Posttranscriptional upregulation of Na(+)-K(+)-ATPase activity in newborn guinea pig renal cortex

1997 ◽  
Vol 273 (2) ◽  
pp. F254-F263 ◽  
Author(s):  
E. N. Guillery ◽  
D. J. Huss ◽  
A. A. McDonough ◽  
L. C. Klein
Keyword(s):  
Guinea Pig ◽  
Atpase Activity ◽  
Renal Cortex ◽  
Specific Activity ◽  
Mrna Abundance ◽  
Beta Subunit ◽  
Protein Abundance ◽  
Developmental Patterns ◽  
Protein Levels ◽  
Subunit Mrna

We measured Na(+)-K(+)-adenosinetriphosphatase (Na(+)-K(+)-ATPase) activity and subunit abundance in renal cortical homogenates and basolateral membranes (BLM) from fetal, newborn, and adult guinea pigs. Pump specific activity increased four- to fivefold in cortical homogenates and BLM during the transition from fetus to newborn. Immunoblots of BLM showed that alpha- and beta-subunit abundance increased four- to seven- and fourfold, respectively, during the transition from fetus to newborn. Immunoblots of cortical homogenates revealed similar developmental patterns, with newborns having 3.5-fold (alpha) and 2.3-fold (beta) greater subunit abundances than fetuses. Therefore, the bulk of the postnatal increase in BLM-Na(+)-K(+)-ATPase abundance resulted from increased pump production or decreased pump degradation, rather than from redistribution of pumps from intracellular pools. Despite the developmental increase in alpha- and beta-subunit protein levels, newborn guinea pig kidneys had only 1.4- to 2.1-fold greater alpha 1-subunit mRNA abundance and only a 1.5-fold greater beta 1-subunit mRNA abundance than fetal kidneys. These results demonstrate large increases in renal cortical Na(+)-K(+)-ATPase specific activity and protein abundance immediately after birth. These increases, which appear to result largely from posttranscriptional upregulation, may play an important role in mediating the rapid postnatal increase in tubular NaCl reabsorption.

Thyroid hormone induction of rat myocardial Na(+)-K(+)-ATPase: alpha 1-, alpha 2-, and beta 1-mRNA and -protein levels at steady state

1992 ◽  
Vol 262 (2) ◽  
pp. C484-C492 ◽  
Author(s):  
C. B. Hensley ◽  
K. K. Azuma ◽  
M. J. Tang ◽  
A. A. McDonough
Keyword(s):  
Steady State ◽  
Thyroid Hormone ◽  
Atpase Activity ◽  
Cardiac Myocytes ◽  
Protein Abundance ◽  
Mrna Levels ◽  
Protein Levels ◽  
Subunit Mrna ◽  
Time Courses ◽  
Rate Limiting

In this study, we measured the time courses of change in Na(+)-K(+)-ATPase alpha 1-, alpha 2-, and beta 1-subunit mRNA and protein abundance in cardiac myocytes isolated from euthyroid, hypothyroid, and hyperthyroid (hypothyroids injected daily with 1 microgram T3/g body wt) rats. In hypothyroids, alpha 1-, alpha 2-, and beta 1-protein levels were decreased to 0.55, 0.42, and 0.57 of euthyroids, predicting the decrease in Na(+)-K(+)-ATPase activity to 0.53 of control. There was no change in these subunits' mRNA levels, indicating that the decreases in protein levels were not due to decreased subunit transcription rates. In hyperthyroids, the alpha 1-mRNA increased to a steady state of threefold over hypothyroid by 1 day of T3 treatment, and the alpha 1-protein levels increased to twofold over hypothyroid by 4 days of T3. alpha 2-mRNA increased to 5-fold over hypothyroid by 2 days, whereas the alpha 2-protein levels increased to 14-fold above hypothyroid by 4 days of T3. Beta 1-mRNA increased to 12-fold above hypothyroid by 1 day of T3 treatment, whereas beta 1-protein increased to only 2.5-fold over hypothyroid by 4 days of T3. The discoordinate changes in alpha 2- and beta 1-mRNA vs. protein can be reconciled with the hypothesis that beta 1 is rate limiting for assembly in eu- and hypothyroids, and favors assembly with alpha 1, while excess unassembled alpha 2 is degraded. In the hyperthyroids we predict beta 1 is not rate limiting and there is increased alpha 2 beta 1-assembly. We calculate that T3 decreases the alpha 1-to-alpha 2 ratio from 24:1 in hypothyroid to 3.4:1 in hyperthyroid cardiomyocytes.

Thyroid hormone specifically regulates skeletal muscle Na(+)-K(+)-ATPase alpha 2- and beta 2-isoforms

1993 ◽  
Vol 265 (3) ◽  
pp. C680-C687 ◽  
Author(s):  
K. K. Azuma ◽  
C. B. Hensley ◽  
M. J. Tang ◽  
A. A. McDonough
Keyword(s):  
Skeletal Muscle ◽  
Thyroid Hormone ◽  
State Level ◽  
Mrna Abundance ◽  
Northern Analysis ◽  
Protein Abundance ◽  
Constant Amount ◽  
Protein Levels ◽  
Subunit Expression ◽  
Beta 2

The purpose of this study was to determine the pattern of thyroid hormone (triiodothyronine, T3) regulation of the Na(+)-K(+)-adenosinetriphosphatase (Na(+)-K(+)-ATPase) alpha- and beta-subunit expression in skeletal muscle, which expresses alpha 1-, alpha 2-, beta 1-, and beta 2-subunits, and compare it with that seen in kidney, which expresses only alpha 1 and beta 1. Three steady states were studied: hypothyroid, euthyroid, and hyperthyroid (hypothyroids injected daily with 1 microgram T3/g body wt for 2-16 days). Protein and mRNA abundance, determined by Western and Northern analysis, were normalized to a constant amount of homogenate protein and total RNA, respectively. In skeletal muscle, there was no change in alpha 1- or beta 1-mRNA or protein levels in the transition from hypothyroid to hyperthyroid. However, alpha 2 was highly regulated; mRNA reached a new steady-state level of fivefold over hypothyroid by 8 days of T3 treatment and protein abundance increased threefold. In addition, beta 2-mRNA and protein were detected in skeletal muscle and were also highly regulated by T3; beta 2-mRNA increased nearly fourfold over hypothyroid level, and beta 2-protein abundance increased over twofold. In kidney in the transition from hypothyroid to hyperthyroid, there were coordinate 1.6-fold increases in both alpha 1- and beta 1-mRNA abundance that predicted the observed changes in alpha 1- and beta 1-protein levels and Na(+)-K(+)-ATPase activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Abundance of Na(+)-K(+)-ATPase mRNA is regulated by glucocorticoid hormones in infant rat kidneys

1991 ◽  
Vol 260 (2) ◽  
pp. F192-F197 ◽  
Author(s):  
G. Celsi ◽  
A. Nishi ◽  
G. Akusjarvi ◽  
A. Aperia
Keyword(s):  
Atpase Activity ◽  
Renal Cortex ◽  
Rapid Onset ◽  
Mrna Abundance ◽  
Adult Rats ◽  
Postnatal Maturation ◽  
Before And After ◽  
Intraperitoneal Dose ◽  
Old Rats ◽  
Ouabain Inhibition

The administration of glucocorticoid hormone (GC) accelerates the postnatal maturation of renal Na(+)-K(+)-ATPase activity. This study examines the role of GC for the regulation of the Na(+)-K(+)-ATPase mRNA abundance in renal cortex during development. In 12- to 14-day-old rats an upsurge in serum GC concentration was accompanied by an increase in Na(+)-K(+)-ATPase activity and by an apparent increase in mRNA abundance. In 10-day-old rats injected with a single intraperitoneal dose of betamethasone (T) or diluent (C) the abundances of alpha 1- and beta-mRNAs were 1.8- to 2-fold higher in T than in C rats after 20 min. The mRNA abundance of both subunits was threefold higher after 1 h (P less than 0.01), and it was six- to sevenfold higher after 6 h (P less than 0.01). In any given sample there was a coordinate change in alpha 1- and beta-mRNAs relative to C rats. GC did not appear to induce the expression of any alternative catalytic subunit. The alpha 2-mRNA was not detectable in any experimental protocol. Furthermore, the ouabain inhibition of the Na(+)-K(+)-ATPase, partially purified from the renal cortex, was the same before and after GC. In adult rats injected with betamethasone neither the alpha 1- nor the beta-mRNA abundance was different at any time after injection from those in adult C rats. The rapid onset of the GC effect on mRNA abundance in infant rats suggests that the hormone directly activates the gene for Na(+)-K(+)-ATPase alpha 1-subunit, as well as beta-subunit in the developing kidney, and that GC thereby plays an important role for the postnatal maturation of the kidney.

