experiment in midbrain primary cultures, enriched in dopaminergic neurons

To identify novel Nurr1 target genes we have used microarrays strategies in rat midbrain primary cultures, enriched in dopaminergic neurons, by the action of basic fibroblast growth factor (bFGF, 20ng/ml) and Sonic hedgedog (SHH), following upregulation of Nurr1 expression by depolarization.To this aim we have treated the cultures after 9 days in vitro for 2h with high KCl and collected 30 min or 2 h after the end of depolarization (2h + 30 min or 2h + 2h). With this experimental protocol we have identify a putative Nurr1 regulator and Nurr1 target Keywords: depolarization, microarrays, time course. cRNA triplicates of cultures treated for 2h with high KCl and collected 30 min or 2 h after the end of depolarization (2h + 30 min or 2h + 2h) and control was hybridized on triplicate microarrays. The extrapolation of expression values from the images was carried out using different statistical filters and were compared up- and down-regulated probesets showing significant change between depolarized and control samples after 30 min or 2 h after the end of depolarization.