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The Journal of Neuroscience, December 1, 1998, 18(23):9573-9584

Single-Cell RT-PCR and Functional Characterization of Ca²⁺ Channels in Motoneurons of the Rat Facial Nucleus

T. D. Plant, C. Schirra, E. Katz, O. D. Uchitel, and A. Konnerth

I. Physiologisches Institut, Universität des Saarlandes, 66421 Homburg, Germany

Voltage-dependent Ca²⁺ channels are a major pathway for Ca²⁺ entry in neurons. We have studied the electrophysiological, pharmacological, and molecular properties of voltage-gated Ca²⁺ channels in motoneurons of the rat facial nucleus in slices of the brainstem. Most facial motoneurons express both low voltage-activated (LVA) and high voltage-activated (HVA) Ca²⁺ channel currents. The HVA current is composed of a number of pharmacologically separable components, including 30% of N-type and ~5% of L-type. Despite the dominating role of P-type Ca²⁺ channels in transmitter release at facial motoneuron terminals described in previous studies, these channels were not present in the cell body. Remarkably, most of the HVA current was carried through a new type of Ca²⁺ channel that is resistant to toxin and dihydropyridine block but distinct from the R-type currents described in other neurons.

Using reverse transcription followed by PCR amplification (RT-PCR) with a powerful set of primers designed to amplify all HVA subtypes of the ₁-subunit, we identified a highly heterogeneous expression pattern of Ca²⁺ channel ₁-subunit mRNA in individual neurons consistent with the Ca²⁺ current components found in the cell bodies and axon terminals. We detected mRNA for _1A in 86% of neurons, _1B in 59%, _1C in 18%, _1D in 18%, and _1E in 59%. Either _1A or _1B mRNAs (or both) were present in all neurons, together with various other ₁-subunit mRNAs. The most frequently occurring combination was _1A with _1B and _1E. Taken together, these results demonstrate that the Ca²⁺ channel pattern found in facial motoneurons is highly distinct from that found in other brainstem motoneurons.

Key words: motoneuron; facial nucleus; calcium channel; calcium current; -conotoxin-GVIA; -agatoxin-IVA; -conotoxin-MVIIC; single-cell RT-PCR; ₁-subunit

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Copyright © 1998 Society for Neuroscience  0270-6474/98/18239573-12$05.00/0

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