Granulosa cells in horses are specialized cells that form a part of the ovarian follicles in the equine reproductive system. These cells are involved in the production of sex steroids and are integral to the process of folliculogenesis and oocyte maturation. Granulosa cells facilitate the growth and development of the oocyte by providing necessary nutrients and signaling molecules. They also produce hormones such as estrogen, which are vital for the regulation of the estrous cycle in mares. Changes in granulosa cell function or structure can impact fertility and reproductive health in horses. This page compiles peer-reviewed research studies and scholarly articles that explore the biology, function, and implications of granulosa cells in equine reproduction.
Catandi GD, Fresa KJ, Cheng MH, Whitcomb LA, Broeckling CD, Chen TW, Chicco AJ, Carnevale EM.Obesity is a growing concern in human and equine populations, predisposing to metabolic pathologies and reproductive disturbances. Cellular lipid accumulation and mitochondrial dysfunction play an important role in the pathologic consequences of obesity, which may be mitigated by dietary interventions targeting these processes. We hypothesized that obesity in the mare promotes follicular lipid accumulation and altered mitochondrial function of oocytes and granulosa cells, potentially contributing to impaired fertility in this population. We also predicted that these effects could be mitigated ...
Ren W, Wang J, Zeng Y, Wang T, Meng J, Yao X.The Kazakh horse, renowned for its excellence as a breed, exhibits distinctive reproductive traits characterized by early maturity and seasonal estrus. While normal reproductive function is crucial for ensuring the breeding and expansion of the Kazakh horse population, a noteworthy decline in reproductive capabilities is observed after reaching 14 years of age. Unassigned: In this study, ovarian granulosa cells (GCs) were meticulously collected from Kazakh horses aged 1, 2, 7, and above 15 years old (excluding 15 years old) for whole transcriptome sequencing. Unassigned: The analysis identifie...
Pompermayer E, Ysebaert MP, Vinardell T, Oikawa MA, Jonhson JP, Fernandes T, David F.A two-year-old Arabian horse presented for abnormal external genitalia and dangerous stallion-like behavior was diagnosed with disorder of sexual development (DSD), also known as intersex/hermaphroditism. Standing 1-stage surgical procedure performed under sedation, and local anesthesia to concurrently eliminate stallion-like behavior, risk of neoplastic transformation of intraabdominal gonads, and to replace ambiguous external genital with a functional, and cosmetically more acceptable anatomy. Step-1) Laparoscopic abdominal exploration and gonadectomy; Step-2) Rudimentary penis resection and...
Tarko A, Fabová Z, Kotwica J, Valocký I, Alrezaki A, Alwasel S, Harrath AH, Sirotkin AV.The influence of environmental contaminant toluene and of plant fennel (Foeniculum vulgare Mill.) on reproduction are reported, but the mechanisms of their action and the protective effect of fennel on contaminant influence remain to be elucidated. In this study, we hypothesized that toluene and fennel directly affects basic ovarian cell functions, and that fennel can be used as an appropriate natural protective agent against the potential adverse effects of toluene. This study aimed to examine the action of toluene (20 μg/mL) and fennel extract (0, 1, 10, 100 μg/mL), and assess their comb...
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...
Gee EK, Dicken M, Archer RM, Herdan CL, Pauwels FE, Drayton BM.A 14-year-old Thoroughbred mare was presented for evaluation of a rapidly enlarging ovary at 73 days gestation. The mare's owner reported behavioural changes in the mare since she returned from stud, notably aggression towards other horses. Results: Transrectal palpation and ultrasonography of the reproductive tract revealed a viable pregnancy, an enlarged, multiloculated right ovary, and a normal-sized left ovary with a corpus luteum present. Concentrations of inhibin and testosterone in serum were 13.8 ng/mL and 1,150 pg/mL, respectively, being markedly higher than reference ranges. Concentr...
Almeida J, Ball BA, Conley AJ, Place NJ, Liu IK, Scholtz EL, Mathewson L, Stanley SD, Moeller BC.Anti-Müllerian hormone (AMH), a member of the transforming growth factor β superfamily of growth and differentiation factors, is expressed in granulosa cells of preantral and small antral ovarian follicles. In humans, AMH appeared to regulate recruitment and growth of small ovarian follicles. Furthermore, circulating AMH concentrations were elevated in women with granulosa-cell tumors (GCT). In the horse, GCTs are the most common tumor of the ovary, and a variety of endocrine assays have been used to diagnose presumptive GCTs. The objectives of the present study were to validate a heterologo...
Goodin JT, Rodgerson DH, Gomez JH.To review the efficacy and safety of unilateral ovariectomy by use of a standing hand-assisted laparoscopic approach and evaluate the effect of ovary size on posttransection hemorrhage after application of a linear stapling device. Methods: Case series. Methods: Horses (n=65) aged 2-20 years. Methods: Medical records of mares that had ovariectomy performed by use of a standing hand-assisted laparoscopic approach were reviewed. Data retrieved were signalment, ovary removed (left, right), ovarian size, ovarian pathology, intraoperative and postoperative complications, and length of hospitalizati...
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...
Ball BA, Conley AJ, MacLaughlin DT, Grundy SA, Sabeur K, Liu IK.Anti-Müllerian hormone (AMH), also known as Müllerian inhibiting substance (MIS), is expressed by granulosa cells in females of many mammalian species, and circulating AMH concentrations have been used to monitor granulosa-cell tumors (GCT) in women. The objective was to characterize expression of AMH in equine GCT, and in normal equine ovaries, based upon immunohistochemistry (IHC), using a polyclonal primary antibody directed against human AMH. Equine GCT (n=27) and normal equine ovaries (n=10) were examined by IHC. In addition, sera from four mares with GCT were characterized for AMH bioa...
Shimada M, Yanai Y, Okazaki T, Yamashita Y, Sriraman V, Wilson MC, Richards JS.During ovulation, granulosa cells and cumulus cells synthesize and secrete a wide variety of factors including members of the IL cytokine family via the process of exocytosis. Exocytosis is controlled by the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex consisting of proteins residing in the vesicle membrane and the plasma membrane. One of the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor proteins, synaptosomal-associated protein (SNAP)25, is expressed abundantly in neuronal cells and is also induced transiently in the rat...
Minervini F, Giannoccaro A, Fornelli F, Dell'Aquila ME, Minoia P, Visconti A.The mycotoxin zearalenone (ZEA) and its derivatives, alpha and beta-zearalenol (alpha and beta-ZOL), synthesized by genera Fusarium, often occur as contaminants in cereal grains and animal feeds. The importance of ZEA on reproductive disorders is well known in domestic animals species, particularly in swine and cattle. In the horse, limited data are available to date on the influence of dietary exposure to ZEA on reproductive health and on its in vitro effects on reproductive cells. The aim of this study was to evaluate the effects of ZEA and its derivatives, alpha and beta-ZOL, on granulosa c...
Zelli R, Sylla L, Monaci M, Stradaioli G, Sibley LE, Roser JF, Munro C, Liu IK.Granulosa-theca cell tumors (GTCTs) are able to secrete variable amounts of sex steroids and immunoreactive inhibin (ir-INH). Although the pituitary appears to be affected by the presence of a GTCT, pituitary responsiveness to exogenous GnRH has not been examined. The aims of the present study were to: (i) assess the plasma hormone concentrations of ir-INH, gonadotropins and sex steroids in eight mares with GTCT and (ii) assess the responsiveness of pituitary gonadotroph cells to exogenous GnRH stimulus both before and after tumor removal. In seven mares, the contralateral ovary was firm, smal...
Sayasith K, Bouchard N, Boerboom D, Brown KA, Doré M, Sirois J.Ovulation is accompanied by a marked infiltration of leukocytes into thecal layers after the gonadotropin surge. P-selectin is known to play a critical role in the initial steps of leukocyte recruitment from the bloodstream during inflammation. Thus, the objective was to investigate the potential regulation of P-selectin by gonadotropins in equine preovulatory follicles. The full-length equine P-selectin cDNA was cloned by a combination of reverse transcription-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends. Results showed that equine P-selectin cDNA encodes...
Carson-Dunkerley SA, Hanson RR.To describe the short- and long-term survival rates in horses undergoing ovariectomy for granulosa cell tumors by use of the diagonal paramedian approach. Methods: Retrospective case study. Methods: 12 horses with granulosa cell tumors. Methods: A diagonal paramedian approach for unilateral ovariectomy was used for removal of each mare's granulosa cell tumor. Information about complications and outcomes was analyzed. Results: Only minimal complications were detected postoperatively when the diagonal paramedian approach was used, regardless of the preferred technique for ovarian pedicle ligatio...
Hurwitz A, Finci-Yeheskel Z, Dushnik M, Milwidsky A, Shimonovitz S, Yagel S, Adashi EY, Mayer M.This study examines the effects of interleukin-1 (IL-1) on plasminogen activator (PA) activity and prostaglandin (PG) E production in pregnant mare serum gonadotropin (PMSG)-primed granulosa cells and the potential involvement of PGE in the regulation of ovarian plasminogen activation. Methods: Granulosa cells were obtained from PMSG-primed rat (27-day-old) ovaries and cultured in serum-free conditions for 48 hours in the absence or presence of IL-1 beta (10 ng/mL) with and without transforming growth factor-beta 1 (10 ng/mL). Cellular PA activity was measured through the conversion of plasmin...
Hinrichs K, Martin MG, Schmidt AL, Friedman PP.Two experiments were conducted to evaluate the effect of follicular components on the maintenance of meiotic arrest in horse oocytes. In Expt 1, oocytes were incubated for 24 h with follicular fluid, or with granulosa cells suspended either in medium or in follicular fluid at 25 x 10(6) cells ml-1. None of the treatments resulted in significant maintenance of the germinal vesicle stage over that of non-suppressive control. Culture with follicular fluid plus granulosa cells resulted in a significantly higher proportion of oocytes at metaphase I compared with controls. In Expt 2, oocytes were di...
Hinrichs K, Hunt PR.The ultrasonic appearance of seven cases of granulosa cell tumour is described, and compared with an ovarian haematoma and an ovarian serous cystadenoma. The granulosa cell tumours varied from being uniformly dense to having one or several large fluid filled cysts. Some tumours resembled the haematoma or cystadenoma and also, in some aspects, normal ovarian structures such as corpora haemorrhagica or follicles in early pregnancy. There was no typical ultrasonographic appearance of the granulosa cell tumours which enabled definitive diagnosis. However, ultrasound may be a diagnostic aid when us...
Allen WE.The situations and conditions that can disrupt the mares normal oestrous cycle are described. Season of the year is a major influence; maximum reproductive efficiency does not totally coincide with the artificially defined "breeding season". Other abnormalities are associated with spontaneously persistent luteal function, psychological influences over behavioural activity, the presence of endometritis, multiple ovulation, reproductive behavior after pregnancy failure and granulosa cell tumours.
Shimada M, Yanai Y, Okazaki T, Yamashita Y, Sriraman V, Wilson MC, Richards JS.During ovulation, granulosa cells and cumulus cells synthesize and secrete a wide variety of factors including members of the IL cytokine family via the process of exocytosis. Exocytosis is controlled by the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex consisting of proteins residing in the vesicle membrane and the plasma membrane. One of the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor proteins, synaptosomal-associated protein (SNAP)25, is expressed abundantly in neuronal cells and is also induced transiently in the rat...
Ball BA, Conley AJ, MacLaughlin DT, Grundy SA, Sabeur K, Liu IK.Anti-Müllerian hormone (AMH), also known as Müllerian inhibiting substance (MIS), is expressed by granulosa cells in females of many mammalian species, and circulating AMH concentrations have been used to monitor granulosa-cell tumors (GCT) in women. The objective was to characterize expression of AMH in equine GCT, and in normal equine ovaries, based upon immunohistochemistry (IHC), using a polyclonal primary antibody directed against human AMH. Equine GCT (n=27) and normal equine ovaries (n=10) were examined by IHC. In addition, sera from four mares with GCT were characterized for AMH bioa...
Almeida J, Ball BA, Conley AJ, Place NJ, Liu IK, Scholtz EL, Mathewson L, Stanley SD, Moeller BC.Anti-Müllerian hormone (AMH), a member of the transforming growth factor β superfamily of growth and differentiation factors, is expressed in granulosa cells of preantral and small antral ovarian follicles. In humans, AMH appeared to regulate recruitment and growth of small ovarian follicles. Furthermore, circulating AMH concentrations were elevated in women with granulosa-cell tumors (GCT). In the horse, GCTs are the most common tumor of the ovary, and a variety of endocrine assays have been used to diagnose presumptive GCTs. The objectives of the present study were to validate a heterologo...
