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J. Biol. Chem., Vol. 269, Issue 44, 27216-27223, Nov, 1994

Regulation of acetylcholinesterase mRNA stability by calcium during differentiation from myoblasts to myotubes

Z Luo, ME Fuentes and P Taylor
Department of Pharmacology, University of California, San Diego, La Jolla 92093.

The expression of acetylcholinesterase (AChE), nicotinic acetylcholine receptors (nAChR), and their corresponding mRNAs increases dramatically during the conversion of myoblasts to myotubes in C2-C12 cells. The increase in expression of nAChR arises from transcriptional activation of the genes encoding the receptor subunits, whereas stabilization of labile transcripts is primarily responsible for enhanced AChE expression. In a search for the signaling pathways responsible for stabilization of the AChE mRNA, we found that ryanodine, synthetic ryanodine receptor antagonists and L-type, but not N-type, Ca2+ channel blockers inhibit the differentiation-induced expression of AChE mRNA, but not the nAChR mRNA. Selective inhibition of increased expression of AChE is also evident. Inhibition by ryanodine and nifedipine is additive suggesting different target sites for the two Ca2+ channel ligands. Ryanodine binding sites can be detected in both myoblasts and myotubes, but they increase substantially during differentiation. Rates of AChE gene transcription are not altered by ryanodine and nifedipine, indicating that decreased Ca2+ availability prevents stabilization of the mRNA normally seen with differentiation. Muscle cells still undergo elongation and fusion in the presence of ryanodine or L-type Ca2+ channel antagonists. Ryanodine block is fully reversible, indicating functional integrity of the cellular expression system after the drug treatment. These findings indicate that intracellular ryanodine- sensitive calcium channels and extracellular L-type Ca2+ channels link to play an important role in stabilizing AChE mRNA and suggest that transient increases in intracellular Ca2+ may be critical for the commitment of AChE expression during myogenesis.
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