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Vol. 299, Issue 3, 901-907, December 2001

Ca2+ Influx through Nonselective Cation Channels Plays an Essential Role in Noradrenaline-Induced Arachidonic Acid Release in Chinese Hamster Ovary Cells Expressing alpha 1A-, alpha 1B-, or alpha 1D-Adrenergic Receptors

Yoshifumi Kawanabe , Nobuo Hashimoto, Tomoh Masaki and Soichi Miwa

Department of Pharmacology (Y.K., T.M.) and Neurosurgery (Y.K., N.H.), Kyoto University Faculty of Medicine, Kyoto, Japan; and Department of Pharmacology (S.M.), Hokkaido University Faculty of Medicine, Sapporo, Japan

We constructed Chinese hamster ovary (CHO) cells stably expressing alpha 1A-, alpha 1B-, or alpha 1D-adrenergic receptors (CHO-alpha 1A, CHO-alpha 1B, or CHO-alpha 1D, respectively) and compared the Ca2+ channels activated by noradrenaline (NA) in these cells using whole-cell recordings and monitoring of the intracellular free Ca2+ concentration ([Ca2+]i). We also investigated the involvement of Ca2+ channels in the NA-induced arachidonic acid release. In all three cell types, NA at concentrations >= 10 nM induced a sustained increase in [Ca2+]i attributable to extracellular Ca2+ influx in [Ca2+]i monitoring and an inward current in whole-cell recording. The current-voltage relationships were linear, and their reversal potentials were close to 0 mV. The reversal potential of the currents was not affected by a change in the concentration of Cl- in the bath solution. Moreover, a current could be induced in a bath solution containing only Ca2+ as the movable cation. LOE 908, a receptor-operated Ca2+ channel blocker, inhibited the sustained increase in [Ca2+]i and inward currents in a concentration-dependent manner, and complete inhibition was observed at concentrations >=  3 µM. NA induced arachidonic acid release in all three cell types. This release was entirely dependent on extracellular Ca2+ influx. Moreover, LOE 908 at concentrations >=  3 µM blocked the NA-induced increase in arachidonic acid release. These results indicate that 1) NA activates LOE 908-sensitive Ca2+-permeable nonselective cation channels (NSCCs) in CHO-alpha 1A, CHO-alpha 1B, and CHO-alpha 1D, and 2) the Ca2+ influx through NSCCs may play an important role in the NA-induced enhancement of arachidonic acid release in these cells.

0022-3565/01/2993-0901$03.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2001 by The American Society for Pharmacology and Experimental Therapeutics


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