Modulation of Ca(2+)-activated Cl- currents in rabbit portal vein smooth muscle by an inhibitor of mitochondrial Ca2+ uptake.
1. The effects of carbonyl cyanide m-chlorophenyl hydrazone (CCCP), an inhibitor of mitochondrial Ca2+ uptake, was investigated on the properties of Ca(2+)-activated chloride currents (ICl(Ca)) in rabbit portal vein smooth muscle cells using the perforated patch whole-cell voltage-clamp technique to ascertain whether this Ca2+ uptake process influences the time course of the subsarcolemmal Ca2+ signal that activates ICl(Ca). 2. In cells bathed in either physiological calcium (2 mM Cao2+) or high calcium (10 mM Cao2+) external solutions, application of CCCP (1-2 microM) evoked an inward current and prolonged the exponential decay time constant (tau) of Ca(2+)-activated Cl- 'tail' currents (Itail) evoked by Ca2+ influx through voltage-dependent calcium channels (VDCCs). The effect of CCCP on tau was greater in cells where the amplitude of Itail was relatively large and, in different cells, the effect of CCCP on tau was positively correlated with the amplitude of Itail. 3. CCCP abolished spontaneously occurring transient Ca(2+)-activated Cl- currents (STICs), but did not alter their time course before complete block. 4. Thapsigargin and cyclopiazonic acid (inhibitors of the sarcoplasmic Ca(2+)-ATPase) inhibited STICs, but did not affect the decay of Itail or STICs. 5. In conclusion, when Ca2+ enters the cell through VDCCs, the time course of the consequent Ca2+ signal in the subsarcolemmal domain containing Ca(2+)-activated chloride channels appears to be regulated by Ca2+ uptake into mitochondria. In contrast, inhibition of Ca2+ uptake by the sarcoplasmic reticulum ATPase does not seem to influence the time course of ICl(Ca).
Greenwood
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Department of Pharmacology and Clinical Pharmacology, St George's Hospital Medical School, London, UK. I.A.Greenwood:i.greenwood@sghms.ac.uk
Helliwell
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Large
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D000818:Animals; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology*; D018118:Chloride Channels / Q000378:metabolism*; D004594:Electrophysiology; D004791:Enzyme Inhibitors; D005260:Female; D007211:Indoles / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D008931:Mitochondria, Muscle / Q000187:drug effects / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011169:Portal Vein; D011817:Rabbits; D019284:Thapsigargin / Q000494:pharmacology
D015220:Calcium Channels; D018118:Chloride Channels; D004791:Enzyme Inhibitors; D007211:Indoles; D007476:Ionophores; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D002118:Calcium; C000543:cyclopiazonic acid