Screening of FGF target genes in Xenopus by microarray: temporal dissection of the signalling pathway using a chemical inhibitor.
Microarray is a powerful tool for analysing gene expression patterns in genome-wide view and has greatly contributed to our understanding of spatiotemporal embryonic development at the molecular level. Members of FGF (fibroblast growth factor) family play important roles in embryogenesis, e.g. in organogenesis, proliferation, differentiation, cell migration, angiogenesis, and wound healing. To dissect spatiotemporally the versatile roles of FGF during embryogenesis, we profiled gene expression in Xenopus embryo explants treated with SU5402, a chemical inhibitor specific to FGF receptor 1 (FGFR1), by microarray. We identified 38 genes that were down-regulated and 5 that were up-regulated in response to SU5402 treatment from stage 10.5-11.5 and confirmed their FGF-dependent transcription with RT-PCR analysis and whole-mount in situ hybridization (WISH). Among the 43 genes, we identified 26 as encoding novel proteins and investigated their spatial expression pattern by WISH. Genes whose expression patterns were similar to FGFR1 were further analysed to test whether any of them represented functional FGF target molecules. Here, we report two interesting genes: one is a component of the canonical Ras-MAPK pathway, similar to mammalian mig6 (mitogen-inducible gene 6) acting in muscle differentiation; the other, similar to GPCR4 (G-protein coupled receptor 4), is a promising candidate for a gastrulation movement regulator. These results demonstrate that our approach is a promising strategy for scanning the genes that are essential for the regulation of a diverse array of developmental processes.
Chung
Hyeyoung A
HA
Department of Developmental Biology, National Institute for Basic Biology, Okazaki, 444-8585 Japan.
Hyodo-Miura
Junko
J
Kitayama
Atsushi
A
Terasaka
Chie
C
Nagamune
Teruyuki
T
Ueno
Naoto
N
D000818:Animals; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology / Q000187:drug effects / Q000378:metabolism; D005326:Fetal Proteins / Q000502:physiology; D005346:Fibroblast Growth Factors / Q000502:physiology*; D005775:Gastrula / Q000502:physiology; D020869:Gene Expression Profiling; D017403:In Situ Hybridization; D009132:Muscles / Q000196:embryology; D020411:Oligonucleotide Array Sequence Analysis; D011758:Pyrroles / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D051496:Receptor, Fibroblast Growth Factor, Type 1; D017468:Receptors, Fibroblast Growth Factor / Q000037:antagonists & inhibitors; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D020825:T-Box Domain Proteins / Q000502:physiology; D014981:Xenopus / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D029867:Xenopus Proteins / Q000235:genetics / Q000502:physiology*; D000098908:Brachyury Protein
D005326:Fetal Proteins; D005346:Fibroblast Growth Factors; D011758:Pyrroles; D012333:RNA, Messenger; D020794:Receptor Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; D017468:Receptors, Fibroblast Growth Factor; D043562:Receptors, G-Protein-Coupled; D020825:T-Box Domain Proteins; D029867:Xenopus Proteins; D000098908:Brachyury Protein; C490468:ERRFI1 protein, Xenopus; C105686:SU 5402; C490469:mgc69520 protein, Xenopus