In mammals distinct steroid hormones termed mineralocorticoids (MCs) and glucocorticoids (GCs) regulate hydromineral balance and the stress response, respectively. In contrast, it is thought that a single corticosteroid, 1[alpha]-hydroxycorticosterone (1[alpha]-B) serves as both a GC and MC in elasmobranchs. I investigated the putative dual MC and GC roles of 1[alpha]-B by examining ex vivo regulation of interrenal 1[alpha]-B synthesis by osmoregulatory and stress hormones in the euryhaline stingray Dasyatis sabina. A commercial enzyme-linked immunoassay was adapted for the quantification of 1[alpha]-B. I also isolated cDNA sequences encoding two rate-limiting steroidogenic enzymes, the steroidogenic acute regulatory protein (StAR) and P450 cholesterol side-chain cleavage (P450scc), and characterized the steroidogenic activity of the encoded proteins using a heterologous expression system. Both the stress hormone adrenocorticotropic hormone (ACTH) and the antinatriuretic peptide angiotensin II (ANG II) were potently steroidogenic in ex vivo interrenal cultures, whereas C-type natriuretic peptide (CNP) inhibited 1[alpha]-B synthesis. StAR and P450scc mRNA levels were increased by 24 h incubation with ACTH and decreased by both ANG II and CNP. To examine changes in osmoregulatory hormone systems that impinge upon 1[alpha]-B synthesis, I also isolated the cDNA sequences of the ANG II and CNP receptors, AT and NPR-B. Both AT and NPR-B mRNA levels were significantly elevated in osmoregulatory tissues of freshwater (FW; Lake Monroe, FL) versus saltwater (SW; Corpus Christi Bay, TX) populations of D. sabina. Interrenal StAR and NPR-B mRNA levels were also significantly higher in FW individuals. The physiological roles of 1[alpha]-B were further investigated in vivo by examining the effects of stress and FW transfer on interrenal synthesis of 1[alpha]-B. Plasma 1[alpha]-B and glucose were significantly elevated by hook-and-line capture stress, indicating that 1[alpha]-B acts in classical GC fashion to facilitate the stress response. In contrast, 1[alpha]-B was significantly decreased 24 h after SW-FW transfer. In light of the osmotic strategy of euryhaline elasmobranchs, this result is consistent with a MC role for 1[alpha]-B. Taken together, the results of this research strongly support a dual role for 1[alpha]-B in facilitating both hydromineral balance and the stress response in elasmobranchs. / text

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