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REDUCED LEVELS OF ADRENAL STEROIDOGENIC ACUTE REGULATORY PROTEIN (StAR) DO NOT LIMIT STEROIDOGENESIS IN ALDOSTERONE PRODUCING CELLS Songül Süren Castillo Istanbul University, Faculty of Science, Department of Biology, Zoology Section, 34459-Verneciler, Istanbul, Turkey. E-mail: [email protected] KEY WORDS: Adrenal gland, insulin-like growth factors, insulin receptors, steroidogenic acute regulatory protein, serotonin, serotonin receptors ABSTRACT: The insulin-like growth factors (IGFs) actions are thought to be mainly due to the activation of the type I IGF receptor, which is homologous to the insulin receptor (IR) in structure1. Mature IGF-I and IGF-II are composed of four domains and share extensive sequence and structural similarity. The IGF-II C and D domains confer high-affinity binding to the IR-A and IR-B, whereas the C and D domains of IGF-I do not2. The sole structural determinant for the differential ability of IGF-I and IGF-II to induce autophosphorylation of specific IR tyrosine residues and activate downstream signalling molecules is the C domain1. On the basis of this knowledge, frogs (Rana ridibunda) were injected with IGF-II C-peptide (2.5 mg/0.2 ml), whereas control animals were injected with Ringer solution (0.2 ml). The adrenal glands were removed at 12 and 24 h after injection and fixed, embedded in paraffin wax and Epon, and examined by immunohistochemistry and transmission electron microscopy to investigate the regulation of frog adrenal steroidogenic cell activity. Sections were stained with hematoxylin and eosin for overall tissue analysis and, in parallel, serotonin and steroidogenic acute regulatory (StAR) protein were localized using the streptavidin– biotin complex technique. At 12 and 24 h after injection of IGF-II C-peptide, a decrease of serotonin immunoreactivity in chromaffin cells (which secrete serotonin) and a decrease of StAR protein immunoreactivity in steroidogenic cells were observed. Electron microscopic observations show an increase in the activity of the steroidogenic cells producing aldosterone. Besides its action on the secretory activity of the adrenal gland, IGF-II C-peptide causes proliferation of steroidogenic cells at 24 h. Therefore, the reduced levels of StAR protein are not a limiting factor of steroidogenesis for aldosterone producing cells. In conclusion, in addition to its direct effect on aldosterone production, a possible indirect effect of IGF-II C-peptide through activation of chromaffin cells in frog adrenal glands may occur. These observations suggest that IGFII C-peptide may play an important role in the regulation of steroidogenic cell activity. REFERENCES 1.Denley A, Brierley GV, Carroll JM, Lindenberg A, Booker GW, Cosgrove LJ, Wallace JC, Forbes BE, Roberts CTJr. Differential activation of insulin receptor isoforms by insulin-like growth factors is determined by the C Domain. Endocrinology 147, 1029-36 (2006). 2.Denley A, Bonython ER, Booker GW, Cosgrove LJ, Forbes BE, Ward CW, Wallace JC. Structural determinants for high-affinity binding of insulin-like growth factor II to insulin receptor (IR)-A, the exon 11 minus isoform of the IR. Mol. Endocrinol. 18, 2502-12 (2004).