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Scientific article
English

RNA-seq Profiling of Small Numbers of Drosophila Neurons

Published inMethods in enzymology, vol. 551, p. 369-386
Publication date2015
Abstract

Drosophila melanogaster has a robust circadian clock, which drives a rhythmic behavior pattern: locomotor activity increases in the morning shortly before lights on (M peak) and in the evening shortly before lights off (E peak). This pattern is controlled by ~75 pairs of circadian neurons in the Drosophila brain. One key group of neurons is the M-cells (PDF(+) large and small LNvs), which control the M peak. A second key group is the E-cells, consisting of four LNds and the fifth small LNv, which control the E peak. Recent studies show that the M-cells have a second role in addition to controlling the M peak; they communicate with the E-cells (as well as DN1s) to affect their timing, probably as a function of environmental conditions (Guo, Cerullo, Chen, & Rosbash, 2014). To learn about molecules within the M-cells important for their functional roles, we have adapted methods to manually sort fluorescent protein-expressing neurons of interest from dissociated Drosophila brains. We isolated mRNA and miRNA from sorted M-cells and amplified the resulting DNAs to create deep-sequencing libraries. Visual inspection of the libraries illustrates that they are specific to a particular neuronal subgroup; M-cell libraries contain timeless and dopaminergic cell libraries contain ple/TH. Using these data, it is possible to identify cycling transcripts as well as many mRNAs and miRNAs specific to or enriched in particular groups of neurons.

eng
Keywords
  • Cell sorting
  • Cycling
  • Morning cells
  • RNA-seq
  • MiRNAs
Citation (ISO format)
ABRUZZI, Katharine et al. RNA-seq Profiling of Small Numbers of <i>Drosophila</i> Neurons. In: Methods in enzymology, 2015, vol. 551, p. 369–386. doi: 10.1016/bs.mie.2014.10.025
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ISSN of the journal0076-6879
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