Gastric inhibitory polypeptide stimulates glucocorticoid secretion in rats, acting through specific receptors coupled with the adenylate cyclase-dependent signaling pathway.

Gastric inhibitory polypeptide (GIP) is a 42-amino acid peptide, belonging to the VIP-secretin-glucagon superfamily, some members of this group are able to regulate adrenocortical function. GIP-receptor mRNA has been detected in the rat adrenal cortex, but investigations on the effect of GIP on steroid-hormone secretion in this species are lacking. Hence, we have investigated the distribution of GIP binding sites in the rat adrenal gland and the effect of their activation in vivo and in vitro. Autoradiography evidenced abundant [125I]GIP binding sites exclusively in the inner adrenocortical layers, and the computer-assisted densitometric analysis of autoradiograms demonstrated that binding was displaced by cold GIP, but not by either ACTH or the selective ACTH-receptor antagonist corticotropin-inhibiting peptide (CIP). The intraperitoneal (IP) injection of GIP dose-dependently raised corticosterone, but not aldosterone plasma concentration: the maximal effective dose (10 nmol/rat) elicited a twofold increase. GIP did not affect aldosterone and cyclic-AMP release by dispersed zona glomerulosa cells. In contrast, GIP enhanced basal corticosterone secretion and cyclic-AMP release by dispersed inner adrenocortical cells in a concentration-dependent manner, and the maximal effective concentration (10(-7) M) evoked 1.5- and 2.4-fold rises in corticosterone and cyclic-AMP production, respectively. GIP (10(-7) M) did not display any additive or potentiating effect on corticosterone and cyclic-AMP responses to submaximal or maximal effective concentrations of ACTH. The corticosterone secretagogue action of 10(-7) M GIP was abolished by the protein kinase A (PKA) inhibitor H-89 (10(-5)M), and unaffected by CIP (10(-6)M). Collectively, these findings indicate that GIP exerts a moderate but statistically significant stimulatory effect on basal glucocorticoid secretion in rats, acting through specific receptors coupled with the adenylate cyclase/PKA-dependent signaling pathway.

Mechanisms and receptor subtypes involved in the stimulatory action of endothelin-1 on rat adrenal zona glomerulosa.

Endothelin (ET)-1 is the prototype of a family of 21-amino acid residue hypertensive peptides, acting through two subtypes of receptors, named ETA and ETB. ETs and their receptors are expressed in the adrenal cortex and medulla, and ET-1 enhances both corticosteroid and catecholamine release. ET-1 concentration-dependently (from 10(-11) to 10(-8) M) increased aldosterone secretion of both dispersed rat zona glomerulosa (ZG) cells and adrenal slices containing a core of medullary chromaffin tissue, but the response of the latter preparations was significantly more intense than that of the formers. The stimulatory effect of 10(-8) M ET-1 on dispersed ZG cells was blocked by the ETB-receptor antagonist BQ-788 (10(-7) M), but not by the ETA-receptor antagonist BQ-123 (10(-7) M); conversely, both ET-receptors antagonists counteracted aldosterone response of adrenal slices to ET-1. The -adrenoceptor antagonist l-alprenolol (10(-6) M) did not affect aldosterone response of dispersed ZG cells to ET-1 (10(-8) M), but it significantly lowered that of adrenal slices. l-Alprenolol also counteracted the aldosterone response of adrenal slices to the pure activation of ETB or ETA receptors, as obtained by using the selective ETB-receptor agonist BQ-3020 (10(-8) M) or ET-1 (10(-8) M) plus BQ-788 (10(-7) M). ET-1 concentration-dependently (from 10(-9) to 10(-8)/10(-7) M) stimulated catecholamine release by adrenal slices, and the effect was counteracted by both BQ-123 and BQ-788 (10(-7) M). Collectively, our findings suggest that, when the integrity of adrenal tissue is preserved, a two-fold mechanism underlies the aldosterone secretagogue action of ET-1 in the rat: i) a direct mechanism mediated by ETB receptors located on ZG cells; and ii) an indirect mechanism involving the ETA and ETB receptor-mediated local release of catecholamines, which in turn stimulate ZG cells in a paracrine manner.

Endothelin-1 enhances thymocyte proliferation in monolaterally adrenalectomized rats with contralateral adrenocortical regeneration.

