The purpose of this research is to understand the role Tbx6 plays in the determination of left/right body axis asymmetry of the mouse embryo. Mutations in genes that regulate body axis asymmetry lead to congenital cardiac malformations as well as other serious health issues related to organ situs. Tbx6 homozygous mutant embryos demonstrate abnormalities in heart looping, embryo turning and vitelline vessel placement at midgestation, all of which are key morphological aspects of left/right asymmetry leads to the hypothesis that Tbx6 plays an important role in the initial events that usher in the breaking of bilateral symmetry. Deviations from bilateral symmetry initially occur in the mouse embryonic node, a bundle of cells located at the anterior most region of the primitive streak. Cilia present in the node all beat in the same direction, creating a unidirectional flow of extra-cellular fluid. This asymmetric flow sets up a cascade of gene expression events, which leads to asymmetric organogenesis. Studies show morphological defects in the nodal cilia of Tbx6 mutant embryos even though Tbx6 is not expressed in the node at the time left/right asymmetry is established. The main hypothesis to be tested is that Tbx6 is involved in left-right asymmetry determination by regulating the formation of the node and nodal cilia and the transfer of asymmetric signals from the embryonic node to the left lateral plate mesoderm (LPM) by affecting different downstream target genes. A Tbx6 lineage tracer will be developed in order to investigate whether the effect of Tbx6 on the embryonic node is direct or indirect. In Situ hybridization will be used to examine the expression pattern of genes involved in the non-canonical Wnt signaling pathway and Nodal signal transduction pathway in Tbx6 mutants to determine whether Tbx6 regulates left-right determination by affecting these genes. Also immunofluorescence will be used to examine whether Tbx6 mutants have altered expression of proteins that are known to phenocopy the Tbx6 mutant cilia defects. Finally, using bacterial artificial chromosomes and molecular cloning a transgenicTbx6ERCre allele will be created in order to conduct lineage trace studies.
Mutations that affect left-right patterning can lead to congenital cardiac malformations, vascular anomalies, and other serious health problems such as anomalies of the spleen and the gastrointestinal system. Therefore, it is critical to investigate genes that when mutated perturb this process in order to understand how organ situs is established. The proposed study on Tbx6 will further our understanding of how mammals genetically regulate body axis asymmetry.
| Concepcion, Daniel; Papaioannou, Virginia E (2014) Nature and extent of left/right axis defects in T(Wis) /T(Wis) mutant mouse embryos. Dev Dyn 243:1046-53 |
| Nowotschin, Sonja; Ferrer-Vaquer, Anna; Concepcion, Daniel et al. (2012) Interaction of Wnt3a, Msgn1 and Tbx6 in neural versus paraxial mesoderm lineage commitment and paraxial mesoderm differentiation in the mouse embryo. Dev Biol 367:1-14 |
