Adult muscle is highly vascularised, with blood vessels being
essential for adequate oxygenation of the tissue and for supporting
increased metabolic demands. Whether this is the case during muscle
development has not been examined. Resin histology was used to map the
muscle splitting process and conventional transmission electron
microscopy to examine early muscle differentiation at the midlimb
level or later at the mid radius/ulna level in the chick wing bud
from stages 24 (4.5 d) to 36 (10 d) (Hamburger & Hamilton, 1951).
Microinjection of India Ink into the extra-embryonic vasculature was
used to visualise the patent muscle microcirculation. The results
showed that the premuscle masses are present at stage 24 and initial
splitting of the muscle masses commences at stage 28. The final muscle
pattern is not established until stage 36. At stage 26 the cells
within the premuscle masses exhibited a mesenchymal morphology, but at
stage 28 overt muscle differentiation was evident with myofibrils
present within myoblasts. Undifferentiated mononucleated cells were
interspersed with the differentiating myoblasts. The ratio of
mononucleated cells[ratio ]myoblasts decreased and the myoblasts
became plumper and increasingly packed with myofibrils with age. There
was no evidence of secondary myotube formation at any of the stages
examined. Vascular invasion of the limb occurred at stage 35 just
prior to the establishment of the final muscle pattern. This was
surprising as it was assumed that myogenic differentiation would be
both oxygen and nutrient dependent. The results of this study provide
descriptions of the splitting of the premuscle masses through to the
establishment of the final muscle pattern at the midlimb or mid
radius/ulna level of the chick wing bud together with the
differentiation of the myogenic cells within the developing muscles.
However, the relationship between muscle patterning at the tissue
level and muscle differentiation at the cellular level with
vascularisation remains unclear. It is hoped that the results of the
study may provide the basis for future investigations into mechanisms
involved in muscle patterning and the signalling mechanisms for
vascular invasion.