Effect of high extracellular K+ on Na-K-ATPase in cultured canine kidney cells

1990 ◽  
Vol 259 (2) ◽  
pp. F227-F232 ◽  
Author(s):  
M. A. Manuli ◽  
I. S. Edelman
Keyword(s):  
Atpase Activity ◽  
Northern Analysis ◽  
Beta Subunits ◽  
Cell Monolayers ◽  
High K ◽  
Confluent Cell ◽  
K Transport ◽  
Darby Canine Kidney ◽  
Canine Kidney ◽  
Specific Mrna

The Madin-Darby canine kidney (MDCK) cell line was used to evaluate the influence of high extracellular K+, independent of hormonal effects, on renal Na-K-adenosinetriphosphatase (ATPase) activity and abundance. Confluent cell monolayers were incubated in control (5 mM) or high K+ (7.5 mM) medium for 24 h. Exposure to high K+ elicited a 46% rise in Na-K-ATPase activity and a 55% increase in ouabain-sensitive 86Rb uptake. Na-K-ATPase abundance, estimated from the number of ouabain-binding sites, also increased 63% over control in cells exposed to 7.5 mM K+, and as a consequence there was no statistically significant change in the catalytic turnover number. Northern blot analysis using rat cDNA probes for the alpha 1- and beta-subunits showed no corresponding changes in subunit-specific mRNA abundances at 24 h. We conclude that chronic exposure to high extracellular K+ produces a rise in renal epithelial Na-K-ATPase activity and active K+ transport, independent of changes in aldosterone, renal blood flow, or extracellular Na+ concentration. This effect is due to an increase in enzyme abundance rather than a change in catalytic turnover rate. The results of Northern analysis suggest that regulation of Na-K-ATPase activity and abundance by high K+ may involve translational or posttranslational mechanisms, but further study with cDNA probes of canine origin is needed to resolve this issue.

Extracellular pH modifies adaptive response to high K+ in cultured canine kidney cells

1992 ◽  
Vol 262 (5) ◽  
pp. F897-F901
Author(s):  
M. A. Manuli ◽  
J. M. Lorenz
Keyword(s):  
Atpase Activity ◽  
Intact Cell ◽  
Ph Control ◽  
Extracellular Ph ◽  
K Uptake ◽  
Acid Ph ◽  
High K ◽  
Effects Of Ph ◽  
Confluent Cell ◽  
Canine Kidney

The chronic interactive and independent effects of extracellular pH and K+ on renal Na(+)-K(+)-adenosinetriphosphatase (ATPase) activity and active K+ transport were studied in the Madin-Darby canine kidney (MDCK) cell line. Confluent cell monolayers were incubated for 24 h in control (4 mM) or high (7.5 mM) K+ medium at acid (6.8) or neutral (7.4) pH. Under acid pH conditions, exposure to high K+ elicited a rise of 133% in maximum Na(+)-K(+)-ATPase activity and 66% in active K+ uptake. In contrast, high K+ had no effect on enzyme activity or K+ uptake at neutral pH. Detergent-activated Na(+)-K(+)-ATPase assay demonstrated a latent pool of enzyme at acid pH-control K+, which seemed to account entirely for the increase in Na(+)-K(+)-ATPase activity after exposure to high K+. The effects of pH appeared unrelated to HCO3- and Cl- concentration in the extracellular environment. We conclude that the upregulatory effect of high K+ on renal Na(+)-K(+)-ATPase is pH dependent. The data suggest that a pool of catalytically inactive enzyme exists only at acid extracellular pH at K+ concentrations in the normal physiological range and that K+ adaptation, at least initially, is the result of recruitment of this latent intracellular pool. In the intact cell extracellular K+ and luminal pH may interact to modify catalytic turnover rate as well as bioavailability of Na(+)-K(+)-ATPase.

scholarly journals Absorption mechanism of oxymatrine in cultured Madin–Darby canine kidney cell monolayers

2016 ◽  
Vol 54 (10) ◽  
pp. 2168-2175 ◽  
Author(s):  
Xiao-Hong Xiong ◽  
Li-Hua Huang ◽  
Yun-Ming Zhong ◽  
Xuan-Ge Cheng ◽  
Mei-Feng Cen ◽  
...  
Keyword(s):  
Kidney Cell ◽  
Madin Darby Canine Kidney ◽  
Absorption Mechanism ◽  
Cell Monolayers ◽  
Darby Canine Kidney ◽  
Canine Kidney

scholarly journals Investigation of Transport Mechanism of Exendin-4 across Madin Darby Canine Kidney Cell Monolayers

2012 ◽  
Vol 35 (5) ◽  
pp. 745-752 ◽  
Author(s):  
Mengshu Wang ◽  
Bingxue Sun ◽  
Jiao Feng ◽  
Haihong Zhang ◽  
Bin Liu ◽  
...  
Keyword(s):  
Kidney Cell ◽  
Transport Mechanism ◽  
Madin Darby Canine Kidney ◽  
Cell Monolayers ◽  
Darby Canine Kidney ◽  
Canine Kidney

scholarly journals Nanoencapsulated capsaicin changes migration behavior and morphology of madin darby canine kidney cell monolayers

