Document Detail


Regulation of Na+,K+-ATPase activity in MDCK kidney epithelial cell cultures: role of growth state, cyclic AMP, and chemical inducers of dome formation and differentiation.
MedLine Citation:
PMID:  6090479     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Na+,K+-ATPase activity was monitored in MDCK kidney epithelial cell monolayers and in cell extracts as a function of cell density, cAMP elevation, and exposure to hexamethylene bisacetamide (HMBA) and dimethylsulfoxide (Me2SO). Ouabain-sensitive Na+,K+-ATPase and 86Rb+ uptake activities, and the number of [3H]-ouabain binding sites were maximal in subconfluent cultures and decreased accompanying the development of a confluent monolayer. A sodium pump density of 8 X 10(7) pumps/cell was estimated for subconfluent cultures, declining to 9 X 10(5) pumps/cell at confluence. Previous studies have shown that dibutyryl cyclic AMP (Bt2cAMP), 1-methyl-3-isobutylxanthine (IBMX), or the differentiation inducers HMBA and Me2SO, which also caused cAMP elevation, all stimulated dome formation, a visible manifestation of active transepithelial Na+ and water transport (Lever, 1979). In the present study, all of these inducers were found to elevate intracellular Na+ content, implicating this variable in control of induction of dome formation. Operationally, inducers could be divided into two classes. HMBA and Me2SO partially inhibited ouabain-sensitive 86Rb+ influx. Ouabain, at a concentration that caused partial sodium pump inhibition and increased intracellular Na+ content, was also effective as an inducer. The second class, exemplified by IBMX and Bt2cAMP caused a furosemide-sensitive increase in intracellular Na+ content. This class of inducers stimulated ouabain-sensitive 86Rb+ uptake, presumably by substrate effects due to increased Na+ levels. The Na+ or ATP activation of Na+,K+-ATPase activity assayed in cell-free extracts, the affinity of the transport system for Rb+ in intact cells and intracellular ATP levels were unchanged by inducer treatment. Elevation of intracellular Na+ concentration, either by cAMP-stimulated, furosemide-sensitive mechanisms or by partial inhibition of the sodium pump may stimulate the induction of dome formation in MDCK cells.
Authors:
B G Kennedy; J E Lever
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of cellular physiology     Volume:  121     ISSN:  0021-9541     ISO Abbreviation:  J. Cell. Physiol.     Publication Date:  1984 Oct 
Date Detail:
Created Date:  1984-11-14     Completed Date:  1984-11-14     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0050222     Medline TA:  J Cell Physiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  51-63     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Acetamides / pharmacology
Adenosine Triphosphate / metabolism
Animals
Cell Differentiation
Cell Division
Contact Inhibition
Cyclic AMP / metabolism
Dimethyl Sulfoxide / pharmacology
Dogs
Kidney / cytology,  metabolism*
Kinetics
Rubidium / diagnostic use
Sodium / metabolism
Sodium-Potassium-Exchanging ATPase / metabolism*
Grant Support
ID/Acronym/Agency:
AM 00768-02/AM/NIADDK NIH HHS; AM 27400-03/AM/NIADDK NIH HHS
Chemical
Reg. No./Substance:
0/Acetamides; 3073-59-4/hexamethylene bisacetamide; 56-65-5/Adenosine Triphosphate; 60-92-4/Cyclic AMP; 67-68-5/Dimethyl Sulfoxide; 7440-17-7/Rubidium; 7440-23-5/Sodium; EC 3.6.3.9/Sodium-Potassium-Exchanging ATPase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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