Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Jul 1;99(1 Pt 1):332–335. doi: 10.1083/jcb.99.1.332

Denervation supersensitivity in skeletal muscle: analysis with a cloned cDNA probe

PMCID: PMC2275635  PMID: 6547444

Abstract

Motor neurons regulate the acetylcholine sensitivity of the muscles they innervate: denervated muscle fiber become "supersensitive" to acetylcholine, due to insertion of newly synthesized acetylcholine receptors (AChRs) in the plasma membrane. We used hybridization analysis with a cloned cDNA specific for AChR alpha-subunit to compare the abundance of AChR mRNA in innervated and denervated adult mouse muscles. Within 3 d of denervation, levels of AChR mRNA increased 100- fold; levels of actin mRNA changed little. The increase in AChR mRNA level was sufficiently large and rapid to account for denervation supersensitivity.

Full Text

The Full Text of this article is available as a PDF (413.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AXELSSON J., THESLEFF S. A study of supersensitivity in denervated mammalian skeletal muscle. J Physiol. 1959 Jun 23;147(1):178–193. doi: 10.1113/jphysiol.1959.sp006233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berg D. K., Hall Z. W. Loss of alpha-bungarotoxin from junctional and extrajunctional acetylcholine receptors in rat diaphragm muscle in vivo and in organ culture. J Physiol. 1975 Nov;252(3):771–789. doi: 10.1113/jphysiol.1975.sp011169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berg D. K., Kelly R. B., Sargent P. B., Williamson P., Hall Z. W. Binding of -bungarotoxin to acetylcholine receptors in mammalian muscle (snake venom-denervated muscle-neonatal muscle-rat diaphragm-SDS-polyacrylamide gel electrophoresis). Proc Natl Acad Sci U S A. 1972 Jan;69(1):147–151. doi: 10.1073/pnas.69.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brockes J. P., Hall Z. W. Acetylcholine receptors in normal and denervated rat diaphragm muscle. I. Purification and interaction with [125I]-alpha-bungarotoxin. Biochemistry. 1975 May 20;14(10):2092–2099. doi: 10.1021/bi00681a008. [DOI] [PubMed] [Google Scholar]
  5. Brockes J. P., Hall Z. W. Synthesis of acetylcholine receptor by denervated rat diaphragm muscle. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1368–1372. doi: 10.1073/pnas.72.4.1368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caravatti M., Minty A., Robert B., Montarras D., Weydert A., Cohen A., Daubas P., Buckingham M. Regulation of muscle gene expression. The accumulation of messenger RNAs coding for muscle-specific proteins during myogenesis in a mouse cell line. J Mol Biol. 1982 Sep;160(1):59–76. doi: 10.1016/0022-2836(82)90131-0. [DOI] [PubMed] [Google Scholar]
  7. Changeux J. P. The acetylcholine receptor: an "allosteric" membrane protein. Harvey Lect. 1979 1980;75:85–254. [PubMed] [Google Scholar]
  8. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  9. Claudio T., Ballivet M., Patrick J., Heinemann S. Nucleotide and deduced amino acid sequences of Torpedo californica acetylcholine receptor gamma subunit. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1111–1115. doi: 10.1073/pnas.80.4.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  11. Devillers-Thiery A., Giraudat J., Bentaboulet M., Changeux J. P. Complete mRNA coding sequence of the acetylcholine binding alpha-subunit of Torpedo marmorata acetylcholine receptor: a model for the transmembrane organization of the polypeptide chain. Proc Natl Acad Sci U S A. 1983 Apr;80(7):2067–2071. doi: 10.1073/pnas.80.7.2067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Devreotes P. N., Fambrough D. M. Turnover of acetylcholine receptors in skeletal muscle. Cold Spring Harb Symp Quant Biol. 1976;40:237–251. doi: 10.1101/sqb.1976.040.01.025. [DOI] [PubMed] [Google Scholar]
  13. Fambrough D. M. Control of acetylcholine receptors in skeletal muscle. Physiol Rev. 1979 Jan;59(1):165–227. doi: 10.1152/physrev.1979.59.1.165. [DOI] [PubMed] [Google Scholar]
  14. Hall Z. W., Reiness C. G. Electrical stimulation of denervated muscles reduces incorporation of methionine into the ACh receptor. Nature. 1977 Aug 18;268(5621):655–657. doi: 10.1038/268655a0. [DOI] [PubMed] [Google Scholar]
  15. Hall Z. W., Roisin M. P., Gu Y., Gorin P. D. A developmental change in the immunological properties of acetylcholine receptors at the rat neuromuscular junction. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 1):101–108. doi: 10.1101/sqb.1983.048.01.013. [DOI] [PubMed] [Google Scholar]
