to values observed in thin mares. Nothing is known regarding the long-term effects of differential feed intake on the IGF system. The objective of this experiment was to quantify IGFs, IGFBPs, GH, glucose, insulin, gonadotropin and progesterone (P4) in blood and in preovulatory follicular fluid (FF) in relation to feeding levels in mares. Methods: Three years prior to the...
Bøgh IB, Bézard J, Duchamp G, Baltsen M, Gérard N, Daels P, Greve T.In the mare, rates of fertilization and development are low in oocytes matured in vitro, and a closer imitation of in vivo conditions during oocyte maturation might be beneficial. The aims of the present study were, therefore, to investigate whether (1) equine oocytes can be matured in vitro in pure equine preovulatory follicular fluid, (2) priming of the follicular fluid donor with crude equine gonadotrophins (CEG) before aspiration of preovulatory follicular fluid promotes the in vitro maturation rate, (3) the in vitro maturation rate differs between oocytes aspirated during estrus and those...
Szóstek AZ, Galvão AM, Ferreira-Dias GM, Skarzynski DJ.This study aimed to evaluate the influence of ovarian steroids on equine endometrial epithelial and stromal cells, specifically i) prostaglandin (PG) production in a time-dependent manner, ii) specific PG synthases mRNA transcription and protein expression, and iii) cell proliferation. After passage I, cells were exposed to vehicle, oxytocin (OT, positive control, 10(-7) M), progesterone (P4, 10(-7) M), 17β estradiol (E2, 10(-9) M), or P4+E2 for 12, 24, 48, or 72 h. Following treatment, PG concentration was determined using the direct enzyme immunoassay (EIA) method. Alterations in PG synth...
Smith LJ, Mair TS.Twelve horses underwent standing laparoscopic ovariectomy using electrocoagulation and fine dissection as the only means of achieving haemostasis of the severed ovarian pedicle. Four mares had bilateral ovariectomy performed as a treatment for aggressive behaviour thought to be associated with the oestrous cycle. Eight mares had unilateral ovariectomy performed for removal of a granulosa thecal cell tumour (GCT). Electrocoagulation provided an effective means of haemostasis in both normal and pathological ovaries. Only one case (removal of a GCT) had mild haemorrhage following electrocoagulati...
Bashir ST, Ishak GM, Gastal MO, Roser JF, Gastal EL.Changes in intrafollicular growth factors and hormones were evaluated in vivo in postdeviation and impending ovulation follicles. Mares (n = 30) were randomly assigned to five experimental groups based on target diameters of 25, 30, 35, 40 mm, and impending signs of ovulation. Furthermore, data belonging to two or more proximal diameter groups that were not different were combined and regrouped for each factor separately. Follicular fluid-free insulin-like growth factor 1 was highest (P 0.05) among groups. Additionally, follicular fluid activin A tended (P < 0.06) to be higher in i...
Kelley DE, Gibbons JR, Smith R, Vernon KL, Pratt-Phillip SE, Mortensen CJ.The objectives were to evaluate the effects of exercise on ovarian folliculogenesis and related hormones in mares. Mares (n = 11) were randomly assigned into a control (non-exercised) or treatment (exercised) group. Treatment mares (n = 5) were moderately exercised for 30 min, 6 d/wk. All mares underwent daily transrectal ultrasonographic examinations and ovarian follicles > 6 mm were measured. Blood samples were collected during the first (Cycle 1) and last (Cycle 4) cycle, and serum concentrations of cortisol, LH, and FSH were determined. Mean cortisol concentrations were elevated (P <...
Ginther OJ.During an ovulatory follicular wave in the monovulatory species of heifers, mares, and women, the two largest follicles deviate in diameter at the end of a common follicle growth phase. The largest follicle before deviation becomes the future ovulatory follicle in most ovulatory waves. In 10-30% of the ovulatory waves, the destiny of the two follicles switches just before or at deviation so that the second-largest follicle becomes the future ovulatory follicle, and the largest follicle becomes a subordinate. In FSH-driven switching in heifers, mares, and women, the wave-stimulating FSH surge d...
Totorikaguena L, Olabarrieta E, Lolicato F, Romero-Aguirregomezcorta J, Smitz J, Agirregoitia N, Agirregoitia E.The cannabinoid (CB) system has been involved in many aspects of reproduction and it is known that the systemic chronic use of exogenous CBs are deleterious to reproductive processes. Even so, it is not known what happens in relation to the physiology of the ovary when CB receptors are absent. The present study investigated the effect of the lack of CB1 and CB2 receptors in mice ovarian morphology, folliculogenesis, oocyte retrieval, and oocyte maturation and evaluated the use of Δ9-tetrahydrocannabinol (THC) on oocyte in vitro maturation (IVM) by comparing classical IVM and two-step IVM by a...
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 ...
Nolan MB, Bertschinger HJ, Roth R, Crampton M, Martins IS, Fosgate GT, Stout TA, Schulman ML.An important determinant in the selection of any contraceptive agent is the impact on ovarian function, both in the short and longer term. In this study, ovarian activity was monitored in mares immunised with one of the following vaccine formulations; native porcine zona pellucida (pZP), recombinant zona pellucida proteins ZP3 and ZP4 (reZP), pZP and reZP combined or a commercially available anti-GnRH vaccine. The ZP antigens were prepared in an adjuvant formulation consisting of 6% polymeric adjuvant (Montanide™ PetGel A, Seppic, France) and 500 μg polyinosinic-polycytidylic acid - TLR3-...
Pollard CL, Gibb Z, Clulow J, Ruiz A, Sheridan A, Bahrami M, Swegen A, Grupen CG.A deficiency in NAD has previously been linked with increased occurrences of congenital abnormalities and embryonic death in humans and mice. Early embryonic death is a major factor involved in pregnancy loss in mares, and very little is known regarding the NAD requirements for optimum reproductive function in horses. The aim of this study was to determine the effect of supplementing the diet of mares with nicotinic acid (NA) on the composition of NAD metabolites in the blood and follicular fluid. Vehicle alone or NA (3 g per os) were administered to seven mares over a minimum of 3 consecutive...
Ginther OJ, Gastal MO, Gastal EL, Jacob JC, Beg MA.An ovulatory follicular wave was induced by ablation of follicles > or =6mm and treatment with prostaglandin F2alpha (PGF) on Day 10 (ovulation=Day 0). Follicle and hormone dynamics of the induced waves were compared among three age groups: young (5-6 y, n=14 waves), intermediate (10-14 y, n=16), and old (> or =18, n=15). During the common-growth phase of the induced wave (Days 12-17), diameter of the future ovulatory follicle was not different among ages, but the young group had more (P or =10mm. The number was correlated (r=+0.7; P<0.0001) within mares between consecutive interovula...
Haag KT, Magalhães-Padilha DM, Fonseca GR, Wischral A, Gastal MO, King SS, Jones KL, Figueiredo JR, Gastal EL.The aims of this study in mares were to: (1) compare preantral follicle parameters between in vitro Biopsy Pick-Up (BPU) and scalpel blade collection methods and between histological and mechanical isolation processing (experiment 1); (2) histologically evaluate preantral follicles (experiment 2); and (3) compare histological analysis with a previously established mechanical isolation technique using a tissue chopper (experiment 3) for ovarian cortical fragments obtained in vivo using a BPU instrument. In experiment 1, preantral follicles were analyzed (N = 220; 90% primordial and 10% primary)...
Ginther OJ.Heifers have two or three major follicular waves per interovulatory interval (IOI). In mares and women, the ovulatory wave is the only major wave in most (75%) IOI. The beginning of diameter deviation during follicle selection of the future dominant follicle (DF) is followed by continued growth of DF and decreasing growth of the future subordinate follicles. Diameter deviation in Bos taurus heifers, mares, and women begins when the future DF is a mean of 8.5, 22.5, and 10.5 mm, respectively. Selection of the ovulatory follicle occurs more frequently from right ovary (RO) in heifers and women...
McCoy DJ.Both ovaries in a 10-year-old American Saddle Horse mare were determined to be larger than normal. The mare had problems with infertility during the preceding 3 years. Clinical evaluation revealed that the mare was polyuric, polydypsic, hyperinsulinemic, and hyperglycemic, and had reduced tolerance to exogenous glucose. Bilateral granulosa cell tumors were identified on histologic examination of the ovaries. The precise mechanism for the secondary diabetes mellitus was not determined; however, the cause was suspected to be a result of the synthesis and release of a hormonal substance by the tu...
Aguiar FLN, Gastal GDA, Alves KA, Alves BG, Figueiredo JR, Gastal EL.During the reproductive lifespan of a female, only a limited quantity of oocytes are naturally ovulated; therefore, the mammalian ovary possesses a substantial population of preantral follicles available to be handled and explored in vitro. Hence, the manipulation of preantral follicles enclosed in ovarian tissue aims to recover a considerable population of oocytes of high-value animals for potential application in profitable assisted reproductive technologies (ARTs). For this purpose, the technique of preantral follicle in vitro culture (IVC) has been the most common research tool, achievin...
Rodriguez Hurtado I, Stewart AJ, Wolfe DF, Caldwell FJ, Harrie M, Whitley EM.Hyaluronan (HA), a glycosaminoglycan, is a major component of the pericellular matrix which envelopes mammalian cells. Binding of hyaluronan to one of its specific receptors, CD44, modulates transduction of intracellular signals which direct a variety of processes, including embryogenesis, wound healing, inflammation, and neoplasia. Since regulation of these processes is critical to equine reproductive success, localization of constitutive CD44 expression was evaluated by immunohistochemical methods in ovarian, oviductal, and uterine tissues from healthy mares. Ovarian stroma contained thecal ...
