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Localization of voltage-sensitive sodium channels on the extrasynaptic membrane surface of mouse skeletal muscle by autoradiography of scorpion toxin binding sites

  • Published:
Journal of Neurocytology

Summary

Voltage-dependent sodium channels (Na+ channels) were localized by autoradiography on mouse skeletal muscle using both light and electron microscopy.125I-scorpion toxins (ScTx) of both the a and β type were used as probes. The specificity of labelling was verified by competitive inhibition with unlabelled toxin and by inhibition of αScTx labelling in depolarizing conditions. Under light microscopy, the labelling of the myocyte surface appeared randomly distributed with both the a and β toxins. No difference in the labelling density obtained with βScTx was observed between a 2 mm central segment of the fibre containing the endplate and an adjacent segment not containing the endplate. At the endplate, however, the βScTx binding site density was about seven fold higher at the edge of the synaptic primary clefts. This density decreased with distance from the synaptic cleft reaching the extrasynaptic value at 30–40 μm. An analysis of myocyte labelling using electron microscopy provided evidence for a specific, but very low labelling of the myocyte interior which can be attributed to the T-tubules. These results confirm a relatively high density of Na+ channels in a perijunctional zone about 50 μm in width, which could ensure the initial spread of the surface depolarization with a high safety factor, and a homogeneous distribution over the remaining surface with a low density evaluated at 5–10 per μm2. However, the very low labelling of T-tubules could be attributed mainly to a low density of tubular Na+ channels.

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Authors and Affiliations

  1. Laboratoires de Biologie Cellulaire, INSERM U172 et CNRS UA1179, Faculté de Médecine-Secteur Nord, 13326, Marseille Cedex 15, France

    T. Le Treut, J. -L. Boudier, E. Jover & P. Cau

  2. Biochimie, INSERM U172 et CNRS UA1179, Faculté de Médecine-Secteur Nord, 13326, Marseille Cedex 15, France

    E. Jover

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  1. T. Le Treut
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  4. P. Cau
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Le Treut, T., Boudier, J.L., Jover, E. et al. Localization of voltage-sensitive sodium channels on the extrasynaptic membrane surface of mouse skeletal muscle by autoradiography of scorpion toxin binding sites. J Neurocytol 19, 408–420 (1990). https://doi.org/10.1007/BF01188407

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  • DOI: https://doi.org/10.1007/BF01188407

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