Uncovering the Physiological Mechanisms Underlying the Roe Deer (Capreolus capreolus) Testicular Cycle: Analyses of Gelatinases and VEGF Patterns and Correlation with Testes Weight and Testosterone

SIMPLE SUMMARY: The roe deer (Capreolus capreolus) is a wild, small ruminant common in Europe and Asia with a peculiar reproductive cycle. The adult male shows a complete suspension of spermatozoa production during the winter that start again in spring with the peak of sexual activity in summer (July–August). The physiological mechanism underlying such a cycle is yet to be clarified. The work aimed at the analysis of some molecules, like gelatinases (MMP) and vascular growth factor, physiologically involved in tissue remodeling, in roe deer samples collected before (June–July) and after (August–September) the rutting period. Samples were provided by hunters of the Bologna Apennines area (Italy) according to the regional hunting plan. The result showed a post-rut reduction of testicular weight and testosterone, indicative of testes involution, correlated with an increase in pro-MMP2, the latent gelatinase capable of sustaining spermatogenesis once activated. It can be assumed that gelatinases are involved in the testicular cycle and start accumulating after the rutting period to be then activated in preparation of the next reproductive season inducing spermatozoa development and migration. Future studies on this pathway during all seasonal testicular cycles will provide more information about the interesting reproductive physiology of roe deer. ABSTRACT: The roe deer (Capreolus capreolus) represents a spontaneous model of testicular inactivation: During winter, bucks show a suspension of spermatogenesis that starts again in spring and peaks during the breeding season (July–August). The underlying mechanisms to the regulation of the cyclic testicular changes are still not fully clear but seem to be imputable to the spermatogenic cell line since other testicular cell populations remain stable without apoptotic phenomena. The aim of the study was to investigate apoptosis, gelatinases (MMP2 and 9), their inhibiting factors (TIMP 1-2), and two isoforms of vascular endothelial growth factor (VEGF121 and 165) with its receptors (VEGFR1-2) in testes collected during pre- and post-rut periods, and to correlate them with testicular weight (TW) and testosterone (TEST). Testes from 18 adult sexually mature bucks were collected in Bologna Apennines (Italy). Samples were weighed and parenchyma collected. Radioimmunoassay, real-time PCR, and zymography were performed. The results showed a post-rut decrease in TW and TEST and an increase in proMMP2, also highlighting a correlation between the gelatinases and the testicular functionality. The VEGF pattern did not show modifications nor correlation with TW and TEST. Overall, gelatinases and their inhibitors, described herein for the first time in roe deer testes, seem to play an important role in the testicular cycle.