Abstract
The level of acetylcholine receptor alpha-subunit mRNA in primary cultures of chicken myotubes increases when the spontaneous electrical activity of the myotube is suppressed by the sodium channel blocker tetrodotoxin. This increase was prevented by two translational inhibitors: cycloheximide and anisomycin. Neither the basal level of alpha-subunit mRNA nor that of muscle-specific creatine phosphokinase mRNA was affected by these inhibitors. In contrast, cycloheximide potentiated the limited increase of alpha-subunit mRNA levels evoked by the neuropeptide calcitonin gene-related peptide. The high level of alpha-subunit mRNA elicited by tetrodotoxin treatment did not persist after subsequent addition of cycloheximide in the presence of tetrodotoxin, indicating that the continuous synthesis of protein factor(s) is necessary for this regulation. Moreover, cycloheximide decreased the high level of alpha-subunit mRNA present at early stages of in vitro maturation of muscle primary cultures without blocking the further increase of the muscle-specific creatine phosphokinase mRNA. Implications of the requirement for constant synthesis of protein factors on the induction of muscle-specific gene expression by blocking myotube electrical activity are discussed.
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- Almendral J. M., Sommer D., Macdonald-Bravo H., Burckhardt J., Perera J., Bravo R. Complexity of the early genetic response to growth factors in mouse fibroblasts. Mol Cell Biol. 1988 May;8(5):2140–2148. doi: 10.1128/mcb.8.5.2140. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Betz H., Changeux J. P. Regulation of muscle acetylcholine receptor synthesis in vitro by cyclic nucleotide derivatives. Nature. 1979 Apr 19;278(5706):749–752. doi: 10.1038/278749a0. [DOI] [PubMed] [Google Scholar]
- Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
- Curran T., Franza B. R., Jr Fos and Jun: the AP-1 connection. Cell. 1988 Nov 4;55(3):395–397. doi: 10.1016/0092-8674(88)90024-4. [DOI] [PubMed] [Google Scholar]
- Fontaine B., Changeux J. P. Localization of nicotinic acetylcholine receptor alpha-subunit transcripts during myogenesis and motor endplate development in the chick. J Cell Biol. 1989 Mar;108(3):1025–1037. doi: 10.1083/jcb.108.3.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fontaine B., Klarsfeld A., Changeux J. P. Calcitonin gene-related peptide and muscle activity regulate acetylcholine receptor alpha-subunit mRNA levels by distinct intracellular pathways. J Cell Biol. 1987 Sep;105(3):1337–1342. doi: 10.1083/jcb.105.3.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fontaine B., Klarsfeld A., Hökfelt T., Changeux J. P. Calcitonin gene-related peptide, a peptide present in spinal cord motoneurons, increases the number of acetylcholine receptors in primary cultures of chick embryo myotubes. Neurosci Lett. 1986 Oct 30;71(1):59–65. doi: 10.1016/0304-3940(86)90257-0. [DOI] [PubMed] [Google Scholar]
- Fontaine B., Sassoon D., Buckingham M., Changeux J. P. Detection of the nicotinic acetylcholine receptor alpha-subunit mRNA by in situ hybridization at neuromuscular junctions of 15-day-old chick striated muscles. EMBO J. 1988 Mar;7(3):603–609. doi: 10.1002/j.1460-2075.1988.tb02853.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldman D., Brenner H. R., Heinemann S. Acetylcholine receptor alpha-, beta-, gamma-, and delta-subunit mRNA levels are regulated by muscle activity. Neuron. 1988 Jun;1(4):329–333. doi: 10.1016/0896-6273(88)90081-5. [DOI] [PubMed] [Google Scholar]
- Greenberg M. E., Hermanowski A. L., Ziff E. B. Effect of protein synthesis inhibitors on growth factor activation of c-fos, c-myc, and actin gene transcription. Mol Cell Biol. 1986 Apr;6(4):1050–1057. doi: 10.1128/mcb.6.4.1050. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hökfelt T., Holets V. R., Staines W., Meister B., Melander T., Schalling M., Schultzberg M., Freedman J., Björklund H., Olson L. Coexistence of neuronal messengers--an overview. Prog Brain Res. 1986;68:33–70. doi: 10.1016/s0079-6123(08)60230-7. [DOI] [PubMed] [Google Scholar]
- Katz A., Kahana C. Transcriptional activation of mammalian ornithine decarboxylase during stimulated growth. Mol Cell Biol. 1987 Jul;7(7):2641–2643. doi: 10.1128/mcb.7.7.2641. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kerr L. D., Holt J. T., Matrisian L. M. Growth factors regulate transin gene expression by c-fos-dependent and c-fos-independent pathways. Science. 1988 Dec 9;242(4884):1424–1427. doi: 10.1126/science.2462278. [DOI] [PubMed] [Google Scholar]
- Klarsfeld A., Changeux J. P. Activity regulates the levels of acetylcholine receptor alpha-subunit mRNA in cultured chicken myotubes. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4558–4562. doi: 10.1073/pnas.82.13.4558. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klarsfeld A., Daubas P., Bourachot B., Changeux J. P. A 5'-flanking region of the chicken acetylcholine receptor alpha-subunit gene confers tissue specificity and developmental control of expression in transfected cells. Mol Cell Biol. 1987 Feb;7(2):951–955. doi: 10.1128/mcb.7.2.951. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klarsfeld A., Laufer R., Fontaine B., Devillers-Thiéry A., Dubreuil C., Changeux J. P. Regulation of muscle AChR alpha subunit gene expression by electrical activity: involvement of protein kinase C and Ca2+. Neuron. 1989 Mar;2(3):1229–1236. doi: 10.1016/0896-6273(89)90307-3. [DOI] [PubMed] [Google Scholar]
