Deletion of synapsins I and II genes alters the size of vesicular pools and rabphilin phosphorylation.

Previous studies established that genetic deletion of synapsins, synaptic vesicle-associated phosphoproteins that regulate neurotransmitter release, decreases the number of synaptic vesicles in nerve terminals. To investigate whether these changes affect the release properties of the remaining synaptic vesicles, we used a radioactive labeling technique to measure release independently of the total number of synaptic vesicles. 3H-glutamate and 14C-gamma-amino-butyric-acid (GABA) release from isolated nerve terminals prepared from the neocortex of synapsins I and II double knock-out mice (DKO) was assayed and compared to wild-type preparations. Hyperosmotic shock-evoked 3H-glutamate was reduced by 20+/-3% from DKO nerve terminals and potassium depolarization-evoked glutamate release was also decreased by 28+/-2%. Surprisingly, sucrose or potassium depolarization-evoked release of 14C-GABA was increased by 32+/-4% and 29+/-5%, respectively. The basal efflux of both 3H-glutamate and 14C-GABA increased by 17+/-2% and 12+/-2% from DKO nerve terminals. As lack of synapsins I and II, major phosphoproteins of synaptic vesicles, may lead to deregulation of phosphorylation events, we compared phosphorylation state of another synaptic vesicle protein, rabphilin. In DKO nerve terminals, membrane-associated rabphilin level was reduced by approximately 0.28-fold, its phosphorylation at 234serine was increased by approximately 1.61-fold whereas cytosolic rabphilin levels showed both more dramatic reduction in abundance, approximately 16.5-fold, and increase in phosphorylation, approximately 4.8-fold. Collectively, these data suggest that deletion of major synapsin isoforms leads to (1) deregulation of basal neurotransmission causing "leaky" basal release, (2) changes in either the size or mobilization of releasable or reserve pools, and (3) a decrease in rabphilin abundance accompanied by an increase in basal phosphorylation of the remaining rabphilin.