Endothelins (ETs) are a family of vasoactive peptides widely distributed in the body systems, where they exert pleiotropic biological effects, acting through two main subtypes of receptors, named ETA and ETB. Evidence indicates that ET-1 plays a permissive role in the development of neural crest-derived structures, among which are the epithelial cells of the thymus. These cells are known to control proliferation and differentiation of thymocytes, a process requiring adequate levels of glucocorticoids. Therefore, we have investigated the effects of ET-1, that binds both ETA and ETB receptors, on thymocyte proliferation in monolaterally adrenalectomized rats with contralateral enucleated adrenal at day 4 and 8 of regeneration, when glucocorticoid production is very low and, respectively, rather normal. Metaphase index (percentage of metaphase arrested cells) of thymocytes is the lowest at day 4 of regeneration, and markedly rose at day 8, thereby confirming the need of sizable levels of circulating glucocorticoid for the maintenance of a normal rate of thymocyte proliferation. ET-1 markedly increased the mitotic index of thymocytes at both times of adrenal regeneration. At day 8 of regeneration, the ETA-receptor antagonist BQ-123 markedly lowered mitotic index of thymocytes, and annulled its ET-1-evoked raise. Conversely, the ETB-receptor antagonist BQ-788 was ineffective. Collectively, these findings clearly indicate that endogenous ETs, through the activation of ETA receptors, are involved in the maintenance and stimulation of thymocyte proliferation in the adult rat, thereby playing a possibly important role in the modulation of the immune-system functions.

Neuropeptide-Y family of peptides in the autocrine-paracrine regulation of adrenocortical function.

Neuropeptide-Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP) are a family of 36-amino-acid peptides, which are widely distributed in the body and act through several subtypes of G-protein-coupled Y receptors. The three members of the NPY-family peptides are contained in the adrenal gland, where they exert various autocrine/paracrine regulatory functions. Binding sites for NPY are present in adrenal medulla and zona glomerulosa (ZG), where also several NPY-ergic fibers end. Binding sites for PYY and PP are prevalently located in the inner adrenocortical zones and adrenal medulla. NPY and PYY inhibit aldosterone secretion of dispersed ZG cells, but this effect has probably to be considered nonspecific and toxic in nature. On the contrary, there are indications that NPY may indirectly stimulate ZG cells, by eliciting the release of catecholamines, which in turn enhance aldosterone secretion. Evidence is also available that NPY modulates the secretory response of ZG cells to their main agonists. PP is able to raise glucocorticoid secretion acting directly on the inner adrenocortical cells. The physiological relevance of these effects of NPY-family peptides remains to be addressed by future experimental studies employing more selective Y receptor agonists and antagonists. In contrast, indirect evidence is available that endogenous NPY-family peptides might play an important role in the modulation of adrenocortical secretory action under paraphysiological or pathological conditions such as like aging, hypoglycemic stress and pheochromocytomas.

Galanin stimulates glucocorticoid secretion in rats through a receptor-dependent activation of the adenylate cyclase/protein kinase A-dependent signaling pathway.

Galanin, a 29-amino acid peptide widely distributed in the central and peripheral nervous systems, was found to induce a concentration-dependent increase in corticosterone secretion and cyclic-AMP release by dispersed rat inner adrenocortical cells (maximal effective concentration, 10(-7) M). The effect of 10(-7) M galanin was blocked by 10(-6) M galantide, a specific antagonist of galanin receptors. Galanin (10(-7) M) also enhanced corticosterone and cyclic-AMP responses of dispersed cells to submaximal but not maximal (10(-9) M) effective concentrations of ACTH, and again this effect was reversed by galantide. The ACTH-receptor antagonist corticotropin-inhibiting peptide (10(-6) M) blocked corticosterone response of dispersed cells to 10(-9) M ACTH but not to 10(-7) M galanin; conversely, the specific protein kinase A inhibitor H-89 (10(-5) M) annulled the secretory response to both ACTH and galanin. In light of these findings, we conclude that galanin stimulates adrenal glucocorticoid secretion in rats, acting through specific receptors, coupled, like those of ACTH, with the adenylate cyclase/protein kinase A-dependent signaling pathway.

A local immuno-endocrine interaction may mediate rat adrenal glucocorticoid response to bacterial endotoxins.

