Abstract
The effects of pharmacological agents on the T-type Ca2+current were studied in dissociated spermatogenic cells from the mouse. Ca2+ currents were elicited by depolarization in 10 mm Ca2+ and recorded in the whole-cell configuration of the patch clamp technique. The T-type current was inhibited by the following compounds: PN200–110 (IC50 = 4 × 10−8 M) > nifedipine (IC50 = 4 × 10−7 M) > pimozide (IC50 = 4.6 × 10−7 M) > mibefradil (IC50 = 5 × 10−6 M) > Ni2+ (IC50 = 3.4 × 10−5 M) > verapamil (IC50 = 7 × 10−5 M) > amiloride (IC50 = 2.4 × 10−4 M) > Cd2+ (IC50 = 2.8 × 10−4 M). However, the agents differed in the reversibility and the use dependence of their effects. Currents recovered rapidly and completely after removal of Ni2+, Cd2+, amiloride, or mibefradil, whereas recovery from verapamil block was rapid but incomplete. In contrast, we observed little recovery after the removal of pimozide and of the dihydropyridines (PN200–110, nifedipine). Moreover, mibefradil and pimozide exhibit a strongly use-dependent inhibition of current that is due to selective interaction of these drugs with the open state and the inactivated state of the channel, respectively, rather than with the resting state. These properties of the spermatogenic T-type Ca2+ channel differ from those of somatic cell T channels and suggest a molecular diversity of low voltage-activated Ca2+ channels.
Footnotes
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Send reprint requests to: Dr. Christophe Arnoult, Laboratoire de Biophysique Moléculaire et Cellulaire, CEA/Grenoble-DBMS/BMC, 17 rue des Martyrs, 38054 Grenoble Cedex 9, France. E-mail: arnoult{at}dsvgre.cea.fr
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This work was supported in part by “Association Française contre les Myopathies” (C.A.) and Fondation pour la Recherche Médicale (C.A.). H.M.F. was supported by National Institutes of Health Grant HD32177, and C.A. and M.V. are supported by the Direction des Sciences du Vivant, C.E.A., France.
- Abbreviations:
- EGTA
- ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid
- HEPES
- 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- Received December 17, 1997.
- Accepted February 20, 1998.
- The American Society for Pharmacology and Experimental Therapeutics
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