Organization, sequence and functional analysis of a mouse BDNF promoter.

To examine the content of the 5' flanking region of the mouse BDNF gene a mouse library was screened using oligonucleotides corresponding to the rat exon I untranslated region. A 6-kb genomic fragment containing exons I and II and flanking regions was isolated and sequenced. The structure of the 5' end of the mouse gene is similar to that of rat, exons I and II are 2 small untranslated regions clustered within 500 bp of each other at the 5' end of the gene. The nucleotide sequence homology between rat and mouse is 93%. Analysis for transcription factor-binding sites show a predominance of AP1 and C/EBP elements which are conserved between the 2 species. Deleted fragments of the 5' flanking region of exons I and II were fused to the luciferase reporter gene and transcriptional activity was analyzed by transient expression in primary cortico-hippocampal cultures. We found that a fragment of 266 bp from exon I transcription start is sufficient for promoter activity in basal conditions. Following experimental stimulation by treatment with kainic acid, we determined that regulatory elements responsive to kainic acid are located within 989 bp of the transcriptional start of exon I.