Assessing the contributions of gene products to the form-shaping events of neurulation: a transgenic approach in chick.
Most of our current knowledge on the tissue and cellular basis of neurulation in amniotes has been gained using the chick embryo as an experimental model system. Gene manipulation during chick neurulation has been difficult, greatly limiting our ability to assess the contribution of gene products to the tissue and cellular behaviors of neurulation. Using electroporation, we have developed a simple and reliable method for expressing transgenes in the ectoderm of the neural folds of chick embryos developing in whole-embryo culture. Sense- or antisense-expressing plasmids are electroporated, resulting in gain or loss of gene function, respectively. The morphogenesis of transgenic tissues was compared to the morphogenesis of contralateral wildtype tissues as neurulation was taking place. As a proof of principle, we present a functional analysis of the chick gene encoding Cartilage Linking Protein 1 (CRTL1), identified as a candidate neurulation gene using subtractive hybridization. This experimental approach provides a much-needed innovation for studying the mechanisms by which genes influence neurulation and reveals here important contributions of CRTL1 to the formation of the neural folds.
Colas
Jean-François
JF
Department of Neurobiology and Anatomy, and Children's Health Research Center, University of Utah School of Medicine, Salt Lake City, UT, USA. jfcolas@snv.jussieu.fr
Schoenwolf
Gary C
GC
D000818:Animals; D019521:Body Patterning / Q000235:genetics; D018929:Cell Culture Techniques; D002642:Chick Embryo; D018274:Electroporation; D004627:Embryonic Induction; D016326:Extracellular Matrix Proteins; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D009024:Morphogenesis; D009420:Nervous System / Q000196:embryology*; D010957:Plasmids; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011509:Proteoglycans; D019076:Transgenes; D000099249:Proteoglycan Link Protein
D016326:Extracellular Matrix Proteins; D011506:Proteins; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein