Caffeine-induced immobilization of gating charges in isolated guinea-pig ventricular heart cells.
The effects of 10 mM caffeine (CAF) on intramembrane charge movements (ICM) were studied in isolated guinea-pig ventricular heart cells with the whole-cell patch-clamp technique. In the presence of CAF, the properties (voltage dependence, maximum Q(ON) [Q(max)], availability with voltage) of Q(ON) charge activated from -110 mV were barely affected. Following a 100 ms prepulse to -50 mV to decrease the participation of charges originating from Na channels, the voltage dependence of Q(ON) was shifted by 5 mV (negative component) and by 10 mV (positive component) towards negative potentials, and Q(max) was depressed by 16.5%. CAF drastically reduced in a time- and voltage-dependent manner Q(OFF) on repolarization to -50 mV, the effects being greater at positive potentials. CAF-induced Q(OFF) immobilization could be almost entirely removed by repolarization to voltages as negative as -170 mV. In these conditions, the voltage-dependence of Q(OFF) (repolarization to +30 to -170 mV) was shifted by 17 mV (negative component) and 30 mV (positive component) towards negative potentials, suggesting an interconversion into charge 2. Most of CAF effects were suppressed when the sarcoplasmic reticulum (SR) was not functional or when the cells were loaded with BAPTA-AM. We conclude that CAF effects on ICM are likely due to Ca(2+) ions released from the SR, and which accumulate in the subsarcolemmal fuzzy spaces in the vicinity of the Ca channels. Because CAF effects were more pronounced on Q(OFF) than on Q(ON) the channels have likely to open before Ca(2+) ions could affect their gating properties. It is speculated that such an effect on gating charges might contribute to the Ca-induced inactivation of the Ca current.
Leroy
Jérôme
J
CNRS UMR 6542, Physiologie des Cellules Cardiaques et Vasculaires, Faculté des Sciences, Parc de Grandmont, 37200 Tours, France.
Lignon
Jacques M
JM
Gannier
François
F
Argibay
Jorge A
JA
Malécot
Claire O
CO
D000818:Animals; D002110:Caffeine / Q000494:pharmacology*; D015220:Calcium Channels / Q000378:metabolism; D002469:Cell Separation; D006168:Guinea Pigs; D015640:Ion Channel Gating / Q000187:drug effects* / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects* / Q000502:physiology; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010477:Perfusion; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D016277:Ventricular Function, Left / Q000187:drug effects* / Q000502:physiology*
D015220:Calcium Channels; D002110:Caffeine