The testes in horses are reproductive organs responsible for the production of sperm and the secretion of hormones such as testosterone. Located in the scrotum, the testes play a vital role in male fertility and reproductive behavior. They consist of seminiferous tubules where spermatogenesis occurs, and interstitial cells, also known as Leydig cells, which produce testosterone. The function and health of equine testes can be influenced by factors such as age, season, and overall health status. This page compiles peer-reviewed research studies and scholarly articles that explore the anatomy, physiology, and pathology of the testes in horses, providing insights into their role in equine reproduction and breeding management.
DAVIES J, DEMPSEY EW, WISLOCKI GB.A fetal horse ovary at the 6th month of gestation and a fetal ovary and testis at the 9th month have been studied histochemically with special reference to the cytoplasmic lipids of the interstitial cells. At least two lipid fractions appeared to be present, one which was soluble in acetone and was mainly responsible for the positive "plasmal" reaction, and another, insoluble in acetone, which was responsible for the sudanophilic, Ashbel-Seligman and periodic acid-Schiff positive material remaining after acetone extraction. The interstitial cell lipids in the older ovary and testis were also a...
Serum anti-Müllerian hormone (AMH), a marker of equine cryptorchidism, is detectable in intact and cryptorchid stallions but not in geldings because it is secreted from Sertoli cells. A 4-year-old uncastrated Thoroughbred racehorse had no visible testes; therefore, the horse was considered a bilateral cryptorchidism. However, the serum AMH was undetectable (<0.08 ng/ml). Human chorionic gonadotrophin (hCG) stimulating test result indicated that the horse was a gelding. The results of sex chromosomal analysis and sequence analysis of SRY gene suggested that the horse was a genetically-intac...
Oh R, Tamaoki BI.The microsomal fraction (10 000–105 000 × g precipitate) of equine testes was fractionated into the smooth- and the rough-surfaced microsomal subfractions by a sucrose density-gradient centrifugation in the presence of CsCl. The validity of this fractionating procedure was confirmed by electron microscopic examination and also by chemical analysis of the RNA contents in these subfractions. The aromatizing enzyme system (19-hydroxylase and aromatase) which was concentrated in the microsomal fractions among the organellae was found to be localized in the smoothsurfaced microsomal fraction. Th...
Hoshiba H, Sinowatz F.Spermadhesins are proteins with various functions in sperm capacitation and zona pellucida binding. In this study the cellular localization of the spermadhesin AWN-1 has been examined in the equine male genital tract. Results obtained by immunohistochemical methods reveal that in the horse AWN-1 is synthesized in spermatogonia, in the rete testis, the ductus epididymidis and the seminal vesicles. These findings indicate that the cellular origin of spermadhesins is species-specific.
Talluri TR, Jhamb D, Paul N, Singh J, Dedar RK, Mehta SC, Legha RA, Pal Y.The recovery of spermatozoa from the cauda epididymis may be the only option to obtain genetic material from elite stallions that had undergone castration or sudden death due to colic or severe injury. Objective: To evaluate two different protocols for retrieval of stallion epididymal spermatozoa and to evaluate different cryoprotectants on the freezability of the epididymal spermatozoa. Methods: Six epididymides from three stallions were collected immediately after routine castration under general anesthesia. In the first experiment, each epididymis (of two testes) of the same stallion were p...
Bucci R, De Amicis I, Parrillo S, Robbe D, Carluccio A.Good sperm production is a notable requirement for subjects intended for reproduction, particularly in endangered species, and it has been demonstrated that in horse stallions, this is correlated to testicular volume. The present study, which involved Martina Franca jacks, aimed to determine whether, also in this endangered breed, there is a correlation between the total sperm number (TSN) and testicular volume. Testes were measured with both ultrasound and a caliper. Testicular volume was calculated using two different formulas: one representing the volume of an ellipsoid and one developed to...
Cinone M, Albrizio M, Guaricci AC, Lacitignola L, Desantis S.Heat shock proteins are the most evolutionarily conserved protein families induced by stressors including hyperthermia. In the context of pathologies of the male reproductive tract, cryptorchidism is the most common genital defect that compromises the reproductive potential of the male because it induces an increase in intratesticular temperature. In equine species, cryptorchidism affects almost 9 % of newborns and few studies have been carried out on the molecular aspects of the retained testis. In this study, the expression pattern of HSP60, 70, and 90 in abdominal and inguinal testes, in t...
Shakeel M, Choi Y, Yoon M.Cryptorchidism affects spermatogenesis and testis development, often resulting in stallion subfertility/infertility. This study aims to identify the specific germ cells impacted by cryptorchism in stallions. In a previous study, we found that PGP9.5 and VASA are molecular markers expressed in different germ cells within stallions. Herein, we assessed the heat stress-induced response of spermatogonial stem cells (SSCs) in the seminiferous tubules (ST) of cryptorchid stallion testes (CST) and normal stallion testes (NST). This goal was accomplished by comparing PGP9.5 and VASA expression pattern...
