Association of plasma concentrations of oxytocin, vasopressin, and serotonin with docility and friendliness of horses.

Oxytocin (OXT) and vasopressin (AVP) have been associated with social interaction and affiliative behavior in animals. Additionally, AVP is known to affect socially aggressive behavior. In addition, serotonin has an association with aggressive behaviors. The objectives of this study were (1) to evaluate OXT, AVP, and serotonin concentrations in the plasma of horses of different breeds, sexes, and ages and (2) to determine if the neurotransmitters are associated with horse docility and friendliness. This study was performed at Sangju International Equestrian Center. Blood samples were collected from 23 horses, including 6 Thoroughbreds (11 to 18 yr), 6 Warmbloods (15 to 26 yr), 6 ponies (8 to 17 yr), and 5 Quarter Horses (4 to 12 yr). The group of horses consisted of 13 mares and 10 geldings. The plasma concentrations of OXT and AVP were measured by enzyme-linked immunosorbent assay, and the serum concentration of serotonin was measured by high-performance liquid chromatography. The characteristics of each horse were surveyed by 3 horse trainers. The effects of breed, sex, and age on the concentration of each neurotransmitter were assessed by a 3-way ANOVA with LSD post-hoc analysis. Linear regression analysis was performed to determine if the concentration of neurotransmitters is related to the docility and friendliness of horses. As a result, the concentrations of OXT and AVP did not vary with the breed, sex, or age of horses. However, the serotonin concentration varied depending on the breed and age of horses. Interestingly, there was a trend toward the existence of a correlation between docility and OXT in Thoroughbred horses. However, AVP and serotonin concentrations had no correlation with the docility and friendliness of horses. In conclusion, the docility and friendliness of Thoroughbred might be related with the blood OXT concentration.

Effects of Hemicastration on Testes and Testosterone Concentration in Stallions.

The endocrine system is critical to the maintenance of testicular function. The homeostasis of sex hormone levels is orchestrated by positive and negative feedback systems controlled by the hypothalamic-pituitary-gonadal axis. This study investigated the long-term effects of hemicastration on testicular size and function in stallions. Four Thoroughbred stallions, 4-6 years of age, were included in this study. Several parameters, including testicular weight and volume, plasma testosterone concentrations, VASA-positive germ cell populations and cross-sectional areas of the seminiferous tubules were compared in stallions that underwent two hemicastrations, approximately 11 months apart. The weights and volumes of testes harvested at the second hemicastration were significantly higher than those of testes collected at the first hemicastration. However, VASA-positive germ cell populations and the cross-sectional areas of seminiferous tubules were not significantly different between testes harvested at the first and second hemicastrations. Similarly, plasma testosterone concentrations measured weekly for 3 weeks before the first hemicastration, 3 weeks after the first hemicastration, and 3 weeks before the second hemicastration were not significantly different. Our results suggest that hemicastration results in compensatory enlargement of the remaining testis and compensatory steroidogenesis to maintain normal reproductive function in stallions.

The Lin28 Expression in Stallion Testes.

The molecular markers for specific germ cell stages can be utilized for identifying, monitoring, and separating a particular stage of germ cells. The RNA-binding protein Lin28 is expressed in gonocytes of human fetal testes. The Lin28 expression is restricted to a very small population of spermatogonial cells in human, mice, and monkey. The main objective of this study was to investigate the expression pattern of Lin28 in stallion testes at different reproductive stages. Based on the presence or absence of full spermatogenesis and lumina in seminiferous tubules, the testicular samples were categorized into two reproductive stages pre-pubertal and post-pubertal. We performed a reverse transcription polymerase chain reaction to confirm the presence of Lin28 mRNA in the testicular tissues and a western blot analysis to verify the cross-reactivity of rabbit Lin28 antibody with horse testicular tissue. For immunohistochemistry, Lin28 (rabbit anti-human), GATA4 (goat anti-human) or DAZL (goat anti-human) antibodies were used. The results of RT-PCR confirmed the expression of Lin28 mRNA in the stallion testes. The western blot analysis showed that the expression of 28 kDa Lin28 protein was localized in the cytoplasm of spermatogonia at both reproductive stages. The numbers of Lin28-positive germ cells per 1000 Sertoli cells in pre- and post-pubertal stages were 253 ± 8.66 and 29.67 ± 2.18, respectively. At both reproductive stages, all Lin28 positive cells showed no co-stained with GATA4 antibody, whereas only some of the Lin28-positive germ cells showed co-staining with DAZL antibody. The results from whole-mount staining showed that the Lin28 expression was limited to Asingle (As) and Apaired (Apr) spermatogonia. In conclusion, Lin28 might be utilized as a molecular marker for undifferentiated spermatogonial stem cells when used with DAZL antibody.