Isolation of differentially expressed genes from wild-type and Twist mutant mouse limb buds

In the mouse, Twist is required for normal limb and craniofacial development. We show that the aristaless‐like transcription factors, Alx3 and Alx4 are downregulated in the Twist−/− mutant and may be potential targets of Twist. By suppression subtractive hybridization we isolated 31 and 18 unique cl...

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Bibliographic Details
Published in:Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2002-07, Vol.33 (3), p.103-113
Main Authors: Loebel, David A.F., O'Rourke, Meredith P., Steiner, Kirsten A., Banyer, Joanne, Tam, Patrick P.L.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
bHLH transcription factor
Conserved Sequence
DNA-Binding Proteins
Down-Regulation
Gene Expression Profiling
Gene Expression Regulation, Developmental
Homeodomain Proteins - genetics
In Situ Hybridization
Limb Buds - embryology
Limb Buds - metabolism
limb development
Mice
Molecular Sequence Data
Mutation - genetics
Nuclear Proteins - genetics
Proteins - genetics
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger - genetics
RNA, Messenger - metabolism
Sequence Homology, Nucleic Acid
suppression-subtractive hybridisation
Transcription Factors
Twist
Twist-Related Protein 1
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Summary:In the mouse, Twist is required for normal limb and craniofacial development. We show that the aristaless‐like transcription factors, Alx3 and Alx4 are downregulated in the Twist−/− mutant and may be potential targets of Twist. By suppression subtractive hybridization we isolated 31 and 18 unique clones representing mRNAs that are putatively downregulated and upregulated respectively in Twist−/− forelimb buds. These included genes encoding cytoskeletal components, metabolic enzymes, hemoglobin molecules, membrane transport proteins, components of transcription and translation complexes, protein modification enzymes and proteins related to cell proliferation and apoptosis. Differential expression of selected clones was validated by whole mount in situ hybridization to E10.5 wild‐type and Twist−/− embryos. We show that four novel clones are expressed in the Twist‐expressing craniofacial tissues and paraxial mesoderm and downregulated in Twist−/− embryos, raising the possibility that they are, in addition to genes of the Alx family, downstream targets of Twist. genesis 33:103–113, 2002. © 2002 Wiley‐Liss, Inc.
ISSN:1526-954X
1526-968X
DOI:10.1002/gene.10091