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

Cell reports methods (2023-06-16)
Michael J Parisi, Michael A Aimino, Timothy J Mosca
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.

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Sigma-Aldrich
Anti-α-tubulina monoclonale, clone DM1A, ascites fluid
Sigma-Aldrich
Anticorpo anti-α-tubulina, monoclonale murino, clone DM1A, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-V5 tag antibody produced in mouse, clone GT1071, affinity isolated antibody