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.

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.

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.

Measurement of intracellular potassium in dispersed adrenal cortical cells

1975 ◽  
Vol 6 (3-4) ◽  
pp. 377-382 ◽  
Author(s):  
F.A.O. Mendelsohn ◽  
C. Mackie ◽  
M.S.R. Mee
Keyword(s):  
Cortical Cells ◽  
Intracellular Potassium ◽  
Adrenal Cortical Cells

Ultrastructural Cytopiasmic Changes of Adrenal Cortex During Pregnancy

1969 ◽  
Vol 27 ◽  
pp. 298-299
Author(s):  
T. M. Murad ◽  
Karen Israel ◽  
Jack C. Geer
Keyword(s):  
Adrenal Gland ◽  
Steroid Hormones ◽  
Adrenal Cortex ◽  
Lipid Droplets ◽  
Plasma Cholesterol ◽  
Adrenal Steroids ◽  
Dynamic Changes ◽  
Cortical Cells ◽  
Acetyl Coenzyme A ◽  
Adrenal Cortical Cells

Adrenal steroids are normally synthesized from acetyl coenzyme A via cholesterol. Cholesterol is also shown to enter the adrenal gland and to be localized in the lipid droplets of the adrenal cortical cells. Both pregnenolone and progesterone act as intermediates in the conversion of cholesterol into steroid hormones. During pregnancy an increased level of plasma cholesterol is known to be associated with an increase of the adrenal corticoid and progesterone. The present study is designed to demonstrate whether the adrenal cortical cells show any dynamic changes during pregnancy.

"STEROID-CELL" ANTIBODY IN ENDOCRINE DISEASES

1974 ◽  
Vol 76 (4) ◽  
pp. 729-740 ◽  
Author(s):  
Peter Kamp ◽  
Per Platz ◽  
Jørn Nerup
Keyword(s):  
Leydig Cells ◽  
Addison’S Disease ◽  
Addison's Disease ◽  
Hormone Production ◽  
Cortical Cells ◽  
Endocrine Diseases ◽  
Cell Antibody ◽  
Indirect Immunofluorescence Technique ◽  
Males And Females ◽  
Adrenal Cortical Cells

ABSTRACT By means of an indirect immunofluorescence technique, sera from 116 patients with Addison's disease, an equal number of age and sex matched controls and 97 patients with other endocrine diseases were examined for the occurrence of antibody to steroid-producing cells in ovary, testis and adrenal cortex. Fluorescent staining was observed in the theca cells of growing follicles, the theca lutein cells, testicular Leydig cells and adrenal cortical cells, i. e. cells which contain enzyme systems used in steroid hormone production. The "steroid-cell" antibody was present in 24 % of the patients with idiopathic Addison's disease, equally frequent in males and females, and in 17 % of the patients with tuberculous Addison's disease, but was rarely found in controls, including patients with other endocrine diseases. Female hypergonadotrophic hypogonadism made an exception, since the "steroid-cell" antibody was found in about half the cases with this condition.

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.

An Investigation of the Electrogenic Sodium Pump in Snail Neurones, Using the Constant-Field Theory

1969 ◽  
Vol 51 (1) ◽  
pp. 181-201
Author(s):  
R. B. MORETON
Keyword(s):  
Field Equation ◽  
Sodium Pump ◽  
Membrane Resistance ◽  
Resting Potential ◽  
Constant Field ◽  
Sodium Ions ◽  
Intracellular Potassium ◽  
Electrogenic Sodium Pump ◽  
Snail Neurones ◽  
Intracellular Potassium Concentration

1. Sodium ions injected into giant neurones of Helix aspersa by diffusion from low-resistance microelectrodes caused hyperpolarization of the cells. Under these conditions the behaviour of the resting potential could be described by a modified ‘constant-field’ equation, including a term representing the effect of a potassiumsensitive, electrogenic sodium pump. 2. Exposure to potassium-free solution, ouabain or cyanide abolished the hyperpolarization, and caused a gradual fall in the intracellular potassium concentration, as estimated from the constant-field equation. 3. Assuming that this fall was due to replacement of intracellular potassium by injected sodium ions, it was possible to calculate the rates of injection and pumping of sodium ions, and, using the measured membrane resistance of the cell, the hyperpolarization which the sodium pump could cause, if it were electrogenic. 4. This was related to the observed hyperpolarization, supporting the view that the latter was caused by stimulation of the electrogenic sodium pump.

scholarly journals Platelet Water Content is Decreased by Gel-Filtration

1977 ◽  
Author(s):  
J.S. Wiley ◽  
G. Wray ◽  
I.A. Cooper
Keyword(s):  
Water Content ◽  
Gel Filtration ◽  
Normal Subjects ◽  
Intracellular Water ◽  
Total Water ◽  
Initial Water ◽  
Mean Cell Volume ◽  
Cell Pellet ◽  
The Mean ◽  
Water Space

One approach to platelet sizing is to measure the intracellular water space of platelets with 3H-H2O since the % water content of platelets remains constant in states with different platelet sizes. Fresh citrated blood was centrifuged for 10 min at 150 'g' to obtain PRP. Aliquots of PRP were briefly incubated with either 3H-H2O or 14C-sucrose then layered over 0.3 ml dibutylphthalate and spun 4 min at 8000 'g'. The cell pellet was solubilized and counted to enable spaces to be calculated. The extracellular (sucrose) space was subtracted from the total water space of the pellet to give a mean intracellular water space of 0.56 ± 0.12 μ1/108 platelets (n =19). Assuming a water content of 7 5% and a density of 1.04, the mean platelet volume for normal subjects is 7.2 fl. Gel-filtration of platelets (GFP) on Sepharose-2B reduced their mean water space by 0.12 μl/108 platelets. However the amount of shrinkage on gel-filtration depended on the initial water space of the platelets in PRP and there was a linear relation between these two variables (r = 0.82). Shrinkage was 40% for an initial platelet water space of 0.70 μl/108 platelets but there was almost no shrinkage below a water space of 0.40 μl/108 platelets. Recovery of platelets from each column averaged 8 0% and showed no relation to the reduction in the mean cell water space. The lower water space of GFP may indicate a reduction in mean cell volume due to gel-filtration.

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