Identification of newt connective tissue growth factor as a target of retinoid regulation in limb blastemal cells.
In order to analyse target genes regulated by retinoic acid in urodele limb regeneration, we have used pseudotyped retroviruses to obtain stably transfected newt limb blastemal (progenitor) cells in culture which express chimeric retinoic acid/thyroid hormone receptors delta1 or delta2. After treatment with thyroid hormone to activate the chimeric receptors, we used a polymerase chain reaction (PCR)-based subtraction method to identify target genes which are retinoid regulated. Newt connective tissue growth factor, a secreted protein recognised in several vertebrates, has been identified in this way and found to be expressed in the limb blastema and regulated by retinoic acid. This approach should permit a systematic analysis of retinoid target genes in limb regeneration.
Cash
D E
DE
Department of Biochemistry and Molecular Biology, University College London, Gower Street, London WC1E 6BT, UK.
Gates
P B
PB
Imokawa
Y
Y
Brockes
J P
JP
D000818:Animals; D003001:Cloning, Molecular; D055513:Connective Tissue Growth Factor; D005121:Extremities / Q000254:growth & development; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006133:Growth Substances / Q000235:genetics*; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D009673:Notophthalmus viridescens / Q000235:genetics*; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism; D011988:Receptors, Thyroid Hormone / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012038:Regeneration / Q000235:genetics*; D012176:Retinoids / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D013234:Stem Cells; D014212:Tretinoin / Q000378:metabolism
D006133:Growth Substances; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D018168:Receptors, Retinoic Acid; D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins; D012176:Retinoids; D055513:Connective Tissue Growth Factor; D014212:Tretinoin