Isolation of the chicken Lmbr1 coding sequence and characterization of its role during chick limb development.
In the developing amniote limb, anteroposterior (A/P) patterning is controlled through secretion of the Sonic Hedgehog (SHH) protein by cells in the zone of polarizing activity (ZPA) located in the posterior mesoderm. In the chicken mutant oligozeugodactyly (ozd), Shh is expressed normally in the entire embryo with the exception that it is undetectable in the developing limbs; this results in the loss of specific bones in wings and legs. The ozd phenotype is similar to that of humans affected with acheiropodia (ACHR), and the ACHR mutation has been mapped to a deletion of exon 4 and portions of introns 3 and 4 in the LMBR1 gene. We have cloned the chick ortholog of LMBR1, Lmbr1, and report that, in chick, Lmbr1 is expressed within the ZPA. Although the ozd phenotype is similar to ACHR, the open reading frame of Lmbr1 is normal in ozd. Sequence analysis of Lmbr1 intron 3 demonstrated that this particular genomic region segregates with the ozd phenotype. In addition, overexpression of Lmbr1 throughout the developing limb mesoderm resulted in morphologically normal limbs. Collectively, these data suggest that the Lmbr1 coding sequence is not required for normal chick limb development. We propose that the ozd mutation is linked to the genomic region containing Shh and Lmbr1.
Maas
Sarah A
SA
Department of Biomolecular Chemistry, University of Wisconsin, Madison, Wisconsin, USA.
Fallon
John F
JF
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D015152:Blotting, Northern; D002642:Chick Embryo; D003001:Cloning, Molecular; D018274:Electroporation; D020224:Expressed Sequence Tags; D005121:Extremities / Q000196:embryology*; D005822:Genetic Vectors; D016678:Genome; D053823:Hedgehog Proteins; D006801:Humans; D017403:In Situ Hybridization; D007438:Introns; D008565:Membrane Proteins / Q000096:biosynthesis*; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D012333:RNA, Messenger / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D015534:Trans-Activators / Q000378:metabolism
D053823:Hedgehog Proteins; C485955:LMBR1 protein, human; C417920:Lmbr1 protein, mouse; D008565:Membrane Proteins; D012333:RNA, Messenger; C509239:SHH protein, human; D015534:Trans-Activators