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. 1987 May 1;104(5):1337–1341. doi: 10.1083/jcb.104.5.1337

Quantitation of an alpha subunit splicing intermediate: evidence for transcriptional activation in the control of acetylcholine receptor expression in denervated chick skeletal muscle

PMCID: PMC2114465  PMID: 3032988

Abstract

We have investigated the mechanisms responsible for the increase in acetylcholine receptor subunit mRNAs during the induction of denervation supersensitivity in skeletal muscle. Using a cRNA probe specific for exon 7 (224 nucleotides; with flanking intron sequences of 105 nucleotides on the 3' end, and of 70 nucleotides on the 5' end) of the alpha subunit of the chicken muscle acetylcholine receptor gene, we were able to quantitate the concentration of mature alpha subunit mRNA and its precursor. In 3-wk-old chicks, the concentration of alpha subunit message in leg muscle was found to be 4.0 attomoles per microgram total RNA, and to increase 40-fold within 1 wk after section of the sciatic nerve. The molar ratio of precursor/mature mRNA, which was approximately 0.023 in innervated as well as denervated muscle, transiently rose to 0.047 at the beginning of the second postoperative day when mature message content increased 20-fold; the rise in precursor level preceded the increase in mature message content. These findings suggest that an accelerated rate of transcription of the message coding for the alpha subunit causes increased message content and the stimulation of receptor synthesis characteristic of denervated muscle.

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Selected References

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

  1. Buonanno A., Merlie J. P. Transcriptional regulation of nicotinic acetylcholine receptor genes during muscle development. J Biol Chem. 1986 Sep 5;261(25):11452–11455. [PubMed] [Google Scholar]
  2. Carlin B. E., Lawrence J. C., Jr, Lindstrom J. M., Merlie J. P. Inhibition of acetylcholine receptor assembly by activity in primary cultures of embryonic rat muscle cells. J Biol Chem. 1986 Apr 15;261(11):5180–5186. [PubMed] [Google Scholar]
  3. Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
  4. Fambrough D. M. Acetylcholine sensitivity of muscle fiber membranes: mechanism of regulation by motoneurons. Science. 1970 Apr 17;168(3929):372–373. doi: 10.1126/science.168.3929.372. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Feramisco J. R., Smart J. E., Burridge K., Helfman D. M., Thomas G. P. Co-existence of vinculin and a vinculin-like protein of higher molecular weight in smooth muscle. J Biol Chem. 1982 Sep 25;257(18):11024–11031. [PubMed] [Google Scholar]
  7. Goldman D., Boulter J., Heinemann S., Patrick J. Muscle denervation increases the levels of two mRNAs coding for the acetylcholine receptor alpha-subunit. J Neurosci. 1985 Sep;5(9):2553–2558. doi: 10.1523/JNEUROSCI.05-09-02553.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. 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]
  11. Olson E. N., Glaser L., Merlie J. P., Sebanne R., Lindstrom J. Regulation of surface expression of acetylcholine receptors in response to serum and cell growth in the BC3H1 muscle cell line. J Biol Chem. 1983 Nov 25;258(22):13946–13953. [PubMed] [Google Scholar]
  12. Pezzementi L., Schmidt J. Rapid modulation of acetylcholine receptor synthesis. FEBS Lett. 1981 Nov 30;135(1):103–106. doi: 10.1016/0014-5793(81)80953-2. [DOI] [PubMed] [Google Scholar]
  13. Pumplin D. W., Fambrough D. M. Turnover of acetylcholine receptors in skeletal muscle. Annu Rev Physiol. 1982;44:319–335. doi: 10.1146/annurev.ph.44.030182.001535. [DOI] [PubMed] [Google Scholar]
  14. Ryffel G. U., Wyler T., Muellener D. B., Weber R. Identification, organization and processing intermediates of the putative precursors of Xenopus vitellogenin messenger RNA. Cell. 1980 Jan;19(1):53–61. doi: 10.1016/0092-8674(80)90387-6. [DOI] [PubMed] [Google Scholar]
  15. Salpeter M. M., Loring R. H. Nicotinic acetylcholine receptors in vertebrate muscle: properties, distribution and neural control. Prog Neurobiol. 1985;25(4):297–325. doi: 10.1016/0301-0082(85)90018-8. [DOI] [PubMed] [Google Scholar]
  16. Schmidt J., Raftery M. A. A simple assay for the study of solubilized acetylcholine receptors. Anal Biochem. 1973 Apr;52(2):349–354. doi: 10.1016/0003-2697(73)90036-5. [DOI] [PubMed] [Google Scholar]
  17. Wang G. K., Schmidt J. Primary structure and binding properties of iodinated derivatives of alpha-bungarotoxin. J Biol Chem. 1980 Dec 10;255(23):11156–11162. [PubMed] [Google Scholar]

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