Minervini F, Giannoccaro A, Fornelli F, Dell'Aquila ME, Minoia P, Visconti A.The mycotoxin zearalenone (ZEA) and its derivatives, alpha and beta-zearalenol (alpha and beta-ZOL), synthesized by genera Fusarium, often occur as contaminants in cereal grains and animal feeds. The importance of ZEA on reproductive disorders is well known in domestic animals species, particularly in swine and cattle. In the horse, limited data are available to date on the influence of dietary exposure to ZEA on reproductive health and on its in vitro effects on reproductive cells. The aim of this study was to evaluate the effects of ZEA and its derivatives, alpha and beta-ZOL, on granulosa c...
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 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...
Tarko A, Fabová Z, Kotwica J, Valocký I, Alrezaki A, Alwasel S, Harrath AH, Sirotkin AV.The influence of environmental contaminant toluene and of plant fennel (Foeniculum vulgare Mill.) on reproduction are reported, but the mechanisms of their action and the protective effect of fennel on contaminant influence remain to be elucidated. In this study, we hypothesized that toluene and fennel directly affects basic ovarian cell functions, and that fennel can be used as an appropriate natural protective agent against the potential adverse effects of toluene. This study aimed to examine the action of toluene (20 μg/mL) and fennel extract (0, 1, 10, 100 μg/mL), and assess their comb...
Carson-Dunkerley SA, Hanson RR.To describe the short- and long-term survival rates in horses undergoing ovariectomy for granulosa cell tumors by use of the diagonal paramedian approach. Methods: Retrospective case study. Methods: 12 horses with granulosa cell tumors. Methods: A diagonal paramedian approach for unilateral ovariectomy was used for removal of each mare's granulosa cell tumor. Information about complications and outcomes was analyzed. Results: Only minimal complications were detected postoperatively when the diagonal paramedian approach was used, regardless of the preferred technique for ovarian pedicle ligatio...
Hinrichs K, Martin MG, Schmidt AL, Friedman PP.Two experiments were conducted to evaluate the effect of follicular components on the maintenance of meiotic arrest in horse oocytes. In Expt 1, oocytes were incubated for 24 h with follicular fluid, or with granulosa cells suspended either in medium or in follicular fluid at 25 x 10(6) cells ml-1. None of the treatments resulted in significant maintenance of the germinal vesicle stage over that of non-suppressive control. Culture with follicular fluid plus granulosa cells resulted in a significantly higher proportion of oocytes at metaphase I compared with controls. In Expt 2, oocytes were di...
Hurwitz A, Finci-Yeheskel Z, Dushnik M, Milwidsky A, Shimonovitz S, Yagel S, Adashi EY, Mayer M.This study examines the effects of interleukin-1 (IL-1) on plasminogen activator (PA) activity and prostaglandin (PG) E production in pregnant mare serum gonadotropin (PMSG)-primed granulosa cells and the potential involvement of PGE in the regulation of ovarian plasminogen activation. Methods: Granulosa cells were obtained from PMSG-primed rat (27-day-old) ovaries and cultured in serum-free conditions for 48 hours in the absence or presence of IL-1 beta (10 ng/mL) with and without transforming growth factor-beta 1 (10 ng/mL). Cellular PA activity was measured through the conversion of plasmin...
Sayasith K, Bouchard N, Boerboom D, Brown KA, Doré M, Sirois J.Ovulation is accompanied by a marked infiltration of leukocytes into thecal layers after the gonadotropin surge. P-selectin is known to play a critical role in the initial steps of leukocyte recruitment from the bloodstream during inflammation. Thus, the objective was to investigate the potential regulation of P-selectin by gonadotropins in equine preovulatory follicles. The full-length equine P-selectin cDNA was cloned by a combination of reverse transcription-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends. Results showed that equine P-selectin cDNA encodes...
Hinrichs K, Hunt PR.The ultrasonic appearance of seven cases of granulosa cell tumour is described, and compared with an ovarian haematoma and an ovarian serous cystadenoma. The granulosa cell tumours varied from being uniformly dense to having one or several large fluid filled cysts. Some tumours resembled the haematoma or cystadenoma and also, in some aspects, normal ovarian structures such as corpora haemorrhagica or follicles in early pregnancy. There was no typical ultrasonographic appearance of the granulosa cell tumours which enabled definitive diagnosis. However, ultrasound may be a diagnostic aid when us...