Abstract 050: mir-192-5p in the Kidney is Protective Against the Development of Hypertension

Hypertension ◽  
2016 ◽  
Vol 68 (suppl_1) ◽  
Author(s):  
Maria Angeles Baker ◽  
Pengyuan Liu ◽  
Yong Liu ◽  
Alison Kriegel ◽  
Kevin Regner ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Renal Cortex ◽  
Human Kidney ◽  
Protein Abundance ◽  
Sequencing Analysis ◽  
Hypertensive Nephrosclerosis ◽  
Arterial Blood ◽  
Renal Tubular Reabsorption ◽  
Reduced Salt ◽  
Renal Tubular

MicroRNAs (miRs) are short RNAs that primarily reduce protein abundance by base-paring with their target mRNA. The role of most miRs in the development of hypertension remains unknown. We performed a deep sequencing analysis of miR expression in human kidney biopsies with hypertensive nephrosclerosis or without any significant injury. miR-192-5p was one of the most abundant miRs detected and was down-regulated in hypertensive nephrosclerosis. Previous studies have shown that miR-192-5p targets the beta 1 subunit of Na/K-ATPase which drives renal tubular reabsorption. We hypothesized that miR-192-5p in the kidney protects against hypertension. We used the Dahl salt sensitive (SS) rat and a congenic rat SS.13BN26 (L26) with reduced salt sensitivity as well as Mir192 knockout mice (KO) to test this hypothesis. SS rats had a decreased level of abundance of miR-192-5p in the renal cortex compared to the L26 rats (n=9, p<0.05). The protein abundance of the beta 1 subunit of Na/K-ATPase was higher in the SS rat compared to the L26 rat (n= 3, p<0.05). Treatment with anti-miR-192-5p, delivered directly into the kidney through renal artery injection, in uninephrectomized L26 rats significantly exacerbated hypertension. Mean arterial blood pressure (MAP) of L26 rats treated with anti-miR-192-5p reached 151+/-5 mmHg at day 14 post anti-miR treatment and 4% NaCl (HS) diet, which was significantly higher than L26 rats treated with a control anti-miR and HS (135+/-5 mmHg, n=6 and 8, p<0.05). Tissues were collected in additional groups of rats at 9 days after anti-miR injection, which was just before MAP became significantly different between the two groups, for analysis of Na/K-ATPase activity. Na/K-ATPase activity was increased in the renal cortex of rats treated with anti-miR-192-5p compared to control anti-miR (9.8 +/-1.8μmole/min/μg vs 7.2+/-1.3μmole/min/μg, n=5 and 6, p<0.05). Furthermore, Mir192 KO mice treated with 1μg/Kg/min of Angiotensin II and HS for 14 days exhibited an increased MAP compared to wild-type (WT) mice (190+/-4 mmHg vs 167+/-12 mmHg; n= 3 WT and 5 KO, p<0.05). In conclusion, miR-192, particularly miR-192-5p in the kidney, confers significant protection against the development of hypertension.

Ontogeny of rabbit renal cortical NHE3 and NHE1: effect of glucocorticoids

1995 ◽  
Vol 268 (5) ◽  
pp. F815-F820 ◽  
Author(s):  
M. Baum ◽  
D. Biemesderfer ◽  
D. Gentry ◽  
P. S. Aronson
Keyword(s):  
Proximal Tubule ◽  
Mammalian Cells ◽  
Renal Cortex ◽  
Apical Membrane ◽  
Basolateral Membrane ◽  
Subcutaneous Administration ◽  
Mrna Abundance ◽  
Protein Abundance ◽  
Twofold Increase ◽  
Threefold Increase