The effects of the bacterial endotoxin lipopolysaccharide (LPS) and interleukin (IL)-1beta on corticosterone secretion has been studied in vivo by employing the technique of in situ perfusion of the isolated rat left adrenal gland. Both LPS and IL-1beta dose-dependently raised corticosterone output, the response peaking at 60 and 90 min, respectively. IL-1 receptor antagonist dose-dependently reversed the effect of LPS and IL-1beta. The IL-1beta converting enzyme (ICE) inhibitor Ac-YVAD-CMK annulled the adrenal response to LPS, but did not affect that to IL-1beta. Collectively, these findings provide evidence that LPS, by enhancing adrenal production of IL-1beta, is able to evoke a sizable glucocorticoid response in the rat, thereby suggesting that local immuno-endocrine interactions may be operative in the adrenal gland of this species.

Distribution and functional significance of angiotensin-II AT1- and AT2-receptor subtypes in the rat adrenal gland.

The distribution and the functional significance of angiotensin-II (ANG-II) receptor subtypes, AT1 and AT2, in the rat adrenal gland has been investigated in vitro. Autoradiographic assessment of the selective displacement of [125I]ANG-II binding by selective ligands of the two receptor subtypes indicated that zona glomerulosa (ZG) was provided with both AT1 and AT2, and adrenal medulla (AM) almost exclusively with AT2 receptors. ANG-II (10(-9) M) evoked a marked rise in the secretion of aldosterone by dispersed ZG cells and catecholamines by AM fragments. The selective AT1-receptor antagonist DuP753 blocked aldosterone response to ANG-II, while the selective AT2-receptor antagonist PD123319 was ineffective. Catecholamine response to ANG-II was inhibited by PD123319 and only moderately affected by high concentrations of DuP753. The selective AT2-receptor agonist CGP42112 did not change basal aldosterone release of ZG cells, but concentration-dependently enhanced basal catecholamine release by AM fragments. In light of these findings the conclusion is drawn that in the rat the aldosterone secretagogue effect of ANG-II is exclusively mediated by the AT1 receptors present in the ZG, while the catecholamine secretagogue action preminently involves the activation of AT2 receptor located on medullary chromaffin cells.

The AT2 receptor-mediated stimulation of adrenal catecholamine release may potentiate the AT1 receptor-mediated aldosterone secretagogue action of angiotensin-II in rats.

The role played by AT1 and AT2 receptors in the mediation of angiotensin-II (ANG-II) aldosterone secretagogue action has been investigated in vitro using different types of rat adrenal preparations. ANG-II enhanced aldosterone secretion of dispersed zona glomerulosa (ZG) cells in a concentration-dependent manner (EC50, 3 x 10(-10) M), and its effect was annulled by the AT1-receptor antagonist DuP753 and unaffected by the AT2-receptor antagonist PD123319. ANG-II was significantly more effective in stimulating aldosterone secretion when capsule-ZG and adrenal slices containing medullary chromaffin cells were used (EC50, 1 x 10(-11) M and 7 x 10(-12) M, respectively); moreover, both DuP753 and PD123319 caused partial reversals (intense and moderate, respectively) of the responses to ANG-II, and when added together annulled them. The beta-adrenoceptor antagonist l-alprenolol did not affect aldosterone response to ANG-II of dispersed ZG cells, but exerted a PD123319-like effect on the responses of capsule-ZG and adrenal slices. In light of these findings we conclude that, when the integrity of adrenal tissue is preserved, ANG-II stimulates aldosterone secretion by activating both AT1 and AT2 receptors, the major role being played by AT1 receptors located on ZG cells. The activation of AT2 receptors probably elicits the local release of catecholamines, which in turn enhance aldosterone secretion in a paracrine manner acting through the beta-adrenoceptors with which ZG cells are provided.

Paracrine control of steroid hormone secretion by chromaffin cells in the adrenal gland of lower vertebrates.

The adrenal glands of lower vertebrates display a notable intermingling between steroidogenic and chromaffin tissues, which increases from Pisces to Aves. As in mammals, adrenal chromaffin cells contain and release, in addition to catecholamines, serotonin and several peptides, which may affect the secretory activity of steroidogenic cells in a paracrine manner. Stimulatory molecules include serotonin, arginine-vasotocin, tachykinins, vasoactive intestinal peptide, pituitary adenylate cyclase-activating peptide and calcitonin gene-related peptide; inhibitory molecules are dopamine, somatotropic hormone-release inhibiting hormone and galanin. Epinephrine and norepinephrine appear to stimulate steroid secretion in Aves and to inhibit it in Pisces, while their action in Amphibia is controversial. Likewise, atrial natriuretic peptide exerts an anti-secretagogue action in Amphibia and a marked secretagogue effect in Pisces and Aves. The effects of opioids (enkephalins and endorphins) have scarcely been investigated and the findings obtained are highly questionable. Compared with the amazing mass of investigations carried out in mammals, studies in lower vertebrates are few, and in large part performed in Amphibia and Aves. It appears that much further work has to be done by comparative endocrinologists to fully clarify the physiological relevance of the functional interactions between chromaffin and steroidogenic cells in the adrenal glands of lower vertebrates.

