Intracellular Potassium and Steroidogenesis of Isolated Rat Adrenal Cells: Effect of Potassium Ions and Angiotensin II on Purified Zona Glomerulosa Cells

1977 ◽  
Vol 53 (3) ◽  
pp. 289-296 ◽  
Author(s):  
C. M. Mackie ◽  
E. R. Simpson ◽  
M. S. R. Mee ◽  
S. A. S. Tait ◽  
J. F. Tait
Keyword(s):  
Angiotensin Ii ◽  
Potassium Concentration ◽  
Zona Glomerulosa ◽  
Intracellular Water ◽  
Potassium Content ◽  
Percentage Increase ◽  
Intracellular Potassium ◽  
External Potassium ◽  
Glomerulosa Cells ◽  
Intracellular Potassium Concentration

1. A method involving centrifugation through phthalate ester oils is described for the rapid separation of isolated cells from suspending medium. Intracellular potassium concentrations and intracellular water space values for isolated adrenal zona glomerulosa and zona fasciculata cells are given. 2. The effects of angiotensin II and a high external potassium concentration (8·4 mmol/l) on intracellular potassium contents, intracellular water spaces and hence intracellular potassium concentrations of purified glomerulosa cells have been investigated. Corticosterone secretion was also measured. Previously reported results showing a significant increase in potassium content with high external potassium were confirmed. However, angiotensin II slightly, but significantly, decreased potassium content. No significant change in intracellular water space or intracellular potassium concentration was observed with either stimulus. 3. Both stimuli significantly increased corticosterone production. There was no correlation between the percentage increase in potassium content or concentration and the percentage increase in steroid output after stimulation with high external potassium. 4. It is concluded that angiotensin II does not stimulate steroidogenesis in adrenal glomerulosa cells by raising the intracellular potassium concentration. The small, but statistically insignificant, increase caused by high-potassium medium is unlikely to be causally linked to steroidogenesis.

Relation of Intracellular K+ and Steroidogenesis in Isolated Adrenal Zona Glomerulosa and Fasciculata Cells

1975 ◽  
Vol 49 (1) ◽  
pp. 13-26
Author(s):  
F. A. Mendelsohn ◽  
C. Mackie
Keyword(s):  
Potassium Concentration ◽  
Zona Glomerulosa ◽  
Zona Fasciculata ◽  
High Potassium ◽  
External Potassium ◽  
Glomerulosa Cells ◽  
Unit Gravity Sedimentation ◽  
Zona Glomerulosa Cells ◽  
Stimulation Of

1. Intracellular K+ content, water spaces and corticosterone output were measured in isolated zona glomerulosa and zona fasciculata-reticularis cell suspensions of rat adrenal cortex, after incubations in vitro under conditions designed to alter steroidogenesis. 2. Intracellular K+ of unpurified zona glomerulosa cells was not altered after stimulation of corticosterone output with serotonin. Similarly, with zona glomerulosa cells purified by unit gravity sedimentation, no change in intracellular K+ was detected after stimulation of steroidogenesis with serotonin or angiotensin II. 3. In high-potassium medium (final concentration 84 mmol/l), parallel increases in intracellular K+ and corticosterone output were observed with both unpurified and purified zona glomerulosa cells. However, a similar increase in intracellular K+ also occurred in high-potassium medium with zona fasciculata cells, whose steroid output is unresponsive to external potassium concentration ([K+]). 4. Ouabain at 10−5 mol/l depressed the intracellular [K+] of glomerulosa cells but did not alter basal or stimulated corticosterone output. Similar results were obtained with fasciculata cells. 5. Ouabain at 5×10−4 mol/l further depressed intracellular [K+] of glomerulosa cells and inhibited basal and stimulated corticosterone output. However, this concentration of ouabain also inhibited steroidogenesis in fasciculata cells. 6. These results demonstrate a variety of situations where changes in intracellular [K+] are dissociated from those in corticosterone output and indicate that intracellular [K+] cannot be the sole mechanism regulating steroidogenesis under these conditions.