The neonatal proximal tubule has a lower rate of bicarbonate absorption and Na+/H+ antiporter activity than the proximal tubule of adult animals. Two isoforms of the Na+/H+ antiporter have been localized to the proximal tubule. NHE3 is located on the apical membrane, whereas NHE1, the isoform found on most mammalian cells, is present on the basolateral membrane. The Na+/H+ antiporter isoforms that increase with renal maturation are unknown. The purpose of the present study was to examine the maturation of rabbit renal cortical NHE3 and NHE1 mRNA and protein abundance and to determine whether the rate of maturation of these isoforms was affected by glucocorticoids. Renal cortex from neonatal rabbits (1 wk) had approximately one-fourth the NHE3 mRNA and protein abundance as that from adult animals. Renal cortical NHE1 mRNA and protein abundance did not change significantly during maturation. Glucocorticoids have been shown to accelerate the maturation of neonatal bicarbonate absorption and apical membrane Na+/H+ antiporter activity. Daily subcutaneous administration of dexamethasone starting at 4 days of age (10 micrograms/100 g body wt) for 3 days and 2 h before being killed resulted in a twofold increase in NHE3 mRNA abundance and a threefold increase in NHE3 protein abundance. NHE1 mRNA and protein abundance were unaffected. These data show that there is selective maturation of NHE3 during renal cortical development, which can be accelerated by administration of glucocorticoids.

Low K+ increases Na(+)-K(+)-ATPase alpha- and beta-subunit mRNA and protein abundance in cultured renal proximal tubule cells

1992 ◽  
Vol 263 (2) ◽  
pp. C436-C442 ◽  
Author(s):  
M. J. Tang ◽  
A. A. McDonough
Keyword(s):  
Atpase Activity ◽  
Proximal Tubule ◽  
Beta Subunit ◽  
Beta Subunits ◽  
Mrna Levels ◽  
Proximal Tubule Cells ◽  
Subunit Mrna ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells ◽  
Low K

Studies from this laboratory demonstrate that LLC-PK1/Cl4 cells, a cultured renal cell line, respond to incubation in low-K+ medium by coordinately increasing abundance of both alpha- and beta-subunits of Na(+)-K(+)-ATPase but increase only beta- and not alpha-mRNA levels (Lescale-Matys et al. J. Biol. Chem. 265: 17935-17940, 1990) and that alpha-abundance is likely increased as a result of increased efficiency of alpha-mRNA translation (L. Lescale-Matys and A. A. McDonough. J. Cell Biol. 111: 311A, 1990). The aim of this report was to determine if nontransformed kidney cells would respond to low K+ in a similar manner. We incubated primary cultures of rat proximal tubule cells in low K+ (0.25 mM) for up to 24 h to address this aim. Na(+)-K(+)-ATPase activity, measured enzymatically, and abundance of alpha- and beta-subunits, measured by immunoblot, were increased significantly and coordinately by 8 h of low K+, and, by 24 h of low K+, these parameters were increased to 2.17 +/- 0.34 (activity), 2.03 +/- 0.21 (alpha), and 2.39 +/- 0.48 (beta)-fold over control. Pretranslationally, beta-mRNA, measured by Northern blot analysis, increased to 1.76 +/- 0.35 after 3 h of low K+ and to 3.4 +/- 0.75-fold over control after 24 h of low K+. The increase in alpha-mRNA was smaller and delayed compared with the beta-mRNA response, but it was sufficient to account for the observed increase in alpha-protein and Na(+)-K(+)-ATPase activity at steady state: alpha-mRNA increased to 1.27 +/- 0.09 after 6 h and to 1.91 +/- 0.41-fold over control after 24 h in low K+. We conclude that the accumulation of sodium pumps in cultured renal proximal tubule cells, unlike LLC-PK1 cells, can be accounted for by increases in both alpha- and beta-subunit mRNA levels.