Goodin JT, Rodgerson DH, Gomez JH.To review the efficacy and safety of unilateral ovariectomy by use of a standing hand-assisted laparoscopic approach and evaluate the effect of ovary size on posttransection hemorrhage after application of a linear stapling device. Methods: Case series. Methods: Horses (n=65) aged 2-20 years. Methods: Medical records of mares that had ovariectomy performed by use of a standing hand-assisted laparoscopic approach were reviewed. Data retrieved were signalment, ovary removed (left, right), ovarian size, ovarian pathology, intraoperative and postoperative complications, and length of hospitalizati...
Gee EK, Dicken M, Archer RM, Herdan CL, Pauwels FE, Drayton BM.A 14-year-old Thoroughbred mare was presented for evaluation of a rapidly enlarging ovary at 73 days gestation. The mare's owner reported behavioural changes in the mare since she returned from stud, notably aggression towards other horses. Results: Transrectal palpation and ultrasonography of the reproductive tract revealed a viable pregnancy, an enlarged, multiloculated right ovary, and a normal-sized left ovary with a corpus luteum present. Concentrations of inhibin and testosterone in serum were 13.8 ng/mL and 1,150 pg/mL, respectively, being markedly higher than reference ranges. Concentr...
Zelli R, Sylla L, Monaci M, Stradaioli G, Sibley LE, Roser JF, Munro C, Liu IK.Granulosa-theca cell tumors (GTCTs) are able to secrete variable amounts of sex steroids and immunoreactive inhibin (ir-INH). Although the pituitary appears to be affected by the presence of a GTCT, pituitary responsiveness to exogenous GnRH has not been examined. The aims of the present study were to: (i) assess the plasma hormone concentrations of ir-INH, gonadotropins and sex steroids in eight mares with GTCT and (ii) assess the responsiveness of pituitary gonadotroph cells to exogenous GnRH stimulus both before and after tumor removal. In seven mares, the contralateral ovary was firm, smal...
Allen WE.The situations and conditions that can disrupt the mares normal oestrous cycle are described. Season of the year is a major influence; maximum reproductive efficiency does not totally coincide with the artificially defined "breeding season". Other abnormalities are associated with spontaneously persistent luteal function, psychological influences over behavioural activity, the presence of endometritis, multiple ovulation, reproductive behavior after pregnancy failure and granulosa cell tumours.
Pompermayer E, Ysebaert MP, Vinardell T, Oikawa MA, Jonhson JP, Fernandes T, David F.A two-year-old Arabian horse presented for abnormal external genitalia and dangerous stallion-like behavior was diagnosed with disorder of sexual development (DSD), also known as intersex/hermaphroditism. Standing 1-stage surgical procedure performed under sedation, and local anesthesia to concurrently eliminate stallion-like behavior, risk of neoplastic transformation of intraabdominal gonads, and to replace ambiguous external genital with a functional, and cosmetically more acceptable anatomy. Step-1) Laparoscopic abdominal exploration and gonadectomy; Step-2) Rudimentary penis resection and...
Ren W, Wang J, Zeng Y, Wang T, Meng J, Yao X.The Kazakh horse, renowned for its excellence as a breed, exhibits distinctive reproductive traits characterized by early maturity and seasonal estrus. While normal reproductive function is crucial for ensuring the breeding and expansion of the Kazakh horse population, a noteworthy decline in reproductive capabilities is observed after reaching 14 years of age. Unassigned: In this study, ovarian granulosa cells (GCs) were meticulously collected from Kazakh horses aged 1, 2, 7, and above 15 years old (excluding 15 years old) for whole transcriptome sequencing. Unassigned: The analysis identifie...
Catandi GD, Fresa KJ, Cheng MH, Whitcomb LA, Broeckling CD, Chen TW, Chicco AJ, Carnevale EM.Obesity is a growing concern in human and equine populations, predisposing to metabolic pathologies and reproductive disturbances. Cellular lipid accumulation and mitochondrial dysfunction play an important role in the pathologic consequences of obesity, which may be mitigated by dietary interventions targeting these processes. We hypothesized that obesity in the mare promotes follicular lipid accumulation and altered mitochondrial function of oocytes and granulosa cells, potentially contributing to impaired fertility in this population. We also predicted that these effects could be mitigated ...
