Neuronal activity-induced late-onset alternative splicing and H3K9me3 modification. Mus musculus

Activity-induced alternative splicing has emerged as an important mechanism for the dynamic modification of neuronal function. To examine whether activity could cause transcriptional programs alteration in alternative splicing and associative H3K9me3 in the specific genomic loci, we performed whole genome RNA-sequencing (RNA-seq) in cultured cortical neurons at 24 hours after membrane depolarization by exposure to high extracellular KCl (50mM, 10min). A transient neuronal activation could induce alternation of splice isoform choice in many genes at 24hrs after stimuli. To detect the correlation of activity-induced H3K9me3 accumulation with the splicing events, we performed ChIP-seq in hippocmapus of mice, which were subjected to enriched environment for 24 hours and kept in homecage for 1 week. Overall design: Total RNA from primary cortical neurons at 24 hr after KCl (50mM, 10min) stimulation was used for RNA-seq to detect activity induced alternative splicing events. To detect neural activation-induced long-term alternations on H3K9me3 in neurons, H3K9me3 ChIP-seq was performed in mice hippocampal extracts at 1 week after enriched enrironment training.