Inhibition of electrical coupling in pairs of murine pancreatic acinar cells by OAG and isolated protein kinase C.
Gap junctional coupling was studied in pairs of murine pancreatic acinar cells using the double whole-cell patch-clamp technique. During stable electrical coupling, addition of OAG (1-oleoyl-2-acetyl-sn-glycerol) induced a progressive reduction of the junctional conductance to the detectable limit (approximately 3 pS). Prior to complete electrical uncoupling, various discrete single channel conductances between 20 and 100 pS could be observed. Polymyxin B, a potent inhibitor of the protein kinase C (PKC) system, completely suppressed OAG-stimulated electrical uncoupling. Dialysis of cell pairs with solutions containing PKC, isolated from rat brain, also caused electrical uncoupling. The presence of 0.1 mM dibutyryl cyclic AMP and 5 mM ATP in the pipette solution, which serves to stabilize the junctional conductance, did not suppress the effects of OAG or isolated PKC. We conclude that an increase of protein kinase C activity leads to the closure of gap junction channels, presumably via a PKC-dependent phosphorylation of the junctional peptide, and that this mechanism is dominant over cAMP-dependent upregulatory effects in the experimental time range (less than or equal to 1 hr). A correlation of the observed single channel conductances with the appearance of channel subconductance states or various channel populations is discussed.
University of Konstanz, Faculty of Biology, Federal Republic of Germany.
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R
Somogyi
A
A
Batzer
H A
HA
Kolb
D000818:Animals; D002450:Cell Communication / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D004075:Diglycerides / Q000494:pharmacology*; D004553:Electric Conductivity; D005989:Glycerides / Q000494:pharmacology*; D007365:Intercellular Junctions / Q000187:drug effects; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008815:Mice, Inbred Strains; D010179:Pancreas / Q000166:cytology* / Q000187:drug effects; D011493:Protein Kinase C / Q000302:isolation & purification / Q000494:pharmacology*
D004075:Diglycerides; D005989:Glycerides; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C