Electrical activity suppresses nicotinic acetylcholine receptor gamma subunit promoter activity

Nicotinic acetylcholine receptors are pentameric, transmembrane, ligand-gated ion channels critical for neuromuscular signal transmission. Prior to innervation, the genes encoding these receptors are expressed in nuclei throughout the muscle fiber. Muscle innervation leads to a dramatic decrease in expression of these genes in extrasynaptic nuclei. This reduction in gene expression can be reversed by muscle denervation. The effects of denervation on receptor gene expression can be blocked by electrical stimulation of muscle using extracellular electrodes. The molecular mechanisms by which muscle electrical activity leads to altered patterns of nicotinic acetylcholine receptor gene expression are not well understood. Using an in vitro electrical stimulation paradigm to induce muscle activity, we have been able to mimic the effect of innervation on extrasynaptic acetylcholine receptor gene expression. We have found that a 93-bp region of 5'-flanking DNA, spanning nucleotides -150 to -57 relative to the transcription start site of the gamma subunit gene, is required for the suppression of gene expression in response to muscle activity. Sequences downstream of this region are transcriptionally active but are not responsive to muscle activity. However, these downstream sequences become responsive to muscle activity when placed under the control of the gamma subunit muscle-specific enhancer.