Involvement of the calcium channel β3 subunit in olfactory signal transduction

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Abstract

Despite the expression of voltage-dependent Ca2+ channels in nasal turbinate epithelium, their role in odorant chemosensation has remained obscure. Therefore, we investigated olfactory neurotransduction in β3-deficient mice. RT-PCR and Western blots confirmed the expression of various types of Ca2+ channels in the nasal turbinate. Electrophysiological evaluations revealed that β3-null mice had a 60% reduction in the high-voltage-dependent Ca2+ currents in olfactory receptor neurons due to reduced N- and L-type channel currents. The β3-null mice showed increased olfactory neuronal activity to triethylamine, and this effect was mimicked by the perfusion of the specific N-type Ca2+ channel inhibitor ω-conotoxin GVIA in the electro-olfactogram. Diluted male urine odors induced higher Fos immunoreactivity in the main olfactory bulbs of β3-deficient mice, indicating enhanced signal transduction of odor information in these mice. Our data indicate the involvement of voltage-dependent Ca2+ channels and importance of the β3 subunit in olfactory signal transduction.

Section snippets

Methods

The β3-deficient mouse (β3−/−) was constructed as previously reported and back-crossed for more than 20 generations [10]. The wild-type (β3+/+) and β3-deficient (β3−/−) mice were 12- to 16-week-old, experimentally naive mice. The animals were maintained at 22 ± 0.5 °C under a 12-h light–dark cycle. All experiments were conducted during the light phase of the cycle and carried out in accordance with the Guidelines for the Use of Laboratory Animals of the Graduate School of Pharmaceutical Sciences,

RNA transcription

In analyzing the RNA transcripts of the α1 and β subunits, 204-, 273-, 234-, 242-, 417-, 249-, 219-, 187-, and 195-bp fragments corresponding to the CaV2.1 (α1A), CaV2.2 (α1B), CaV1.2 (α1C), CaV1.3 (α1D), CaV2.3 (α1E), β1, β2, β3, and β4 subunit-specific fragments, respectively, were generated by PCR (Fig. 1A). Because CaV2.1, CaV2.2, and CaV2.3 are thought to compose the P/Q-, N-, and R-type Ca2+ channels, our results indicate the presence of these Ca2+ channels in olfactory tissue and suggest

Acknowledgments

We thank Drs. Veit Flockerzi, David Stevens, and Bernd Lindemann for their help.

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