CORTICOSTERONE, 18-OH-DEOXYCORTICOSTERONE, DEOXYCORTICOSTERONE AND ALDOSTERONE SECRETION IN TISSUE CULTURE OF FOETAL RAT ADRENALS IN THE PRESENCE AND THE ABSENCE OF ACTH

1976 ◽  
Vol 83 (4) ◽  
pp. 781-793 ◽  
Author(s):  
M. Salmenperä ◽  
A. I. Kahri
Keyword(s):  
Tissue Culture ◽  
Cortical Cell ◽  
Secretory Activity ◽  
Zona Glomerulosa ◽  
Zona Fasciculata ◽  
Gas Liquid Chromatography ◽  
Aldosterone Secretion ◽  
Foetal Rat ◽  
Rat Adrenals ◽  
Differentiation Stage

ABSTRACT A method for simultaneous analysis of the main corticosteroids secreted by the rat adrenal is described. Purification is performed using previously established steps involving dichloromethane extraction, ethanol-cyclohexane partition and Sephadex-LH-20 column chromatography. Deoxycorticosterone and aldosterone are then quantitated with radioimmunoassay and corticosterone and 18-hydroxy-deoxycorticosterone with gas liquid chromatography as their 0-methyloxime-trimethylsilyl ethers. The coefficient of variation of the method for these steroids in concentrations found in the tissue culture of foetal rat adrenals varies between 3.9–10.9% The secretion of these steroids in the tissue culture of foetal rat adrenals is studied. The steroid secretory pattern is correlated with a differentiation stage of the cultures as detected by electron microscopy. The initial secretory activity declines considerably after 15 days of cultivation to a low level. This is the case also with respect to aldosterone although the cortical cell population changed from the mixed population of the differentiated (zona fasciculata-like) and undifferentiated (zona glomerulosa-like) cells to homogenous undifferentiated growth. Addition of ACTH increased deoxycorticosterone secretion rapidly. Corticosterone, 18-hydroxy-deoxycorticosterone and aldosterone secretion was observed only after the differentiation of the cells (especially changes in their mitochondrial compartment) was evident morphologically.

The properties of adrenal zona glomerulosa cells after purification by gravitational sedimentation

1974 ◽  
Vol 185 (1081) ◽  
pp. 375-407 ◽  
Keyword(s):  
Angiotensin Ii ◽  
Cyclic Amp ◽  
Microscopic Examination ◽  
Zona Glomerulosa ◽  
Maximal Response ◽  
Zona Fasciculata ◽  
Gravitational Sedimentation ◽  
Rat Adrenals ◽  
Glomerulosa Cells ◽  
Zona Glomerulosa Cells

The densities of latex spheres and biological cells can be reliably determined from their sedimentation rate in an albumin gradient under unit gravitational force. The densities of zona glomerulosa and fasciculata cells of rat adrenals were found to be 1.072 ± 0.004 and 1.040 ± 0.001 respectively. Purified zona glomerulosa cells of rat adrenals can be prepared by gravitational sedimentation of dispersed cells from capsule strippings of the gland, which originally contain 3 to10% zona fasciculata contamination. Electron and phase microscopic examination of the sedimented glomerulosa cells and their steroidogenic response to ACTH and cyclic AMP indicate that they are reasonably free of contamination from zona fasciculata cells. Electron microscopic examination of the purified glomerulosa cells indicates that most of them are reasonably normal in structure. Their basal production of corticosterone is decreased after sedimentation. However, their maximal response of corticosterone output to serotonin and potassium and their response to all potassium concentrations is not significantly altered, indicating normal function for the cells producing steroids. Their maximal responses to ACTH, valine angiotensin II and cyclic AMP are decreased, but, at the doses used, steroidogenesis by the zona fasciculata contamination in the unfractionated preparation would be stimulated by these substances. Purified zona glomerulosa cells have about the same maximal response of corticosterone output (about twofold) to potassium, valine and isoleucine angiotensin II, serotonin and ACTH. The maximal response of the purified zona glomerulosa cells to cyclic AMP is similar to that elicited by valine and isoleucine angiotensin II, potassium, serotonin or ACTH. This indicates that if these stimuli act by increasing cyclic AMP output, then the maximal response of corticosterone output (about twofold) is defined by the limited response of the biosynthetic pathways to cyclic AMP.

scholarly journals Quantitative assessment of CYP11B1 and CYP11B2 expression in aldosterone-producing adenomas

