Doc2b Ca²⁺ binding site mutants enhance synaptic release at rest at the expense of sustained synaptic strength.

ABSTRACT: Communication between neurons involves presynaptic neurotransmitter release which can be evoked by action potentials or occur spontaneously as a result of stochastic vesicle fusion. The Ca²⁺-binding double C₂ proteins Doc2a and -b were implicated in spontaneous and asynchronous evoked release, but the mechanism remains unclear. Here, we compared wildtype Doc2b with two Ca²⁺ binding site mutants named DN and 6A, previously classified as gain- and loss-of-function mutants. They carry the substitutions D218,220N or D163,218,220,303,357,359A respectively. We found that both mutants bound phospholipids at low Ca²⁺ concentrations and were membrane-associated in resting neurons, thus mimicking a Ca²⁺-activated state. Their overexpression in hippocampal primary cultured neurons had similar effects on spontaneous and evoked release, inducing high mEPSC frequencies and increased short-term depression. Together, these data suggest that the DN and 6A mutants both act as gain-of-function mutants at resting conditions.

SUBMITTER: Bourgeois-Jaarsma Q 

PROVIDER: S-EPMC6783474 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature