Abstract
Memory-relevant neuronal plasticity is believed to require local translation of new proteins at synapses. Understanding this process requires the visualization of the relevant mRNAs within these neuronal compartments. Here, we used single-molecule fluorescence in situ hybridization to localize mRNAs at subcellular resolution in the adult Drosophila brain. mRNAs for subunits of nicotinic acetylcholine receptors and kinases could be detected within the dendrites of co-labeled mushroom body output neurons (MBONs) and their relative abundance showed cell specificity. Moreover, aversive olfactory learning produced a transient increase in the level of CaMKII mRNA within the dendritic compartments of the γ5β'2a MBONs. Localization of specific mRNAs in MBONs before and after learning represents a critical step towards deciphering the role of dendritic translation in the neuronal plasticity underlying behavioral change in Drosophila.
Highlights
Memories are believed to be encoded as changes in the efficacy of specific synaptic connections
Single-molecule fluorescence in situ hybridization enables cellular mRNAs to be imaged at single-molecule resolution through the hybridization of a set of complementary oligonucleotide probes, each labeled with a fluorescent dye
Mammalian calmodulin-dependent protein kinase II (CaMKII) mRNA is transported to neuronal dendrites, where it is locally translated in response to neuronal activity (Bagni et al, 2000; Miller et al, 2002; Ouyang et al, 1999)
Summary
Memories are believed to be encoded as changes in the efficacy of specific synaptic connections. We demonstrate localization of mRNAs in the 3D volumes of MBON dendrites by registering smFISH signals with co-labeled neurons using a custom image analysis pipeline. We find that aversive learning transiently elevates dendritic CaMKII transcript levels within g5b’2a MBONs. Results and discussion mRNA localization in the intact adult Drosophila brain
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