Objective: The aim of this report was to set up an effective experimental model of cocultures between cells from spinal cord explants and myotubes from adductor muscle.
Methods: We obtained neuronal cells from chick spinal cord explants at embryonic day 5 (ED5) by means of an enzymatic digestion. Small spinal cord fragments were added in cultured muscle cells committed to the differentiative program. Myoblasts were isolated from the chick adductor muscle at ED12.
Results and discussion: The validation of the experimental model was confirmed by a remarkable spreading pattern of neuronal cells, labeled with a NF200 antibody, and high concentration of myotubes, marked by alpha-actinin antibody. The indication of neuronal contacts was highlighted by the alpha-bungarotoxin. This communication reports one of the few morphologic description of muscular and neuronal coculture preparation, performed on chick embryos.
Conclusion: The experimental model presented in this work might be a useful tool to study the cascade of myogenic positive and negative signals activated by paracrine neuronal factors.