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.
Cuervo-Arango J, Claes AN, Beitsma M, Stout TAE.The in vitro production of embryos by ovum pickup (OPU) and intracytoplasmic sperm injection (ICSI) is gaining popularity among horse breeders and veterinarians. Various collection media are available for flushing follicles during OPU. The objective of this study was to determine whether the type of flushing media used to aspirate follicles and collect oocytes influences the outcome of a commercial equine OPU-ICSI program. Two commercial embryo flushing media (EFM1 and EFM2) supplemented with heparin were compared with a flushing media designed specifically for the collection of oocytes (oocy...
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...
Devick IF, Hendrickson DA.To describe a standing left paralumbar fossa approach combined with mesocolon fenestration for bilateral equine ovariectomy. Methods: Prospective experimental clinical study. Methods: Four healthy client-owned mares and 1 client-owned Molly mule. Methods: Mares were restrained in standing stocks and sedated with an α -agonist continuous infusion to effect. The abdominal cavity was approached via a left paralumbar fossa approach, and the contralateral right ovary was accessed through an incision in the mesocolon. The location of the window was adjusted to allow triangulation, adequate visualiz...
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...
Grady ST, Watts AE, Thompson JA, Penedo MCT, Konganti K, Hinrichs K.This study aims to determine if intra-ovarian injection of bone marrow-derived mesenchymal stem cells (MSCs) improves or restores ovarian function in aged females. Methods: Prospective randomized study of eight aged mares and six young mares receiving intra-ovarian injection of MSCs or vehicle. Main outcome measures were antral follicle count and serum anti-Müllerian hormone (AMH) (aged and young mares), and for aged mares, oocyte meiotic and developmental competence; gross and histological ovarian assessment; evaluation of presence of chimerism in recovered granulosa cells and in ovarian tis...
Uliani RC, Conley AJ, Corbin CJ, Friso AM, Maciel LFS, Alvarenga MA.Anti-Müllerian hormone (AMH) is used as a marker of follicle population numbers and potential fertility in several species including horses but limited data exist across the lifespan. No one has decreased ovarian reserve experimentally to investigate whether a corresponding, quantitative decrease in AMH results. Concentrations of AMH across the lifespan were compiled from 1101 equine females sampled from birth to >33 years of age. Young and old mares (averaging 6 and 19 years) were hemi-ovariectomized and circulating AMH was assessed before and daily thereafter for 15 days. The remaining o...
Ginther OJ, Dangudubiyyam SV.Effect of side (left ovary, LO; right ovary, RO) and intraovarian presence of the regressing corpus luteum (CL) on the preovulatory follicle (PF) and ovulation were studied using ultrasonic- records of complete interovulatory intervals (IOI) for 500 IOI in heifers and 485 IOI in mares. Only IOI with a recording of side of a single PF and a single CL at each end of the IOI were used. In heifers, ovulation at the end of the IOI occurred more frequently (P < 0.0001) from RO (n = 285, 57%) than LO (n = 215, 43%). In mares, frequency of ovulation did not differ between RO (n = 250. ...
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...
Max MC, Bizarro-Silva C, Búfalo I, González SM, Lindquist AG, Gomes RG, Barreiros TRR, Lisboa LA, Morotti F, Seneda MM.Folliculogenesis is a process of development and maturation of the ovarian follicles, being essential for the maintenance of fertility. In in vivo conditions, 99.9% of the follicles of an ovary do not ovulate and undergo atresia. In order to minimize this loss and to clarify the existing mechanisms, a technique was developed that allows for the in vitro follicular development. The objective of this study was to evaluate the effects of different epidermal growth factor (EGF) concentrations on the in vitro culturing of equine preantral follicles. Ovaries (n = 10) were collected from a local ...
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...
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-...
Ishak GM, Bashir ST, Dutra GA, Gastal GDA, Gastal MO, Cavinder CA, Feugang JM, Gastal EL.In vivo studies involving molecular markers of the follicle wall associated with follicular fluid (FF) milieu are crucial for a better understanding of follicle dynamics. The inability to obtain in vivo samples of antral follicle wall (granulosa and theca cells) without jeopardizing ovarian function has restricted advancement in knowledge of folliculogenesis in several species. The purpose of this study in mares was to develop and validate a novel, minimally invasive in vivo technique for simultaneous collection of follicle wall biopsy (FWB) and FF samples, and repeated collection from the sam...
