Regulation of muscle AChR α subunit gene expression by electrical activity: Involvement of protein kinase C and Ca 2+

Using primary cultures of chicken myotubes, we investigated the involvement of protein kinase C and Ca2+ in the repression of nicotinic acetylcholine receptor (AChR) biosynthesis by electrical activity. Treatment with the Ca2+ channel blocker verapamil or the Na+ channel blocker tetrodotoxin increased alpha subunit mRNA levels 11.5- to 15-fold. The effect of tetrodotoxin was abolished in the presence of the Ca2+ ionophore A23187. Dantrolene, which blocks Ca2+ efflux from the sarcoplasmic reticulum, caused only a 1.7-fold increase in alpha subunit mRNA levels. Down regulation of protein kinase C by prolonged exposure to the phorbol ester TPA or inhibition of protein kinase C by staurosporine led to 8- to 10-fold increases in alpha subunit mRNA levels. Mature and precursor forms of AChR alpha subunit mRNA were found to vary in parallel throughout all of these treatments, suggesting that protein kinase C and Ca2+ ions may modulate AChR alpha subunit biosynthesis at the transcriptional level.

[1]  S. Berman,et al.  Phorbol esters inhibit the synthesis of acetylcholine receptors in cultured muscle cells , 1988, Biology of the cell.

[2]  J. Schmidt,et al.  Ryanodine alters the rate of acetylcholine receptor synthesis in chick skeletal muscle cell cultures. , 1981, The Journal of biological chemistry.

[3]  H. Smilowitz,et al.  Regulation of the number of α‐bungarotoxin binding sites in cultured chick myotubes by a 1,4 dihydropyridine calcium channel antagonist , 1988, Journal of neuroscience research.

[4]  M. Sheng,et al.  Calcium and growth factor pathways of c-fos transcriptional activation require distinct upstream regulatory sequences , 1988, Molecular and cellular biology.

[5]  J. Changeux,et al.  Activity regulates the levels of acetylcholine receptor alpha-subunit mRNA in cultured chicken myotubes. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[6]  A. Ullrich,et al.  Protein kinase C , 1987, Journal of cellular physiology. Supplement.

[7]  J. McManaman,et al.  Inhibitors of membrane depolarization regulate acetylcholine receptor synthesis by a calcium-dependent, cyclic nucleotide-independent mechanism. , 1982, Biochimica et biophysica acta.

[8]  J. Sanes,et al.  Denervation supersensitivity in skeletal muscle: analysis with a cloned cDNA probe , 1984, The Journal of cell biology.

[9]  Miriam M. Salpeter,et al.  Nicotinic acetylcholine receptors in vertebrate muscle: Properties, distribution and neural control , 1985, Progress in Neurobiology.

[10]  J P Changeux,et al.  Calcitonin gene-related peptide and muscle activity regulate acetylcholine receptor alpha-subunit mRNA levels by distinct intracellular pathways , 1987, The Journal of cell biology.

[11]  R. Steinhardt,et al.  Depletion of intracellular calcium stores by calcium ionophore A23187 induces the genes for glucose-regulated proteins in hamster fibroblasts. , 1987, The Journal of biological chemistry.

[12]  S. Heinemann,et al.  Acetylcholine receptor α-, β-, γ-, and δ-subunit mRNA levels are regulated by muscle activity , 1988, Neuron.

[13]  J. Putney.  Phosphoinositides and receptor mechanisms , 1986 .

[14]  J. Changeux,et al.  Regulation of muscle acetylcoline receptor synthesis in vitro by cyclic nucleotide derivatives , 1979, Nature.

[15]  S. Burden,et al.  Development of the neuromuscular junction in the chick embryo: the number, distribution, and stability of acetylcholine receptors. , 1977, Developmental biology.

[16]  T. Tamaoki,et al.  Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase. , 1986, Biochemical and biophysical research communications.

[17]  S. Heinemann,et al.  Muscle denervation increases the levels of two mRNAs coding for the acetylcholine receptor alpha-subunit , 1985, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[18]  J P Changeux,et al.  Localization of nicotinic acetylcholine receptor alpha-subunit transcripts during myogenesis and motor endplate development in the chick , 1989, The Journal of cell biology.

[19]  J. Changeux,et al.  Evolution of cholinergic proteins in developing slow and fast skeletal muscles in chick embryo. , 1980, The Journal of physiology.

[20]  G. Fischbach,et al.  Acetylcholine receptor-inducing factor from chicken brain increases the level of mRNA encoding the receptor alpha subunit. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[21]  E. Leys,et al.  Dihydrofolate reductase gene expression in cultured mouse cells is regulated by transcript stabilization in the nucleus , 1984, The Journal of cell biology.

[22]  J P Changeux,et al.  Detection of the nicotinic acetylcholine receptor alpha‐subunit mRNA by in situ hybridization at neuromuscular junctions of 15‐day‐old chick striated muscles. , 1988, The EMBO journal.

[23]  S. Moss,et al.  Differential expression of nicotinic acetylcholine receptor genes in innervated and denervated chicken muscle. , 1987, The EMBO journal.

[24]  J. Schmidt,et al.  Quantitation of an alpha subunit splicing intermediate: evidence for transcriptional activation in the control of acetylcholine receptor expression in denervated chick skeletal muscle , 1987, The Journal of cell biology.

[25]  J. Schmidt,et al.  Acetylcholine receptor synthesis rate and levels of receptor subunit messenger RNAs in chick muscle , 1988, Neuroscience.

[26]  J. Merlie,et al.  Inhibition of acetylcholine receptor assembly by activity in primary cultures of embryonic rat muscle cells. , 1986, The Journal of biological chemistry.

[27]  L. Pezzementi,et al.  Trifluoperazine Stimulates Acetylcholine Receptor Synthesis in Cultured Chick Myotubes , 1984, Journal of neurochemistry.

[28]  S. Heinemann,et al.  Muscle acetylcholine receptor biosynthesis. Regulation by transcript availability. , 1987, The Journal of biological chemistry.

[29]  T. Maniatis,et al.  Regulation of inducible and tissue-specific gene expression. , 1987, Science.

[30]  P. Narayan,et al.  Stabilization of a specific nuclear mRNA precursor by thyroid hormone , 1985, Molecular and cellular biology.

[31]  H. Pollard,et al.  Characterization of protein kinase C and its role in catecholamine secretion from bovine adrenal-medullary cells. , 1985, The Biochemical journal.

[32]  A. Shainberg,et al.  Decrease of acetylcholine receptor synthesis in muscle cultures by electrical stimulation , 1976, Nature.

[33]  W. Anderson,et al.  Protein kinase C: subcellular redistribution by increased Ca2+ influx. Evidence that Ca2+-dependent subcellular redistribution of protein kinase C is involved in potentiation of beta-adrenergic stimulation of pineal cAMP and cGMP by K+ and A23187. , 1988, The Journal of biological chemistry.

[34]  M. Karin,et al.  Multiple cis- and trans-acting elements mediate the transcriptional response to phorbol esters , 1987, Nature.

[35]  E. Richter,et al.  Contraction‐associated translocation of protein kinase C in rat skeletal muscle , 1987, FEBS letters.

[36]  G. Dubyak,et al.  Effects of 12-O-tetradecanoyl-phorbol-13-acetate on Myofibril integrity and Ca2+ content in developing myotubes. , 1982, Developmental biology.