Johnson SA, Sikes KJ, Johnson JW, Van Zeeland E, Wist S, Santangelo KS, King MR, Frisbie DD.Low load exercise training with blood flow restriction (BFR) has become increasingly used by human physical therapists to prescribe controlled exercise following orthopaedic injury; its effects on the equine superficial digital flexor tendon (SDFT), however, are unknown. Objective: To investigate outcomes of pressure specific BFR walking exercise on uninjured equine SDFT biomechanics and histomorphology. Methods: Controlled in vivo experiment. Methods: Four forelimbs of four horses were exposed to 40 BFR-walk sessions (10-min interval walking) on a treadmill over a 56-day study period with the...
Yi M, Asgenbaatar N, Wang X, Ulaangerel T, Shen Y, Wen X, Du M, Dong X, Dugarjav M, Bou G.DNA methyltransferases (DNMTs) are important epigenetic modification during spermatogenesis. To further evaluate the pattern of DNMTs in horse testes during development, we investigated the expression and localization of DNMT1, DNMT3a and DNMT3b at different time points. The qRT-PCR results showed that DNMT1 expression was maintained in testes tissue from 6-month-old (0.5y) to 2-year-old (2y) of age and decreased after 3-year-old (3y) (P < 0.01). The expression levels of DNMT3a and DNMT3b peaked in testes tissue at 3y (P < 0.01). At 4-year-old (4y), the expression of DNMT3a and DNMT3b was ...
Liu Y, Du M, Li X, Zhang L, Zhao B, Wang N, Dugarjaviin M.This study aimed to investigate differences in testicular tissue morphology, gene expression, and marker genes between sexually immature (1-year-old) and sexually mature (10-year-old) Mongolian horses. The purposes of our research were to provide insights into the reproductive physiology of male Mongolian horses and to identify potential markers for sexual maturity. The methods we applied included the transcriptomic profiling of testicular cells using single-cell sequencing techniques. Our results revealed significant differences in tissue morphology and gene expression patterns between the tw...
Shakeel M, Yoon M.Spermatogonial stem cells (SSCs) are essential for the maintenance of male fertility and survival of species. Environmental conditions, notably heat stress, have been identified as important causes of male infertility and have a negative impact on SSCs. Animals with cryptorchid testes (CT) are optimal models for the study of long-term heat stress-related changes in germ cells. The effect of heat stress on germ cells differs depending on the spermatogenesis stage. Thus, verifying whether the specific phase of spermatogenesis is dependent or independent of heat stress in stallions is important. ...
Liu Y, Du M, Zhang L, Wang N, He Q, Cao J, Zhao B, Li X, Li B, Bou G, Zhao Y, Dugarjaviin M.Testicular development and spermatogenesis are tightly regulated by both coding and non-coding genes, with mRNA and lncRNA playing crucial roles in post-transcriptional gene expression regulation. However, there are significant differences in regulatory mechanisms before and after sexual maturity. Nevertheless, the mRNAs and lncRNAs in the testes of Mongolian horses have not been systematically identified. In this study, we first identified the testicular tissues of sexually immature and sexually mature Mongolian horses at the tissue and protein levels, and comprehensively analyzed the express...
Asgenbaatar N, Yi M, Wang X, Ulaangerel T, Shen Y, Wen X, Du M, Dong X, Mengkh Y, Dugarjav M, Bou G.The preparation of paraffin sections is an important experimental technique in animal histological research, and key factors that determine the quality of a section include the dehydration time, waxing time, and drying temperature of the paraffin section. Paraffin sections obtained from testis tissue of adult horses exhibited higher quality with clear tissue structure and complete cell morphology after they underwent gradient dehydration for 6 hours, were immersed in wax for 60 minutes, and were dried in a 75-degree oven for 15 minutes. The detailed, optimized procedures that are developed in ...
Jung H, Yoon M.Spermatogonial stem cells originate from gonocytes and undergo self-renewal and differentiation to generate mature spermatozoa via spermatogenesis in the seminiferous tubules of the testis in male mammals. Owing to the unique capacity of these cells, the spermatogonial stem cell transplantation technique, which enables the restoration of male fertility by transfer of germlines between donor and recipient males, has been developed. Thus, spermatogonial stem cell transplantation can be used as an important next-generation reproductive and breeding tool in livestock production. However, in large ...
Posastiuc FP, Rizzoto G, Constantin NT, Nicolae G, Chiers K, Diaconescu AI, Șerban AI, Van Soom A, Codreanu MD.Anti-Müllerian hormone (AMH) has become a pivotal subject in the study of testicular descent, maturation, integrity, and male fertility. Recent studies explored its roles and implications across various domestic species. A prominent approach involved the understanding of the modulation of AMH in reproductive disorders, including cryptorchidism. While substantial findings have been reported in dogs, ruminants, swine, and horses, data on AMH in feline cryptorchidism remains limited. Here, we aimed to bridge this gap by comparing AMH serum levels among cryptorchid, healthy intact, and castrated ...
Pozor MA, Macpherson ML, Kelleman AA, Smith H, McNaughten JW.This retrospective study aimed to validate the usefulness of spectral Doppler ultrasonography in diagnosing testicular dysfunction in stallions. The first part of the study included a pony stallion with age-related testicular degeneration (18 - 21 yrs. old). Testicular dimensions, spectral Doppler parameters of testicular blood flow, and semen parameters were obtained numerous times from this pony over four consecutive years. While testicular volume decreased dramatically, and the stallion became azoospermic, there were no changes in any of the spectral Doppler parameters of testicular blood f...
Vitoria A, Barrachina L, Romero A, Fuente S, de Blas I, Gil L, Vázquez FJ.A new surgical technique using a polyether ether ketone (PEEK) anchoring device for testicle-sparing laparoscopic inguinal hernioplasty in stallions was described in 2023 and is known as the PEEK harpoon technique (PHT). In breeding stallions, it is essential that the surgery is effective in preventing inguinal hernia but also that it does not impair the testicular function. This study aims to evaluate whether the PHT may affect testicular function. To achieve that, changes in the testicular blood flow, sperm production and motility characteristics were assessed 28 days after use of the PHT. S...
Khan IU, Khairullah AR, Khan AY, Rehman AU, Mustofa I.This review explores advanced strategies for enhancing fertility and optimizing reproductive outcomes in equine breeding programs. Horses, being seasonal breeders, present unique reproductive challenges influenced by environmental and physiological factors such as photoperiods, hormone cycles, and aging. Key approaches discussed include hormonal therapies, artificial light manipulation, and nutritional supplementation to improve ovulation and conception rates during the breeding season. Specific hormones such as gonadotropin-releasing hormone analogs, equine follicle-stimulating hormone, and p...
Sinovich M, Monné Rodriguez J, Pieńkowska-Schelling A, Schelling C, Kelly PG.Introduction Monorchidism is a rarely described condition in the horse and is not to be confused with cryptorchidism. The diagnosis is challenging and confirmed by surgery and histology in combination with hormonal assays. This report describes, to the best of the author's knowledge, the first case of monorchidism and abdominal cryptorchidism of the developed testicle in a horse. Methods An Irish Cob underwent laparoscopic castration for removal of bilateral cryptorchid testicles. At surgery the horse was diagnosed as a monorchid with the testicle retained intra-abdominally. Histopathological,...
Giorgio SD, Monti S, Palmieri V, Marino G, Napoli E, Vullo C, Catone G.A 3-year-old Quarter horse stallion with unilateral left cryptorchidism was referred for the absence of the left testis in the scrotal bursa and an enlarged right testis. The ultrasonographic examination revealed the presence of the left testis at the abdominal level. Two small hyperechogenic lines were observed running parallel and resembling a cylindrical cavitary foreign body, within a non-homogeneous area with blurred margins. The testis was removed by laparoscopy and macroscopically, the testis's cut section revealed the presence of two live adult nematodes emerging from 2 mm blood-filled...
Baatar T, Song D, Weng Y, Wang G, Jin L, Guo R, Li B, Dugarjaviin M.The epigenetic regulation of gene expression through the covalent modification of histones is crucial for developing germline cells. To study the regulatory role of alternative splicing (AS) of euchromatic histone lysine methyltransferase 2 (EHMT2/G9A) in spermatogenesis in Mongolian horses, this study first examines the localization of the EHMT2 gene in testicular support cells and then predicts the higher-order structures of sequences with and without AS. Two types of lentiviral vectors for overexpression were subsequently constructed for the EHMT2 gene, one with AS and one without, to infec...
Lu Z, Wen M, Yao X, Meng J, Wang J, Zeng Y, Li L, Ren W.The reproductive performance of horses is closely associated with testicular development, which involves a complex network of gene regulation. The molecular mechanisms underlying the reproductive traits of Kazakh horses are yet to be fully understood. In this study, transcriptomic analysis was performed on testicular tissues from Kazakh horses at different developmental stages using whole-transcriptome sequencing, aiming to investigate differential gene expression and regulatory mechanisms during testicular development. The results revealed significant differential expression of 7678 mRNAs, in...
Liu J, Yang Y, Wen L, Wen M, Zeng Y, Ren W, Yao X.The Kazakh horse is an outstanding dual-purpose dairy and meat breed in China, characterized by early maturity, tolerance to coarse feed, and strong stress resistance. Previous studies have examined gene expression patterns in the testicular tissues of Kazakh horses at different age stages, but the molecular mechanisms regulating testicular sexual maturation remain unclear. To address this gap, this study conducted HE staining and in-depth transcriptome sequencing analysis of Kazakh horse testicular tissue before and after sexual maturity. HE staining showed that the G3 group had well-formed s...
de Castro Sasahara TH, Gomes SP, Abdala FCM, Schimming BC, Miglino MA.Sexual differentiation in the equine fetus involves coordinated morphogenetic processes that shape both the gonads and the genital ducts. Although the formation of testes and ovaries has been relatively well documented, the temporal dynamics and morphometric patterns of the mesonephric (Wolffian) and paramesonephric (Müllerian) ducts remain insufficiently characterized. This study aimed to detail the timeline of gonadal and ductal differentiation in male and female equine fetuses during the critical developmental window between 60 and 80 days of gestation. Gross anatomical evaluation, histo...
