Glucocorticoid effects on Na-K-ATPase in rabbit nephron segments

1985 ◽  
Vol 248 (4) ◽  
pp. F487-F491
Author(s):  
L. C. Garg ◽  
N. Narang ◽  
C. S. Wingo

We determined the effect of dexamethasone on Na-K-ATPase activity in six nephron segments of the adrenalectomized rabbit. Treatment consisted of 1.4 micrograms dexamethasone X 100 g body wt-1 X day-1 for 7 days prior to the study of the nephron segments. Enzyme activity was determined in individual nephron segments by a microfluorometric assay. There was 40-50% less activity of Na-K-ATPase in the S1 portion of the proximal convoluted tubule (PCT, S1), the medullary thick ascending limb (MTAL), and the distal convoluted tubule (DCT) of adrenalectomized rabbits compared with that of control (sham-operated) animals. There was no significant difference in the enzyme activity in proximal straight tubules (PST, S2 and S3) and cortical thick ascending limb (CTAL) of adrenalectomized and control animals. Dexamethasone treatment produced a dexamethasone concentration of 5 +/- 0.8 nM in the plasma and increased Na-K-ATPase activity in PCT (S1), MTAL, and DCT of the adrenalectomized animals to the control levels without significantly affecting the enzyme activity in the PST (S2, S3) or CTAL. The concentration of dexamethasone in the plasma was such that the hormone should bind mainly to dexamethasone receptors (Kd = 5 nM) and very little to aldosterone receptors (Kd greater than 60 nM). Thus, glucocorticoids probably stimulate Na-K-ATPase in PCT, MTAL, and DCT through glucocorticoid (Type II) receptors and not through mineralocorticoid (Type I) receptors.

1985 ◽  
Vol 249 (6) ◽  
pp. F863-F869 ◽  
Author(s):  
L. C. Garg ◽  
N. Narang ◽  
S. McArdle

Na-K-ATPase activity was determined in seven specific nephron segments of 5- and 12-wk-old spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto (WKY) controls. The enzyme activity in proximal convoluted tubule (PCT) and proximal straight tubule (PST) was significantly higher in 5-wk-old SHR than in WKY. However, Na-K-ATPase activity in medullary thick ascending limb (MTAL), cortical thick ascending limb (CTAL), and distal convoluted tubule (DCT) was significantly lower in 5-wk-old SHR than in WKY. There were no significant differences in the enzyme activity in PCT, PST, MTAL, CTAL, and DCT in 12-wk-old SHR and WKY. Furthermore, there were no significant differences in Na-K-ATPase activity in collecting duct segments of 5- or 12-wk-old SHR and age-matched WKY. The possible role of the abnormal pattern of Na-K-ATPase activity in PCT, PST, MTAL, CTAL, and DCT in 5-wk-old SHR in generation of hypertension in this strain remains to be determined.


1992 ◽  
Vol 262 (4) ◽  
pp. F583-F590 ◽  
Author(s):  
C. Khadouri ◽  
S. Marsy ◽  
C. Barlet-Bas ◽  
L. Cheval ◽  
A. Doucet

An N-ethylmaleimide (NEM)-sensitive adenosinetriphosphatase (ATPase) displaying the kinetic and pharmacological properties of an electrogenic proton pump has been described in the different segments of rat nephron, where it mediates part of the active tubular proton secretion. This study was therefore designed to evaluate whether changes in urinary acidification observed during metabolic acidosis or alkalosis were associated with alterations of the activity of tubular NEM-sensitive ATPase, and if so, to localize the nephron segments responsible for these changes. Within 1 wk after the onset of ammonium chloride treatment, rats developed a metabolic acidosis, and NEM-sensitive ATPase activity was markedly increased in the medullary thick ascending limb of Henle's loop and outer medullary collecting tubule, and slightly increased in the cortical collecting tubule. Conversely, treatment with sodium bicarbonate induced a metabolic alkalosis that was accompanied by decreased NEM-sensitive ATPase activity in medullary thick ascending limb and outer medullary collecting tubule. NEM-sensitive ATPase activity was not altered in any other nephron segment tested in alkalotic and acidotic rats, i.e., the proximal tubule and the cortical thick ascending limb of Henle's loop. Changes qualitatively similar were observed as soon as 3 h after the onset of NaHCO3 or NH4Cl-loading. In the medullary collecting tubule, alterations of NEM-sensitive ATPase activity are in part due to hyperaldosteronism observed in both acidotic and alkalotic rats.(ABSTRACT TRUNCATED AT 250 WORDS)


1986 ◽  
Vol 251 (3) ◽  
pp. F506-F512 ◽  
Author(s):  
S. K. Mujais ◽  
N. A. Kurtzman