- Lassar A. B., Thayer M. J., Overell R. W., Weintraub H. Transformation by activated ras or fos prevents myogenesis by inhibiting expression of MyoD1. Cell. 1989 Aug 25;58(4):659–667. doi: 10.1016/0092-8674(89)90101-3. [DOI] [PubMed] [Google Scholar]
- Laufer R., Changeux J. P. Activity-dependent regulation of gene expression in muscle and neuronal cells. Mol Neurobiol. 1989 Spring-Summer;3(1-2):1–53. doi: 10.1007/BF02935587. [DOI] [PubMed] [Google Scholar]
- Laufer R., Changeux J. P. Calcitonin gene-related peptide elevates cyclic AMP levels in chick skeletal muscle: possible neurotrophic role for a coexisting neuronal messenger. EMBO J. 1987 Apr;6(4):901–906. doi: 10.1002/j.1460-2075.1987.tb04836.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lenardo M. J., Baltimore D. NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control. Cell. 1989 Jul 28;58(2):227–229. doi: 10.1016/0092-8674(89)90833-7. [DOI] [PubMed] [Google Scholar]
- Lillie J. W., Green M. R. Transcription activation by the adenovirus E1a protein. Nature. 1989 Mar 2;338(6210):39–44. doi: 10.1038/338039a0. [DOI] [PubMed] [Google Scholar]
- Linzer D. I., Wilder E. L. Control of proliferin gene expression in serum-stimulated mouse cells. Mol Cell Biol. 1987 Jun;7(6):2080–2086. doi: 10.1128/mcb.7.6.2080. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merlie J. P., Isenberg K. E., Russell S. D., Sanes J. R. Denervation supersensitivity in skeletal muscle: analysis with a cloned cDNA probe. J Cell Biol. 1984 Jul;99(1 Pt 1):332–335. doi: 10.1083/jcb.99.1.332. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merlie J. P., Kornhauser J. M. Neural regulation of gene expression by an acetylcholine receptor promoter in muscle of transgenic mice. Neuron. 1989 Apr;2(4):1295–1300. doi: 10.1016/0896-6273(89)90067-6. [DOI] [PubMed] [Google Scholar]
- New H. V., Mudge A. W. Calcitonin gene-related peptide regulates muscle acetylcholine receptor synthesis. 1986 Oct 30-Nov 5Nature. 323(6091):809–811. doi: 10.1038/323809a0. [DOI] [PubMed] [Google Scholar]
- Osterlund M., Fontaine B., Devillers-Thiery A., Geoffroy B., Changeux J. P. Acetylcholine receptor expression in primary cultures of embryonic chick myotubes--I. Discoordinate regulation of alpha-, gamma- and delta-subunit gene expression by calcitonin gene-related peptide and by muscle electrical activity. Neuroscience. 1989;32(2):279–287. doi: 10.1016/0306-4522(89)90078-x. [DOI] [PubMed] [Google Scholar]
- Piette J., Klarsfeld A., Changeux J. P. Interaction of nuclear factors with the upstream region of the alpha-subunit gene of chicken muscle acetylcholine receptor: variations with muscle differentiation and denervation. EMBO J. 1989 Mar;8(3):687–694. doi: 10.1002/j.1460-2075.1989.tb03427.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shainberg A., Burstein M. Decrease of acetylcholine receptor synthesis in muscle cultures by electrical stimulation. Nature. 1976 Nov 25;264(5584):368–369. doi: 10.1038/264368a0. [DOI] [PubMed] [Google Scholar]
- Shieh B. H., Ballivet M., Schmidt J. Quantitation of an alpha subunit splicing intermediate: evidence for transcriptional activation in the control of acetylcholine receptor expression in denervated chick skeletal muscle. J Cell Biol. 1987 May;104(5):1337–1341. doi: 10.1083/jcb.104.5.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tamaoki T., Nomoto H., Takahashi I., Kato Y., Morimoto M., Tomita F. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase. Biochem Biophys Res Commun. 1986 Mar 13;135(2):397–402. doi: 10.1016/0006-291x(86)90008-2. [DOI] [PubMed] [Google Scholar]
- Tsay H. J., Schmidt J. Skeletal muscle denervation activates acetylcholine receptor genes. J Cell Biol. 1989 Apr;108(4):1523–1526. doi: 10.1083/jcb.108.4.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Usdin T. B., Fischbach G. D. Purification and characterization of a polypeptide from chick brain that promotes the accumulation of acetylcholine receptors in chick myotubes. J Cell Biol. 1986 Aug;103(2):493–507. doi: 10.1083/jcb.103.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Villar M. J., Huchet M., Hökfelt T., Changeux J. P., Fahrenkrug J., Brown J. C. Existence and coexistence of calcitonin gene-related peptide, vasoactive intestinal polypeptide- and somatostatin-like immunoreactivities in spinal cord motoneurons of developing embryos and post-hatch chicks. Neurosci Lett. 1988 Mar 21;86(1):114–118. doi: 10.1016/0304-3940(88)90193-0. [DOI] [PubMed] [Google Scholar]
- Wall R., Briskin M., Carter C., Govan H., Taylor A., Kincade P. A labile inhibitor blocks immunoglobulin kappa-light-chain-gene transcription in a pre-B leukemic cell line. Proc Natl Acad Sci U S A. 1986 Jan;83(2):295–298. doi: 10.1073/pnas.83.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wang Y., Xu H. P., Wang X. M., Ballivet M., Schmidt J. A cell type-specific enhancer drives expression of the chick muscle acetylcholine receptor alpha-subunit gene. Neuron. 1988 Aug;1(6):527–534. doi: 10.1016/0896-6273(88)90183-3. [DOI] [PubMed] [Google Scholar]