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Volume 16, Number 23, Issue of December 1, 1996 pp. 7557-7565
Copyright ©1996 Society for Neuroscience

Expression and Subunit Interaction of Voltage-Dependent Ca²⁺ Channels in PC12 Cells

Received July 8, 1996; revised Sept. 18, 1996; accepted Sept. 24, 1996.

Hongyan Liu², Ricardo Felix¹, Christina A. Gurnett¹, Michel De Waard¹, Derrick R. Witcher¹, and Kevin P. Campbell¹

¹ Department of Physiology and Biophysics, The Howard Hughes Medical Institute, and ² Program in Neuroscience, University of Iowa College of Medicine, Iowa City, Iowa 52242

Nerve growth factor (NGF)-induced differentiation in PC12 cells is accompanied by changes in the expression of voltage-dependent Ca²⁺ channels. Ca²⁺ channels are multimeric complexes composed of at least three subunits (₁, , and ₂) and are involved in neuronal migration, gene expression, and neurotransmitter release. Although attempts have been undertaken to elucidate NGF regulation of Ca²⁺ channel expression, the changes in subunit composition of these channels during differentiation still remain uncertain. In the present study, patch-clamp recordings show that in addition to the previously documented L-type and N-type Ca²⁺ currents, undifferentiated PC12 cells also express an -agatoxin-IVA-sensitive (P/Q-type) component. In addition, the corresponding mRNA encoding the pore-forming ₁ subunits for these channels (C, B, and A, respectively) was detected. Likewise, mRNA for three distinct auxiliary  subunits (1, 2, 3) were also found, ₃ protein being dominantly expressed. Immunoprecipitation experiments show that the N-type Ca²⁺ channel is associated with either a ₂ or ₃ subunit and that NGF increases the channel expression without affecting its  subunit association. These results (1) indicate that the diversity of Ca²⁺ currents in PC12 cells arise from the expression of three distinct ₁ and three different  subunit genes; (2) support a model for heterogenous  subunit association of the N-type Ca²⁺ channel in a single cell type; and (3) suggest that the regulation of the N-type Ca²⁺ channel during NGF-mediated differentiation involves an increase in the number of functional channels with no apparent changes in subunit composition.

Key words: calcium channels; _1B subunit; subunit; PC12; nerve growth factor; P/Q-type; N-type; -agatoxin-IVA; -conotoxin GVIA

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