HrT is required for cardiovascular development in zebrafish.
The recently identified zebrafish T-box gene hrT is expressed in the developing heart and in the endothelial cells forming the dorsal aorta. Orthologs of hrT are expressed in cardiovascular cells from Drosophila to mouse, suggesting that the function of hrT is evolutionarily conserved. The role of hrT in cardiovascular development, however, has not thus far been determined in any animal model. Using morpholino antisense oligonucleotides, we show that zebrafish embryos lacking hrT function have dysmorphic hearts and an absence of blood circulation. Although the early events in heart formation were normal in hrT morphant embryos, subsequently the hearts failed to undergo looping, and late onset defects in chamber morphology and gene expression were observed. In particular, we found that the loss of hrT function led to a dramatic upregulation of tbx5, a gene required for normal heart morphogenesis. Conversely, we show that overexpression of hrT causes a significant downregulation of tbx5, indicating that one key role of hrT is to regulate the levels of tbx5. Secondly, we found that HrT is required to inhibit the expression of the blood lineage markers gata1 and gata2 in the most posterior lateral plate mesoderm. Finally, we show that HrT is required for vasculogenesis in the trunk, leading to similar vascular defects to those observed in midline mutants such as floating head. hrT expression in the vascular progenitors depends upon midline mesoderm, indicating that this expression is one important component of the response to a midline-derived signal during vascular morphogenesis.
Szeto
Daniel P
DP
Department of Biochemistry, Box 357350, University of Washington, Seattle 98195-7530, USA.
Griffin
Kevin J P
KJ
Kimelman
David
D
D000818:Animals; D002319:Cardiovascular System / Q000196:embryology*; D018507:Gene Expression Regulation, Developmental; D006330:Heart Defects, Congenital / Q000196:embryology / Q000235:genetics; D006410:Hematopoiesis / Q000235:genetics / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics / Q000502:physiology; D006801:Humans; D017403:In Situ Hybridization; D051379:Mice; D009154:Mutation; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics; D020825:T-Box Domain Proteins / Q000235:genetics* / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*; D029961:Zebrafish Proteins
D018398:Homeodomain Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D011993:Recombinant Fusion Proteins; D020825:T-Box Domain Proteins; C103172:T-box transcription factor 5; C420984:TBX20 protein, human; C486013:Tbx20 protein, mouse; D014157:Transcription Factors; D029961:Zebrafish Proteins; C096660:noto protein, zebrafish; C486014:tbx20 protein, zebrafish