2002 ◽  
pp. 795-802 ◽  
Author(s):  
F Fallo ◽  
V Pezzi ◽  
L Barzon ◽  
P Mulatero ◽  
F Veglio ◽  
...  
Keyword(s):  
Real Time ◽  
Secretory Activity ◽  
Zona Glomerulosa ◽  
Zona Fasciculata ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Pathophysiological Role ◽  
Aldosterone Production

BACKGROUND: The presence and pathophysiological role of CYP11B1 (11beta-hydroxylase) gene in the zona glomerulosa of human adrenal cortex is still controversial. METHODS: In order to specifically quantify CYP11B1, CYP11B2 (aldosterone synthase) and CYP17(17alpha-hydroxylase) mRNA levels, we developed a real-time RT-PCR assay and examined the expression in a series of adrenal tIssues, including six normal adrenals from patients adrenalectomized for renal cancer and twelve aldosterone-producing adenomas (APA) from patients with primary aldosteronism. RESULTS: CYP11B1 mRNA levels were clearly detected in normal adrenals, which comprised both zona glomerulosa and fasciculata/reticularis cells, but were also measured at a lower range (P<0.05) in APA. The levels of CYP11B2 mRNA were lower (P<0.005) in normal adrenals than in APA. CYP17 mRNAlevels were similar in normal adrenals and in APA. In patients with APA, CYP11B2 and CYP11B1 mRNA levels were not correlated either with basal aldosterone or with the change from basal aldosterone in response to posture or to dexamethasone. No correlation between CYP11B1 mRNA or CYP11B2 mRNA and the percentage of zona fasciculata-like cells was observed in APA. CONCLUSIONS: Real-time RT-PCR can be reliably used to quantify CYP11B1 and CYP11B2 mRNA levels in adrenal tIssues. Expression of CYP11B1 in hyperfunctioning zona glomerulosa suggests an additional formation of corticosterone via 11beta-hydroxylase, providing further substrate for aldosterone biosynthesis. CYP11B1 and CYP11B2 mRNA levels in APA are not related to the in vivo secretory activity of glomerulosa cells, where post-transcriptional factors might ultimately regulate aldosterone production.

ADRENAL POTASSIUM AND SODIUM IN EXPERIMENTAL HYPER- AND HYPOALDOSTERONISM IN THE RAT. DETERMINATION BY ELECTRON PROBE X-RAY MICROANALYSIS

1975 ◽  
Vol 80 (1) ◽  
pp. 114-125 ◽  
Author(s):  
Katalin Sz. Szalay ◽  
Ernö Bácsy ◽  
Ervin Stark
Keyword(s):  
Electron Probe ◽  
Sodium Content ◽  
Zona Glomerulosa ◽  
Potassium Content ◽  
Zona Fasciculata ◽  
Aldosterone Secretion ◽  
X Ray ◽  
Intracellular Potassium

ABSTRACT Potassium and sodium contents in the various adrenal zones were determined in experimental hyper- and hypoaldosteronism in the rat by electron probe X-ray microanalysis. The analysis aimed at revealing intracellular values. There was no change in the potassium content of the zona glomerulosa, zona fasciculata and medulla neither in hyperaldosteronism, induced by Na-deficiency, nor in hypoaldosteronism, elicited by Na-rich diet. The sodium content in the zona glomerulosa and zona fascicularis was increased in the Na-loaded rats, while that of the medulla was not changed. Our data are not consistent with the hypothesis that a change of adrenal intracellular potassium would act as a final stimulus in the regulation of aldosterone secretion.

scholarly journals Aldosterone secretion in patients with septic shock: a prospective study

2013 ◽  
Vol 57 (8) ◽  
pp. 636-641 ◽  
Author(s):  
Rafael Barberena Moraes ◽  
Gilberto Friedman ◽  
Marina Verçoza Viana ◽  
Tiago Tonietto ◽  
Henrique Saltz ◽  
...  
Keyword(s):  
Septic Shock ◽  
Serum Levels ◽  
Zona Glomerulosa ◽  
High Dose ◽  
Zona Fasciculata ◽  
Cortisol Secretion ◽  
Aldosterone Secretion ◽  
Acth Stimulation ◽  
Aldosterone Production ◽  
A Prospective Study

