Candidate EDA targets revealed by expression profiling of primary keratinocytes from Tabby mutant mice.
EDA, the gene mutated in anhidrotic ectodermal dysplasia, encodes ectodysplasin, a TNF superfamily member that activates NF-kB mediated transcription. To identify EDA target genes, we have earlier used expression profiling to infer genes differentially expressed at various developmental time points in Tabby (Eda-deficient) compared to wild-type mouse skin. To increase the resolution to find genes whose expression may be restricted to epidermal cells, we have now extended studies to primary keratinocyte cultures established from E19 wild-type and Tabby skin. Using microarrays bearing 44,000 gene probes, we found 385 preliminary candidate genes whose expression was significantly affected by Eda loss. By comparing expression profiles to those from Eda-A1 transgenic skin, we restricted the list to 38 "candidate EDA targets", 14 of which were already known to be expressed in hair follicles or epidermis. We confirmed expression changes for 3 selected genes, Tbx1, Bmp7, and Jag1, both in keratinocytes and in whole skin, by Q-PCR and Western blotting analyses. Thus, by the analysis of keratinocytes, novel candidate pathways downstream of EDA were detected.
Laboratory of Genetics, National Institute on Aging, NIH Biomedical Research Center, 251 Bayview Boulevard, Suite 100, Baltimore, MD 21224, USA.
Diana
D
Esibizione
Chang-Yi
CY
Cui
David
D
Schlessinger
D000818:Animals; D055419:Bone Morphogenetic Protein 7 / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D053331:Ectodysplasins / Q000096:biosynthesis / Q000235:genetics*; D004817:Epidermis / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D018859:Hair Follicle / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D000072100:Jagged-1 Protein; D015603:Keratinocytes / Q000166:cytology*; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009154:Mutation; D016328:NF-kappa B / Q000378:metabolism; D000072099:Serrate-Jagged Proteins; D012867:Skin / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000378:metabolism
D055419:Bone Morphogenetic Protein 7; D002135:Calcium-Binding Proteins; D053331:Ectodysplasins; C506019:Eda protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; C000606274:Jag1 protein, mouse; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D016328:NF-kappa B; D000072099:Serrate-Jagged Proteins; D020825:T-Box Domain Proteins; C491573:Tbx1 protein, mouse; C525820:bmp7 protein, mouse