Formation of electrical synapses between isolated, cultured Helisoma neurons requires mutual neurite elongation

Our previous experiments have suggested the hypothesis that conjoint active neuronal outgrowth may be necessary for formation of new electrical synapses between identified neurons of adult Helisoma buccal ganglia. This growth dependence hypothesis now has been tested by examining the responses of individual pairs of neurons in isolation from the influences of the ganglionic environment. Isolated cell culture of identified neurons (neuron 5) showed that: (i) neurons growing in cell culture undergo a predictable sequence of morphological changes culminating in a stable morphological state (i.e., growth stops); (ii) contact between actively growing neurons in cell culture results in the formation of electrical connections, just as in ganglia; and (iii) when an actively growing neuron encounters a neuron that is morphologically stable, electrical connections do not form or are very weak, even though strong connections are made between pairs of actively growing neurons in the same culture. These results establish that processes closely associated with growth are required for formation of electrical synapses between these neurons.