OBJECTIVE: To assess serum levels of the main factors that regulate the activation of the zona glomerulosa and aldosterone production in patients with septic shock, as well as their response to a high-dose (250 µg) adrenocorticotropic hormone (ACTH) stimulation test. SUBJECTS AND METHODS: In 27 patients with septic shock, baseline levels of aldosterone, cortisol, ACTH, renin, sodium, potassium, and lactate were measured, followed by a cortrosyn test. RESULTS: Renin correlated with baseline aldosterone and its variation after cortrosyn stimulation. Baseline cortisol and its variation did not correlate with ACTH. Only three patients had concomitant dysfunction of aldosterone and cortisol secretion. CONCLUSIONS: Activation of the zona glomerulosa and zona fasciculata are independent. Aldosterone secretion is dependent on the integrity of the renin-angiotensin-aldosterone system, whereas cortisol secretion does not appear to depend predominantly on the hypothalamic-pituitary-adrenal axis. These results suggest that activation of the adrenal gland in critically ill patients occurs by multiple mechanisms.

Electrical responses in cat adrenal cortex: possible relation to aldosterone secretion.

1979 ◽  
Vol 237 (2) ◽  
pp. E158 ◽  
Author(s):  
E Natke ◽  
E Kabela
Keyword(s):  
Membrane Potential ◽  
Angiotensin Ii ◽  
Adrenal Cortex ◽  
Adrenal Glands ◽  
Membrane Depolarization ◽  
Zona Glomerulosa ◽  
Membrane Potentials ◽  
Zona Fasciculata ◽  
Aldosterone Secretion ◽  
Stimulus Secretion Coupling

The effects of secretagogues for aldosterone release were studied on the membrane potential of cells in the adrenal cortex of the cat. Adrenal glands were excised, sliced, and continuously superfused. Membrane potentials were recorded from both zona glomerulosa and zona fasciculata-reticularis. Secretagogues, angiotensin II (1 microgram/ml) and 20 mM KCl, were found to depolarize cells rapidly. Ouabain (10(-5) M) also depolarized the membrane potential although the response was sluggish. Samples of the superfusate were collected and analyzed by radioimmunoassay for their aldosterone and cortisol content. Depolarizing concentrations of angiotensin II, KCl, and ouabain seemed to increase aldosterone release. Cortisol output was more variable. Saralasin blocked the effects of angiotensin II on the membrane potential. These experiments suggest that membrane depolarization plays a role in the stimulus-secretion coupling of mineral corticoids.

THE HISTOGENESIS OF THE ADRENAL CORTEX IN THE FOETAL SHEEP

1979 ◽  
Vol 91 (1) ◽  
pp. 134-149 ◽  
Author(s):  
Peter M. Robinson ◽  
Elisabeth J. Rowe ◽  
E. Marelyn Wintour
Keyword(s):  
Endoplasmic Reticulum ◽  
Steroid Hormones ◽  
Adrenal Glands ◽  
Smooth Endoplasmic Reticulum ◽  
Rapid Development ◽  
Light And Electron Microscopy ◽  
Cortical Cell ◽  
Outer Zone ◽  
Zona Glomerulosa ◽  
Zona Fasciculata

ABSTRACT The cortex of sheep foetal adrenal glands from 25 days gestation until newborn (term equals 147 ± 3 days) were examined by light and electron microscopy. Three stages of development are of particular importance in relating structure to function: 1) from 35 to 60 days, 2) from 60 to 120 days and 3) from 120 days to term. Between 35 and 60 days one cortical cell type predominated. It contained mitochondria with lamellar and vesicular cristae, scattered long strands of granular endoplasmic reticulum and only small amounts of smooth endoplasmic reticulum. After about 60 days two zones were apparent in the cortex and chromaffin cells became concentrated in the medulla. After 80 days the outer zone contained cells which resembled mature zona glomerulosa cells and the cells in the inner zone remained like those seen between 35 and 60 days, except they contained even less smooth endoplasmic reticulum. However, after about 90 days a small number of deep inner zone cells contained mitochondria with vesicular cristae which thus resemble mitochondria in the mature zona fasciculata. From about 120 days there was an increase in the number of cells in the inner zone that contained mitochondria with vesicular cristae. These cells also contained substantial quantities of smooth endoplasmic reticulum. At term most inner zone cells have this mature appearance. Thus there is no "foetal cortex" in the sheep analogous to that found in human adrenal development, i. e. there is no prominent zone of cells containing large amounts of smooth endoplasmic reticulum which is present throughout most of the foetal period of development, and which regresses at birth. The structure of the cells present between 35 and 60 days was unexpected because it has been shown previously that sheep foetal adrenals of this age are capable of producing relatively large quantities of steroid hormones. However, the appearance of cells resembling mature zona glomerulosa cells at about 80 days correlates with the previously demonstrated ability of sheep adrenal glands of this age to produce relatively large quantities of aldosterone. The rapid development of numbers of mature cells in the last 3 weeks of gestation correlates with the previously described ability of near term sheep foetal adrenals to produce very large quantities of steroid hormones.