Na+,K+-ATPase Expression in Maleic-Acid-Induced Fanconi Syndrome in Rats

1997 ◽  
Vol 92 (3) ◽  
pp. 247-253 ◽  
Author(s):  
Esther Castaño ◽  
PAU Marzabal ◽  
F. Javier Casado ◽  
Antonio Felipe ◽  
Marçal Pastor-Anglada
Keyword(s):  
Atpase Activity ◽  
Fanconi Syndrome ◽  
Membrane Vesicles ◽  
Transport Systems ◽  
Kidney Cortex ◽  
Protein Levels ◽  
Subunit Mrna ◽  
Low Activity ◽  
Α1 Subunit ◽  
Hsp60 Protein

1. Na+,K+-ATPase activity and its α1 subunit protein and mRNA in kidney cortex were monitored in rats developing Fanconi syndrome after the administration of maleate. Na+,K+-ATPase activity was significantly lower than in saline-injected controls, although this was partially mediated by a general, non-specific decrease in the cortex protein content. 2. The low activity of the sodium pump correlated with low abundance of α1 subunit mRNA and protein levels. Hsp60 protein levels were also decreased in kidney cortex from maleate-treated rats. 3. Kidney cortex brush-border membrane vesicles from maleate-treated rats showed a marked decrease in Na+-dependent alanine and glucose transport, which was not dependent on the Na+-transmembrane gradient itself, a finding which is consistent with a more stable effect at the plasma membrane level. 4. The effect of maleate may be partially nonspecific and involve a great variety of proteins, but seems to be restricted to selected tissues because α1 subunit Na+,K+-ATPase and hsp60 protein amounts were not significantly modified in livers from rats developing Fanconi syndrome. 5. These results show that maleate administration induces a low activity of selected concentrative transport systems and a decrease in Na+,K+-ATPase activity and expression. The combination of both effects may explain the increased excretion of most organic solutes present in rats developing Fanconi syndrome.

scholarly journals Differential regulation of Na,K-ATPase alpha 1, alpha 2, and beta subunit mRNA and protein levels by thyroid hormone.

1990 ◽  
Vol 265 (24) ◽  
pp. 14308-14314
Author(s):  
B. Horowitz ◽  
C.B. Hensley ◽  
M. Quintero ◽  
K.K. Azuma ◽  
D. Putnam ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Differential Regulation ◽  
Beta Subunit ◽  
Protein Levels ◽  
Subunit Mrna

Hypokalemia decreases Na(+)-K(+)-ATPase alpha 2- but not alpha 1-isoform abundance in heart, muscle, and brain

1991 ◽  
Vol 260 (5) ◽  
pp. C958-C964 ◽  
Author(s):  
K. K. Azuma ◽  
C. B. Hensley ◽  
D. S. Putnam ◽  
A. A. McDonough
Keyword(s):  
Skeletal Muscle ◽  
Atpase Activity ◽  
Heart Muscle ◽  
Similar Pattern ◽  
Protein Abundance ◽  
Large Decrease ◽  
Ouabain Binding ◽  
Protein Levels ◽  
K Deficiency

K+ deficiency has been linked to a loss of K+ from muscle associated with a decrease in ouabain binding and K(+)-dependent phosphatase activity. This study aimed to quantitate the Na(+)-K(+)-ATPase alpha- and beta-isoform-specific responses to hypokalemia in vivo in heart, skeletal muscle, and brain at pre- and posttranslational levels. Two-week dietary K+ restriction resulted in decreases in alpha 2-mRNA in heart and skeletal muscle to 0.60 and 0.65, and in alpha 2-protein abundance to 0.38 and 0.18 of control, respectively. The decrease in alpha 2-protein was greater than the decrease in mRNA in both tissues, suggesting translational and/or posttranslational mechanism(s) of regulation as well as pretranslational regulation in response to hypokalemia. K(+)-dependent p-nitrophenyl phosphatase (pNPPase) activity decreased in heart and skeletal muscle to 0.67 and 0.58, respectively. There were no changes in alpha 1-. or beta-mRNA or protein levels in skeletal muscle or heart. In brain, there was a similar pattern of regulation. While brain alpha 2-mRNA did not change in hypokalemia, protein levels decreased to 0.72 of control. In conclusion, hypokalemia is associated with a large decrease in expression of the alpha 2-isoform of Na(+)-K(+)-ATPase. These results support the hypothesis that in skeletal and heart muscle hypokalemia induces a decrease in Na(+)-K(+)-ATPase activity (measured as K(+)-dependent pNPPase activity) by specifically decreasing the expression of the alpha 2-isoform of Na(+)-K(+)-ATPase.(ABSTRACT TRUNCATED AT 250 WORDS)

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