Aplysia hemolymph promotes neurite outgrowth and synaptogenesis of identified Helix neurons in cell culture

Hemolymph of adult Aplysia californica significantly affects neurite outgrowth of identified neurons of the land snail Helix pomatia. The metacerebral giant cell (MGC) and the motoneuron C3 from the cerebral ganglion and the neuron B2 from the buccal ganglion of H. pomatia were isolated by enzymatic and mechanical dissociation and plated onto poly-L-lysine-coated dishes either containing culture medium conditioned by Helix ganglia, or pre-treated with Aplysia hemolymph. To determine the extent of neuronal growth we measured the neurite elongation and the neuritic field of cultured neurons at different time points. Aplysia hemolymph enhances the extent and rate of linear outgrowth and the branching domain of Helix neurons. With the hemolymph treatment the MGC neuron more consistently forms specific chemical synapses with its follower cell B2, and these connections are more effective than those established in the presence of the conditioned medium.