polyA+ RNA expression profiles of Drosophila melanogaster male and female brains after treatment with Sodium salicylate, Chlorpromazine, or Trifluoperazine2HCl

To explore some of the sex differential responses to drug treatment, we exposed 3 day post-eclosion virgin Drosophila melanogaster (genotype DGRP-900, FlyBase line ID: FBst0028261) to 3 FDA approved drugs and characterized transcript abundance among male and female brains after 24 hours of drug exposure. The 3 tested drugs are water-soluble group N (Nervous system) compounds according to The Anatomical Therapeutic Chemical (ATC) classification system. Two (Chlorpromazine and Trifluoperazine2HCl) are primarily used to treat psychotic disorders, while the third (Sodium salicylate) is used as an analgesic and antipyretic. Following drug treatment, we dissected fly brains, isolated polyA+ RNA and performed high-throughput RNA-Seq analysis. We conducted quantification and statistical analysis of gene expression difference between tested drugs, sexes and their interaction. Overall design: We analyzed 48 brain samples in 6 independent biological replicates with 8 samples each. Eight samples of each replicate included 2 sexes (female and male) in 4 different conditions: (1) control, (2) Chlorpromazine, (3) Trifluoperazine2HCl, (4) Sodium Salicylate. Replicates were performed on different days. Each sample was a pool of 5 individual brains. Brains were dissected from DGRP-900 virgin 3 days post-eclosion flies after 24 hours of drug exposure. We isolated total RNA from the samples, purified polyA+ mRNA, and generated multiplexed stranded polyA+ libraries. We added external RNA controls during library preparation to quality check and performed 75bp single end sequencing on Illumina Hiseq 2500.