Comprehensive identification of the Pet-1-target genes by chromatin immunoprecipitation assays in RN46A cells derived from the rat raphe nucleus.

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To identify Pet-1 (serotonergic neuron-specific transcription factor)-binding sites in rat raphe nucleus-derived RN46A cells, chromatin-immunoprecipitation (ChIP) assays were performed. It was revealed that Pet-1 binding was inhibited through the neuron-restrictive silencer factor(NRSF)-NRSF binding element(NRSE) system or the potential repressor proteins independent of NRSF. Additionally, the detection of Pet-1 binding to target genes was occasionally difficult even under conditions where Pet-1 was overexpressed. Taken together, to exclude the function of repressor proteins including NRSF may be necessary to identify the Pet-1 binding to its target genes by ChIP assays in RN46A cells. To overcome these difficulties should be required to use RN46A cells as a proper model of central serotonergic neurons and characterize the Pet-1-mediated transcriptional network in serotonergic neurons.