This study has examined the temporal profile and the segmental localization along the rat nephron of the increase in Na-K-ATPase produced by uninephrectomy, and the role of the adrenal gland in the generation of the increase in enzyme activity. In adrenal-intact rats, an increase in Na-K-ATPase activity in the cortical collecting tubule (CCT) was observed at 1 wk (140 +/- 13% of sham, P less than 0.05) and sustained at 2 wk (140 +/- 8% of sham, P less than 0.05). In contrast, the enhancement of enzyme activity in the proximal convoluted tubule (PCT) was transient (at 1 wk: 164 +/- 20% of sham, P less than 0.05; and at 2 wk: 97 +/- 9% of sham, P greater than 0.5). No changes in Na-K-ATPase activity were observed in the other nephron segments studied: pars recta, medullary thick ascending limb, cortical thick ascending limb, distal convoluted tubule, and medullary collecting tubule. In adrenalectomized rats, CCT enzyme activity was lower than in adrenal-intact rats (761 +/- 84 vs. 1,984 +/- 276 pmol X mm-1 X h-1, P less than 0.001) and was not altered by uninephrectomy (849 +/- 91 pmol X mm-1 X h-1, NS). We conclude that the increase in Na-K-ATPase activity following uninephrectomy is restricted to two segments of the nephron and follows a distinctive pattern in each. In the PCT a transient enhancement in enzyme activity is observed, whereas in the CCT the increase in Na-K-ATPase is sustained and requires the presence of an intact adrenal gland.


1990 ◽  
Vol 259 (2) ◽  
pp. F246-F250 ◽  
Author(s):  
C. Barlet-Bas ◽  
L. Cheval ◽  
C. Khadouri ◽  
S. Marsy ◽  
A. Doucet

The sensitivity of Na(+)-K(+)-ATPase to Na was determined in single segments of rabbit nephron isolated by microdissection. In the cortical collecting tubule (CCT), Na(+)-K(+)-ATPase was threefold more sensitive to Na (apparent K0.5 approximately 3 mM) than in proximal convoluted tubule and cortical thick ascending limb (apparent K0.5 approximately 10 mM). Furthermore, increasing K concentration from 5 to greater than 100 mM markedly reduced the affinity of the pump for Na in all three nephron segments. In fact, the main shift in Na affinity occurred when K changed from 100 to 120 mM; in the CCT, increasing K concentration from 100 to 120 mM while maintaining Na concentration at 10 mM reduced Na(+)-K(+)-ATPase activity by greater than 35%. These findings confirm that, in kidney cells as in other cells, intracellular Na limits the rate of Na(+)-K(+)-ATPase. Thus any alteration of intracellular Na concentration modifies the pump activity in a way that contributes to the restoration of intracellular Na homeostasis. This adaptive property is particularly efficient in the collecting tubule in which the apparent K0.5 of the pump for Na is close to normal intracellular Na concentration. Furthermore, changes in intracellular K concentration, which usually accompany those of Na so as to maintain the total cation concentration constant, potentiate the regulatory role of Na through modifications of its affinity for the pump.


1987 ◽  
Vol 252 (5) ◽  
pp. F838-F843 ◽  
Author(s):  
M. E. Chamberlin ◽  
L. J. Mandel

Na+-K+-ATPase activity was measured in a suspension of rabbit medullary thick ascending limb tubules under oxygenated and anoxic conditions. Oxygenated, K-depleted tubules rapidly take up added extracellular potassium accompanied by a simultaneous increase in oxygen consumption. The ATP/O2 ratio was 12.5 +/- 0.7, suggesting a tight coupling between oxidative metabolism (6 ATP/O2) and Na+-K+-ATPase activity (2 K/ATP). On reaching anoxia, the tubules released potassium into the medium, but this rate was accelerated by the addition of ouabain, which indicated that the Na+-K+-ATPase was still operative in anoxia. Because 10 min of anoxia led to only a 15.7% decline in potassium content, a new steady state of potassium uptake and leakage must be reached during anoxia. Anaerobic metabolism maintained 73% of cellular ATP during 10 min of anoxia. Exposure of anoxic tubules to iodoacetate produced a 57% decline in ATP levels and a 33% decline in potassium content, which indicated that glycolysis is an important pathway in supplying energy during anaerobiosis.


1990 ◽  
Vol 127 (3) ◽  
pp. 377-382 ◽  
Author(s):  
J. A. Charlton ◽  
P. H. Baylis

ABSTRACT Previous studies have indicated that ornithine decarboxylase (ODC) may be involved in the stimulation of Na+/K+-ATPase activity by arginine vasopressin (AVP) in the rat renal medullary thick ascending limb of Henle's loop. The present study was aimed at establishing the role of the polyamines, the conversion products of ODC activity, in the stimulation of Na+/K+-ATPase by AVP. Using cytochemical methods, we have demonstrated an increase in Na+/K+-ATPase activity after stimulation with putrescine, spermidine and spermine (each 1 mmol/l) for 2·5,2 and 1·5 min respectively. The specific inhibitors of spermidine and spermine synthase, bis-cyclohexylammonium sulphate and N-alkylated-1,3-diaminopropane respectively, inhibited the stimulation of Na+/K+-ATPase by AVP, this inhibition being reversed by spermine. These findings suggest that polyamines are involved in the stimulus-response coupling of the hormone-mediated response. Journal of Endocrinology (1990) 127, 377–382


1987 ◽  
Vol 252 (5) ◽  
pp. F910-F915 ◽  
Author(s):  
P. Scherzer ◽  
H. Wald ◽  
M. M. Popovtzer

To evaluate the effect of furosemide on kidney function, glomerular filtration rate (GFR), urinary Na excretion (UNaV), Na reabsorption (NAR), and Na+-K+-ATPase in isolated nephron segments were measured in 1) rats treated with furosemide 10 mg X 100 g-1 X 24 h-1 ip for 7 days, and 2) rats receiving an oral Na load for 12 days. In furosemide-treated rats, GFR rose from 0.61 +/- 0.03 (mean +/- SD) to 0.83 +/- 0.06 ml/min (P less than 0.01), UNaV rose from 904 +/- 71 to 1,402 +/- 85 mueq/day (P less than 0.001), and net NAR rose from 87.5 +/- 3.7 to 116.7 +/- 9.0 mueq/min (P less than 0.01). Na+-K+-ATPase remained unchanged in the proximal convoluted tubule (PCT), proximal straight tubule (PST), cortical thick ascending limb of Henle's loop (cTALH), and medullary thick ascending limb of Henle's loop (mTALH), but was increased in the distal convoluted tubule (DCT) and in cortical collecting duct (CCD) from 48.5 +/- 1.2 to 75.3 +/- 0.7 (P less than 0.001) and from 18.6 +/- 0.7 to 27.1 +/- 2.7 (P less than 0.02) X 10(-11) mol X mm-1 X min-1, respectively. In Na-loaded rats GFR rose from 0.61 +/- 0.04 to 0.86 +/- 0.03 ml/min (P less than 0.001), UNaV rose from 1,064 +/- 118 to 18,532 +/- 2,045 mueq/day (P less than 0.001), net NAR from 88.1 +/- 3.0 to 107.8 +/- 3.9 mueq/min and Na-K-ATPase in the mTALH rose from 40.3 +/- 1.4 to 56.2 +/- 2.11 (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


1990 ◽  
Vol 127 (2) ◽  
pp. 213-216 ◽  
Author(s):  
J. A. Charlton ◽  
P. H. Baylis

ABSTRACT In previous studies, we have demonstrated that 1–10 fmol arginine vasopressin (AVP)/l maximally stimulates the activity of the enzyme Na+/K+-ATPase in the rat renal medullary thick ascending limb (MTAL) of Henle's loop after 4 or 10 min of stimulation when measured using a cytochemical bioassay. We have tested the hypothesis that this stimulation is mediated by the V2 receptor in the MTAL. A cytochemical bioassay was used to investigate the effect of specific V1 and V2/V1 antagonists and a synthetic V2 agonist [1-deamino,8-d-arginine]-vasopressin (dDAVP), on the activity of Na+/K+-ATPase. There was no effect of the V1 antagonist (1 fmol-1 μmol/l) in inhibiting the activity of Na+/K+-ATPase stimulated by 1 fmol AVP/l. In contrast, 100 pmol of the V2/V1 antagonist/l significantly (P < 0·001) inhibited the stimulation of Na+/K+-ATPase activity by 1 fmol AVP/l from 55·5±4·3 (s.e.m.) to 31·9±1·6 mean integrated extinction (MIE) after 4 min of stimulation and from 67·0±3·2 to 36·9±0·7 MIE after 10 min of stimulation. Similarly, the stimulation of Na+/K+-ATPase by 10 fmol dDAVP/l was inhibited by the V2/V1 antagonist from 55·1±1·0 to 26·1±0·5 MIE after 4 min of stimulation. We conclude that the stimulation of Na+/K+-ATPase by AVP is mediated by the V2 receptor in the rat renal MTAL. Journal of Endocrinology (1990) 127, 213–216


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