Visualization of cellular mechanisms regulating differential neuronal synapse formation

Over thirty years ago electrical coupling was observed in embryonic cells prior to chemical communication. This temporal relationship of electrical coupling preceding functional chemical neurotransmission occurs throughout neurogenesis, prompting the idea that gap junctional coupling synchronizes the synaptogenic establishment of functional neural networks. Helisoma neuronal pairs treated with trophic factors exhibit increased electrical coupling and subsequently delay the formation of inhibitory chemical connections. Studies in this thesis addressed the mechanism regulating this inverse relationship between electrotonic and chemical communication. Synaptogenesis between two neurons from the Helisoma buccal ganglia, B110 and B19, were examined using alternative culturing conditions that were either exposed to or deprived of trophic factors. Incubating neuronal pairs in trophic factors induced transient electrical synapses and postponed the formation of chemical connections. In electrically coupled neuronal pairs, presynaptic secretory vesicles were recruited to the sites of presynaptic contact, but did not respond to calcium elevation (i.e., photolytic release of calcium from NP-EGTA) with neurotransmitter release. These and other studies demonstrated that transient electrical coupling does not disrupt calcium handling or postsynaptic responsiveness. Rather, electrotonic coupling delays chemical synaptic transmission by imposing a functional block between the accumulation of presynaptic calcium and the synchronized vesicular release of neurotransmitter.