mini-INTACT reveals social isolation-induced epigenetic changes in Drosophila dopaminergic neurons. mini-INTACT reveals social isolation-induced epigenetic changes in Drosophila dopaminergic neurons

We introduce a more sensitive method for purifying specific cell-types from small numbers of fly heads for epigenomic analysis: a method we call ‘mini-INTACT’. We modified the INTACT method in Drosophila [Henry G. L. et al., Nucleic Acids Research, 2012, 1–14] to enable purification of genetically tagged rare cell types from Drosophila brain requiring ~50-100 fold less material. We used male fruit flies as a model to examine the effects of social-isolation vs. social-enrichment on the epigenome and transcriptome of dopaminergic neurons defined by expression of the TH-GAL4 driver. Using ChIP-seq on mini-INTACT purified nuclei, RNA-seq and bioinformatic analysis, we identify regulation of the transcriptome and behavior by Activity Related Genes who’s expression is induced by social enrichment. Overall design: Genome wide occupancy profiling for histone modification marks by high throughput sequencing. Two or three biological replicates were used for each condition. Corresponding input DNA was used as control (ChIP-seq). Expression profiling by high throughput sequencing on FACS purified dopaminergic neurons isolated from group-housed and single-housed fruit fly adult male brains. Three biological replicates were used for each condition (RNA-seq).