A comparative transcriptomics approach unveils networks of activity-driven neuronal gene expression

Primary cultured hippocampal neurons were infected with lentiviral particles bearing constitutively active versions of the transcription factors CREB, SRF, EGR1 and FOS to define the downstream genetic program associated with each of these paradigmatic transcription factors in neurons. For CREB, we also extracted samples at a shorter infection time to try to dissect the different components of the wider genetic response observed upon expression of a constitutively active version of this transcription factor. In order to further explore the contribution of these transcriptional regulators to activity-driven gene expression, we also treated cultured hippocampal neurons infected with GFP-only expressing lentiviral particles with different drugs (bicuculline, forskokine, and BDNF) and profiled the associated transcriptional response. Finally, in order to further explore the role of CREB in forskolin-mediated gene expression, we also used A-CREB, a dominant negative form of this transcription factor. Specifically, we stimulated neurons with forskolin in the presence or absence of lentiviral particles expressing A-CREB and carried out genome-wide transcriptional profiling.