RESUMO
Horse transport is considered a cause of stress in animals and is known to affect the 5-HT concentrations in both the brain and other tissues. The aim of this research was to investigate the effect of horse transportation and slaughter stress on plasma serotonin's concentration and the expression levels of the related 5-HT1B and 5-HT2A receptors in PBMCs. Furthermore, the IL-12 levels and a variety of blood parameters, including triglycerides, total cholesterol, glucose, aspartate aminotransferase, creatine phosphokinase, lactate dehydrogenase, were also considered. This research was carried out on 32 horses submitted to short road transport of 40 km to slaughter. Blood samples were collected in baseline conditions (T0) and 24 h later, after they were slaughtered (T1). The results showed a significantly increased expression of 5-HT1B and 5-HT2A receptors and a significantly decreased expression of IL-12 in PBMCs at T1 vs. T0. Furthermore, a significant increase in cortisol and glucose concentrations, and LDH activity was observed at T1. In contrast, a significantly lower circulating 5-HT concentration was observed at T1 vs. T0. These results indicate that the stress induced by transport and slaughter stimuli led to the serotoninergic system's activation, suggesting that the expression of serotonin receptors could be used as a pivotal marker of stress, with potential applications for the improvement of elective protocols to observe the guidelines relating to transported horses.
RESUMO
Serotonin (5-HT) is a neurohormone transported from plasma into platelets and leukocytes by a specific transporter (SERT). While it is known that the brain 5-HT system is modulated by physical exercise, the peripheral serotoninergic response to exercise is not yet fully elucidated. In particular, this study aimed to evaluate changes in plasma 5-HT levels and equine leukocyte SERT expression in response to treadmill exercise in untrained horses. Analyses were carried out pre- and post-treadmill exercise. 5-HT plasma levels were analysed by HPLC. Leukocytes and platelets were isolated to perform Real Time PCR for the evaluation of SERT mRNA levels. Western blot was conducted for the detection of SERT protein levels. The presence of SERT in leukocytes was analysed by flow cytometry. The functionality of SERT on leukocytes was investigated by using paroxetine as inhibitor of 5-HT reuptake. Results showed a significant decrease in SERT levels after exercise in both leukocytes and platelets and a significant increase in plasma 5-HT levels. Flow cytometry revealed that SERT is functional in one specific horse leukocyte subpopulation, still not identified, and paroxetine was able to block 5-HT reuptake into leukocytes. The exercise may have induced an increased mobilization of free-tryptophan and a release of 5-HT from the stores in the blood. High concentrations of plasma 5-HT could have caused a reduction in SERT expression affecting cellular 5-HT storage/uptake. The increase of cortisol levels after treadmill exercise was not significant. Exercise modulates the peripheral serotonin metabolism. More research is needed to assess its physiological implications.