Multiple N-cadherin enhancers identified by systematic functional screening indicate its Group B1 SOX-dependent regulation in neural and placodal development

Neural plate and sensory placodes share the expression of N-cadherin and Group B1 Sox genes, represented by Sox2. A 219-kb region of the chicken genome centered by the N-cadherin gene was scanned for neural and placodal enhancers. Random subfragments of 4.5 kb average length were prepared and insert...

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Bibliographic Details
Published in:Developmental biology 2005-10, Vol.286 (2), p.601-617
Main Authors: Matsumata, Miho, Uchikawa, Masanori, Kamachi, Yusuke, Kondoh, Hisato
Format: Article
Language:English
Subjects:
Animals
Binding Sites
Cadherins - genetics
Chick Embryo
Chickens
Conserved sequence
DNA-Binding Proteins - genetics
Ectoderm - cytology
Electroporation
Enhancer Elements, Genetic
Enhancers
Genome scan
Group B1 Sox
HMGB Proteins - genetics
Lens, Crystalline - embryology
N-cadherin
Prosencephalon - embryology
Prosencephalon - growth & development
Retina - embryology
Sox2
SOXB1 Transcription Factors
tkEGFP
Transcription Factors - genetics
Transfection
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Summary:Neural plate and sensory placodes share the expression of N-cadherin and Group B1 Sox genes, represented by Sox2. A 219-kb region of the chicken genome centered by the N-cadherin gene was scanned for neural and placodal enhancers. Random subfragments of 4.5 kb average length were prepared and inserted into tkEGFP reporter vector to construct a library with threefold coverage of the region. Each clone was then transfected into N-cadherin-positive (lens, retina and forebrain) or -negative embryonic cells, or electroporated into early chicken embryos to examine enhancer activity. Enhancers 1–4 active in the CNS/placode derivatives and non-specific Enhancer 5 were identified by transfection, while electroporation of early embryos confirmed enhancers 2–4 as having activity in the early CNS and/or sensory placodes but with unique spatiotemporal specificities. Enhancers 2–4 are dependent on SOX-binding sites, and misexpression of Group B1 Sox genes in the head ectoderm caused ectopic development of placodes expressing N-cadherin, indicating the involvement of Group B1 Sox functions in N-cadherin regulation. Enhancers 1, 2 and 4 correspond to sequence blocks conserved between the chicken and mammalian genomes, but enhancers 3 and 5 are unique to the chicken.
ISSN:0012-1606
1095-564X
DOI:10.1016/j.ydbio.2005.08.005