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.
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...
Seidel GE.Principles for selecting future research projects include interests of investigators, fundability, potential applications, ethical considerations, being able to formulate testable hypotheses and choosing the best models, including selection of the most appropriate species. The following 10 areas of assisted reproduction seem especially appropriate for further research: efficacious capacitation of bovine spermatozoa in vitro; improved in vitro bovine oocyte maturation; decreasing variability and increasing efficacy of bovine superovulation; improved fertility of sexed semen; improving equine IV...
Alves KA, Alves BG, Rocha CD, Visonná M, Mohallem RF, Gastal MO, Jacomini JO, Beletti ME, Figueiredo JR, Gambarini ML, Gastal EL.Regardless of species, advances in preantral follicle culture and cryopreservation and transplant of ovarian tissue techniques are dependent on the number and density of preantral follicles in the ovary. This study tested the effect of different histological section thicknesses on number, classification, and density of equine preantral follicles. An ovarian fragment was obtained from 5- to 10-year-old mares (n = 14) after slaughter, and each fragment was submitted to three histological section thickness treatments: 3, 5, and 7 μm. The area (cm(2)) of each ovarian fragment was measured, and...
Turna Yilmaz O, Gunduz MC, Evkuran Dal G, Kurban I, Erzengin OM, Ucmak M.Doppler ultrasonography is a noninvasive technique which enables us to follow the physiologic and physiopathologic changes in blood flow in tissues. It is becoming an essential tool in veterinary medicine, especially in theriogenology. Twenty-seven Arabian mares were grouped by age ('young', 3-10 y, n=15; 'old', 19-23 y, n=12). The uterine arteries of the mares were examined using Doppler ultrasonography when an ovarian follicle ≥35mm was visible (Day -1). After these measurements, human chorionic gonadotropin (hCG; 2500IU) was administered to 14 mares selected randomly. One day later (Day 0...
Sayasith K, Sirois J, Lussier JG.The luteinizing hormone preovulatory surge stimulates several signal pathways essential for ovulation, and the regulator of G-protein signaling protein-2 (RGS2) is thought to be involved in this process. The objectives of this study were to characterize the regulation of RGS2 transcripts in equine and bovine follicles prior to ovulation and to determine its transcriptional control in bovine granulosa cells. To assess the regulation of equine RGS2 prior to ovulation, RT-PCR was performed using total RNA extracted from equine follicles collected at various times after human chorionic gonadotropi...
Beltman ME, Walsh SW, Canty MJ, Duffy P, Crowe MA.The objective was to characterise the hormonal composition of follicular fluid from mares with distinct anovulatory-cystic follicles. Follicular fluid was aspirated from six mares that presented with cystic follicles and from pre-ovulatory follicles of five normal mares (controls). Differences in progesterone, oestradiol, testosterone, IGF-I and IGF binding were analysed using Fisher's exact test. There were greater (P < 0.03) follicular fluid oestradiol concentrations in normal follicles and the testosterone concentration of the cystic fluid was greater (P < 0.05) than that of t...
Claes A, Ball BA, Scoggin KE, Esteller-Vico A, Kalmar JJ, Conley AJ, Squires EL, Troedsson MH.Anti-Müllerian hormone (AMH) is a granulosa-cell-derived glycoprotein, which plays an important inhibitory role during folliculogenesis. Concentrations of AMH are highly correlated with antral follicle counts (AFCs) in other species, which in turn are related to follicular reserve. Relatively little is known about AMH and AFC in the mare. Objective: To determine plasma AMH concentrations and AFCs in mares of different ages, to measure the repeatability of AMH concentrations and AFCs within and across oestrous cycles and to assess the relationship between plasma AMH concentrations and AFCs wit...
Donadeu FX, Fahiminiya S, Esteves CL, Nadaf J, Miedzinska K, McNeilly AS, Waddington D, Gérard N.Several aspects of equine ovarian physiology are unique among domestic species. Moreover, follicular growth patterns are very similar between horses and humans. This study aimed to characterize, for the first time, global gene expression profiles associated with growth and preovulatory (PO) maturation of equine dominant follicles. Granulosa cells (GCs) and theca interna cells (TCs) were harvested from follicles (n = 5) at different stages of an ovulatory wave in mares corresponding to early dominance (ED; diameter ≥22 mm), late dominance (LD; ≥33 mm) and PO stage (34 h after administration...
