Expression of orexin B and its receptor 2 in rat testis

The peptides orexin A (OxA) and orexin B (OxB) deriving from a common precursor molecule, prepro-orexin, by proteolytic cleavage, bind the two G-coupled OX₁ and OX₂ receptors. While OX₁ selectively binds OxA, OX₂ shows similar affinity for both orexins. Firstly discovered in the hypothalamus, orexins and their receptors have been found in other brain regions as well as in peripheral tissues of mammals, thus resulting involved in the regulation of a broad variety of physiological functions. While the functional localization of OxA and OX₁ in the mammalian genital tract has been already described, the expression of OxB and OX₂ and their potential role in the reproductive functions remain to be explored. Here, we investigated the presence of OxB and OX₂ in the rat testis by immunohistochemical and biochemical analyses. The results definitely demonstrated the localization of OxB and OX₂ in pachytene and second spermatocytes as well as in spermatids at all stages of the cycle of the seminiferous epithelium. The expression of both OX₂ mRNA and protein in the rat testis was also established by RT-PCR and Western blotting, respectively. The analysis of the molecular mechanism of action of OxB in the rat testis showed that OxB, in contrast with OxA, is unable to promote steroidogenesis. These results translate into the regulation of diverse biological actions by OxA and OxB in the male gonad.