Data on Na,K-ATPase in primary cultures of renal proximal tubule cells treated with catecholamines

This data article is concerned with chronic regulation of Na,K-ATPase by catecholamines. After a chronic treatment, inhibition of Na,K-ATPase activity was observed in cultures with dopamine, while a stimulation was observed in cultures treated with norepinephrine. Following a chronic incubation with guanabenz, an α adrenergic agonist, an increase in Na,K-ATPase α and β subunit mRNAs was observed. This data supports the research article entitled, “Renal proximal tubule Na, K-ATPase is controlled by CREB regulated transcriptional coactivators as well as salt inducible kinase 1” (Taub et al. 2015) [1].


a b s t r a c t
This data article is concerned with chronic regulation of Na,K-ATPase by catecholamines. After a chronic treatment, inhibition of Na,K-ATPase activity was observed in cultures with dopamine, while a stimulation was observed in cultures treated with norepinephrine. Following a chronic incubation with guanabenz, an α adrenergic agonist, an increase in Na,K-ATPase α and β subunit mRNAs was observed. This data supports the research article entitled, "Renal proximal tubule Na, K-ATPase is controlled by CREB regulated transcriptional coactivators as well as salt inducible kinase 1" (Taub et al. 2015) [1].
& 2015 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Subject area
Biology More specific subject area

Renal transport regulation
Type of data Figure  How data  Value of the data This data will have an impact on therapies using catecholamines for blood pressure regulation. The data can be compared with other studies of transcriptional regulation of the genes encoding for each of these subunits.

Data
The data shown in this report measures changes both in Na,K-ATPase activity and Na,K-ATPase mRNA levels following a chronic incubation of renal proximal tubule cells with catecholamines.

Rubidium uptake studies
Primary cultures of rabbit kidney proximal tubule cells, were prepared as described previously [1,2]. Rabbits employed to obtain the primary cultures were used by procedures approved by the University at Buffalo Institutional Animal Care and Use Committee. The primary cultures were grown Fig. 1. The effect of PGE 1 , norepinephrine and dopamine on transport. A. Primary RPT cells were incubated 30 min with either 280 nM PGE 1 , 1 mM norepinephrine or untreated (and þ / À ouabain), followed by a 20 min uptake period with 1 mM 86 Rb þ . Uptake values are averages ( þ/ À SEM) of ouabain-sensitive Rb þ uptake relative to the untreated control. The ouabain-sensitive component of Rb þ uptake was calculated by subtracting the Rb þ uptake observed in the presence of ouabain from total Rb þ uptake. The results were divided by the untreated control value. B. Primary RPT cells were incubated 30 min with either 10 mM dopamine þ / À 5 mM monensin, or untreated (þ / À 5 mM monensin). Uptake studies were conducted both in the presence and in the absence of 1 mM ouabain for each of the 4 conditions, followed by a 20 min uptake period with 1 mM 86 Rb þ . The ouabain-sensitive component of Rb þ uptake was determined as described in part A. *po 0.05 relative to untreated Control. in a 50:50 mixture of Dulbecco's Modified Eagle's medium and Ham's F12 (DME/F12) supplemented with 5 mg/ml bovine insulin, 5 mg/ml human transferrin and 5 Â 10 À 8 M hydrocortisone (i.e. Medium RK-1) [2]. 86 Rb þ uptake studies were conducted, as described previously, in K þ -free DME/F12 supplemented with insulin, transferrin and indomethacin [3,4]. Agonists (PGE 1 , norepinephrine, Fig. 1A, and dopamine, Fig. 1B) were added after a 30 min pre-incubation in this supplemented, K þ -free DME/ F12 ( þ/ À 1 mM ouabain). Subsequently, agonists were added, followed by a 30 min incubation, the addition of 86 Rb þ (1 mM), and a 20 min uptake period. Uptake values were corrected for zero time uptake, and standardized with respect to protein. The ouabain-sensitive component of Rb þ uptake was calculated, and divided by the value obtained for ouabain-sensitive Rb þ uptake with untreated controls (Fig. 1). Differences were considered significant if po 0.05 relative to the untreated control value.