Roux's archives of developmental biology

, Volume 200, Issue 4, pp 188–192 | Cite as

Vital labelling of somite-derived myogenic cells in the chicken limb bud

  • Kensuke Hayashi
  • Eijiro Ozawa
Original Articles


In order to understand how myogenic cells migrate in the limb bud, it is indispensable to distinguish undifferentiated myogenic cells from other mesenchymal cells. Thus, a suitable method for this purpose has been sought. A method to exchange the somites of a chicken and a quail microsurgically has widely been used, since the nuclei of the two species are morphologically distinguishable. However, microsurgery is accompanied by disturbances at the operated locus, and introducing cells of different species might induce unexpected effects. We report a new method for labelling chicken myogenic cells without transplantational operations, and describe their migration pattern in limb buds. Injection of a fluorescent carbocyanine dye into the somite lumen intensely labelled the somitic cells. Myogenic cells derived from the somite were clearly detected in limb buds. Before stage 20, the labelled cells were diffusely distributed in the proximal region of the limb bud. At about stage 21 in both wing and leg buds, labelled cells began to form dorsal and ventral masses. The label was followed until the cells differentiated and expressed myosin. This vital labelling method has advantages over the somite transplantation method: it does not include surgical operations that may disturb the normal development, and the cells are labelled intensely enough to be detected in a whole mount preparation.

Key words

Myogenesis Migration Muscle Vital staining Limb morphogenesis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bader DM, Masaki T, Fishman DA (1982) Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro. J Cell Biol 95:763–770Google Scholar
  2. Chevallier A (1978) Etude de la migration des cellules somitiques dans le mesoderme somatopleural de l'ebauche de l'aile. Rouxs Arch Dev Biol 184:57–73Google Scholar
  3. Chevallier A, Kieny M (1982) On the role of the connective tissue in the patterning of the chick limb musculature. Roux's Arch Dev Biol 191:277–280Google Scholar
  4. Chevallier A, Kieny M, Mauger A (1977) Limb-somite relationship: origin of the limb musculature. J Embryol Exp Morphol 41:245–258Google Scholar
  5. Christ B, Jacob HJ, Jacob M (1977) Experimental analysis of the origin of the wing musculature in avian embryos. Anat Embryol 150:171–186Google Scholar
  6. Christ B, Jacob M, Jacob HJ (1983) On the origin and development of the ventrolateral abdominal muscles in the avian embryo (an experimental and ultrastructural study). Anat Embryol 166:87–101Google Scholar
  7. Gumpel-Pinot M (1974) Contribution du mesoderme somitique a la genese du membre chez l'embryon d'Oiseau. CR Acad Sci Paris 279:1305–1309Google Scholar
  8. Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:44–92Google Scholar
  9. Honig MG, Hume RI (1986) Fluorescent carbocyanine dyes allow living neurons of identical origin to be studied in long term cultures. J Cell Biol 103:171–187Google Scholar
  10. Honig MG, Hume RI (1989) DiI and DiO: versatile fluorescent dyes for neuronal labelling and pathway tracing. Trends Neurosci 12:333–341Google Scholar
  11. Jacob M, Christ B, Jacob HJ (1979) The migration of myogenic cells from the somites into the leg region of avian embryos (an ultrastructural study). Anat Embryol 157:291–309Google Scholar
  12. Lance-Jones C (1988) The somitic level of origin of embryonic chick hindlimb muscles. Dev Biol 126:394–407Google Scholar
  13. Mauger A, Kieny M (1980) Migratory and organogenetic capacities of muscle cells in bird embryos. Roux's Arch Dev Biol 189:123–134Google Scholar
  14. Newman SA, Pautou M-P, Kieny M (1981) The distal boundary of myogenic primordia in chimeric avian limb buds and its relation to an accessible population of cartilage progenitor cells. Dev Biol 84:440–448Google Scholar
  15. Schramm C, Solursh M (1990) The formation of premuscle masses during chick wing bud development. Anat Embryol 182:235–247Google Scholar
  16. Serbedzija GN, Bronner-Fraser M, Fraser SE (1989) A vital dye analysis of the timing and pathways of avian trunk neural crest cell migration. Development 106:809–816Google Scholar
  17. Solursh M, Drake C, Meier S (1987) The migration of myogenic cells from the somites at the wing level in avian embryos. Dev Biol 121:389–396Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Kensuke Hayashi
    • 1
  • Eijiro Ozawa
    • 1
  1. 1.Division of Cell BiologyNational Institute of NeuroscienceKodaira, TokyoJapan

Personalised recommendations