Transport axonal de protéines marquées dans le motoneurone géant de l'écrevisse

Isolated abdomens of crayfish were maintained in vitro and lysine (-³H) was intracellularly injected into one of the giant motoneuron of the ventral cord ganglia by iontophoresis. Membrane potentials ranging between -60 and -70 mV were recorded all along the experiment. Light microscope radioautographs showed an intense reaction over the injected nerve cell body and the initial segment of its axon; most of the surrounding tissues were free of radioactivity when the diffusion of the injected lysine (-³H) was prevented by adding cold lysine to the bathing medium. Some exchange of label was however noted with electrically coupled axons and glial sheaths. No radioactive protein could be traced in the numerous nerve endings of the neuromuscular junction. Nonetheless when a ligature was placed on the nerve root, the amount of accumulated radioactivity was increased from 3 to 6 h. in the axon of the injected motoneuron only. Electron microscope radioautographs indicate that the fast transported proteins were mainly associated with the endoplasmic reticulum and the axonal membrane. It is concluded that the visualization of the nerve endings was limited by the dispersion of the label into the numerous thin terminal branchs of the axon; however the combination of iontophoretic injection and radioautography permits to trace endogenous protein along the axon and to study molecular exchanges with other cells.