Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation.
We have developed a simple approach for large-scale transgenesis in Xenopus laevis embryos and have used this method to identify in vivo requirements for FGF signaling during gastrulation. Plasmids are introduced into decondensed sperm nuclei in vitro using restriction enzyme-mediated integration (REMI). Transplantation of these nuclei into unfertilized eggs yields hundreds of normal, diploid embryos per day which develop to advanced stages and express integrated plasmids nonmosaically. Transgenic expression of a dominant negative mutant of the FGF receptor (XFD) after the mid-blastula stage uncouples mesoderm induction, which is normal, from maintenance of mesodermal markers, which is lost during gastrulation. By contrast, embryos expressing XFD contain well-patterned nervous systems despite a putative role for FGF in neural induction.
Kroll
K L
KL
Department of Cell and Molecular Biology, University of California at Berkeley, 94720, USA. kkroll@bcmp.med.harvard.edu
Amaya
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D000818:Animals; D030801:Animals, Genetically Modified; D002454:Cell Differentiation; D002467:Cell Nucleus; D017690:Cell Transplantation; D004171:Diploidy; D005260:Female; D005775:Gastrula / Q000378:metabolism / Q000502:physiology*; D017353:Gene Deletion; D008297:Male; D008648:Mesoderm / Q000502:physiology; D008845:Microinjections; D012313:RNA; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D013094:Spermatozoa; D014982:Xenopus laevis / Q000196:embryology* / Q000378:metabolism / Q000502:physiology
D017468:Receptors, Fibroblast Growth Factor; D012313:RNA