Bastos HB, Kretzmann NA, Santos GO, Esmeraldino AT, Rechsteiner SF, Mattos RC, Neves AP.The period from the emergence of a dominant follicle until its formation requires tissue remodeling. Enzymes promoting collagen lysis, such as matrix metalloproteinases (MMPs), are fundamental for the process of extracellular matrix remodeling, which allows changes in ovarian tissue architecture during follicular growth. It has been suggested that the production of these enzymes may be affected by the rise in circulating concentrations of LH, which acts on the ovarian surface epithelium (OSE). The aim of this study was to determine the expression of MMP-1, MMP-2, and LH receptor (LHR) in the o...
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...
da Silveira JC, Carnevale EM, Winger QA, Bouma GJ.Ovarian follicle growth and maturation requires extensive communication between follicular somatic cells and oocytes. Recently, intercellular cell communication was described involving cell-secreted vesicles called exosomes (50-150 nm), which contain miRNAs and protein, and have been identified in ovarian follicular fluid. The goal of this study was to identify a possible role of exosomes in follicle maturation. Methods: Follicle contents were collected from mares at mid-estrous (~35 mm, before induction of follicular maturation) and pre-ovulatory follicles (30-34 h after induction of folli...
Lund CM, Ragle CA, Lutter JD, Farnsworth KD.To describe the use of a motorized morcellator for elective bilateral laparoscopic ovariectomy in standing equids and to evaluate long-term outcome. Methods: Retrospective case series. Methods: 30 equids (20 horses, 9 mules, and 1 pony). Methods: Medical records of equids undergoing elective bilateral laparoscopic ovariectomy from 2007 to 2013 were evaluated. Cases were selected on the basis of use of a motorized morcellator for ovary extraction. Data collected included age, breed, reason for surgery, surgery date, surgical approach, intraoperative complications, surgery and morcellation times...
Sayasith K, Sirois J.The interaction between stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) has been implicated in leukocyte attraction, tissue remodeling and angiogenesis. The objective of the present study was to characterize the expression and regulation of SDF1 and CXCR4 in equine follicles during the ovulatory process. Equine preovulatory follicles were isolated during estrus 0-39h after hCG treatment. Follicle wall preparations (theca interna with attached granulosa cells) and isolated preparations of granulosa cells and theca interna were obtained, and total RNA extracts wer...
Collins CW, Monfort SL, Vick MM, Wolfe BA, Weiss RB, Keefer CL, Songsasen N.To date, there has been limited research on manipulation of the estrous cycle in endangered equids. The objectives of this study were to assess the efficacy of using combinations of: (a) oral altrenogest and PGF2α, and (b) injectable altrenogest and PGF2α for manipulation of ovarian activity in Przewalski's mares. Reproductive cycles were monitored by assessing follicular changes with rectal ultrasound and changes in urinary steroid hormones. In Study 1, five cycling mares were treated with oral altrenogest (n=11 cycles) for 14 days. In Study 2, cycling mares were treated with oral altrenoge...
Iacono E, Merlo B, Rizzato G, Mislei B, Govoni N, Tamanini C, Mari G.The aim of the present study was to verify how repeated ovum pick-up (OPU), performed in anestrous and cyclic mares, affect ovarian activity, measured by progesterone (P4) and 17ß-estradiol (E2) plasma levels. Ovum pick-up of all visible follicles was performed every 9 to 12 days, and four sessions were carried out during anestrous (A) and breeding season (BS). The number of aspirated follicles per mare at each session was not significantly different between the two periods (BS: 6.1 ± 2.4; A: 7.5 ± 4.4; P > 0.05), but the mean follicular diameter was significantly higher during BS (16.0 ...