PLoS ONE ◽  
2017 ◽  
Vol 12 (11) ◽  
pp. e0187497 ◽  
Author(s):  
Mathias Kaiser ◽  
Luisa Pohl ◽  
Steffi Ketelhut ◽  
Lena Kastl ◽  
Christian Gorzelanny ◽  
...  
Keyword(s):  
Kidney Cell ◽  
Migration Behavior ◽  
Madin Darby Canine Kidney ◽  
Cell Monolayers ◽  
Darby Canine Kidney ◽  
Canine Kidney

scholarly journals The α Isoform of Protein Kinase C Is Involved in Signaling the Response of Desmosomes to Wounding in Cultured Epithelial Cells

2000 ◽  
Vol 11 (3) ◽  
pp. 1077-1092 ◽  
Author(s):  
Sarah Wallis ◽  
Susan Lloyd ◽  
Irene Wise ◽  
Grenham Ireland ◽  
Tom P. Fleming ◽  
...  
Keyword(s):  
Cell Periphery ◽  
Kidney Cells ◽  
Madin Darby Canine Kidney ◽  
Cell Sheets ◽  
Wound Edge ◽  
Calcium Dependence ◽  
Confluent Cell ◽  
Rapid Transition ◽  
Darby Canine Kidney ◽  
Canine Kidney

Initiation of reepithelialization upon wounding is still poorly understood. To enhance this understanding, we focus here on changes in the adhesive state of desmosomes of cultured Madin-Darby canine kidney cells in response to wounding of confluent cell sheets. Previous results show that desmosomal adhesion in Madin-Darby canine kidney cells changes from a calcium-dependent state to calcium independence in confluent cell sheets. We show that this change, which requires culture confluence to develop, is rapidly reversed upon wounding of confluent cell sheets. Moreover, the change to calcium dependence in wound edge cells is propagated to cells hundreds of micrometers away from the wound edge. Rapid transition from calcium independence to calcium dependence also occurs when cells are treated with phorbol esters that activate PKC. PKC inhibitors, including the conventional isoform inhibitor Gö6976, cause rapid transition from calcium dependence to calcium independence, even in subconfluent cells. The cellular location of the α isoform of PKC correlates with the calcium dependence of desmosomes. Upon monolayer wounding, PKCα translocates rapidly to the cell periphery, becomes Triton X-100 insoluble, and also becomes concentrated in lamellipodia. The PKCα translocation upon wounding precedes both the increase in PKC activity in the membrane fraction and the reversion of desmosomes to calcium dependence. Specific depletion of PKCα with an antisense oligonucleotide increases the number of cells with calcium-independent desmosomes. These results show that PKCα participates in a novel signaling pathway that modulates desmosomal adhesion in response to wounding.

scholarly journals The lateral mobility of the (Na+,K+)-dependent ATPase in Madin-Darby canine kidney cells.

1986 ◽  
Vol 102 (4) ◽  
pp. 1256-1263 ◽  
Author(s):  
A J Jesaitis ◽  
J Yguerabide
Keyword(s):  
Plasma Membrane ◽  
Developmental Stage ◽  
Developmental Stages ◽  
Mdck Cells ◽  
Lateral Mobility ◽  
Madin Darby Canine Kidney ◽  
Confluent Cell ◽  
Restricted Mobility ◽  
Darby Canine Kidney ◽  
Canine Kidney

Fluorescence microphotolysis (recovery after photobleaching) was used to determine the lateral mobility of the (Na+,K+)ATPase and a fluorescent lipid analogue in the plasma membrane of Madin-Darby canine kidney (MDCK) cells at different stages of development. Fluorescein-conjugated Fab' fragments prepared from rabbit anti-dog (Na+,K+)ATPase antibodies (IgG) and 5-(N-hexadecanoyl)aminofluorescein (HEDAF) were used to label the plasma membrane of confluent and subconfluent cultures of MDCK cells. Fractional fluorescence recovery was 50% and 80-90% for the protein and lipid probes, respectively, and was independent of developmental stage. The estimated diffusion constants of the mobile fraction were approximately 5 X 10(-10) cm2/s for the (Na+,K+)ATPase and approximately 2 X 10(-9) cm2/s for HEDAF. Only HEDAF diffusion showed dependency on developmental stage in that D for confluent cells was approximately twice that for subconfluent cells. These results indicate that (Na+,K+)ATPase is 50% immobilized in all developmental stages, whereas lipids in confluent MDCK cells are more mobile than in subconfluent cells. They suggest, furthermore, that the degree of immobilization of the (Na+,K+)ATPase is insufficient to explain its polar distribution, and they support restricted mobility of the ATPase through the tight junctions as the likely mechanism for preventing the diffusion of this protein into the apical domain of the plasma membrane in confluent cell cultures.

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