Abstract
The ultrastructure of presynaptic areas of lamprey reticulospinal axons was studied before, during, and after periods of elevated transmitter release produced either by repetitive action potential activity or depolarization by elevated extracellular potassium. Controls for possible effects of these procedures per se were done by replacing extracellular Ca with Mg to block transmitter release. In some experiments the time course of ultrastructural changes during K depolarization and subsequent recovery were studied by fixing tissue samples at various times. Transmitter release produced by action potential activity (20/sec for 15 min) in the presence of extracellular Ca significantly and reversibly decreased the number of synaptic vesicles, the area occupied by the vesicles, and the density of synaptic vesicles. An unexpected finding was a reversible decrease in the length of the differentiated membrane during periods of increased transmitter release. Transmitter release significantly and reversibly increased the number of coated vesicles, expanded the presynaptic membrane, and increased the number of pleomorphic vesicles. K depolarization (50 mM K for 15 min) produced identical, reversible effects, except that the expansion of the presynaptic membrane, although significant, was relatively small and there was no change in the number of pleomorphic vesicles. Raising the temperature of the saline from 2 degrees C (K depolarization experiments) or 7 degrees C (action potential experiments) to 20 degrees C did not change the results qualitatively but did produce somewhat larger effects during stimulation and appeared to increase the speed of recovery. Action potential activity or K depolarization in control experiments with the Ca in the saline replaced by Mg had little or no effect on synaptic ultrastructure. Synaptic vesicles in lamprey reticulospinal axons never contacted the axonal membrane anywhere other than at the differentiated membrane. During periods of elevated transmitter release, although the absolute number of vesicles in contact with the differentiated membrane decreased, the percentage of total vesicles in contact with the differentiated membrane increased dramatically. This suggests that the differentiated membrane is the site of vesicle release and there is an active process of vesicle movement to this membrane. In the course of this work it was observed that presynaptic areas closer than approximately 2 mm to the site of axonal transection, regardless of the composition of the saline or the experimental conditions, showed ultrastructural changes typical of increased transmitter release.