Spacek SG, Carnevale EM.Equine follicular fluid (FF) provides autocrine and paracrine factors from theca, granulosa, and cumulus cells, both reflecting and impacting oocyte and follicle maturation. We hypothesized that maturation of oocytes in FF from old versus young mares has a deleterious effect on oocyte maturation and their subsequent developmental potential. Follicular fluid was collected from the large, dominant follicle from young mares (4-13 years) or old mares (21-26 years) and classified as: (1) Noninduced follicular fluid (NFF), FF from noninduced follicle 33 ± 3 mm, or (2) Induced follicular fluid (I...
Alves BG, Alves KA, Gastal GDA, Gastal MO, Figueiredo JR, Gastal EL.Comprehensive studies on spatial distribution of preantral follicles in the ovary are scarce. Considering that preantral follicles represent the main ovarian reserve, harvesting of these follicles is crucial for the development/use of assisted reproductive techniques. Therefore, knowledge on follicle spatial distribution can be helpful for targeting areas with richer number of preantral follicles through biopsy procedures. The aim of this study was to assess the distribution and localization of equine preantral follicles according to: (i) age, (ii) ovarian portion (lateral and intermediary) an...
Scarlet D, Wulf M, Kuhl J, Köhne M, Ille N, Conley AJ, Aurich C.Anti-Müllerian hormone (AMH) has gained increasing interest as a biomarker for assessment of gonadal activity. The ability to predict the ovarian follicular reserve of prepubertal female horses (fillies) or to identify stallions with testicular pathologies already during their prepubertal life has not been analyzed so far. Both would help to select fertile horses and reduce costs associated with keeping animals. The objectives of the present study were to (1) assess AMH, LH, FSH, progesterone (females) and testosterone (males) dynamics in prepubertal horses from birth onwards and (2) determin...
Oberhaus EL, Thompson DL, Foster BA, Pinto CR.This experiment assessed the hormonal production, secretory aspects, and changes in luteinizing hormone (LH) receptor gene expression of early induced ovulatory-sized follicles relative to the first ovulatory-sized follicles occurring naturally in the spring. Anovulatory mares were treated on January 21 with (1) 50 mg of estradiol cypionate (ECP, n = 8) alone or (2) with ECP followed by two 3-g sulpiride injections (n = 8), 5 and 12 days later. Half of each group also received complete follicle ablation via transvaginal aspiration before ECP treatment. Ovaries were scanned regularly until ...
Ginther OJ, Domingues RR, Dangudubiyyam SV, Araujo ER.Diameter deviation or follicle selection during a follicular wave begins with continuous growth rate of F1 (dominant or selected follicle; DF) and decreasing growth rate of F2 (largest subordinate). Intraovarian patterns based on presence or absence of the DF and CL are DF-CL, DF, CL, and devoid (neither DF nor CL). The DF and CL relationships in a pair of ovaries are ipsilateral (DF-CL pattern and devoid pattern) and contralateral (DF pattern and CL pattern). The effects of patterns and relationships on F1, F2, and FSH during deviation were determined in 21 mares. Diameters of F1 and F2 at ex...
Easley JT, McGilvray KC, Hendrickson DA, Bruemmer J, Hackett ES.To determine the temperature of a vessel sealer and divider device during unilateral paralumbar laparoscopic ovariectomy in standing, sedated mares. Methods: Prospective study. Methods: Fifteen healthy research mares. Methods: Healthy mares with normal ovarian palpation and ultrasonographic appearance were enrolled. Horses were restrained in standing stocks and sedated. A right or left paralumbar ovariectomy was performed with a laparoscopic portal and 2 instrument portals. Ovaries were excised with traumatic forceps and a blunt tip vessel sealer and divider. Temperatures of the vessel sealer ...
Joonè CJ, Schulman ML, Fosgate GT, Claes ANJ, Gupta SK, Botha AE, Human A, Bertschinger HJ.Circulating anti-Müllerian hormone concentration (AMH) is positively correlated to the number of small growing follicles in the mare and may reflect ovarian function. Dynamics of AMH during immunocontraception have not previously been investigated. This study aimed to compare serum AMH in mares following treatment with native porcine zona pellucida (pZP), recombinant pZP3 and pZP4 (reZP) or gonadotrophin releasing hormone (GnRH) vaccines, and saline-treated controls. Stored sera collected during two previous studies examining ovarian activity in mares during zona pellucida (ZP) or GnRH immuno...
Gomes RG, Silva CB, González SM, Oliveira RL, Max MC, Lisboa LA, Barreiros TRR, Santos MM, Sarapião FD, Gastal EL, Seneda MM.The effect of different concentrations of alpha lipoic acid (ALA) on the development and morphology of preantral follicles, as well as the proliferative activity of granulosa cells, was assessed after short-term culture. Ovaries (n = 5) of five seasonal anestrous mares were harvested in a local abattoir. At the laboratory, nine ovarian fragments (5 × 5 × 1 mm) from each animal were used. One fragment was immediately fixed in Bouin and subjected to histological and immunohistochemistry (proliferating cell nuclear antigen, PCNA) analyses (noncultured group; D0 = day 0). The other eigh...
Hall SE, Upton RMO, McLaughlin EA, Sutherland JM.The mare ovary is unique in its anatomical structure; however, the signalling pathways responsible for physiological processes, such as follicular activation, remain uncharacterised. This provided us with the impetus to explore whether signalling molecules from important folliculogenesis pathways, phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) and Janus kinase/signal transducer and activator of transcription (JAK/STAT), are conserved in the mare ovary. Messenger RNA expression of six genes important in follicle development was measured using quantitative polymerase chain reaction and pr...
Douet C, Parodi O, Martino NA, Lacalandra GM, Nicassio M, Reigner F, Deleuze S, Dell'Aquila ME, Goudet G.Most wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) techn...
Ishak GM, Bashir ST, Gastal MO, Gastal EL.Color Doppler ultrasonography was used to study the temporal relationships between pre-ovulatory follicle (POF) and corpus luteum (CL) diameter and blood flow, with systemic progesterone (P4) concentration during two transitional ovulatory seasons in mares. Variables of POF and CL/P4 were evaluated for 6days before and 17days after ovulation, respectively. Evaluations were performed during two consecutive estrous cycles in spring and fall seasons, and during the last estrous cycle of the season. There were significant correlations among POF and CL variables, and P4 concentration that ranged fr...
Paulini F, Chaves SB, Rôlo JLJP, Azevedo RB, Lucci CM.Visualization and clear understanding of the ovarian structures are important in determining the stage of oestrus, helping to diagnose several pathologies and supporting advances in reproductive technologies. In this research, computerized microtomography (microCT) was used to explore and characterize the ovarian structure of seven mammalian species. Ovaries of rats, female dog, queens, cows, mares, sows and a female donkey were used. After microCT scanning, the same samples were prepared for histologic evaluation, used here as a validation criterion. It was possible to distinguish regions of ...
Pinto-Bravo P, Galvão A, Rebordão MR, Amaral A, Ramilo D, Silva E, Szóstek-Mioduchowska A, Alexandre-Pires G, Roberto da Costa R, Skarzynski DJ....The oviduct plays important roles in the early reproductive process. The aim of this study was to evaluate gene transcription and protein expression of progesterone receptor (PGR), estrogen receptors 1 (ESR1) and 2 (ESR2); oxytocin receptor (OXTR); prostaglandin F2α synthase (AKR1C3), and prostaglandin E2 synthase (Ptges) in mare oviduct in different estrous cycle stages. Estradiol (E), progesterone (P), oxytocin (OXT), and tumor necrosis factor α (TNF) effect on in vitro PGE and prostaglandin F2α (PGF) secretion by equine oviduct explants or by oviductal epithelial cells (OECs) were also ...
Max MC, Silva CB, González SM, Lindquist AG, Búfalo I, Gomes RG, Morotti F, Costa CB, Barreiros T, Lisboa LA, Seneda MM.This study investigated the effects of different concentrations of FSH (10, 50, 100 and 200 ng/ml) in supplemented MEM+ on the development of equine pre-antral follicles that were cultured in vitro for 2 or 6 days. The ovaries (n = 5) from mares in seasonal anoestrus were collected from a local abattoir. Ten ovarian tissue fragments of approximately 3 × 3 × 1 mm were obtained from each animal. The fragments were cultured in situ for 2 days (D2) or 6 days (D6) in MEM+ or MEM+ supplemented with FSH at four different concentrations, establishing the following 11 groups: control (D0);...
Gonzalez SM, da Silva CB, Lindquist AG, Bufalo I, Morotti F, Lisboa LA, Seneda M.The goal of this study was to determine the distribution of pre-antral follicles in the ovarian parenchyma of mares. For Experiment 1, each ovary was cut longitudinally at the greater curvature, performing two hemiovaries. After that, six fragments from each hemiovary were obtained, resulting in 12 fragments, which were divided into the innermost region of the parenchyma, the middle region and the outermost region. All the three obtained sections were cut transversally to obtain two fragments from each one. For Experiment 2, each ovary also submitted to a longitudinal cut on the greater curvat...
Aguiar FLN, Gastal GDA, Ishak GM, Gastal MO, Teixeira DIA, Feugang JM, Figueiredo JR, Gastal EL.The effect of FSH supplementation on an enriched cultured medium containing insulin (10 ng/mL) and EGF (50 ng/mL) was investigated on in vitro culture of equine ovarian biopsy tissue. Ovarian tissue fragments were collected from mares (n = 10) and distributed in the following treatments: noncultured control, cultured control, and cultured + FSH. Both treated groups were cultured for 7 or 15 days. The end points evaluated were: follicular morphology, estradiol levels in the culture medium, fluorescence intensity for TUNEL, EGFR and Ki-67 detection, and gene expression of GDF-9, BMP-15, a...
Alves KA, Alves BG, Gastal GDA, Haag KT, Gastal MO, Figueiredo JR, Gambarini ML, Gastal EL.The aims of the present study were to: (1) evaluate preantral follicle density in ovarian biopsy fragments within and among mares; (2) assess the effects of mare age on the density and quality of preantral follicles; and (3) determine the minimum number of ovarian fragments and histological sections needed to estimate equine follicle density using a mathematical model. The ovarian biopsy pick-up method was used in three groups of mares separated according to age (5-6, 7-10 and 11-16 years). Overall, 336 preantral follicles were recorded with a mean follicle density of 3.7 follicles per cm. Fol...
Gastal GDA, Aguiar FLN, Alves BG, Alves KA, de Tarso SGS, Ishak GM, Cavinder CA, Feugang JM, Gastal EL.Ovarian tissue cryopreservation allows the preservation of the female fertility potential for an undetermined period. The objectives of this study were to compare the efficiency of cryoprotective agents (CPAs; dimethyl sulfoxide, DMSO; ethylene glycol, EG; and propylene glycol, PROH) using slow-freezing and vitrification methods, and evaluate the viability of cryopreserved equine ovarian tissue after 7 days of culture. Fresh and cryopreserved ovarian fragments were evaluated for preantral follicle morphology, stromal cell density, EGFR, Ki-67, Bax, and Bcl-2 protein expression, and DNA fragmen...
Donadeu FX, Watson ED.The annual reproductive cycle in the horse involves a reduction in ovarian activity during short days. The absence of ovulatory activity during winter has important consequences for an equine industry eager to breed mares early during the year. The anovulatory season results from a reduction in the secretion of pituitary gonadotropin that is in turn triggered by the inhibitory effects of short photoperiod on the hypothalamus-pituitary axis. Recent studies have provided evidence that the response of the ovaries to endocrine stimuli during the anovulatory season is affected not only by circulati...
Barber MR, Lee SM, Steffens WL, Ard M, Fayrer-Hosken RA.A comparative evaluation of the location of immunoreactive porcine zona pellucida (pZP) glycoproteins was performed with polyclonal rabbit anti-pZP antibodies on ovarian sections of the dog, cat, horse, and elephant. For this, formalin (light microscopy) and glutaraldehyde (transmission electron microscopy [TEM]) fixed ovarian sections were incubated with antibodies raised against highly purified pZP. Staining patterns were determined with diaminobenzidine (DAB) at the light level. The dog ZP had a distinct staining distribution that is characterized by intense staining around the periphery of...
Ginther OJ, Gastal EL, Gastal MO, Beg MA.The functional and temporal relationships between circulating gonadotropins and ovarian hormones in mares during Days 7-27 (ovulation = Day 0) was studied using control, follicle ablation, and ovariectomy groups (n = 6 mares/group). In the follicle-ablation group, all follicles > or = 6 mm were ablated on Day 7, and every 2 days thereafter, newly emerging follicles were also ablated. Estradiol concentrations decreased (P < 0.01) similarly in the controls and the follicle-ablation group between Days 7 and 11 and by Day 15 began to increase in the controls and continued to decrease in the ...
Cuervo-Arango J, Newcombe JR.The development of hemorrhagic anovulatory follicles (HAF) involves luteinization and hemorrhage of the follicle. This is observed on ultrasound as an increase in the echogenicity of the granulosa layer and formation of echoic particles in the antrum. The inhibition of prostaglandin synthesis with flunixin meglumine (FM) during the periovulatory period induces ovulatory failure with development of luteinized unruptured follicles (LUF). These two types of anovulatory follicles appear to share similar ultrasound features but they have not been compared critically. The following endpoints: follic...
Kimura J, Hirano Y, Takemoto S, Nambo Y, Ishinazaka T, Himeno R, Mishima T, Tsumagari S, Yokota H.The equine ovary has a very unique structure in terms of its extreme large size, the presence of the ovulation fossa and the inverted location of its cortex and medulla. In the previous study, it was recognized that the application of three-dimensional internal structure microscopy (3D-ISM) to observe the mare ovary is very effective. Three-dimensional reconstruction of serially sliced images made by 3D-ISM was successful in this study with the aid of the sophisticated image processing technique. The rotation of the reconstructed ovary has been carried out with and without the application of t...
Ginther OJ.Individual follicles were monitored by ultrasonography in 15 mares during the transitional period preceding the first ovulation of the year and in 9 mares during the first interovulatory interval. During the transitional period, 7 mares developed 1-3 anovulatory follicular waves characterized by a dominant follicle (maximum diameter greater than or equal to 38 mm) that had growing, static, and regressing phases. The emergence of a subsequent wave (anovulatory or ovulatory) did not occur until the dominant follicle of the previous wave was in the static phase. After the emergence of the subsequ...
Uliani RC, Conley AJ, Corbin CJ, Friso AM, Maciel LFS, Alvarenga MA.Anti-Müllerian hormone (AMH) is used as a marker of follicle population numbers and potential fertility in several species including horses but limited data exist across the lifespan. No one has decreased ovarian reserve experimentally to investigate whether a corresponding, quantitative decrease in AMH results. Concentrations of AMH across the lifespan were compiled from 1101 equine females sampled from birth to >33 years of age. Young and old mares (averaging 6 and 19 years) were hemi-ovariectomized and circulating AMH was assessed before and daily thereafter for 15 days. The remaining o...
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 ...
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...
Hinrichs K.In vitro embryo production is possible in the horse both clinically and for research applications. Oocytes may be collected from excised ovaries post-mortem, or from either immature follicles or stimulated pre-ovulatory follicles in the live mare. In vitro maturation of immature oocytes typically yields approximately 60% mature oocytes. As standard in vitro fertilization is not yet repeatable in the horse, fertilization is performed by intracytoplasmic sperm injection. Embryo culture requires medium with high glucose, at least during blastocyst development, and rates of blastocyst development ...