Szóstek AZ, Adamowski M, Galvão AM, Ferreira-Dias GM, Skarzynski DJ.Tumor necrosis factor-α (TNF) is a cytokine that plays important roles in functions of the endometrium. The aims of this study were to determine whether (i) ovarian steroids modulate TNF production by endometrial cells (Experiment 1); (ii) TNF effects on prostaglandin (PG) production in cultured equine endometrial cells and tissue (Experiment 2). Epithelial and stromal cells were isolated from equine endometrium (Days 2-5 of the estrous cycle; n=20) and treated after passage 1. In Experiment 1, epithelial and stromal cells were exposed to progesterone (P4; 10(-7)M), 17-β estradiol (E2; 10(-9...
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...
Kelley D, LeBlanc MM, Warren LK, Mortensen CJ.Supplementation with L-arginine can increase uterine arterial blood flow and vascular perfusion of the preovulatory follicle in mares. Increased vascular perfusion of the preovulatory follicle has been correlated with successful pregnancy in mares. The objective of this study was to determine if supplemental L-arginine would increase ovarian arterial blood flow, vascular perfusion of the preovulatory follicle, and embryo recovery rates in mares. Mares were blocked by age and breed and assigned at random within block to L-arginine supplementation or control groups. Mares were fed L-arginine beg...
Galli C, Duchi R, Colleoni S, Lagutina I, Lazzari G.Assisted reproductive techniques developed for cattle in the last 25 years, like ovum pick up (OPU), intracytoplasmic sperm injection (ICSI), and somatic cell nuclear transfer, have been transferred and adapted to buffalo and horses. The successful clinical applications of these techniques require both the clinical skills specific to each animal species and an experienced laboratory team to support the in vitro phase of the work. In cattle, OPU can be considered a consolidated technology that is rapidly outpacing conventional superovulation for embryo transfer. In buffalo, OPU represents the ...
Leonel EC, Bento-Silva V, Ambrozio KS, Luna HS, Costa e Silva EV, Zúccari CE.The aim of this study was to test the use of mechanical and mechanical-enzymatic methods, saline solution (SS), and PBS solution for the manipulation and isolation of mare ovarian preantral follicles (PAFs). The ovaries were subjected to mechanical isolation (mixer) alone or in association with enzymatic digestion (collagenase). Incubation times of 10 and 20 min were employed. In the first group, 4.1 ± 4.9 PAFs were harvested with the mechanical-enzymatic method vs 71.1 ± 19.2 with the mechanical procedure, showing a significant difference between methods; using SS and PBS, these number...
Schauer SN, Sontakke SD, Watson ED, Esteves CL, Donadeu FX.Previous evidence from in vitro studies suggests specific roles for a subset of miRNAs, including miR-21, miR-23a, miR-145, miR-503, miR-224, miR-383, miR-378, miR-132, and miR-212, in regulating ovarian follicle development. The objective of this study was to determine changes in the levels of these miRNAs in relation to follicle selection, maturation, and ovulation in the monovular equine ovary. In Experiment 1, follicular fluid was aspirated during ovulatory cycles from the dominant (DO) and largest subordinate (S) follicles of an ovulatory wave and the dominant (DA) follicle of a mid-cycle...
Donadeu FX, Schauer SN.Relatively little is known about the physiological roles of microRNAs (miRNAs) during follicular development. Previous evidence from in vitro studies suggests specific roles for a subset of miRNAs, including miR-21, miR-23a, miR-145, miR-503, miR-224, miR-383, miR-378, miR-132, and miR-212, in regulating ovarian follicle development. The objective of this study was to gain insight on the involvement of these miRNAs during follicle maturation. Follicular fluid was aspirated from dominant follicles (>32 mm) during the ovulatory season (July to October) and the anovulatory season (January to M...
Pereira GR, Lorenzo PL, Carneiro GF, Ball BA, Bilodeau-Goeseels S, Kastelic J, Pegoraro LM, Pimentel CA, Esteller-Vico A, Illera JC, Granado GS....The objectives of this study were to evaluate the effects of equine growth hormone (eGH) on nuclear and cytoplasmic maturation of equine oocytes in vitro, steroid production by cumulus cells, and expression and subcellular localization of eGH-receptors (eGH-R) on equine ovarian follicles. Cumulus-oocyte complexes (COCs) were recovered by aspirating follicles <30 mm in diameter from abattoir-derived ovaries. The COCs were morphologically evaluated and randomly allocated to be cultured in either a control maturation medium or supplemented with 400 ng/mL eGH, for 30 h at 38.5°C in air with 5%...
