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Journal of Neuroscience, Vol 7, 2957-2966, Copyright © 1987 by Society for Neuroscience
Localization of sodium channel subtypes in adult rat skeletal muscle using channel-specific monoclonal antibodies
B Haimovich, DL Schotland, WE Fieles and RL Barchi
Five monoclonal antibodies specific for the 260 kDa subunit of the rat
skeletal muscle sodium channel were used to probe the distribution of this
channel in adult muscle. All the antibodies reacted with the surface
membrane of fast- and slow-twitch fibers in the rat anterior tibial and
soleus muscles. Immunoreactivity was also present in the endplate region;
this was significantly more intense than that in the surrounding
extrajunctional membrane. At the electron microscopic level, this
junctional immunoreactivity could be traced uniformly throughout the
secondary folds of the post-synaptic membrane. Three of the monoclonal
antibodies (A/B2, F/E4, and I/E3) exhibited an additional distinct
immunoreactivity pattern, staining the interior of selected fibers in the
anterior tibial muscle that were subsequently identified as slow-twitch
fibers. An identical reactivity pattern was observed with most of the
soleus muscle fibers. In longitudinal sections of slow fibers examined at
the light microscopic level, transversely oriented, regularly spaced
doublets of fluorescence were localized at the junction of the A and I
bands in each sarcomere. In permeabilized slow fibers exposed to A/B2 and
examined at the electron microscopic level, internal reactivity was
associated exclusively with the membranes of the T-tubular system. A/B2
also strongly stained a transversely oriented pattern within cardiac muscle
fibers exhibiting the characteristics of the T-tubular system in that
tissue. We conclude that at least 3 subpopulations of sodium channels are
present in adult skeletal muscle: those in the sarcolemma of fast and slow
fibers, those in slow-twitch fiber T-tubular membranes, and those in the
T-tubular system of fast fibers. The channels in the slow fiber T-system
apparently share common epitopes with those in the T-system of cardiac
fibers.
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