Micromethods for the Simultaneous Measurement of Intracellular Potassium and Corticosterone Output of Isolated Adrenal Cortical Cells

1975 ◽  
Vol 49 (1) ◽  
pp. 1-12
Author(s):  
F. A. O. Mendelsohn ◽  
R. L. Warren
Keyword(s):  
Simultaneous Measurement ◽  
Intracellular Water ◽  
Potassium Content ◽  
Cortical Cells ◽  
Intracellular Potassium ◽  
Flame Spectrophotometry ◽  
Adrenal Cortical Cells ◽  
Total Potassium ◽  
Water Space ◽  
Intracellular Potassium Concentration

1. A technique is described for the measurement of potassium and water spaces in less than 1 μ1 of packed cells. 2. The total potassium content of cell pellets is measured in a perchloric and nitric acid extract by flame spectrophotometry. The potassium in trapped medium is estimated from the distribution space of hydroxy[14C]methylinulin and subtracted from the total potassium content to give intracellular potassium content. Corticosterone output was measured by radioimmunoassay. 3. The simultaneous measurement of total water from the [3H]water space allows calculation of the intracellular water space. 4. Values obtained for intracellular potassium content, intracellular water space and calculated intracellular potassium concentration are presented for different preparations of isolated adrenal cortical cells.

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.

Internalisation of the type I angiotensin II receptor (ATI) and angiotensin II function in the rat adrenal zona glomerulosa cell.

1994 ◽  
Vol 141 (2) ◽  
pp. R5-R9 ◽  
Author(s):  
G. P. Vinson ◽  
M. M. Ho. ◽  
J.R. Puddefoot ◽  
R. Teja ◽  
S. Barker
Keyword(s):  
Monoclonal Antibody ◽  
Plasma Membrane ◽  
Angiotensin Ii ◽  
Zona Glomerulosa ◽  
Type I ◽  
Specific Monoclonal Antibody ◽  
Angiotensin Ii Receptor ◽  
Cellular Localisation ◽  
Glomerulosa Cells

ABSTRACT Little is known about the cellular localisation of the angiotensin II (AII) type 1 receptor (ATI) in the rat adrenal glomerulosa cell, but some studies have suggested that receptor internalisation and recycling may occur. Using a specific monoclonal antibody (6313/G2) to the first extracellular domain, we show here that most of the receptor is internalised in the unstimulated cell. When viable glomerulosa cells are incubated with 6313/G2, the receptor is transiently concentrated on the cell surface, and aldosterone output is stimulated. This stimulated output is enhanced by neither threshold nor maximal stimulatory concentrations of All amide, although the antibody does not inhibit All binding to the receptor. Conversely, the stimulatory actions of the antibody and those of ACTH are additive. The data suggest that recycling to the plasma membrane is constitutive, or regulated by unknown factors. Retention of the ATI receptor in the membrane is alone enough to allow sufficient G protein interaction to generate maximal stimulatory events.

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.

Regional differences in potassium content of smooth muscle from opossum esophagus.

1978 ◽  
Vol 235 (6) ◽  
pp. E709
Author(s):  
K Schulze ◽  
J J Hajjar ◽  
J Christensen
Keyword(s):  
Smooth Muscle ◽  
Distal Part ◽  
Proximal Part ◽  
Distal Segment ◽  
Intracellular Concentration ◽  
Potassium Content ◽  
Intracellular Potassium ◽  
The Mean ◽  
Intracellular Potassium Concentration ◽  
Space Volume

Strips from the proximal part of the smooth muscle segment of opossum esophagus have a significantly higher potassium content (50 +/- 3 meq/kg) than those from the distal part (38 +/- 3 meq/kg). There are no significant differences between the two regions in content of sodium (65 +/- 4 meq/kg in proximal, 71 +/- 3 meq/kg in distal) or chloride (48 +/- 10 meq/kg in proximal, 42 +/- 5 meq/ kg in distal). The mean [14C]inulin uptake is 240 +/- 10 ml/kg in both proximal and distal strips. [14C]polyethylene glycol uptake is smaller and [14C]sucrose and [14C]mannitol uptake in both areas are larger than that of inulin. Intracellular potassium concentration (based on the inulin uptake as an estimate of the extracellular space volume) is significantly higher proximally (71 +/- 3 mM) than distally (52 +/- mM). Ouabain, 10(-4) M, increases the intracellular concentration of sodium and decreases the intracellular concentration of potassium in both the proximal and distal segment. The efflux of 86Rb, measured by a washout technique, is higher in the distal than in the proximal smooth muscle segment. A difference in membrane permeability to rubidium and hence, potassium between proximal and distal smooth muscle segments may account in part for the different intracellular potassium concentrations.

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