Advertisement

Journal of Muscle Research & Cell Motility

, Volume 26, Issue 1, pp 49–56 | Cite as

In ovo neuromuscular stimulation alters the skeletal muscle phenotype of the chick

  • J.L. HEYWOOD
  • G.M. MCENTEEEmail author
  • N.C. STICKLAND
Article

Abstract

4-aminopyridine (4-AP) is a drug that blocks the potassium channels in neurons and stimulates the release of the neurotransmitter acetylcholine (ACh), enhancing its availability at the synaptic cleft. The effects of 4-AP induced neuromuscular stimulation on skeletal muscle formation and development were investigated in embryonic chicks. Fertile white Leghorn eggs were incubated at 37.5°C and windowed on day three of incubation. On embryonic days (E) 10, 11, 12 and 13 half of the eggs were injected with 100 μl of PBS buffer containing 0.2 μg 4-AP and the control group was administered 100 μl of PBS only. 4-AP treated (T) embryos showed at least a 10% increase in mean body mass relative to the controls (C) (P<0.05) at ages E14, E15 and E16. Tibia and femur lengths in the 4-AP treated embryos were significantly greater than the controls at E15 and E16 (P<0.05). The 4-AP treated animals had a 36.8% greater number of myofibres than the control animals at E20. Nuclear number per cross sectional area in the M. Semitendinosus was significantly greater (P<0.01) at E16 in the treated compared to the control embryos. The 4-AP treated group exhibited a greater percentage area of oxidative fibres in cross sections of M. Semitendinosus than the control group at E16 (P<0.01) and at E20 (P<0.05). It may be concluded from these results that 4-AP induced neuromuscular stimulation has a significant effect on skeletal muscle characteristics, leg bone length and overall body mass.

Keywords

Control Embryo Embryonic Chicken Motor Nerve Terminal Myoblast Proliferation Semitendinosus Muscle 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Notes

Acknowledgements

The authors wish to acknowledge the BBSRC for funding this research. Dr. Stéphanie Bayol is also thanked for her input and advice on the data interpretation.