Role of endothelins in regulation of vascular tone in the in situ perfused rat adrenals.

This study examined the role of endothelins (ETs) and their receptor subtypes ETA and ETB in the regulation of vascular tone in the in situ perfused rat left adrenal gland. Endothelin-1 (ET-1), which binds both ETA and ETB receptors, decreased adrenal flow rate of the perfusion medium, and its effect was reversed by the ETA antagonist BQ-123 and enhanced by the ETB antagonist BQ-788. ET-3, which preferentially binds ETB, and the selective ETB agonist BQ-3020 increased adrenal flow rate of perfusate, and their effects were annulled by BQ-788. BQ-123 magnified the effect of ET-3 and did not affect that of BQ-3020. The ETA-mediated decrease and the ETB-mediated rise in the rate of collection of perfusate were abolished by Ro-31-8220, an inhibitor of protein kinase C (PKC), and by N(G)-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthase (NOS), respectively. Collectively, these findings suggest that ETs can regulate vascular tone in the in situ perfused rat adrenals via both PKC-coupled ETA and NOS-coupled ETB receptors, the activation of which evokes vasoconstriction and vasodilation, respectively.

Angiotensin-II stimulates DNA synthesis in rat adrenal zona glomerulosa cells: receptor subtypes involved and possible signal transduction mechanism.

Using an in situ perfusion technique of isolated left rat adrenal gland, it has been demonstrated that angiotensin-II (ANG-II) increases DNA synthesis in the zona glomerulosa (ZG), but not fasciculata-reticularis cells. The AT1 receptor antagonist DuP753 abolished the effect of ANG-II, while the AT2 receptor antagonist PD 123319 potentiated it. Both Ro31-8220, an inhibitor of protein kinase C (PKC), and tyrphostin-23, an inhibitor of tyrosine kinase (TK), evoked a partial reversal of ANG-II effect, and when added together to the perfusion medium abolished it. In contrast, the phospholipase C inhibitor U-73122 alone was able to induce a complete blockade of ANG-II effect. Neither the phospholipase A2 inhibitor AACOCF3 nor the cyclooxygenase inhibitor indomethacin and the lipoxygenase inhibitor phenidone affected ANG-II-induced stimulation of DNA synthesis, thereby making unlikely the involvement of the arachidonic acid signaling pathways. Our findings suggest that (i) ANG-II stimulates rat ZG cell proliferation acting via AT1 receptors coupled with phospholipase C, which activates both PKC and TK signaling systems; and (ii) the proliferogenic effect of ANG-II is partially counteracted by the activation of the AT2 receptor subtype.

Effects of adrenomedullin on the human adrenal glands: an in vitro study.

Numerous lines of evidence indicate that adrenal medulla exerts a paracrine control on the secretory activity of the cortex by releasing catecholamines and several regulatory peptides. Adrenomedullin (ADM) is contained in adrenal medulla of several mammalian species, including humans. Thus, we investigated whether human ADM1-52 exerts a modulatory action on steroid secretion of human adrenal cortex in vitro. Dispersed adrenocortical cells (obtained from the gland tail deprived of chromaffin cells) and adrenal slices (including both capsule and medulla) were employed. ADM specifically inhibited angiotensin II-stimulated aldosterone secretion of dispersed cells and enhanced basal aldosterone production by adrenal slices, minimal effective concentrations being 10(-7) and 10(-9) mol/L, respectively. These effects of ADM were suppressed by the CGRP1 receptor antagonist CGRP8-37 (10(-5) mol/L). Neither basal and ACTH-stimulated aldosterone secretion of dispersed cells nor agonist-enhanced aldosterone production by adrenal slices were affected by ADM, which also did not alter cortisol secretion of both types of adrenal preparations. ADM (10(-6) mol/L) blunted the aldosterone secretagogue action of the Ca2+ ionophore A23187 (10(-5) mol/L) on dispersed cells and adrenal slices. The beta-adrenoceptor antagonist l-alprenolol (10(-6) mol/L) suppressed aldosterone response of adrenal slices to 10(-7) mol/L isoprenaline and ADM. ADM concentration dependently raised epinephrine and norepinephrine release by adrenal slices, minimal effective concentration being 10(-9) mol/L. Collectively, these findings suggest that ADM, acting via the CGRP1 receptor subtype, exerts a direct inhibitory effect on angiotensin II-stimulated aldosterone secretion, which, when the integrity of adrenal tissue is preserved, is overcome and reversed by an indirect stimulatory action, conceivably involving the release of catecholamines by adrenal chromaffin cells.

Neurotensin stimulates CRH and ACTH release by rat adrenal medulla in vitro.

Neurotensin (NT) is a 13-amino acid peptide, widely distributed in the central and peripheral nervous system, which is able to stimulate the activity of the hypothalamo-pituitary CRH-ACTH system. We investigated by RIA the effect of NT on the release of CRH and ACTH immunoreactivities (ir) by rat adrenal medulla in vitro. NT enhanced the release of both CRH-ir and ACTH-ir, the maximal effective concentration being 10(-8) M. [D-Trp11]-NT, a specific NT receptor antagonist, abrogated the effects of NT. The stimulatory effect of 10(-8) M NT on ACTH-ir release was blocked by alpha-helical-CRH (an antagonist of CRH receptors), thereby suggesting that the enhancement in ACTH secretion is consequent to the stimulation of CRH release. These findings suggest that NT is a stimulator not only of the central (hypothalamo-pituitary), but also of the peripheral (intramedullary), branch of the CRH-ACTH system.

Neuropeptide K and neurokinin A stimulate CRH and ACTH release by rat adrenal medulla in vitro.

Tachykinins are a family of peptides that are able to modulate the activity of the hypothalamo-pituitary CRH-ACTH system. Mammalian tachykinins include neurokinin A (NKA), neurokinin B (NKB), neuropeptide K (NPK), and substance P (SP). We investigated by RIA the effects of tachykinins on the release of CRH and ACTH by rat adrenal medulla in vitro. NKA and NPK concentration-dependently enhanced the release of both CRH and ACTH, NPK being more active than NKA. NKB exerted only a minor stimulatory action exclusively on CRH release, and SP was ineffective. The stimulatory effect of both NKA and NPK on ACTH release was blocked by the CRH receptor antagonist alpha-helical-CRH, thereby suggesting that the increase in ACTH secretion is consequent to the stimulation of CRH release. These findings indicate that NKA and NPK are stimulators not only of the central (hypothalamo-pituitary), but also of the peripheral (intramedullary) branch of the CRH-ACTH system.

Further investigations on the effects of neuropeptide Y on the secretion and growth of rat adrenal zona glomerulosa.

NPY is a regulatory peptide, high levels of which are contained in adrenal glands of several mammals and which is co-released with catecholamines during various stressful conditions. The acute and chronic effects of NPY on adrenocortical secretion and growth were studied in the rat. NPY concentration-dependently increased aldosterone (ALDO), but not corticosterone (B) secretion of adrenal slices (maximal effective concentration was 10(-7) M). Two competitive inhibitors of NPY receptors, named PYX-1 and PYX-2, were found to dose-dependently inhibit ALDO response of adrenal preparations to 10(-7) M NPY; PYX-2 was more efficient than PYX-1, and at a concentration of 10(-5) M completely annulled the effect of 10(-7) M NPY. The acute bolus intraperitoneal (i.p.) injection of NPY (3 nmol/kg) raised plasma ALDO concentration (PAC), but not that of B (PBC); this effect of NPY was blocked by the simultaneous injection of PYX-2 (300 nmol/kg). The prolonged i.p. infusion with NPY (3 nmol/kg/h for 7 days) increased PAC (but not PBC) and induced a marked hypertrophy of the zona glomerulosa (ZG) and its parenchymal cells; dispersed ZG cells obtained from NPY-infused rats displayed a significantly enhanced basal and maximally agonist-stimulated ALDO production. The simultaneous infusion with PYX-2 (300 nmol/kg/h) completely annulled all these effects of NPY. The acute or chronic administration of PYX-2 alone did not evoke any apparent effect on the ZG secretion and growth. In light of these findings the following conclusions can be drawn: (i) NPY is able to stimulate not only the secretion, but also the growth of adrenal ZG in rats, via a receptor-mediated mechanism (since this effect is blocked by PYX-2); (ii) endogenous NPY does not play a prominent role in the physiological maintenance of secretion and growth of rat ZG (since PYX-2 alone is ineffective); (iii) NPY may play a crucial role in the fine tuning of the ZG functions in conditions requiring an increased release of mineralocorticoid hormones.

