title: Mitochondrial uncoupler FCCP activates proton conductance but does not block store-operated Ca(2+) current in liver cells.
abstract: Uncouplers of mitochondrial oxidative phosphorylation, including carbonilcyanide p-triflouromethoxyphenylhydrazone (FCCP) and carbonilcyanide m-cholorophenylhydrazone (CCCP), are widely used in experimental research to investigate the role of mitochondria in cellular function. Unfortunately, it is very difficult to interpret the results obtained in intact cells using FCCP and CCCP, as these agents not only inhibit mitochondrial potential, but may also affect membrane potential and cell volume. Here we show by whole-cell patch clamping that in primary rat hepatocytes and H4IIE liver cells, FCCP induced large proton currents across the plasma membrane, but did not activate any other observable conductance. In intact hepatocytes FCCP inhibits thapsigargin-activated store-operated Ca(2+) entry, but in patch clamping under the conditions of strong Ca(2+) buffering it has no effect on store-operated Ca(2+) current (I(SOC)). These results indicate that there is no direct connection between mitochondria and activation of I(SOC) in liver cells and support the notion of indirect regulation of I(SOC) by mitochondrial Ca(2+) buffering.
authors.affiliation: University of Adelaide, South Australia, Australia.
authors.forename: Minh-Son
authors.initials: MS
authors.lastname: To
authors.forename: Edoardo C
authors.initials: EC
authors.lastname: Aromataris
authors.forename: Joel
authors.initials: J
authors.lastname: Castro
authors.forename: Michael L
authors.initials: ML
authors.lastname: Roberts
authors.forename: Greg J
authors.initials: GJ
authors.lastname: Barritt
authors.forename: Grigori Y
authors.initials: GY
authors.lastname: Rychkov
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology*; D002460:Cell Line; D002478:Cells, Cultured; D016257:Fura-2; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D008099:Liver / Q000166:cytology; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011522:Protons; D051381:Rats; D017208:Rats, Wistar; D014475:Uncoupling Agents / Q000494:pharmacology*
chemical_list: D011522:Protons; D014475:Uncoupling Agents; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D002118:Calcium; D016257:Fura-2