Identification of genes expressed in the mouse limb using a novel ZPA microarray approach.
One well-studied signaling center in the developing vertebrate limb, the zone of polarizing activity (ZPA), produces the morphogen sonic hedgehog that is necessary for normal growth and pattern formation. To identify additional factors expressed in the ZPA of the mouse limb bud, the Shhgfpcre allele was used to purify ZPA cells using fluorescence-activated cell sorting. Microarray technology was then used to identify genes whose expression was elevated in the ZPA compared to the rest of the limb. In situ hybridization confirmed the expression of two known transcription factors, Hlxb9 and Tcfap2b, an uncharacterized EST, and a transmembrane protein of unknown function in domains overlapping the ZPA. The expression of two other genes was confirmed by rtPCR. The methods described in this report will be applicable for identifying genes enriched in Shh-expressing cells throughout development.
Rock
Jason R
JR
Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Cancer and Genetics Research Complex, 1376 Mowry Road, Gainesville, FL 32610, United States.
Lopez
M Cecilia
MC
Baker
Henry V
HV
Harfe
Brian D
BD
D000818:Animals; D005121:Extremities / Q000196:embryology / Q000254:growth & development*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D053823:Hedgehog Proteins / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D017403:In Situ Hybridization; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D051379:Mice; D050656:Transcription Factor AP-2 / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*
D053823:Hedgehog Proteins; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; C509239:SHH protein, human; C493669:Tfap2b protein, mouse; D050656:Transcription Factor AP-2; D014157:Transcription Factors; C084059:Hb9 protein, mouse