  16. Hartzell H. C., Fambrough D. M. Acetylcholine receptors. Distribution and extrajunctional density in rat diaphragm after denervation correlated with acetylcholine sensitivity. J Gen Physiol. 1972 Sep;60(3):248–262. doi: 10.1085/jgp.60.3.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Howe P. R., Livett B. G., Austin L. Increased binding of alpha-bungarotoxin in dystrophic mouse muscle. Exp Neurol. 1976 Apr;51(1):132–140. doi: 10.1016/0014-4886(76)90058-3. [DOI] [PubMed] [Google Scholar]
  18. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  19. Linden D. C., Fambrough D. M. Biosynthesis and degradation of acetylcholine receptors in rat skeletal muscles. Effects of electrical stimulation. Neuroscience. 1979;4(4):527–538. doi: 10.1016/0306-4522(79)90129-5. [DOI] [PubMed] [Google Scholar]
  20. Lomo T., Westgaard R. H. Control of ACh sensitivity in rat muscle fibers. Cold Spring Harb Symp Quant Biol. 1976;40:263–274. doi: 10.1101/sqb.1976.040.01.027. [DOI] [PubMed] [Google Scholar]
  21. MILEDI R. The acetylcholine sensitivity of frog muscle fibres after complete or partial devervation. J Physiol. 1960 Apr;151:1–23. [PMC free article] [PubMed] [Google Scholar]
  22. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  23. Merlie J. P. Biogenesis of the acetylcholine receptor, a multisubunit integral membrane protein. Cell. 1984 Mar;36(3):573–575. doi: 10.1016/0092-8674(84)90335-0. [DOI] [PubMed] [Google Scholar]
  24. Merlie J. P., Heinemann S., Lindstrom J. M. Acetylcholine receptor degradation in adult rat diaphragms in organ culture and the effect of anti-acetylcholine receptor antibodies. J Biol Chem. 1979 Jul 25;254(14):6320–6327. [PubMed] [Google Scholar]
  25. Merlie J. P., Sebbane R., Gardner S., Lindstrom J. cDNA clone for the alpha subunit of the acetylcholine receptor from the mouse muscle cell line BC3H-1. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3845–3849. doi: 10.1073/pnas.80.12.3845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Merlie J. P., Sebbane R., Gardner S., Olson E., Lindstrom J. The regulation of acetylcholine receptor expression in mammalian muscle. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 1):135–146. doi: 10.1101/sqb.1983.048.01.016. [DOI] [PubMed] [Google Scholar]
  27. Metafora S., Felsani A., Cotrufo R., Tajana G. F., Del Rio A., De Prisco P. P., Rutigliano B., Esposito V. Neural control of gene expression in the skeletal muscle fibre: changes in the muscular mRNA population following denervation. Proc R Soc Lond B Biol Sci. 1980 Aug 13;209(1175):257–273. doi: 10.1098/rspb.1980.0094. [DOI] [PubMed] [Google Scholar]
  28. Miledi R., Potter L. T. Acetylcholine receptors in muscle fibres. Nature. 1971 Oct 29;233(5322):599–603. doi: 10.1038/233599a0. [DOI] [PubMed] [Google Scholar]
  29. Noda M., Furutani Y., Takahashi H., Toyosato M., Tanabe T., Shimizu S., Kikyotani S., Kayano T., Hirose T., Inayama S. Cloning and sequence analysis of calf cDNA and human genomic DNA encoding alpha-subunit precursor of muscle acetylcholine receptor. 1983 Oct 27-Nov 2Nature. 305(5937):818–823. doi: 10.1038/305818a0. [DOI] [PubMed] [Google Scholar]
  30. Noda M., Takahashi H., Tanabe T., Toyosato M., Kikyotani S., Furutani Y., Hirose T., Takashima H., Inayama S., Miyata T. Structural homology of Torpedo californica acetylcholine receptor subunits. Nature. 1983 Apr 7;302(5908):528–532. doi: 10.1038/302528a0. [DOI] [PubMed] [Google Scholar]
  31. Olson E. N., Glaser L., Merlie J. P., Lindstrom J. Expression of acetylcholine receptor alpha-subunit mRNA during differentiation of the BC3H1 muscle cell line. J Biol Chem. 1984 Mar 10;259(5):3330–3336. [PubMed] [Google Scholar]
  32. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  33. Sebbane R., Clokey G., Merlie J. P., Tzartos S., Lindstrom J. Characterization of the mRNA for mouse muscle acetylcholine receptor alpha subunit by quantitative translation in vitro. J Biol Chem. 1983 Mar 10;258(5):3294–3303. [PubMed] [Google Scholar]
  34. Sumikawa K., Houghton M., Smith J. C., Bell L., Richards B. M., Barnard E. A. The molecular cloning and characterisation of cDNA coding for the alpha subunit of the acetylcholine receptor. Nucleic Acids Res. 1982 Oct 11;10(19):5809–5822. doi: 10.1093/nar/10.19.5809. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES