Maternal VegT is the initiator of a molecular network specifying endoderm in Xenopus laevis.
During cleavage stages, maternal VegT mRNA and protein are localized to the Xenopus embryo's vegetal region from which the endoderm will arise and where several zygotic gene transcripts will be localized. Previous loss-of-function experiments on this T-box transcription factor suggested a role for VegT in Xenopus endoderm formation. Here, we test whether VegT is required to initiate endoderm formation using a loss of function approach. We find that the endodermal genes, Bix1, Bix3, Bix4, Milk (Bix2), Mix.1, Mix.2, Mixer, Xsox17 alpha, Gata4, Gata5, Gata6 and endodermin, as well as the anterior endodermal genes Xhex and cerberus, and the organizer specific gene, Xlim1, are downstream of maternal VegT. We also find that the TGF beta s, Xnr1, Xnr2, Xnr4 and derrière rescue expression of these genes, supporting the idea that cell interactions are critical for proper endoderm formation. Additionally, inhibitory forms of Xnr2 and Derrière blocked the ability of VegT mRNA injection to rescue VegT-depleted embryos. Furthermore, a subset of endodermal genes was rescued in VegT-depleted vegetal masses by induction from an uninjected vegetal mass. Finally, we begin to establish a gene hierarchy downstream of VegT by testing the ability of Mixer and Gata5 to rescue the expression of other endodermal genes. These results identify VegT as the maternal regulator of endoderm initiation and illustrate the complexity of zygotic pathways activated by VegT in the embryo's vegetal region.
Xanthos
J B
JB
Department of Genetics, Cell Biology, and Development, University of Minnesota, 321 Church Street SE, Minneapolis, MN 55455, USA.
Kofron
M
M
Wylie
C
C
Heasman
J
J
D000818:Animals; D001483:Base Sequence; D002450:Cell Communication; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D004707:Endoderm / Q000166:cytology* / Q000378:metabolism; D005260:Female; D050992:GATA5 Transcription Factor; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D020825:T-Box Domain Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000235:genetics; D029867:Xenopus Proteins; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics / Q000502:physiology
D017931:DNA Primers; D004268:DNA-Binding Proteins; C494752:GATA-5a protein, Xenopus; C494753:GATA-5b protein, Xenopus; D050992:GATA5 Transcription Factor; D018398:Homeodomain Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D020825:T-Box Domain Proteins; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; C103966:VegT protein, Xenopus; D029867:Xenopus Proteins; C405705:mixer protein, Xenopus