Effects of alpha1-adrenergic stimulation on L-type Ca2+ current in rat ventricular myocytes.
The effect of alpha1-adrenergic stimulation on L-type Ca2+ current (ICa,L) in adult rat ventricular myocytes was investigated using three different methods of current recording. During conventional whole-cell recordings with 5 mm-BAPTA included in the pipette solution, phenylephrine (20 microM) did not increase ICa,L after 10 min of application. With nystatin perforated-patch whole-cell recordings, phenylephrine potentiated ICa,L, although there were variations among myocytes. The most frequent response was a transient suppression of peak ICa,L at approximately 2 min of exposure followed by a sustained increase of current amplitude evident after 5-10 min exposure. The relative current amplitude 10 min after phenylephrine application was 1.08+/-0.05 compared to control (n=14 cells,P<0.05). During cell-attached single channel recordings, phenylephrine (1 microM) increased the L-type Ca2+ channel open probability (NPo) by 2.25+/-0.31-fold (n=21,P<0.01). It potentiated NPo by increasing the number of openings per sweep and also by promoting longer openings. These effects developed slowly in approximately 10 min. Phenylephrine had no on unitary current amplitude. The potentiation was also elicited by methoxamine (5 microM) and was blocked by prazosin (1 microM), indicating that it was mediated by alpha1-adrenergic receptor stimulation. The increase in NP(o) was suppressed by chelerythrine, a protein kinase C inhibitor. Our results demonstrate that ICa,L can be enhanced by alpha1-adrenergic stimulation, and stress the importance of not disturbing the intracellular environment during studies of the modulation of cardiac ICa,L by alpha1-adrenergic stimulation.
Zhang
S
S
Department of Cardiovascular Diseases, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113, Japan.
Hiraoka
M
M
Hirano
Y
Y
D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000818:Animals; D015220:Calcium Channels / Q000502:physiology*; D020746:Calcium Channels, L-Type; D002478:Cells, Cultured; D006352:Heart Ventricles; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008729:Methoxamine / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology; D018408:Patch-Clamp Techniques; D010656:Phenylephrine / Q000494:pharmacology*; D011224:Prazosin / Q000494:pharmacology*; D011433:Propranolol / Q000494:pharmacology*; D051381:Rats; D018340:Receptors, Adrenergic, alpha-1 / Q000187:drug effects / Q000502:physiology*
D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D018340:Receptors, Adrenergic, alpha-1; D010656:Phenylephrine; D011433:Propranolol; D008729:Methoxamine; D011224:Prazosin