Reorganization of synaptic ultrastructure at facilitated lobster neuromuscular terminals

Prolonged stimulation of the single excitor axon to the lobster distal accessory flexor muscle in the presence of ouabain caused long-term facilitation at its neuromuscular synapses. Hence the extracellularly recorded synaptic potentials failed less frequently and increased their mean amplitude, compared to the non-facilitated (control) potentials from homologous sites in the contralateral muscle. The fine structure of synaptic terminals between matched pairs of facilitated and control preparations was compared with the aid of serial section electron microscopy. Differences between facilitated and control preparations were similar both when the latter were bathed in normal saline or ouabain-containing saline, suggesting that the changes were related to the electrical stimulation rather than to the presence of ouabain. First, the facilitated terminals were smaller in surface area than the control. Second, the number and size of synaptic contacts in the facilitated terminals resembled those in the control. Third, presynaptic dense bodies or active sites increased in number although their sizes remained unaltered in the facilitated terminal. This increase is attributed to the addition of dense bodies at existing synaptic contacts since synaptic contacts remained constant in number between facilitated and control preparations. Fourth, the number and size of synaptic vesicles were unaffected by prolonged stimulation although there was a redistribution of vesicles such that they appeared to be channelled in distinct streams to synaptic contacts. Fifth, mitochondria increased in number and were situated closer to the dense bodies at facilitated nerve terminals than at control terminals. Overall, these changes denote considerable reorganization of the synaptic terminals associated with elevated transmitter release.