[Morphological and functional correlations of the adrenal cortex].
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To assess sex differences in stress-induced acetylcholine (ACh) release in the hippocampus and corticosterone (CS) release from the adrenal cortex, an in vivo microdialysis study was performed in intact male (n = 6) and cycling female (diestrus 1, n = 5; proestrus, n = 6) rats. Dialysates and blood samples were taken from 11.00 to 16.00 h in freely moving rats, but restraint stress was applied from 12.00 to 13.00 h. Basal ACh release in the hippocampus was low without significant differences between groups. Although ACh release promptly increased with the onset of restraint stress in both sexes, the magnitude of the increase in males was significantly greater than in female rats during diestrus and proestrus (p < 0.01). Basal serum CS concentrations were significantly greater in proestrus than in diestrus (p < 0.01) or in male rats (p < 0.01). Serum CS concentrations significantly increased at the onset of restraint stress in both sexes (p < 0.01), but the magnitude of the increase was significantly greater in female than in male rats (p < 0.01). These results indicate that sex and/or gonadal steroid environment can affect the stress-induced ACh release in the hippocampus and CS release from the adrenal cortex.
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Aberrant adrenocortical function during the first trimester of human fetal development underlies the severe virilization of congenital adrenal hyperplasia due to cytochrome P450 21-hydroxylase (CYP21) deficiency. Although valuable information of human adrenocortical development after 12 weeks gestation is available, less is known earlier in pregnancy. In our studies, the adrenal cortex was first detected in human embryos by hematoxylin and eosin staining at 33 days post-conception (dpc) with distinction between the definitive and fetal zones possible at 52 dpc. Vascular development was apparent within the adrenal gland at 41 dpc. CYP11A and CYP17 were expressed centrally within the fetal zone at 50 dpc and all later time points during the first trimester. Weaker CYP11A immunoreactivity also was visible in the outer region of the adrenal cortex consistent with definitive zone expression. In this location, immunoreactivity was observed for 3beta-hydroxysteroid dehydrogenase and the proliferation marker, Ki67. These data raise the possibility of de novo cortisol biosynthesis during the first trimester of human development and are relevant to the pathophysiology of 46,XX virilization in CYP21 deficiency.
Cytochrome P-450 specific for steroid 17 alpha-hydroxylation (P-450(17 alpha] was immunohistochemically observed in pig adrenal cortex, testis and ovary by the biotin-streptavidin method using a specific antibody against P-450(17 alpha) purified from neonatal pig testis. In the adrenal cortex, P-450(17 alpha) was present in the zona fasciculata and reticularis while no immunoreactivity was observed in the zona glomerulosa, confirming the absence of 17-hydroxylated steroid synthesis in the zona glomerulosa. In the testis, P-450(17 alpha) was present exclusively in Leydig cells and immunoreactivity was absent in seminiferous tubules. In the ovary, immunoreactivity was observed only in the theca interna but not in the membrana granulosa of follicles. Among the tissues examined, the relative intensity of immunoreactivity was greatest in the Leydig cells, and progressively less in theca interna cells, outer fasciculata cells and inner fasciculata and reticularis cells.
Using light and electron microscopy, we have observed the presence of rays containing medullary tissue extending across the cortex of rat adrenal glands. Within these rays chromaffin cells, as well as collagen and nerve fibers, were present. It is suggested that these endocrine cells may have a paracrine function within the cortex, possibly via their secretory product.
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Adrenocorticotropic hormone (ACTH)-like peptide immunocortin (IMC) VKKPGSSVKV, corresponding to the amino acid sequence 11-20 of the variable part of human immunoglobulin G (IgG) 1 heavy chain, at concentrations of 10(-9)-10(-6) I was found to increase the adenylate cyclase activity in adrenal cortex membranes, while intramuscular injection of immunocortin at doses of 10-100 microg/kg was found to stimulate the secretion of 11-oxycorticosteroids (CS) from the adrenals to the bloodstream. Immunocortin was labeled with tritium to specific activity of 22 Ci/mmol. Receptor binding studies revealed that [(3)H]immunocortin ([(3)H]IMC) bound with high affinity and specificity to ACTH receptors on rat adrenal cortex membranes (K(d)=2.1+/-0.2 nM, B(max)=1.1+/-0.1 pmol/mg protein).
Nuclear volumes of cells in rat adrenal cortex were measured. The rats were forced to swim during one month for 3 hours a day. During the experiment, the extension of nuclear hypetrophy in the external zona fasciculata underwent some phasic changes, phases being as follows: 1) increase in hypertrophy up to the maximum (2nd and 5th days of experiment); 2) the stability of hypertrophy close to the maximum (12th, 18th and 24th days), and the decrease in hypertrophy (30th day). The phases are assumed to correspond, respectively, to stages of alarm reaction, resistance and exhaustion caused by chronic stress. The increased nuclear volume of cells was also observed in the internal zona fasciculata. The discrepancy between nuclear volumes of zona reficularis in the experiment and in the control was statistically insignificant.
Effects of ACTH and calcium on cyclic AMP and steroid production by the zona fasciculata-reticularis (the decapsulated fraction) from the rat adrenal cortex have been studied. Increasing concentrations of extracellular calcium enhanced the action of ACTH on cyclic AMP and steroid production. These effects of ACTH with calcium were prevented by lanthanum, but not by tetracaine or verapamil, suggesting that ACTH stimulation may be mediated by calcium through a process not involving the tetracaine- or verapamil-vulnerable step(s) of the calcium current. High concentrations of external calcium itself increased cyclic AMP accumulation without any increase in steroidogenesis. A calcium ionophore, X537A was stimulatory for steroidogenesis but inhibitory with respect to cyclic AMP accumulation. Considered together with the findings of AMP increase, these results suggest that ACTH primarily increases intracellular calcium mobilization thus stimulating directly the steroidogenesis, which is independent of the cyclic AMP system. Relatively high concentrations of ACTH activate the adenylate cyclase, which depends on extracellular calcium to increase cyclic AMP levels and stimulation of steroidogenesis by the decapsulated fractions of the adrenal cortex.
The effect of repeated stress (1 h of daily immobilization for seven consecutive days) on the adrenal cortex of young adult male albino rats was evaluated by morphohistochemical methods and plasma assays; at the same time, testes and major salivary glands, as steroid-producing and -depending organs, respectively, were examined. Morphological and histochemical changes were found in the adrenal cortex, testis and submaxillary gland, though varying in degree and extent depending on the gland examined. Corticosterone and progesterone plasma levels increased, in agreement with the lipid depletion observed in the zona fasciculata, while testosterone and androstenedione decreased, as confirmed by the less marked enzymatic activity in the Leydig cells. The study thus proves that repeated stress, even of temporary duration, is able to influence directly or indirectly the morphofunctional state of the three examined glands, suggesting a functional linkage.
Recently, an ANF-sensitive guanylate cyclase (GC-A) has been cloned from a rat brain cDNA library. Here we studied the stimulation of cyclic GMP accumulation in response to atrial natriuretic factor (ANF), urodilatin and atriopeptin I (AP-1) in a rat glioma C6 cell line permanently transfected with GC-A as well as GC-A activity in membranes from these C6 cells and in membranes from COS-7 cells that were transiently transfected with GC-A. We also measured binding affinities for these natriuretic peptides in the membrane preparations. These characteristics of GC-A were compared to those of membrane preparations from adrenal cortex of bovine and human origin. The order of potency of stimulation of cyclic GMP accumulation in permanently transfected glioma cells was ANF greater than urodilatin greater than AP I; AP I stimulated cyclic GMP accumulation. A similar order of potency was obtained for stimulation of guanylate cyclase activity in membranes from permanently transfected glioma cells as well as from transiently transfected COS-7 cells. In contrast, AP-1 was uneffective to stimulate guanylate cyclase in membrane preparations from adrenal cortex from bovine as well as from human origin. Furthermore, urodilatin was equipotent to ANF in these preparations. Binding affinities were comparable for ANF and urodilatin in membranes from cells transfected with GC-A and in membranes from adrenal cortex of both sources, whereas AP-1 had a weaker affinity in all preparations studied.(ABSTRACT TRUNCATED AT 250 WORDS)
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