Both 5' and 3' flanks regulate Zebrafish brain-derived neurotrophic factor gene expression.
BACKGROUND
Precise control of developmental and cell-specific expression of the brain-derived neurotrophic factor (BDNF) gene is essential for normal neuronal development and the diverse functions of BDNF in the adult organism. We previously showed that the zebrafish BDNF gene has multiple promoters. The complexity of the promoter structure and the mechanisms that mediate developmental and cell-specific expression are still incompletely understood.


RESULTS
Comparison of pufferfish and zebrafish BDNF gene sequences as well as 5' RACE revealed three additional 5' exons and associated promoters. RT-PCR with exon-specific primers showed differential developmental and organ-specific expression. Two exons were detected in the embryo before transcription starts. Of the adult organs examined, the heart expressed a single 5' exon whereas the brain, liver and eyes expressed four of the seven 5' exons. Three of the seven 5' exons were not detectable by RT-PCR. Injection of promoter/GFP constructs into embryos revealed distinct expression patterns. The 3' flank profoundly affected expression in a position-dependent manner and a highly conserved sequence (HCS1) present in 5' exon 1c in a dehancer-like manner.


CONCLUSIONS
The zebrafish BDNF gene is as complex in its promoter structure and patterns of differential promoter expression as is its murine counterpart. The expression of two of the promoters appears to be regulated in a temporally and/or spatially highly circumscribed fashion. The 3' flank has a position-dependent effect on expression, either by affecting transcription termination or post-transcriptional steps. HCS1, a highly conserved sequence in 5' exon 1c, restricts expression to primary sensory neurons. The tools are now available for detailed genetic and molecular analyses of zebrafish BDNF gene expression.
Heinrich
Gerhard
G
Medical Service, VA Northern California Health Care System, Martinez, CA 94553, USA. gheinrich@ucdavis.ed
Pagtakhan
Carl John
CJ
D024509:3' Flanking Region / Q000235:genetics / Q000502:physiology*; D024506:5' Flanking Region / Q000235:genetics / Q000502:physiology*; D017398:Alternative Splicing; D000818:Animals; D030801:Animals, Genetically Modified; D019208:Brain-Derived Neurotrophic Factor / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D004625:Embryo, Nonmammalian; D005091:Exons / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017385:Sequence Homology; D023721:Tetraodontiformes / Q000235:genetics; D015027:Zebrafish / Q000235:genetics*
D019208:Brain-Derived Neurotrophic Factor; D011994:Recombinant Proteins