Voltage-dependent suppression of calcium current by caffeine in single smooth muscle cells of the guinea-pig urinary bladder.
The suppressive action of caffeine on L-type Ca current (Ica) in smooth muscle cells of the guinea-pig urinary bladder was investigated using the whole-cell patch clamp technique. Caffeine (5-30 mM) suppressed Ica, the effect having two phases: a rapid and transient suppression of Ica, which was followed by a sustained suppression. When intracellular Ca2+ was strongly buffered by the Ca2+ chelator EGTA (20 mM) or BAPTA (5 mM) in the patch pipette, the transient suppression of Ica was abolished, whereas the sustained effect remained. Similarly, inclusion of both 10 mM procaine and 1 mg/ml heparin in the patch pipette blocked the transient suppression of Ica, but did not block the sustained effect. The degree of the sustained effect of caffeine on Ica was dose-dependent with a kd of 20 mM. Application of the cyclic AMP analogue, 8-bromo-cyclic AMP (100 microM) or forskolin (10 microM) to the bath failed to mimick the sustained suppression of Ica, suggesting that inhibition of phosphodiesterase activity was not involved in the caffeine action. The steady-state activation curve remained unchanged by 10 mM caffeine but the steady-state inactivation curve was significantly shifted in the negative direction by 15.6 mV in 1.8 mM Ca2+ solution or by 10 mV in 1.8 mM Ba2+ solution. From these results it appears that caffeine inhibits L-type Ica via two mechanisms: (1) it releases Ca2+ from an internal store causing a transient Ca2+ -mediated inactivation of the Ca channel; (2) it inhibits Ca channel via a mechanism that does not require such a Ca2+ release. It is possible that caffeine suppresses Ica through a preferential binding to the inactivated state of L-type Ca channel.
Yoshino
M
M
Department of Physiology, Sapporo Medical University, Japan.
Matsufuji
Y
Y
Yabu
H
H
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D000931:Antidotes / Q000494:pharmacology; D002110:Caffeine / Q000008:administration & dosage / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D000697:Central Nervous System Stimulants / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004533:Egtazic Acid / Q000494:pharmacology; D004558:Electric Stimulation; D006168:Guinea Pigs; D006493:Heparin / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects*; D018408:Patch-Clamp Techniques; D011343:Procaine / Q000494:pharmacology; D001743:Urinary Bladder / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
D000779:Anesthetics, Local; D000925:Anticoagulants; D000931:Antidotes; D015220:Calcium Channels; D000697:Central Nervous System Stimulants; D015124:8-Bromo Cyclic Adenosine Monophosphate; D002110:Caffeine; D011343:Procaine; D004533:Egtazic Acid; D006493:Heparin; D002118:Calcium