The proliferative effect of synthetic N-POMC(1-28) peptides in rat adrenal cortex: a possible role for cyclin E.

Modified synthetic N-POMC(1-28) without disulfide bridges has been shown to act as an adrenal mitogen. Cyclins and their inhibitors are the major cell cycle controls, but in the adrenal cortex the effect of ACTH and N-POMC on the expression of these proteins remains unclear. In this work, we evaluate the effect of different synthetic N-POMC peptides on the S-phase of the cell cycle. In addition, we examine the cyclin E expression in rat adrenal cortex. Rats treated with dexamethasone were injected with ACTH and/or synthetic modified N-POMC and/or synthetic N-POMC with disulfide bridges. DNA synthesis was determined by BrdU incorporation and protein expression was analyzed by immunoblotting and immunohistochemistry. The results showed that similarly to modified N-POMC without disulfide bridges, administration of synthetic N-POMC with disulfide bridges and the combination of ACTH and N-POMC promoted an increase of BrdU-positive nuclei in adrenal cortex. However, the proliferative effect of N-POMC was comparable to that of ACTH only in the zona glomerulosa. An increase in cyclin E expression was observed 6 h after N-POMC treatment in the outer fraction of the adrenal cortex, in agreement with immunohistochemical findings in the zona glomerulosa. In summary, the effect of synthetic N-POMC with disulfide bridges was similar to modified synthetic N-POMC, increasing proliferation in the adrenal cortex, confirming previous evidence that disulfide bridges are not essential to the N-POMC mitogenic effect. Moreover, cyclin E appears to be involved in the N-POMC- and ACTH-stimulated proliferation in the zona glomerulosa of the adrenal cortex.

Mechanisms of action of endothelin-1 in rat adrenal.

Displacement curves of 125I-Endothelim-1 (ET-1) binding to rat adrenal cells with unlabeled ET-1, and the ET-1 receptor-related peptides sarafotoxin and BQ-123, show that rat adrenal cortex possess, as its bovine counterpart, two different receptors to ET-1 named ET-A and ET-B. Binding of ET-1 to its rat adrenal receptors stimulates i) aldosterone production, in vivo and in vitro ii) calcium influx, which is mediated through voltage dependent- and receptor operated- calcium channels, iii) cholesterol uptake, iv) stimulation of Na+/K+-ATPase and iv) diacylglycerol production. While the last effect is mediated through ET-A receptors the others involve binding of ET-1 to ET-B receptors. Finally, ouabain potentiates the ET-1-mediated stimulation of aldosterone production, suggesting that the effect of the peptidic hormone on Na+/K+-ATPase could act as a negative feedback mechanism.

Endothelin-1 stimulation of aldosterone and zona glomerulosa ouabain-sensitive sodium/potassium-ATPase.

Endothelin stimulates the cells of the zona glomerulosa of the adrenal gland and releases aldosterone. While it is a less potent aldosterone secretagogue than angiotensin II endothelin also potentiates the effects of angiotensin II on aldosterone biosynthesis. Two endothelin receptors have been cloned and are expressed in the adrenal zona glomerulosa. Intravenous infusion of endothelin at a rate of 80 ng/kg/min for 30 min into rats produced increases in blood pressure, adrenal content of aldosterone and stimulated the ouabain-sensitive sodium potassium ATPase in the zona glomerulosa, but not in the zona fasciculata, of the adrenal. The simultaneous infusion of the isopeptide specific endothelin receptor A (ETA) antagonist BQ-123 blocked the pressor effects of endothelin, but did not alter the increase in aldosterone content of the zona glomerulosa or the ouabain-sensitive sodium potassium ATPase activity. Infusion of Sarafotoxin 6b, an ETB agonist, also increased the aldosterone content of the adrenal and stimulated the ouabain-sensitive sodium potassium ATPase in the zona glomerulosa, further indicating that the effect of endothelin is probably mediated by ETB or isopeptide non-specific endothelin receptor. The mechanism by which endothelin stimulates the sodium potassium ATPase is unclear as is the relation between a stimulated sodium potassium ATPase and the potentiation of angiotensin II effect on the adrenal.

Stimulation of aldosterone production by hemin in calf adrenal glomerulosa cell cultures.

Aldosterone production from 11-deoxycorticosterone was stimulated by hemin in primary cultures and homogenates of calf adrenal zona glomerulosa, in a time- and dose-dependent fashion. The ferrochelatase inhibitor 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine (DDC) blocked the stimulation of aldosterone mediated by adrenocorticotropin (ACTH). Addition of hemin after treatment with DDC partially restored ACTH action. These results suggest that hemin may play a role in regulation of aldosterone production.

In vivo stimulation of aldosterone biosynthesis by endothelin: loci of action and effects of doses and infusion rate.

Infusion of endothelin-1 (ET-1) into rats increased adrenal mitochondrial synthesis of aldosterone from deoxycorticosterone and the adrenal cytosolic content of aldosterone. The dose-response relationships for these last two effects of ET-1 were found to be biphasic with a maximum (corresponding to 80 to 200% increase) at 50 to 80 ng ET-1/kg/min, and were also dependent on the infusion rate. Plasma aldosterone levels were also increased in a similar ratio. Previous infusion of the converting enzyme inhibitor enalapril did not affect the ET-1-induced increase in steroidogenesis. Finally, pregnenolene production was also increased in incubations of mitochondria from treated rats. These results indicate that ET-1 augments aldosteronogenesis by increasing the early as well as the late pathway. These effects were independent of the formation of angiotensin II. Isolated glomerulosa cells responded to ET-1 increasing aldosterone production in a dose-related fashion. These results confirm a direct effect of ET-1 on the adrenal gland in vivo.

Insulin stimulatory action on amino acid uptake in bovine adrenal cortex or glomerulosa zone.

1. Insulin stimulated the [1-14C] methylaminoisobutyric acid and [1-14C] aminoisobutyric acid uptake in the bovine adrenal cortex or in the glomerulosa zone through the A system. 2. Verapamil nullified the insulin stimulatory action indicating that this hormonal action is probably related to the voltage-dependent Ca2+ channels.