Skip to main content
SpringerLink
Log in

Developmentally regulated expression of calcitonin gene-related peptide at mammalian neuromuscular junction

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Using indirect immunofluorescence techniques, we have found that calcitonin gene-related peptide-like immunoreactivity is present in the neuromuscular junctions of somatic muscles as well as in almost all motor neurons of the lumbar enlargement of 1-week-old rats. It gradually decreases in both motor neurons and motor nerve endings as the animal grows up and completely disappears from the neuromuscular junctions in adult rats, persisting only in the motor nerve endings on the intrafusal fibers. In situ hybridization experiments have shown that the down-regulation of calcitonin generelated peptide-like immunoreactivity is strictly related to a reduction in CGRP mRNA levels in the spinal motor neurons. These results indicate that the expression of CGRP is developmentally regulated in spinal cord α motor neurons. They also suggest that the peptide may play an important role at the immature neuromuscular junction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Amara, S.G., Jonas, V., Rosenfeld, M.G., Ong, E.S., Evans, R.M. (1982). Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products. Nature 298:240–244

    Article  PubMed  CAS  Google Scholar 

  • Arbuthnott, E.R., Ballard, K.J., Boyd, I.A., Gladden, M.H., Sutherland, F.I. (1982). The ultrastructure of cat fusimotor endings and their relationship to foci of sarcomer convergence in intrafusal fibers. J. Physiol. 331:285–309

    PubMed  CAS  Google Scholar 

  • Barthó, L., Lembeck, F., Holzer, P. (1987). Calcitonin gene-related peptide is a potent relaxant of intestinal muscle. Eur. J. Pharmacol. 135:449–451

    Article  PubMed  Google Scholar 

  • Brewer, P.A., Lynch, K. (1986). Stimulation-associated changes in frog neuromuscular junctions. A quantitative ultrastructural comparison of rapid-frozen and chemically fixed nerve terminals. Neuroscience 17:881–895

    Article  PubMed  CAS  Google Scholar 

  • Brodal, A. (1969). Neurological Anatomy. Oxford University Press, London

    Google Scholar 

  • Ceccarelli, B., Hurlbut, P. (1980). Vesicle hypothesis of release of quanta of ACh. Physiol. Rev. 60:396–441

    PubMed  CAS  Google Scholar 

  • Chesselet, M.F., Weiss, L., Wuenschell, C., Tobin, A.J., Affolter, H.U. (1987). Comparative distribution of mRNAs for glutamic acid decarboxylase, tyrosine hydroxylase and tachykinins in the basal ganglia: an in situ hybridization study in the rodent brain. J. Comp. Neurol. 262:125–140

    Article  PubMed  CAS  Google Scholar 

  • Fontaine, B., Klarsfeld, A., Hökfelt, T., Changeux, J.P. (1986). Calcitonin gene-related peptide, a peptide present in spinal cord motoneurones increases the number of acetylcholine receptors in primary cultures of chick embryo myotubes. Neurosci. Lett. 71:59–65

    Article  PubMed  CAS  Google Scholar 

  • Fontaine, B., Klarsfeld, A., Changeux, J.P. (1987). Calcitonin gene-related peptide and muscle activity regulate acetylcholine receptor a-subunit mRNA levels by distinct intracellular pathways. J. Cell. Biol. 105: 1337–1342

    Article  PubMed  CAS  Google Scholar 

  • Fraher, J.P., Kaar, G.F., Bristol, D.C., Rossiter, J.P. (1988). Development of ventral spinal motoneurone fibres: a correlative study of the growth and maturation of central and peripheral segments of large and small fibre class. Prog. Neurobiol. 31:199–239

    Article  PubMed  CAS  Google Scholar 

  • Gibson, S.J., Polak, J.M., Bloom, S.R., Sabate, I.M., Mulderry, P.M., Ghatel, M.A., McGregor, G.P., Morrison, J.F.B., Kelly, J.S., Evans, R.M., Rosenfeld, M.G. (1984). Calcitonin gene-related peptide immunoreactivity in the spinal cord of man and of eight other species. J. Neurosci. 4:3101–3111

    PubMed  CAS  Google Scholar 

  • Green, M.R., Maniatis, T., Melton, D.A. (1983). Human b-globin pre-mRNA synthetized in vitro is accurately spliced in xenopus oocyte nuclei. Cell 32:681–694

    Article  PubMed  CAS  Google Scholar 

  • Gulbenkian, S., Merighi, A., Wharton, J., Varndell, I.M., Polak, J.M. (1986). Ultrastructural evidence for the coexistence of calcitonin gene-related peptide and substance P in secretory vesicles of peripheral nerves in guinea pig. J. Neurocytol. 15:535–542

    Article  PubMed  CAS  Google Scholar 

  • Hardman, V.J., Brown, M.C. (1985). Absence of postnatal death among motoneurones supplying the inferior gluteal nerve of rat. Dev. Brain Res. 19:1–9

    Article  Google Scholar 

  • Klein, R., Lagercrantz, H., Zimmermann, H. (1982). Neurotransmitter Vesicles. Academic Press, New York, 375 pp

    Google Scholar 

  • Kubota, Y., Inagaki, I., Shimada, S., Girgis, S., Zadi, M., MacIntyre, I., Tohyama, M., Kito, S. (1988). Ontogeny of the calcitonin gene-related peptide in the nervous system of rat brainstem: An immunohistochemical analysis. Neuroscience 26:905–926

    Article  PubMed  CAS  Google Scholar 

  • Lundberg, J.M., Hökfelt, T. (1983). Coexistence of peptides and classical neurotransmitter. Trends Neurosci. 6:325–333

    Article  CAS  Google Scholar 

  • Matteoli, M., Haimann, C., Torri-Tarelli, F., Polak, J.M., Ceccarelli, B., De Camilli, P. (1988). Differential effect of alphalatrotoxin on exocytosis from small synaptic vesicles and from large dense core vesicles containing calcitonin gene related peptide at the frog neuromuscular junction. Proc. Natl. Acad. Sci. U.S.A. 85:7366–7370

    Article  PubMed  CAS  Google Scholar 

  • Matteoli, M., Haimann, C., De Camilli, P. (1990). Substance P Immunoreactivity at the frog neuromuscular junction. Neuroscience, in press

  • McCullouch, J., Uddman, R., Kingman, T.A., Edvinsson, L. (1986). Calcitonin gene-related peptide: Function role in cerebrovascular regulation. Proc. Natl. Acad. Sci. U.S.A. 83:5731–5735

    Article  Google Scholar 

  • McHanwell, S., Biscoe, T.J. (1981). The size of motoneurons supplying hindlimb muscles in the mouse. Proc. R. Soc. London Ser. B 213:201–216

    Article  CAS  Google Scholar 

  • Melton, D.A., Krieg, P.A., Rebogliat, M.R., Maniatis, T., Zinn, K., Green, M.R. (1984). Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing bacteriophage SP6 promoter. Nucleic Acids Res. 12:7035–7056

    Article  PubMed  CAS  Google Scholar 

  • Martin, M.R., Caddy, K.W.T., Biscoe, R.J. (1977). Numbers and diameters of motoneurons and myelinated axons in the facial nucleus and nerve of the albino rat. J. Anat. 123:579–587

    PubMed  CAS  Google Scholar 

  • Mora, M., Marchi, M., Polak, J.M., Gibson, S., Cornelio, F. (1989). Calcitonin gene-related peptide immunoreactivity at the human neuromuscular junction. Brain Res. 492:404–407

    Article  PubMed  CAS  Google Scholar 

  • Mulle, C., Benoit, P., Pinset, C., Roa, M., Changeux, J.P. (1988). Calcitonin gene-related peptide enhances the rate of desensitization of the nicotinic acetylcholine receptor in cultured mouse muscle cells. Proc. Natl. Acad. Sci. U.S.A. 85:5728–5732

    Article  PubMed  CAS  Google Scholar 

  • New, H.V., Mudge, A.W. (1986). Calcitonin gene-related peptide regulates muscle acetylcholine receptor synthesis. Nature (London) 323:809–811

    Article  CAS  Google Scholar 

  • Nicolopoulos-Strournaras, S., Iles, J.F. (1983). Motor neuron columns in the lumbar spinal cord of the rat. J. Comp. Neurol. 217:75–85

    Article  Google Scholar 

  • Ohhashi, T., Jacobowitz, D.M. (1988). Effects of calcitonin gene related peptide on neuromuscular transmission in the isolated diaphragm. Peptides 9:613–617

    Article  PubMed  CAS  Google Scholar 

  • Popper, P., Micevych, P.E. (1989). Localization of calcitonin gene-related peptide and its receptors in a striated muscle. Brain Res. 496:180–186

    Article  PubMed  CAS  Google Scholar 

  • Rethelyi, M., Metz, C.B., Lund, P.K. (1989). Distribution of neurons expressing calcitonin gene-related peptide mRNAs in the brain stem, spinal cord and dorsal root ganglia of rat and guinea pig. Neuroscience 29:225–239

    Article  PubMed  CAS  Google Scholar 

  • Rodrigo, J., Polak, J.M., Terenghi, G., Cervantes, C., Ghatei, M.A., Mulderry, P.K., Bloom, S.R. (1985a). Calcitonin gene-related peptide immunoreactive sensory and motor nerves of the mammalian palate. Histochemistry 82:67–74

