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¹ INSERM U413, Laboratory of Cellular and Molecular Neuroendocrinology, European Institute for Peptide Research (IFRMP 23), University of Rouen, 76821 Mont-Saint-Aignan, France ² INSERM U567, CNRS UMR8104, Department of Endocrinology-Metabolism-Cancer, Institut Cochin, Université Paris V-René Descartes, 75014 Paris, France ³ Department of Endocrinology, Institute for Biomedical Research, Rouen University Hospital, 76031 Rouen Cedex, France

(Correspondence should be addressed to H Lefebvre; Email: herve.lefebvre{at}chu-rouen.fr)

Cortisol secretion in ACTH-independent macronodular adrenal hyperplasia (AIMAH) causing Cushing's syndrome can be controlled by illegitimate receptors. The aim of the present study was to characterize the molecular, immunohistochemical, and pharmacological profiles of vasopressin receptors in cells derived from three patients with AIMAH (H1–H3), in order to evaluate the role of ectopic vasopressin receptors in the physiopathology of hypercortisolism. Expression of mRNAs encoding the vasopressin receptor types (V_1a, V_1b, and V₂) were analyzed by RT-PCR in adrenal tissues. The presence of V_1a and V₂ receptors was studied by immunohistochemistry on adrenal sections. The pharmacological profiles of vasopressin receptors involved in the control of cortisol secretion were investigated using the V_1a receptor antagonist SR49059 and the V₂ receptor agonist [deamino-Cys¹, Val⁴, D-Arg⁸]-vasopressin on cultured cells. The V_1a receptor protein was present and functional in H1 and H3 tissues, whereas the V_1b receptor was not expressed in any of the tissues. RT-PCR experiments revealed that V₂ receptor mRNAs were detected in the three tissues. In contrast, immunohistochemical and cell incubation studies showed that the V₂ receptor was involved in the stimulatory effect of AVP on cortisol secretion in H1 and H2, but not in H3 cells. Taken together, these data show that expression of functional ectopic V₂ receptors and repression of eutopic V_1a receptor can coexist in some hyperplastic corticosteroidogenic tissues. They also reveal that immunohistochemical and incubation studies are essential for the characterization of ectopic receptors actually involved in the control of cortisol secretion by AIMAHs.

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