Cavalleri JM, Bienert-Zeit A, Feige K.The article summarizes the relevant clinical pathological assessment of horses with acute colic. A minimal laboratory evaluation should include the patient's haematocrit (or packed cell volume), total protein, and lactate concentration in the blood. Haematocrit and total protein provide an indication of the severity of dehydration (haematocrit < 0.45 l/l is evidence of no to mild dehydration whereas > 0.5 l/l points to a severe dehydration). The degree of dehydration is often associated with the severity of the colic. Additionally, the blood lactate concentration rises with increasing intesti...
Binder AK, Rodriguez KF, Hamilton KJ, Stockton PS, Reed CE, Korach KS.Determining the spatial and temporal expression of genes involved in the ovulatory pathway is critical for the understanding of the role of each estrogen receptor in the modulation of folliculogenesis and ovulation. Estrogen receptor (ER)-β is highly expressed in ovarian granulosa cells, and mice lacking ER-β are subfertile due to inefficient ovulation. Previous work has focused on isolated granulosa cells or cultured follicles and, although informative, provides confounding results due to the heterogeneous cell types present including granulosa and theca cells and oocytes and exposure to in...
Caron JP.Laparoscopic removal of the gonads from male and female horses is regularly performed for a variety of reasons. In female horses, common indications for ovariectomy include behavioral problems and signs of abdominal pain that are linked to estrus. Another indication is an abnormal, enlarged ovary, typically due to ovarian neoplasia, the most common being granulosa thecal cell tumor. Ovariectomy is also a treatment option for ovarian hematomas and other abnormalities. In addition, ovariectomy is used to sterilize mares intended for artificial insemination or embryo transfer programs.
Troedsson MH, Liu IK, Crabo BG.After the deposition of semen in the mare's uterus, spermatozoa must be transported to the site of fertilization, be maintained in the female tract until ovulation occurs, and be prepared to fertilize the released ovum. Sperm motility, myometrial contractions, and a spontaneous post-mating uterine inflammation are important factors for the transport and survival of spermatozoa in the mare's reproductive tract. Fertilizable sperm are present in the oviduct within 4 h after insemination. At this time, the uterus is the site of a hostile inflammatory environment. Our data suggest that spermatozoa...
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...
Gastal EL, Gastal MO, Wiltbank MC, Ginther OJ.By definition, follicle deviation begins on the day the two largest follicles of a wave begin to differ in growth rates. The relationships between follicle deviation and intrafollicular and systemic estradiol concentrations were studied in ponies, using a two-follicle model in which all but the two largest follicles were ablated. A 20-microliter sample of follicular fluid was obtained from each of the two follicles by transvaginal ultrasonography. In experiment 1, the two follicles were sampled when the larger follicle reached 15 mm. No differences (p > 0.05) in post-sampling follicle chara...
Wesson JA, Ginther OJ.Reproductive tracts were collected monthly over a 3-year period from 1,003 nonpregnant ponies at a slaughtering plant in Wisconsin. Ages of the animals were estimated from tooth replacement and wear. Ovaries were examined for follicular and luteal activity. There were no differences between left and right ovaries in size or occurrence of ovulation. The frequency of multiple ovulations, 10%, was higher than previously reported for ponies. The follicular changes during the ovulatory cycle did not support a two-wave theory of follicular growth. Ovarian activity changed seasonally, a finding simil...
Gomes RG, Andrade ER, Lisboa LA, Ciquini A, Barreiros TR, Fonseca NA, Seneda MM.The objective was to evaluate the efficiency of phosphate-buffered saline (PBS) and Minimum Essential Medium (MEM) during the transport of equine preantral and antral follicles at various temperatures and incubation interval. Equine ovaries (n = 10) from an abattoir were cut into 19 fragments; one was immediately fixed in Bouin's solution (control) and the other fragments were placed in PBS or MEM solution at 4, 20, or 39 °C for 4, 12, or 24 h. After the respective incubation periods, all fragments were fixed in Bouin's solution for 24 h and then submitted to standard histologic analysis. In ...