Endocrine control of the distribution of basic fibroblast growth factor, insulin-like growth factor-I and transforming growth factor-β1 mRNAs in adult rat adrenals using non-radioactive in situ hybridization

1995 ◽  
Vol 144 (2) ◽  
pp. 379-387 ◽  
Author(s):  
M M Ho ◽  
G P Vinson
Keyword(s):  
Growth Factor ◽  
Adrenal Cortex ◽  
Transforming Growth Factor ◽  
Zona Glomerulosa ◽  
Dietary Sodium ◽  
Zona Fasciculata ◽  
Sodium Restriction ◽  
Igf I ◽  
Rat Adrenals

Abstract This study located the particular cell types involved in the synthesis of growth factors in adult female rat adrenal glands. Non-isotopic in situ hybridization was used and the cellular localizations of the mRNAs of basic fibroblast growth factor (bFGF), IGF-I, and transforming growth factor-β1 (TGF-β1) were studied in adrenals from control animals and from those treated with ACTH or subjected to dietary sodium restriction. The adrenal medulla was the richest source of both bFGF and IGF-1 mRNA in both control and experimental rat adrenals. In the cortex, bFGF and IGF-I mRNAs were found mainly in the zona fasciculata in control animals, although some transcription was also detected in the zona reticularis and zona glomerulosa. Both ACTH and sodium restriction activated bFGF and IGF-I gene expression in the zona glomerulosa. Since cellular proliferation and differentiation occur primarily in the outer cortex, the data are consistent with the view that bFGF and IGF-I act as an autocrine/paracrine mitogen and differentiation regulator respectively in the rat adrenal cortex. Very small amounts of TGF-β1 mRNA were detected, predominantly in the zona fasciculata of control rats. There were no observable differences in amounts and localization of TGF-β1 mRNA between the adrenals of control rats and those treated with ACTH for 1 day. TGF-β1 mRNA was very weak or undetectable in the adrenals from rats treated with ACTH for three and five days or from sodium-restricted rats. Although TGF-β1 immunoreactive protein has been shown to be present in the zonae fasciculata and reticularis and to modulate negatively the steroidogenic activities in the adrenal cortex of other species, its gene is not actively expressed in rat adrenals. The present results showed that ACTH administration or dietary sodium restriction, both important adrenal mitogens in vivo, significantly altered the spatial patterns of the distribution of bFGF and IGF-I mRNAs and also increased the amount of bFGF mRNA in the adrenal cortex. This suggests that growth and differentiation of the adrenal cortex are partly mediated by bFGF and IGF-I. Journal of Endocrinology (1995) 144, 379–387

scholarly journals Adrenal Capillary Endothelial Cells Stimulate Aldosterone Release through a Protein That Is Distinct from Endothelin1

Endocrinology ◽  
1999 ◽  
Vol 140 (10) ◽  
pp. 4411-4418 ◽  
Author(s):  
Lori J. Rosolowsky ◽  
Craig J. Hanke ◽  
William B. Campbell
Keyword(s):  
Endothelial Cells ◽  
Receptor Antagonist ◽  
Gel Filtration ◽  
Zona Glomerulosa ◽  
Maximal Response ◽  
Zona Fasciculata ◽  
Aldosterone Secretion ◽  
Etb Receptor ◽  
Capillary Endothelial Cells