    Article  PubMed  CAS  Google Scholar 

  • Rodrigo, J., Polak, J.M., Fernandez, L., Ghatei, M.A., Mulderry, P., Bloom, S.R. (1985b). Calcitonin gene related peptide immunoreactive sensory and motor nerves of the rat cat and monkey oesophagus. Gastroenterology 88:441–451

    Google Scholar 

  • Rosenfeld, M.G., Mermod, J., Amara, S.G., Swanson, L.W., Sawchencko, P.E., Rivier, J., Vale, W.W., Evans, R.M. (1983). Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing. Nature (London) 304:129–135

    Article  CAS  Google Scholar 

  • Samuelson, U.E., Dalsgaard, C.J., Lundberg, J.M., Hökfelt, T. (1985). Calcitonin gene related peptide inhibits spontaneous contractions in human uterus and fallopian tube. Neurosci. Lett. 62:225–230

    Article  PubMed  CAS  Google Scholar 

  • Sassoon D., Lyons, G., Wright, W.E., Lin, V., Lassar, A., Weintraub, H., Buckingham, M. (1989). Expression of two myogenic regulatory factors myogenin and MyoD1 during mouse embryogenesis. Nature 341:303–307

    Article  PubMed  CAS  Google Scholar 

  • Sato M., Mizuno, N., Konishi, A. (1977). Postnatal differentiation of cell body volumes of spinal motoneurones innervating slow-twitch and fast-twitch muscles. J. Comp. Neurol. 117:387–398

    Google Scholar 

  • Schiaffino, S., Gorza, L., Pitton, G., Saggin, L., Ausoni, S., Sartore, S., Lomo, T. (1988). Embryonic and neonatal mysoin heavy chain in denervated and paralyzed rat skeletal muscle. Dev. Biol. 127:1–11

    Article  PubMed  CAS  Google Scholar 

  • Skofitsch, G., Jacobowitz, D.M. (1985). Calcitonin generelated peptide: detailed immunohistochemical distribution in the central nervous system. Peptides 6:721–745

    Article  PubMed  CAS  Google Scholar 

  • Takami, K., Kaway, Y., Shisaka, S., Lee, Y., Girgis, S., Hillyard, C.J., Macyntire, J., Emson, P.C., Tohyama, M. (1985a). Immunohistochemical evidence for the coexistence of calcitonin gene related peptide and choline acetyltransferase-like immunoreactivity in neurons of the rat hypoglossal facial and ambiguus nuclei. Brain Res. 328:386–389

    Article  PubMed  CAS  Google Scholar 

  • Takami, K., Kawai, Y., Uchida, S., Tohyama, M., Shiotanj, Y., Yoshida, H., Emson, P.C., Girgis, S., Hillyard, C.J., MacIntyre, I. (1985b). Effect of calcitonin gene-related peptide on contraction of striated muscle in the mouse. Neurosci. Lett. 60:217–230

    Article  Google Scholar 

  • Terenghi, G., Polak, J.M., Rodrigo, J., Mulderry, P.K., Bloom, S.R. (1986). Calcitonin gene-related peptide-immunoreactive nerves in the tongue, epiglottis and pharynx of the rat: Occurrence, distribution and origin. Brain Res. 365:1–14

    Article  PubMed  CAS  Google Scholar 

  • Villar, M.J., Roa, M., Huchet, M., Hökfelt, T., Changeux, J.P., Fahrenkrug, J., Brown, J.C., Epstein, M., Hersh, L. (1989). Immunoreactive calcitonin gene-related peptide, vasoactive intestinal peptide, and somatostatin in developing chicken spinal cord motorneurons. Distribution and role in regulation of cAMP in cultured muscle cells. Eur. J. Neurosci. 1:269–287

    Article  PubMed  Google Scholar 

  • Vitadello, M., Triban, C., Fabris, M., Gorio, A., Schiaffino, S. (1986). Heterogeneity of rat neurofilament polypeptides revealed by a monoclonal antibody. J. Neurochem. 46:665–670

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. CNR Istituto Fisiologia Centri Nervosi, Milan, Italy

    M. Matteoli & M. Vitadello

  2. CNR Center of Cytopharmacology and Department of Medical Pharmacology, Milan, Italy

    S. Balbi, C. Sala, B. Chini & G. Fumagalli

  3. Department of Pharmacology, Urbino, Italy

    M. Cimino

Authors
  1. M. Matteoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Balbi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Sala
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B. Chini
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Cimino
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. Vitadello
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. G. Fumagalli
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Matteoli, M., Balbi, S., Sala, C. et al. Developmentally regulated expression of calcitonin gene-related peptide at mammalian neuromuscular junction. J Mol Neurosci 2, 175 (1990). https://doi.org/10.1007/BF02896842

Download citation

  • DOI: https://doi.org/10.1007/BF02896842

Keywords

Search

Navigation