Brown Christopher B, Wenning Jennifer M, Lu Min Min, Epstein Douglas J, Meyers Erik N, Epstein Jonathan A
Cardiovascular Division, Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Dev Biol. 2004 Mar 1;267(1):190-202. doi: 10.1016/j.ydbio.2003.10.024.
Tbx1 has been implicated as a candidate gene responsible for defective pharyngeal arch remodeling in DiGeorge/Velocardiofacial syndrome. Tbx1(+/-) mice mimic aspects of the DiGeorge phenotype with variable penetrance, and null mice display severe pharyngeal hypoplasia. Here, we identify enhancer elements in the Tbx1 gene that are conserved through evolution and mediate tissue-specific expression. We describe the generation of transgenic mice that utilize these enhancer elements to direct Cre recombinase expression in endogenous Tbx1 expression domains. We use these Tbx1-Cre mice to fate map Tbx1-expressing precursors and identify broad regions of mesoderm, including early cardiac mesoderm, which are derived from Tbx1-expressing cells. We test the hypothesis that fibroblast growth factor 8 (Fgf8) functions downstream of Tbx1 by performing tissue-specific inactivation of Fgf8 using Tbx1-Cre mice. Resulting newborn mice display DiGeorge-like congenital cardiovascular defects that involve the outflow tract of the heart. Vascular smooth muscle differentiation in the great vessels is disrupted. This data is consistent with a model in which Tbx1 induces Fgf8 expression in the pharyngeal endoderm, which is subsequently required for normal cardiovascular morphogenesis and smooth muscle differentiation in the aorta and pulmonary artery.
Tbx1被认为是与DiGeorge/心脏颜面综合征中咽弓重塑缺陷相关的候选基因。Tbx1(+/-)小鼠模拟了DiGeorge表型的某些方面,且具有可变的外显率,而基因敲除小鼠则表现出严重的咽发育不全。在此,我们鉴定了Tbx1基因中通过进化保守且介导组织特异性表达的增强子元件。我们描述了利用这些增强子元件来指导Cre重组酶在Tbx1内源性表达结构域中表达的转基因小鼠的产生。我们使用这些Tbx1-Cre小鼠来追踪表达Tbx1的前体细胞的命运,并确定中胚层的广泛区域,包括早期心脏中胚层,这些区域源自表达Tbx1的细胞。我们通过使用Tbx1-Cre小鼠对Fgf8进行组织特异性失活,来检验成纤维细胞生长因子8(Fgf8)在Tbx1下游发挥作用的假说。所产生的新生小鼠表现出类似DiGeorge的先天性心血管缺陷,累及心脏流出道。大血管中的血管平滑肌分化受到破坏。这些数据与一个模型一致,即Tbx1在咽内胚层中诱导Fgf8表达,而这随后是主动脉和肺动脉正常心血管形态发生和平滑肌分化所必需的。