Smith RL, Vernon KL, Kelley DE, Gibbons JR, Mortensen CJ.The advent of embryo transfer has allowed horses to continue to train and compete during the breeding season. However, the associated stress of exercise may be detrimental to reproduction. The objectives of this study were to evaluate differing exercise protocols on reproductive blood flow and embryonic outcomes in mares. Light-horse mares were randomized into control (n = 4), partial-exercised (n = 6), and full-exercised (n = 6) groups. Partial-exercised mares were moderately exercised 30 min daily during the periovulatory period and rested after ovulation for 7 d. Full-exercised mares were e...
Kerban A, Doré M, Sirois J.In contrast to other species, the histology of the equine follicle during ovulation has not been described. Preovulatory follicles were isolated during oestrus at 0, 12, 24, 30, 33, 36 and 39 h (n = 5-6 follicles per time point) after an ovulatory dose of hCG to characterize the cellular and vascular changes associated with ovulation in mares. Pieces of follicle wall were formalin-fixed and processed for light microscopy to evaluate the general follicular morphology and quantify selected parameters. Marked changes were observed in the histology of equine follicles in the hours before ovulation...
Spicer LJ, Hammond JM.Development of the ovarian follicle(s) destined for ovulation appears to be a process in which antral follicles undergo a recruitment, selection and subsequent dominance phase. Several intraovarian or autocrine/paracrine regulatory mechanisms have been evoked to explain these processes. One of these potential autocrine/paracrine regulators is a catecholestrogen, 2-hydroxy-estradiol (2-OH-E2). Evidence implicating 2-OH-E2 as an autocrine/paracrine regulator of follicular function is reviewed. Studies have shown 2-OH-E2 to be present in nanomolar concentrations in fluid of human and equine folli...
Van Hoogmoed LM, Galuppo LD.To evaluate use of ENDO-GIA staples and ENDO-Catch pouches for ovariectomy in mares and to evaluate the efficacy of epidural morphine analgesia. Methods: Randomized clinical trial. Methods: Mares (n = 10) with normal ovaries. Methods: An Endo-GIA II stapler (United States Surgical Corp., Norwalk, CT) was used to amputate the ovaries bilaterally, in standing mares, and Endo-Catch II pouches (United States Surgical Corp.) were used for retrieval. Epidural morphine was used in 5 mares, and procedure duration, volume of lidocaine required to desensitize the ovarian pedicle, and sedation were compa...
Sessions DR, Vick MM, Fitzgerald BP.Matrix metalloproteinases (MMP) and tissue inhibitors of MMP (TIMP) regulate tissue remodeling events necessary for ovulation. Thus, changes in MMP and TIMP expression and protein enzyme activity were examined in vivo and in vitro during follicular development and atresia in the horse. Equine granulosa cells and follicular fluid from medium (15 to 29 mm) healthy and atretic follicles and from large (>30 mm) healthy and preovulatory follicles were collected by transvaginal aspiration. The cells were either snap-frozen (in vivo study) or cultured for 48 h (in vitro study) to determine gene ex...
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...
Ginther OJ, Bergfelt DR, Scraba ST, Pivonka PR, Nuti LC.The effect on reproduction of the plant derivative 6-methoxybenzoxazolinone (6-MBOA), which stimulates reproductive function in voles, was tested in pony mares, laboratory mice and rats, and mink. There was not a significant effect of intravenous injections of 6-MBOA on the ovarian follicles during the transition between the anovulatory and ovulatory seasons in mares. No significant effect of intraperitoneal injections of 6-MBOA on the weight of uterus or ovaries was found in eight-week-old mice, failing to confirm the results of an earlier report. In immature white rats, 6-MBOA treatment resu...
