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Protein kinase C-α regulation of gallbladder Na + transport becomes progressively more dysfunctional during gallstone formation
Journal article

Protein kinase C-α regulation of gallbladder Na + transport becomes progressively more dysfunctional during gallstone formation

Seth C. Narins, Ramugounder Ramakrishnan, Eun H. Park, Paul B. Bolno, David A. Haggerty, Peter R. Smith, William C. Meyers and Mohammad Z. Abedin
The Journal of laboratory and clinical medicine, v 146(4), pp 227-237
2005
PMID: 16194684

Abstract

J Na ms RT PKC mRNA NHE Gö 6976 bp PDB HOE-694 kb M rS GBEC Doctor of Musical Arts (DMA) cDNA GAPDH PCR
Gallbladder Na + absorption and biliary Ca 2+ are both increased during gallstone formation and may promote cholesterol nucleation. Na +/H +exchange (NHE) is a major pathway for gallbladder Na + transport. Ca 2+-dependent second messengers, including protein kinase C (PKC), inhibit basal gallbladder Na + transport. Multiple PKC isoforms with species- and tissue-specific expression have been reported. In this study we sought to characterize Ca 2+-dependent PKC isoforms in gallbladder and to examine their roles in Na + transport during gallstone formation. Gallbladders were harvested from prairie dogs fed either nonlithogenic chow or 1.2% cholesterol-enriched diet for varying periods to induce various stages of gallstone formation. PKC was activated with the use of phorboldibutyrate, and we assessed gallbladder NHE regulation by measuring unidirectional Na + flux and dimethylamiloride-inhibitable 22Na + uptake. We measured gallbladder PKC activity with the use of histone III-S phosphorylation and used Gö 6976 to determine PKC-α contributions. Gallbladder PKC isoform messenger RNA and protein expression were examined with the use of Northern- and Western-blot analysis, respectively. Prairie dog and human gallbladder expresses PKC-α, βII, and δ isoforms. The PKC activation significantly decreased gallbladder J Na ms and reduced baseline 22Na + uptake by inhibiting NHE. PKC-α mediated roughly 42% of total PKC activity under basal conditions. PKC-α regulates basal gallbladder Na + transport by way of stimulation of NHE isoform NHE-2 and inhibition of isoform NHE-3. PKC-α blockade reversed PKC-induced inhibition of J Na ms and 22Na + uptake by about 45% in controls but was progressively less effective during gallstone formation. PKC-α contribution to total PKC activity is progressively reduced, whereas expression of PKC-α mRNA, and protein increases significantly during gallstone formation. We conclude that PKC-α regulation of gallbladder NHE becomes progressively more dysfunctional and may in part account for the increased Na + absorption observed during gallstone formation.

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Collaboration types
Domestic collaboration
Web of Science research areas
Medical Laboratory Technology
Medicine, Research & Experimental
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