Evidence that both ETA and ETB receptor subtypes are involved in the in vivo aldosterone secretagogue effect of endothelin-1 in rats.

Endothelins (ET) are a family of vasoconstrictor peptides, secreted by vascular endothelium, which act through two main subtypes of receptors: ETA and ETB. ET-1 is known to stimulate aldosterone (ALDO) secretion by adrenal zona glomerulosa (ZG), and in vitro its effect was recently found to be exclusively mediated by ETB receptors. In this study the involvement of ETA and ETB in the mediation of the in vivo acute ALDO secretagogue action of ET-1 was investigated by the use of their selective antagonists BQ-123 and BQ-788, respectively. The bolus intraperitoneal administration of ET-1 dose-dependently raised both basal and angiotensin II (ANG II)-enhanced plasma ALDO concentration (PAC) in rats. Both antagonists counteracted the stimulatory effect of ET-1 on basal PAC, and when administered together completely annulled it. Conversely, only BQ-788 reversed the effect of ET-1 on ANG II-enhanced PAC. ET-1 increased systolic blood pressure (BP) in normal rats, but not in animals simultaneously administered ANG II. The hypertensive effect of ET-1 was completely abolished by BQ-123, and not affected by BQ-788. In light of these findings the following conclusions can be drawn: (i) the in vivo ALDO secretagogue action of ET-1 is mediated by both ETA and ETB, this latter subtype of ET receptors playing a major role; and (ii) the mechanism whereby ETA participates in this in vivo effect of ET-1 is indirect, and probably connected with the ET-1-induced rise in BP and adrenal blood flow.

Adrenomedullin stimulates steroid secretion by the isolated perfused rat adrenal gland in situ: comparison with calcitonin gene-related peptide effects.

Adrenomedullin (ADM), a vasodilatatory peptide contained in adrenal medulla, was found to induce a dose-dependent increase in aldosterone (ALDO) and corticosterone (B) release by the in situ perfused rat adrenal gland, along with a rise in the flow rate of the perfusion medium. The minimal effective dose for ALDO response was three and two orders of magnitude less than those able to evoke B and medium flow rate responses. Calcitonin gene-related peptide (CGRP), another vasodilatatory peptide contained in adrenal medulla and showing a slight homology in its amino acid sequence with ADM, elicited similar effects. CGRP (8-37), a specific antagonist of CGRP1 receptors, annulled all the effects of both ADM and CGRP, whereas l-alprenolol, a beta-adrenoceptor antagonist, partially reversed only ALDO response to the peptides. In light of these findings the following conclusions are drawn: i) ADM and CGRP stimulate rat adrenals in vivo to release B by raising blood flow rate; ii) ADM and CGRP enhance ALDO secretion via an indirect mechanism probably requiring the release of catecholamines by medullary chromaffin cells; and iii) the effects of ADM and CGRP on the rat adrenal gland are mediated by a common receptor of the CGRP1 subtype.

Adrenomedullin and calcitonin gene-related peptide inhibit aldosterone secretion in rats, acting via a common receptor.

Adrenomedullin (ADM) and calcitonin gene-related peptide (CGRP) did not affect either basal or ACTH-stimulated secretion of a1dosterone and corticosterone by dispersed rat capsular and inner adrenocortical cells, respectively. However, both peptides strongly depressed angiotensin-II (ANG- II)-stimulated a1dosterone production by capsular cells, the minimal effective concentration was 10(-7) M. The inhibitory effect of both ADM and CGRP was reversed by CGRP8-37, a specific CGRP1 receptor antagonist; a complete reversal was obtained with a CGRP8-37 concentration of 10(-6) M. Our findings indicate that ADM and CGRP specifically interfere with the intracellular mechanisms transducing the secretagogue signal of ANG-II, and suggest that the ADM effect is mediated by CGRP receptors