Effects of innervation on the distribution of acetylcholine receptors on cultured muscle cells

Abstract

1. Myotomal muscle cells from embryos of Xenopus laevis were cultured as a monolayer either alone or together with neural tube cells from the same embryos. 2. Spontaneous twitching and contractions evoked by electrical stimulation of neural perikarya were observed only in nerve-contacted muscle cells, and could be abolished by curare or alpha-bungarotoxin. 3. Within 2 days in culture muscle cells not contacted by nerve developed one or more discrete patches of acetylcholine (ACh) receptors as revealed by staining with fluorescent conjugates of alpha-bungarotoxin. Similar patches were also seen when staining was carried out after paraformaldehyde fixation, suggesting that they were not induced by the dyetoxin conjugate. 4. Radioautography after labelling with [125I]alpha-bungarotoxin revealed patches with grain densities approximately twenty-five-old greater than over the remainder of the cell. 5. Fluorescent stain on innervated cells was restricted to the path of nerve-muscle contact and sometimes extended for greater lengths than the largest patches seen on non-contacted muscle cells. 6. Similar long bands of stain associated with nerve--muscle contacts were observed when cultures were grown in high concentrations of curare and carbachol which prevented spontaneous twitching. They were also seen in cultures in which the addition of neural tube cells was delayed for 2-3 days. 7. It is concluded that innervation caused receptors to accumulate at sites of nerve-muscle contact and that this process can operate independently of muscle activity.