Mortensen CJ, Choi YH, Hinrichs K, Ing NH, Kraemer DC, Vogelsang SG, Vogelsang MM.The effect of exercise on mare reproductive efficiency was evaluated by comparing rates of embryo recovery from mares assigned to either an exercise regimen or a non-exercise (control) regimen. Exercised mares were worked daily for 30 min under average ambient conditions of >30 degrees C and >50% humidity. Mares were inseminated during estrus and subjected to uterine flush for embryo recovery on d 7 after ovulation for two consecutive cycles. After this, mares were allocated to the opposite group and allowed an estrous cycle without reproductive manipulation; then insemination and uterin...
Kenney RM, Ganjam VK.Focal enlargements of the uterus are palpable through the rectal wall and have various aetiology. Aspects of their diagnosis, possible pathogenesis, significance and treatment are presented. Lymphatic lacunae appear to be quite common in older mares and may occasionally give rise to large endometrial cyst, but more frequently cause widespread change throughout the uterine horns. Three unusual ovarian abnormalities are presented, a large, oestrogen-producing granulosa cell tumour, an oestrogen-producing large follicle in a 4-month-old filly, and an ectopic progesterone-producing adrenal structu...
Trostle SS, White NA, Donaldson L, Freeman LJ, Hendrickson DA.To describe a technique for laparoscopic colopexy in horses. Methods: Description of a technique to perform laparoscopic colopexy in horses. Methods: Six healthy adult horses between 3 and 23 years of age and weighing between 440 and 636 kg. Methods: Horses were positioned in dorsal recumbency, and the abdomen was insufflated to an intraabdominal pressure of 15 mm Hg. A laparoscope portal was placed at the umbilicus, and instrument portals were placed cranial to the laparoscope and lateral to midline. The lateral taenia of the left ventral colon was laparoscopically identified and retracted wi...
Matthews NS, Hartsfield SM, Hague B, Carroll GL, Short CE.To evaluate propofol for induction and maintenance of anesthesia, after detomidine premedication, in horses undergoing abdominal surgery for creation of an experimental intestinal adhesion model. Methods: Prospective study. Methods: Twelve horses (424 +/- 81 kg) from 1 to 20 years of age (5 females, 7 males). Methods: Horses were premedicated with detomidine (0.015 mg/kg i.v.) 20 to 25 minutes before induction, and a propofol bolus (2 mg/kg i.v.) was administered for induction. Propofol infusion (0.2 mg/kg/min i.v.) was used to maintain anesthesia. The infusion rate was adjusted to maintain an...
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...
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...
Jennings MW, Boime I, Daphna-Iken D, Jablonka-Shariff A, Conley AJ, Colgin M, Bidstrup LA, Meyers-Brown GA, Famula TR, Roser JF.The efficacy of a recently engineered single chain recombinant equine follicle stimulating hormone (reFSH) was investigated in estrous cycling mares whose gonadotropins and follicular activity had been suppressed by concurrent treatment with progesterone and estradiol (P&E). Time of estrus was synchronized in 15 estrous cycling mares during the breeding season with prostaglandins F(2alpha) (PGF(2alpha)). The day after ovulation, mares were treated once daily with P&E for 14 days. Mares received a second injection of PGF(2alpha) on day 6 of the synchronized estrous cycle to induce luteolysis. O...
Koskinen E, Kuntsi H, Lindeberg H, Katila T.Follicular growth and hormone determinations were used to predict ovulation in the mare. Thirty Finnhorse mares were used for the investigation and 38 oestrous cycles were studied. The mares were examined by rectal palpation and ultrasonography every 6 hours during late oestrus. Daily blood samples were obtained for progesterone and oestrone sulphate determination. The preovulatory follicle grew 3 mm a day up to 2 days before ovulation. The size then remained constant, before diminishing by 2-3 mm during the last 12 hours. The maximal diameter of the follicle was 43 +/- 4 mm. In 89% of the fol...