References

  1. Bayol S, Jones D, Goldspink G, Stickland NC, 2004. The influence of undernutrition during gestation on skeletal muscle cellularity and on the expression of genes that control muscle growth Brit J Nutr 91(3): 331–339CrossRefPubMedGoogle Scholar
  2. Bever C, Katz E, Tierney D, Johnson K, 1995. Experience with slow release 4-aminopyridine in multiple sclerosis patients: long term tolerability and safety J Neuroimmunol 56–63(1): 58CrossRefGoogle Scholar
  3. Burley ES, Jacobs RS, 1981. Effects of 4-aminopyridine on nerve terminal action potentials J Pharmacol Exp Ther 219: 268–273PubMedGoogle Scholar
  4. Clemmons DR, 1998. Role of insulin-like growth factor binding proteins in controlling IGF actions Mol Cell Endocrinol 140: 19–24CrossRefPubMedGoogle Scholar
  5. Dekkers J, Waters J, Vrbova G, Greensmith L, 2001. Treatment of the neuromuscular junction with 4-aminopyridine results in improved reinnervation following nerve injury in neonatal rats Neuroscience 103(1): 267–274CrossRefPubMedGoogle Scholar
  6. Drachman DB, 1964 Atrophy of skeletal muscles in chick embryos treated with botulinum toxin Science 145: 719–721PubMedCrossRefGoogle Scholar
  7. Edgerton VR, Roy RR, 1991. Regulation of skeletal muscle fiber size, shape and function J Biomech 24 (Suppl 1) 123–133CrossRefPubMedGoogle Scholar
  8. Engert JC, Berglund EB, Rosenthal N, 1996. Proliferation precedes differentiation in IGF-I-stimulated myogenesis J Cell Biol 135(2): 431–440CrossRefPubMedGoogle Scholar
  9. Florini JR, Ewton DZ, Coolican SA, 1996. Growth hormone and the insulin-like growth factor system in myogenesis Endocrinol Rev 17(5): 481–517CrossRefGoogle Scholar
  10. Gollick PD, Parsons D, Riedy M, Moore RL, 1983 Fiber number and size in overloaded chicken anterior latissimus dorsi muscle J Appl Physiol 54(5):1292–1297PubMedGoogle Scholar
  11. Gordon T, Vrbova G, 1975. Changes in chemosensitivity of developing chick muscle fibres in relation to endplate formation Pflugers Arch 360(4):349–364CrossRefPubMedGoogle Scholar
  12. Harvey AL, Marshall IG, 1977 The facilitatory actions of aminopyridines and tetraethylammonium on neuromuscular transmission and muscle contractility in avian muscle Naunyn Schmiedebergs Arch Pharmacol 299(1): 53–60CrossRefPubMedGoogle Scholar
  13. Harris AJ, Fitzsimons RB, McEwan JC, 1989. Neural control of the sequence of expression of myosin heavy chain isoforms in foetal mammalian muscles Development 107(4): 751–769PubMedGoogle Scholar
  14. Keresztes M, Takacs O, Guba F, 1985. The effect of 4-aminopyridine-induced neuromuscular activity on the metabolism of developing muscles in chick embryos Cell Differ 16(2): 133–137CrossRefPubMedGoogle Scholar
  15. Lefeuvre B, Crossin F, Fontaine-Pérus J, Bandman E, Gardahaut M, 1996. Innervation regulates myosin heavy chain isoform expression in developing skeletal muscle fibres Mech Develop 58(1–2): 115–127CrossRefGoogle Scholar
  16. Lundh H, Thesleff S, 1977. The mode of action of 4-aminopyridine and guanidine on transmitter release from motor nerve terminals Eur J Pharmacol 42: 411–412PubMedCrossRefGoogle Scholar
  17. Maltby V, Somaiya A French NA, Stickland NC, 2004 In ovo temperature manipulation influences post-hatch muscle growth in the turkey Brit Poultry Sci 45(4): 491–498CrossRefGoogle Scholar
  18. Misgeld T, Burgess RW, Lewis RM, Cunningham JM, Lichtman JW, Sanes JR, 2002. Roles of neurotransmitter in synapse formation: development of neuromuscular junctions lacking choline acetyltransferase Neuron 36: 271–284CrossRefGoogle Scholar
  19. Mitchell PJ, Johnson SE, Hannon K, 2002. Insulin-like growth factor I stimulates myoblast expansion and myofibre development in the limb Dev Dynam 223(1): 12–23CrossRefGoogle Scholar
  20. Nachlas MM, Tsou KC, Desouza E, Cheng CS, Seligman AM, 1957. Cytochemical demonstration of succinic dehydrogenase by the use of p-nitrophenyl substituted ditetrazolium J Histochem 5: 420–436Google Scholar
  21. Osborne AC (2000) Mechanisms by which movement exerts its essential role on diarthordial joint cavity. PhD Thesis Google Scholar
  22. Perrone CE, Fenwick-Smith D, Vandenburgh HH, 1995. Collagen and stretch modulate autocrine secretion of insulin-like growth factor-1 and insulin-like growth factor binding proteins from differentiated skeletal muscle cells J Biol Chem 270(5): 2099–2106CrossRefPubMedGoogle Scholar
  23. Rehfeldt C, Stickland NC, Fielder I, Wenger J, 1999. Environmental and genetic factors as sources of variation in skeletal muscle fibre number Basic Appl Myol 9: 237–255Google Scholar
  24. Ross JJ Duxson MJ, Harris AJ, 1987. Neural determination of muscle fibre numbers in embryonic rat lumbrical muscles Development 10(3): 395–409Google Scholar
  25. Salmons S, Vrbová G, 1969. The influence of activity on some contractile characteristics of mammalian fast and slow muscles J Physiol 201: 535–549PubMedGoogle Scholar
  26. Smith KJ, Felts PA, John GR, 2000. Effects of 4-aminopyridine on demyelinated axons, synapses and muscle tension Brain 123(1): 171–184CrossRefPubMedGoogle Scholar
  27. Srihari T, Vrbova G, 1978 The role of muscle activity in the differentiation of neuromuscular junctions in slow and fast chick muscles J Neurocytol 7(5): 529–540CrossRefPubMedGoogle Scholar
  28. Vandenburgh HH, Hatfaludy S, Karlisch P, Shansky J, 1989. Skeletal muscle growth is stimulated by intermittent stretch-relaxation in tissue culture American Journal of Physiology. 256(3 Pt 1): C674-C682PubMedGoogle Scholar
  29. Vrbová G, Lowrie M, 1989. Role of activity in developing synapses, search for molecular mechanisms News Physiol Sci 4: 75–78Google Scholar
  30. Walters EH, Osborne A, Pitsillides A, Loughna PT, Stickland NC, 2000. The effect of neuromuscular stimulation during myogenesis on muscle fibre number and myogenic regulatory factor expression Comp Biochem Physiol 126(Part A): S163CrossRefGoogle Scholar
  31. Yang S, Alnaqeeb M, Simpson H, Goldspink G, 1996 Cloning and characterization of an IGF-1 isoform expressed in skeletal muscle subjected to stretch J Muscle Res Cell Motil 17: 487–495CrossRefPubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  1. 1.Southampton Oceanography CentreUniversity of SouthamptonSouthamptonUK
  2. 2.Department of Veterinary Basic SciencesThe Royal Veterinary CollegeLondonUK

Personalised recommendations