Abstract We tested the possibility that bovine adrenal capillary endothelial cells (ECs) stimulate aldosterone secretion from bovine zona glomerulosa (ZG) cells by the release of a transferable factor. In coincubations of ZG cells and ECs in serum-free medium, aldosterone release was stimulated approximately 17-fold, and the stimulation was related to the concentration of ECs. The maximal stimulation by ECs was 75% of the maximal response to ACTH. In contrast, adrenal pericytes and fibroblasts were without effect. ECs incubated alone without ZG cells did not produce aldosterone. Conditioned medium from ECs (EC-CM), but not adrenal fibroblasts, stimulated aldosterone release approximately 3-fold. The stimulation increased with the concentration of EC-CM and the duration of conditioning time. Steroidogenic activity in EC-CM was abolished by pronase treatment, indicating that the active factor was a protein. However, the activity in EC-CM was distinct from that of endothelin-1 (ET-1), an endothelial peptide that also stimulates aldosterone secretion, as it was not blocked by the ETB receptor antagonist PD-145065, it did not alter[ 125I]ET-1 binding to ZG cells, and its release occurred before the release of ET-1. Neither ECs nor EC-CM stimulated the production of cortisol from zona fasciculata cells. The activity of EC-CM was not blocked by an angiotensin II AT1 receptor antagonist or a bradykinin B2 receptor antagonist. EC-CM stimulated increased intracellular calcium in fura-2-loaded ZG cells, but did not increase the production of cAMP. Using gel filtration, this peptide had an approximate molecular mass of 3000 Da and migrated earlier than ET-1. This study demonstrates that ECs in vitro alter steroidogenesis through the release of a transferable substance and suggests the existence of an endothelium-derived steroidogenic factor that is produced by adrenal capillary ECs. This endothelium-derived steroidogenic factor may function in the adrenal gland as a paracrine regulator of aldosterone secretion.

Age-related impairment of K(+)- and ACTH-stimulated aldosterone secretion by rat adrenal capsules

1993 ◽  
Vol 264 (1) ◽  
pp. E82-E89
Author(s):  
K. J. Radke
Keyword(s):  
Zona Glomerulosa ◽  
Threshold Sensitivity ◽  
Zona Fasciculata ◽  
Aldosterone Secretion ◽  
Cortical Function ◽  
Plasma Aldosterone Concentration ◽  
Fischer 344 ◽  
Age Related ◽  
Adrenal Cortical Function

It has been suggested that the decrease in plasma aldosterone concentration with advancing age is due to a direct effect of aging on adrenal cortical function. In this study, aldosterone secretion by adrenal capsules from male Fischer 344 rats that were 3–5, 11–13, and 19–21 mo of age was evaluated. Adrenal capsules, comprising the capsule, zona glomerulosa, and small portions of the zona fasciculata-reticularis, were perifused in vitro. Basal aldosterone secretion progressively decreased with aging. In response to concentrations of KCl that ranged from 5.0 to 9.0 mM and to adrenocorticotropic hormone (ACTH) that ranged from 10(-11) to 10(-8) M, threshold sensitivity of adrenal capsules to these secretagogues decreased with aging. Moreover, the magnitude of change in K(+)- and ACTH-stimulated aldosterone secretion and maximal aldosterone response of the adrenal capsules to KCl decreased with aging. Thus it is concluded that aging directly affects basal aldosterone secretion by rat adrenal capsules and that there is an age-related impairment of threshold sensitivity and responsiveness of adrenal capsules to the secretagogues, KCl and ACTH.

Detection of Na+ and K+ in the Rat Adrenal Cortex with the Electron Microprobe

1977 ◽  
Vol 53 (5) ◽  
pp. 423-430 ◽  
Author(s):  
C. Decorzant ◽  
A. M. Riondel ◽  
M.-J. Philippe ◽  
J. Bertrand ◽  
M. B. Vallotton
Keyword(s):  
Adrenal Cortex ◽  
Electron Microprobe ◽  
Electrolyte Concentration ◽  
Zona Glomerulosa ◽  
Plasma Aldosterone ◽  
Zona Fasciculata ◽  
Aldosterone Secretion ◽  
Dietary Regimen ◽  
High K ◽  
Aldosterone Production

1. In order to demonstrate whether modification of aldosterone secretion is mediated by parallel changes of K+ in the adrenal zona glomerulosa, the total (intracellular + extracellular) Na+ and K+ content of the rat adrenal cortex was determined with the electron microprobe. 2. Groups of rats were submitted to one of the following dietary regimens: standard, low Na+, high K+ or high Na+. 3. Distribution of Na+ and K+ across the zona glomerulosa and zona fasciculata was compared. Standards of known electrolyte concentration were also analysed. 4. The [Na+] was found to be greater in the zona glomerulosa than in the zona fasciculata but K+ was distributed evenly in both zones. This was independent of dietary regimen. 5. Aldosterone production, assessed by plasma aldosterone concentrations, could not be correlated with zona glomerulosa K+ content.

Sign in / Sign up

Export Citation Format

Share Document