Mlodawska W, Slomczynska M.Ovarian steroidogenesis from the neonatal to pubertal period in horses is poorly understood. This study was designed to immunolocalize cytochrome P450 aromatase in the ovarian follicles of slaughtered fillies ages approximately (I) 6-9 mo (5 mm in diameter. Staining intensity was dependent on the size and morphology of the follicle. In nonatretic follicles 5-10 mm in diameter, the reaction was weak and heterogeneous, while most intense staining was observed in preovulatory follicles. In follicles (diameter <20 mm) in the groups <10MF and 1YF, the reaction was less intense than in adult m...
Driancourt MA, Paris A, Roux C, Mariana JC, Palmer E.Five pony and 5 saddle-type mares were used to compare total ovarian follicular populations between breeds. The animals were hemi-ovariectomized at the preovulatory stage during the breeding season and the ovary bearing the large preovulatory follicle was studied using histological techniques. Pony and saddle-type mares did not differ as to mean number of primordial follicles, mean number of growing follicles, the variability of these numbers and follicular distribution into various size classes. No difference was detected either in the initiation processes of follicular growth, oocyte growth,...
Lorenzo PL, Liu IK, Carneiro GF, Conley AJ, Enders AC.Epidermal growth factor (EGF) has been shown to have a positive effect during oocyte in vitro maturation in several species. This study was performed to establish the capacity of equine oocytes to undergo nuclear maturation in the presence of EGF and to localise its receptor in the equine ovary by immunohistochemical methods. Oocytes were obtained by aspiration and subsequent scraping from equine follicles (15-25 mm diameter) and cultured in 3 different treatment groups for 36 h: control Group (modified TCM 199 with 0.003% BSA), EGF Group (TCM-199 supplemented with 50 ng/ml EGF) and EMS Group ...
Mattos MC, Bastos MR, Guardieiro MM, Carvalho JO, Franco MM, Mourão GB, Barros CM, Sartori R.The aim of this study was to evaluate the superovulatory (SOV) response of Sindhi (Bos indicus) donors submitted to an ovarian follicular superstimulatory protocol replacing the last two doses of pFSH by eCG. Forty-eight SOV treatments were performed in a crossover design in 19 nulliparous and primiparous females that were randomly divided into two groups: FSH (n=24), which consisted of eight pFSH injections, or FSH/eCG (n=24), which consisted of six pFSH injections followed by two eCG injections. Each female underwent two or three SOV treatments that consisted of an i.m. injection of 2mg estr...
Kelley DE, Warren LK, Mortensen CJ.L-arginine is an amino acid which can alter pituitary function and increase blood flow to the reproductive tract. The objective was to determine the effect of supplementing 100g of L-arginine on plasma arginine concentrations, follicular dynamics and ovarian and uterine artery blood flow during the estrus that occurs subsequent to foaling. In Experiment 1, mares were fed 100g L-arginine for 1 day during the last 3 weeks of pregnancy and plasma samples taken for every hour for the first 4h and every other hour until 12h.L-arginine supplementation elevated plasma arginine concentrations from 1 t...
Ishak GMA, Dutra GA, Gastal GDA, Gastal MO, Feugang JM, Gastal EL.The inability to obtain in vivo samples of antral follicle wall layers without removing the ovaries or sacrificing the animals has limited more in-depth studies on folliculogenesis. In this study, a novel ultrasound-guided follicle wall biopsy (FWB) technique was used to obtain in vivo follicle wall layers and follicular fluid samples of growing antral follicles. The expression of proliferative, hormonal, angiogenic, and pro-/antiapoptotic receptors and proteins in the follicular wall among three follicle classes were compared during the spring transitional anovulatory (SAN) and spring ovulato...
Agnieszka N, Joanna K, Wojciech W, Adam O.The aim of this study was determine the viability and developmental competence of equine oocytes after IVM and vitrification using the Rapid-I method, as part of an effort to develop an effective equine oocyte vitrification protocol. Equine oocytes were collected by scraping ovarian follicles of slaughtered mares. A total of 1052 ovaries were used in this study, from which 3135 oocytes were obtained. Of the 2853 oocytes retrieved, 2557 underwent in vitro maturation for approximately 36 h. After in vitro culture, 1202 oocytes (47%) had a first polar body. To evaluate the toxicity of the solu...
