Isolation of native Mauthner cell axoplasm and an analysis of organelle movement in non-aqueous and aqueous media.

A method for isolating Mauthner cell (M-cell) axoplasm from goldfish is described. The axon may be isolated in a fluorocarbon oil (e.g., Fluorinert), or in an aqueous bathing medium (ABM). An analysis of motile organelles in axoplasm isolated in Fluorinert or in ABM indicated that the range of particle velocities extended over an order of magnitude, and that there was a significant selective slowing of retrograde particle movement in axons isolated in ABM. No apparent correlation between particle size and velocity was noted. In addition to confirming that calcium was not required for particle transport, it was observed that adenylylimidodiphosphate produced an irreversible blockade of organelle movements, suggesting that force generating mechanisms involved in translocation are probably similar to those proposed for invertebrate axons.