Gastal GDA, Aguiar FLN, Alves BG, Alves KA, de Tarso SGS, Ishak GM, Cavinder CA, Feugang JM, Gastal EL.Ovarian tissue cryopreservation allows the preservation of the female fertility potential for an undetermined period. The objectives of this study were to compare the efficiency of cryoprotective agents (CPAs; dimethyl sulfoxide, DMSO; ethylene glycol, EG; and propylene glycol, PROH) using slow-freezing and vitrification methods, and evaluate the viability of cryopreserved equine ovarian tissue after 7 days of culture. Fresh and cryopreserved ovarian fragments were evaluated for preantral follicle morphology, stromal cell density, EGFR, Ki-67, Bax, and Bcl-2 protein expression, and DNA fragmen...
Boerboom D, Sirois J.The preovulatory LH rise is the physiological trigger of follicular luteinization, a process during which the synthesis of progesterone is markedly increased. To study the control of follicular progesterone biosynthesis in mares, the objectives of this study were to clone and characterize the equine cholesterol side-chain cleavage cytochrome P450 (P450(scc)) and 3 beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4)-isomerase (3 beta-HSD), and describe the regulation and cellular localization of their transcripts in equine follicles during hCG-induced ovulation. Complementary DNA cloning and pr...
Alm H, Torner H, Kanitz W, Becker F, Hinrichs K.The object of this study was to compare 4 different methods of oocyte recovery from mares; 1) transvaginal follicle aspiration in vivo; 2) follicle aspiration in vitro; 3) oocyte recovery by isolation of follicles in vitro and 4) follicle scraping in vitro. Oocyte recovery was highest after follicle scraping (71.1%) and follicle isolation and rupture (61.3%). Follicle aspiration in vitro and in vivo yielded oocytes on 31.2% and 19.3% of occasions, respectively. The output of different types of cumulus-oocyte-complexes was different among the methods; the portion of compact cumulus-oocyte-compl...
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...
Bollwei H, Weber F, Kolberg B, Stolla R.Uterine and ovarian blood flow was investigated in four mares during two consecutive estrous cycles using transrectal color Doppler sonography. The uterine and ovarian arteries of both sides were scanned to obtain waves of blood flow velocity. The pulsatility index (PI) reflected blood flow. There were significant time trends in PI values of all uterine and ovarian blood vessels during the estrous cycle (P < 0.05). PI values did not differ between the uterine arteries ipsi- and contralateral to the corpus luteum or the ovulatory follicle. PI values of the uterine arteries showed a wave shap...
Gerstenberg C, Allen WR, Stewart F.Immunohistochemical detection of the proliferation marker Ki-67 antigen was used to monitor mitotic activity in the endometrium of mares. The monoclonal antibody MIB1 was validated for use on equine tissues by demonstrating its reaction with activated peripheral blood lymphocytes, and endometrial biopsies were recovered from 26 non-pregnant mares at selected stages during the reproductive cycle. The proportion of positively stained nuclei was counted in five random areas on each histological section to determine the percentage and type of proliferating cells. Multiplication rates in the types ...
Max MC, Bizarro-Silva C, Búfalo I, González SM, Lindquist AG, Gomes RG, Barreiros TRR, Lisboa LA, Morotti F, Seneda MM.Folliculogenesis is a process of development and maturation of the ovarian follicles, being essential for the maintenance of fertility. In in vivo conditions, 99.9% of the follicles of an ovary do not ovulate and undergo atresia. In order to minimize this loss and to clarify the existing mechanisms, a technique was developed that allows for the in vitro follicular development. The objective of this study was to evaluate the effects of different epidermal growth factor (EGF) concentrations on the in vitro culturing of equine preantral follicles. Ovaries (n = 10) were collected from a local ...
Watson ED, Sertich PL.This study investigated the sequence of hormonal changes within the preovulatory follicles of mares. Mares were injected i.v. with 2500 IU human chorionic gonadotrophin (hCG) when a preovulatory follicle of 35 mm in diameter was detected. Fluid was aspirated from preovulatory follicles before (0 h), and 12, 24 and 36 h after administration of hCG. Concentrations of progesterone, prostaglandin (PG) E2, PGF, 6-keto-PGF1 alpha and thromboxane B2 in follicular fluid increased significantly (P less than 0.01) between 0 and 36 h. At 36 h, PGE2 was present in highest concentrations, followed by PGF a...
Brown KA, Boerboom D, Bouchard N, Doré M, Lussier JG, Sirois J.Aldo-keto reductases (AKRs) are multifunctional enzymes capable of acting on a wide variety of substrates, including sex steroids. AKRs having 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) activity can reduce progesterone to 20alpha-hydroxy-4-pregnen-3-one (20alpha-DHP), a metabolite with lower affinity for the progesterone receptor. The objective of this study was to investigate the regulation of equine AKR1C23 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine AKR1C23 cDNA was cloned and shown to encode a 322 amino acid protein that is conserved (71-81...
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...
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:...
Christman SA, Bailey MT, Wheaton JE, Troedsson MH, Ababneh MM, Santschi EM.To determine whether concentrations of dimeric inhibin (CaCA) are greater in plasma and tumor fluid from mares with granulosa-theca cell tumors (GTCT), compared with concentrations in plasma and equine follicular fluid (eFF) from control mares. Methods: 6 mares with GTCT and 12 clinically normal mares. Methods: The alphabetaA immunoradiometric assay used 2 antibodies, one against each subunit of inhibin (alpha and betaA subunits). Tumor tissue, tumor fluid, and a single blood sample were collected at the time of surgical removal of the GTCT. A single blood sample was collected from 7 control m...
Lamm CG, Njaa BL.Dozens of red, raised nodules scattered along the serosal surface of the small intestine and the right and left ovaries were observed as incidental findings on gross examination in a 21-year-old Thoroughbred mare euthanatized for severe lameness. Histologically, these nodules were composed of numerous, variably sized, redundant vascular profiles filled with red blood cells and fibrin thrombi. Based on the presence of multiple nodules composed of benign vascular channels scattered within the small intestine and ovary, a diagnosis of angiomatosis is proposed. To the authors' knowledge, this is t...
Traversari J, Aepli H, Knutti B, Lüttgenau J, Bruckmaier RM, Bollwein H.The anti-Müllerian hormone (AMH) plays an inhibitory role during folliculogenesis by regulating the number of follicles entering the growing pool. Antral follicle counts (AFC) are highly correlated with serum AMH concentrations and both appear to be related to the ovarian reserve in several species. Few data on AMH and AFC in mares exist, especially with regard to fertility. Therefore, the objective of the current study was to investigate the interrelationship between antral follicle count, serum AMH concentrations and fertility outcome in mares. One hundred and twenty-seven mares were enroll...
Ginther OJ, Gastal MO, Gastal EL, Jacob JC, Beg MA.A follicular wave and luteolysis were induced in mares by ablation of follicles > or =6 mm and treatment with prostaglandin F(2alpha) (PGF) on Day 10 (where ovulation = Day 0). The incidence of haemorrhagic anovulatory follicles (HAFs) in the induced waves (20%) was greater (P < 0.007) than in preceding spontaneous waves (2%). Hormone and follicle dynamics were compared between induced follicular waves that ended in ovulations (ovulating group; n = 36) v. HAFs (HAF group; n = 9). The day of the first ovulation or the beginning of HAF formation at the end of an induced wave was designated as po...
Ortega-Ferrusola C, Gómez-Arrones V, Martín-Cano FE, Gil MC, Peña FJ, Gaitskell-Phillips G, Da Silva-Álvarez E.Ultrasound technology has led to new lines of research in equine reproduction, and it has helped to greatly improve clinical diagnosis and reproductive outcomes in equine practice. This review aims to discuss the potential clinical uses and new approaches of ultrasonography in equine reproduction. Doppler modalities are usually used to evaluate the vascularization of the follicles, corpus luteum (CL), and the uterus in the mare for diagnostic purposes. Inclusion of Doppler ultrasound in artificial insemination and embryo transfer programs could improve the reproductive outcome of these techniq...
