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J. Biol. Chem., Vol. 278, Issue 36, 33936-33942, September 5, 2003

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The Neuronal ₄ Subunit Increases the Unitary Conductance of L-type Voltage-gated Calcium Channels in PC12 Cells^*

Jessica M. Schjött, Shu-Chan Hsu and Mark R. Plummer 

From the Rutgers University, Department of Cell Biology and Neuroscience, Nelson Laboratories, Piscataway, New Jersey 08854-8082

subunits of voltage-gated calcium channels influence channel behavior in numerous ways, including enhancing the targeting of ₁ subunits to the plasma membrane and shifting the voltage dependence of activation and inactivation. Of the four subunits that have been identified, ₄ is of particular interest because mutation of its ₁ subunit interaction domain produces severe neurological defects. Its differential distribution in the hippocampus prompted us to examine whether this subunit was responsible for the heterogeneity of hippocampal L-type calcium channels. To study the functional effects of the ₄ subunit on native L-type calcium channels, we transfected ₄ cDNA subcloned out of embryonic hippocampal neurons into PC12 cells, a cell line that contains the ₁, ₂, and ₃ subunits but not the ₄ subunit. Cell-attached single-channel recordings of L-type channel activity from untransfected and transfected PC12 cells compared with recordings obtained from hippocampal neurons revealed an effect of the ₄ subunit on single-channel conductance. L-type channels in untransfected PC12 cells had a significantly smaller conductance (19.8 picosiemens (pS)) than L-type channels in hippocampal neurons (22 pS). After transfection of ₄, however, L-type single-channel conductance was indistinguishable between the two cell types. Our data suggest that calcium channel ₄ subunits affect the conductance of L-type calcium channels and that native hippocampal L-type channels contain the ₄ subunit.

Received for publication, February 27, 2003 , and in revised form, June 20, 2003.

^* This study was supported in part by the National Institutes of Health Grant NS 34061. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Rutgers University, Nelson Laboratories, 604 Allison Rd., Piscataway, NJ 08854-8082. Tel.: 732-445-0422; Fax: 732-445-5870; E-mail: mplummer{at}rci.rutgers.edu.

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