Overexpression of regucalcin mitigates the ageing-related changes in oxidative stress and sperm quality

Age-related changes, namely the increase in oxidative stress (OS) with the consequent sperm damage, result in decreased male fertility. Regucalcin (RGN) is a Ca²⁺-binding protein that has been shown to have beneficial effects on spermatogenesis by suppressing OS and chemical/radiation-induced damage. This work aims to evaluate whether RGN overexpression reduces the ageing-associated decline of male reproductive function. Sperm and testicular function analysis were performed in young-adult and senescent transgenic rats overexpressing RGN (Tg-RGN) comparatively with their wild-type (Wt) littermates. The gonadosomatic index (GI), tubular differentiation index and the expression levels of RGN and other proliferation regulators were evaluated. Moreover, the sperm parameters, OS analysis and immunolocalization of RGN were assessed, as well as morphometric evaluation of epididymal tubules. Both GI and sperm counts were reduced in the senescent Wt rats, but maintained in the Tg-RGN. Also, the levels of stem cell factor (SCF), c-Kit, and Akt were maintained in the testis of aged Tg-RGN rats, suggesting that the normal spermatogenic output was preserved over time in these animals, an effect not observed in Wt. Senescent Tg-RGN rats also presented lower sperm lipid peroxidation and total oxidant status relative to the Wt. Furthermore, aged Tg-RGN rats displayed higher sperm viability, higher frequency of sperm with normal morphology, and reduced incidence of head and neck/midpiece defects when compared with Wt, which may be a consequence of the lower OS levels found in the sperm of these animals. Interestingly, RGN expression increased with ageing in sperm, being mainly localized in the acrosome. Altogether, these findings indicate that the modulation of RGN levels may alleviate the age-related decline in sperm quality and testicular function.