A conditional strategy for cell-type-specific labeling of endogenous excitatory synapses in Drosophila

Cell Rep Methods. 2023 May 11;3(5):100477. doi: 10.1016/j.crmeth.2023.100477. eCollection 2023 May 22.

Abstract

Chemical neurotransmission occurs at specialized contacts where neurotransmitter release machinery apposes neurotransmitter receptors to underlie circuit function. A series of complex events underlies pre- and postsynaptic protein recruitment to neuronal connections. To better study synaptic development in individual neurons, we need cell-type-specific strategies to visualize endogenous synaptic proteins. Although presynaptic strategies exist, postsynaptic proteins remain less studied because of a paucity of cell-type-specific reagents. To study excitatory postsynapses with cell-type specificity, we engineered dlg1[4K], a conditionally labeled marker of Drosophila excitatory postsynaptic densities. With binary expression systems, dlg1[4K] labels central and peripheral postsynapses in larvae and adults. Using dlg1[4K], we find that distinct rules govern postsynaptic organization in adult neurons, multiple binary expression systems can concurrently label pre- and postsynapse in a cell-type-specific manner, and neuronal DLG1 can sometimes localize presynaptically. These results validate our strategy for conditional postsynaptic labeling and demonstrate principles of synaptic organization.

Keywords: CNS; Drosophila; PNS; PSD95; circuit; conditional epitope tag; dlg1; olfactory system; postsynaptic; synapse.

MeSH terms

  • Animals
  • Drosophila*
  • Neurons / physiology
  • Receptors, Neurotransmitter / metabolism
  • Synapses* / metabolism
  • Synaptic Transmission / physiology

Substances

  • Receptors, Neurotransmitter