Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Aug 2;130(4):949–957. doi: 10.1083/jcb.130.4.949

Local neurotrophic repression of gene transcripts encoding fetal AChRs at rat neuromuscular synapses

PMCID: PMC2199949  PMID: 7642710

Abstract

The spatio-temporal expression patterns of mRNA transcripts coding for acetylcholine receptor (AChR) subunits and myogenic factors were measured in denervated rat soleus muscle and in soleus muscle chronically paralyzed for up to 12 d by conduction block of the sciatic nerve by tetrodotoxin (TTX). In denervated muscle the AChR alpha-, beta- , gamma-, and delta-subunit mRNAs were elevated with highest expression levels in the former synaptic and the perisynaptic region and with lower levels in the extrasynaptic fiber segments. In muscle paralyzed by nerve conduction block the alpha-, beta-, gamma-, and delta-subunit mRNA levels increased only in extrasynaptic fiber segments. Surprisingly, in the synaptic region the gamma-subunit mRNA that specifies the fetal-type AChR, and alpha-, beta-, delta-subunit mRNAs were not elevated. The expression of the gene encoding the epsilon- subunit, which specifies the adult-type AChR, was always restricted to synaptic nuclei. The mRNA for the regulatory factor myogenin showed after denervation similar changes as the subunit transcripts of the fetal AChR. When the muscle was paralyzed by nerve conduction block the increase of myogenin transcripts was also less pronounced in synaptic regions compared to extrasynaptic fiber segments. The results suggest that in normal soleus muscle a neurotrophic signal from the nerve locally down-regulates the expression of fetal-type AChR channel in the synaptic and perisynaptic muscle membrane by inhibiting the expression of the gamma-subunit gene and that inhibition of the myogenin gene expression may contribute to this down-regulation.

Full Text

The Full Text of this article is available as a PDF (2.8 MB).

Selected References

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

  1. Berberich C., Dürr I., Koenen M., Witzemann V. Two adjacent E box elements and a M-CAT box are involved in the muscle-specific regulation of the rat acetylcholine receptor beta subunit gene. Eur J Biochem. 1993 Sep 1;216(2):395–404. doi: 10.1111/j.1432-1033.1993.tb18157.x. [DOI] [PubMed] [Google Scholar]
  2. Brenner H. R., Herczeg A., Slater C. R. Synapse-specific expression of acetylcholine receptor genes and their products at original synaptic sites in rat soleus muscle fibres regenerating in the absence of innervation. Development. 1992 Sep;116(1):41–53. doi: 10.1242/dev.116.1.41. [DOI] [PubMed] [Google Scholar]
  3. Brenner H. R., Rotzler S., Kues W. A., Witzemann V., Sakmann B. Nerve-dependent induction of AChR epsilon-subunit gene expression in muscle is independent of state of differentiation. Dev Biol. 1994 Oct;165(2):527–536. doi: 10.1006/dbio.1994.1272. [DOI] [PubMed] [Google Scholar]
  4. Brenner H. R., Witzemann V., Sakmann B. Imprinting of acetylcholine receptor messenger RNA accumulation in mammalian neuromuscular synapses. Nature. 1990 Apr 5;344(6266):544–547. doi: 10.1038/344544a0. [DOI] [PubMed] [Google Scholar]
  5. Buonanno A., Apone L., Morasso M. I., Beers R., Brenner H. R., Eftimie R. The MyoD family of myogenic factors is regulated by electrical activity: isolation and characterization of a mouse Myf-5 cDNA. Nucleic Acids Res. 1992 Feb 11;20(3):539–544. doi: 10.1093/nar/20.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cangiano A., Lutzemberger L. Partial denervation affects both denervated and innervated fibers in the mammalian skeletal muscle. Science. 1977 Apr 29;196(4289):542–545. doi: 10.1126/science.850797. [DOI] [PubMed] [Google Scholar]
  7. Cangiano A., Magherini P. C., Pasino E., Pellegrino M., Risaliti R. Interaction of inactivity and nerve breakdown products in the origin of acute denervation changes in rat skeletal muscle. J Physiol. 1984 Oct;355:345–365. doi: 10.1113/jphysiol.1984.sp015423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cleveland D. W., Lopata M. A., MacDonald R. J., Cowan N. J., Rutter W. J., Kirschner M. W. Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980 May;20(1):95–105. doi: 10.1016/0092-8674(80)90238-x. [DOI] [PubMed] [Google Scholar]
  9. Connor E. A., McMahan U. J. Cell accumulation in the junctional region of denervated muscle. J Cell Biol. 1987 Jan;104(1):109–120. doi: 10.1083/jcb.104.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davis R. L., Weintraub H., Lassar A. B. Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell. 1987 Dec 24;51(6):987–1000. doi: 10.1016/0092-8674(87)90585-x. [DOI] [PubMed] [Google Scholar]
  11. Edmondson D. G., Olson E. N. A gene with homology to the myc similarity region of MyoD1 is expressed during myogenesis and is sufficient to activate the muscle differentiation program. Genes Dev. 1989 May;3(5):628–640. doi: 10.1101/gad.3.5.628. [DOI] [PubMed] [Google Scholar]
  12. Eftimie R., Brenner H. R., Buonanno A. Myogenin and MyoD join a family of skeletal muscle genes regulated by electrical activity. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1349–1353. doi: 10.1073/pnas.88.4.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Goldman D., Carlson B. M., Staple J. Induction of adult-type nicotinic acetylcholine receptor gene expression in noninnervated regenerating muscle. Neuron. 1991 Oct;7(4):649–658. doi: 10.1016/0896-6273(91)90377-c. [DOI] [PubMed] [Google Scholar]
  16. Goldman D., Staple J. Spatial and temporal expression of acetylcholine receptor RNAs in innervated and denervated rat soleus muscle. Neuron. 1989 Aug;3(2):219–228. doi: 10.1016/0896-6273(89)90035-4. [DOI] [PubMed] [Google Scholar]
  17. Gundersen K., Sanes J. R., Merlie J. P. Neural regulation of muscle acetylcholine receptor epsilon- and alpha-subunit gene promoters in transgenic mice. J Cell Biol. 1993 Dec;123(6 Pt 1):1535–1544. doi: 10.1083/jcb.123.6.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hall Z. W., Sanes J. R. Synaptic structure and development: the neuromuscular junction. Cell. 1993 Jan;72 (Suppl):99–121. doi: 10.1016/s0092-8674(05)80031-5. [DOI] [PubMed] [Google Scholar]
  19. Huang C. F., Tong J., Schmidt J. Protein kinase C couples membrane excitation to acetylcholine receptor gene inactivation in chick skeletal muscle. Neuron. 1992 Oct;9(4):671–678. doi: 10.1016/0896-6273(92)90030-h. [DOI] [PubMed] [Google Scholar]
  20. Jia H. T., Tsay H. J., Schmidt J. Analysis of binding and activating functions of the chick muscle acetylcholine receptor gamma-subunit upstream sequence. Cell Mol Neurobiol. 1992 Jun;12(3):241–258. doi: 10.1007/BF00712929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. KOELLE G. B., FRIEDENWALD J. A. A histochemical method for localizing cholinesterase activity. Proc Soc Exp Biol Med. 1949 Apr;70(4):617–622. doi: 10.3181/00379727-70-17013. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Kues W. A., Sakmann B., Witzemann V. Differential expression patterns of five acetylcholine receptor subunit genes in rat muscle during development. Eur J Neurosci. 1995 Jun 1;7(6):1376–1385. doi: 10.1111/j.1460-9568.1995.tb01129.x. [DOI] [PubMed] [Google Scholar]
  24. Lømo T., Slater C. R. Control of junctional acetylcholinesterase by neural and muscular influences in the rat. J Physiol. 1980 Jun;303:191–202. doi: 10.1113/jphysiol.1980.sp013280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Martinou J. C., Merlie J. P. Nerve-dependent modulation of acetylcholine receptor epsilon-subunit gene expression. J Neurosci. 1991 May;11(5):1291–1299. doi: 10.1523/JNEUROSCI.11-05-01291.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Miledi R., Parker I., Schalow G. Transmitter induced calcium entry across the post-synaptic membrane at frog end-plates measured using arsenazo III. J Physiol. 1980 Mar;300:197–212. doi: 10.1113/jphysiol.1980.sp013158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mishina M., Takai T., Imoto K., Noda M., Takahashi T., Numa S., Methfessel C., Sakmann B. Molecular distinction between fetal and adult forms of muscle acetylcholine receptor. Nature. 1986 May 22;321(6068):406–411. doi: 10.1038/321406a0. [DOI] [PubMed] [Google Scholar]
  28. Murray M. A., Robbins N. Cell proliferation in denervated muscle: identity and origin of dividing cells. Neuroscience. 1982 Jul;7(7):1823–1833. doi: 10.1016/0306-4522(82)90040-9. [DOI] [PubMed] [Google Scholar]
  29. Neville C. M., Schmidt M., Schmidt J. Response of myogenic determination factors to cessation and resumption of electrical activity in skeletal muscle: a possible role for myogenin in denervation supersensitivity. Cell Mol Neurobiol. 1992 Dec;12(6):511–527. doi: 10.1007/BF00711232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Numberger M., Dürr I., Kues W., Koenen M., Witzemann V. Different mechanisms regulate muscle-specific AChR gamma- and epsilon-subunit gene expression. EMBO J. 1991 Oct;10(10):2957–2964. doi: 10.1002/j.1460-2075.1991.tb07846.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Piette J., Bessereau J. L., Huchet M., Changeux J. P. Two adjacent MyoD1-binding sites regulate expression of the acetylcholine receptor alpha-subunit gene. Nature. 1990 May 24;345(6273):353–355. doi: 10.1038/345353a0. [DOI] [PubMed] [Google Scholar]
  32. Prody C. A., Merlie J. P. The 5'-flanking region of the mouse muscle nicotinic acetylcholine receptor beta subunit gene promotes expression in cultured muscle cells and is activated by MRF4, myogenin and myoD. Nucleic Acids Res. 1992 May 11;20(9):2367–2372. doi: 10.1093/nar/20.9.2367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rhodes S. J., Konieczny S. F. Identification of MRF4: a new member of the muscle regulatory factor gene family. Genes Dev. 1989 Dec;3(12B):2050–2061. doi: 10.1101/gad.3.12b.2050. [DOI] [PubMed] [Google Scholar]
  34. Rotzler S., Schramek H., Brenner H. R. Metabolic stabilization of endplate acetylcholine receptors regulated by Ca2+ influx associated with muscle activity. Nature. 1991 Jan 24;349(6307):337–339. doi: 10.1038/349337a0. [DOI] [PubMed] [Google Scholar]
  35. Salpeter M. M., Marchaterre M., Harris R. Distribution of extrajunctional acetylcholine receptors on a vertebrate muscle: evaluated by using a scanning electron microscope autoradiographic procedure. J Cell Biol. 1988 Jun;106(6):2087–2093. doi: 10.1083/jcb.106.6.2087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sanes J. R., Johnson Y. R., Kotzbauer P. T., Mudd J., Hanley T., Martinou J. C., Merlie J. P. Selective expression of an acetylcholine receptor-lacZ transgene in synaptic nuclei of adult muscle fibers. Development. 1991 Dec;113(4):1181–1191. doi: 10.1242/dev.113.4.1181. [DOI] [PubMed] [Google Scholar]
  37. Simon A. M., Burden S. J. An E box mediates activation and repression of the acetylcholine receptor delta-subunit gene during myogenesis. Mol Cell Biol. 1993 Sep;13(9):5133–5140. doi: 10.1128/mcb.13.9.5133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Witzemann V., Brenner H. R., Sakmann B. Neural factors regulate AChR subunit mRNAs at rat neuromuscular synapses. J Cell Biol. 1991 Jul;114(1):125–141. doi: 10.1083/jcb.114.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Witzemann V., Sakmann B. Differential regulation of MyoD and myogenin mRNA levels by nerve induced muscle activity. FEBS Lett. 1991 May 6;282(2):259–264. doi: 10.1016/0014-5793(91)80490-t. [DOI] [PubMed] [Google Scholar]
  40. Witzemann V., Stein E., Barg B., Konno T., Koenen M., Kues W., Criado M., Hofmann M., Sakmann B. Primary structure and functional expression of the alpha-, beta-, gamma-, delta- and epsilon-subunits of the acetylcholine receptor from rat muscle. Eur J Biochem. 1990 Dec 12;194(2):437–448. doi: 10.1111/j.1432-1033.1990.tb15637.x. [DOI] [PubMed] [Google Scholar]
  41. Yoshioka K., Miyata Y. Changes in the distribution of the extrajunctional acetylcholine sensitivity along muscle fibers during development and following cordotomy in the rat. Neuroscience. 1983 Jun;9(2):437–443. doi: 10.1016/0306-4522(83)90306-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES