Journal of Neuroscience, Vol 10, 247-255, Copyright © 1990 by Society for Neuroscience
Local accumulation of acetylcholine receptors is neither necessary nor sufficient to induce cluster formation
J Stollberg and SE Fraser
Department of Physiology and Biophysics, College of Medicine, University of California, Irvine 92717.
Acetylcholine receptors (AChRs) accumulate at developing neuromuscular
junctions in part via lateral migration of diffusely expressed receptors.
Using a model system--cultured Xenopus muscle cells exposed to electric
fields--we have shown that AChRs, concentrated at the cathode-facing cell
pole, continue to aggregate there after the field is terminated (Stollberg
and Fraser, 1988). These observations are consistent with the possibility
that the field-induced increase in receptor concentration triggers the
aggregation event. Only 2 other molecular events could initiate the
electric field-induced receptor aggregation: (1) a local increase in the
density of some other molecules, or (2) a voltage-sensitive mechanism.
Treatment of muscle cell cultures with neuraminidase changes the cell
surface charge and has been reported to reverse the direction of
electromigration for AChRs and concanavalin A binding sites (Orida and Poo,
1978). Using digitally analyzed fluorescence videomicroscopy, we find that
AChRs in neuraminidase-treated cultures accumulate at both cell poles in an
electric field. After termination of the field, the AChR continues to
aggregate at the cathode-facing pole, as in cells not treated with
neuraminidase. However, receptor density decreases at the anode-facing
pole, indicating that elevated AChR density does not initiate receptor
aggregation. Cells pretreated with neuraminidase and trypsin (which blocks
receptor aggregation) display reversed receptor distributions compared to
untreated controls, indicating that electromigration has indeed been
reversed. The rate at which neuraminidase- and trypsin- treated cells
approach steady-state distributions indicates a receptor diffusion constant
of approximately 1.2 x 10(-9) cm2/sec, consistent with a diffusion trap
mechanism of receptor aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)