Effect of bromodomain inhibitors on dopamine D1R induced gene expression in striatal neurons

The dopamine D1R is Gs coupled GPCR which is expressed in striatal neurons and stimulates gene expression upon activation by dopamine or other agonists. In this work we investigated the role of the BET protein bromodomain containing protein 4 (Brd4) in mediating D1R-dependent gene expression in rat striatal neurons. Here we report the results of 3 RNA sequencing experiments. In Experiment 1 we treated primary striatal neurons with the D1R-selective agonist SKF-81297 alone or in combination with the non-selective BET inhibitor JQ1 for 60 minutes to assess the effect of BET inhibition on acute D1R-dependent gene expression. In Experiment 2, we treated primary striatal neurons with the D1R-selective agonist SKF-81297 alone or in combination with the BET bromodomain 2 selective inhibitor iBD2 for 60 minutes to determine the contribution of BD2 specifically to D1R-dependent gene expression. In Experiment 3 we treated primary striatal neurons with the D1R-selective agonist SKF-81297 alone or in combination with the non-selective BET inhibitor JQ1 for 24 hours to assess the effect of prolonged BET inhibition on basal gene expression and D1R-dependent gene expression. RNA sequencing of striatal cultures treated with varying combinations of D1R agonist and either JQ1 or iBD2, for either 60 minutes or 24 hours.