Formation of renal tubules in both mammals and amphibians requires Wnt pathways. In kidney, aberrant Wnt signaling has been linked to developmental pathologies such as Wilms tumor and cystic diseases, as well as with renal cell carcinoma. Recent studies from our group and others indicate that canonical (?-catenin- mediated) Wnt signaling is essential for induction of nephron development. Additionally, studies suggest that primary cilia are required for a switch from canonical to non-canonical Wnt signaling, and that defects in primary cilia formation or signaling lead to cystogenesis. Varied tissues undergoing morphogenesis employ non-canonical Wnt (PCP: "planar cell polarity") pathways to promote processes including cell polarization and cytoskeletal modulation. Deficiencies in non-canonical Wnt signaling, which contributes to PCP, underlie a number of developmental or later defects, such as polycystic kidney disease and nephronophthisis. Using Xenopus laevis (frog) embryos, supplemented in some contexts with embryos of Danio rerio (zebrafish), this proposal first assesses the roles of non-canonical Wnt PCP components in kidney tubule morphogenesis. Our hypothesis is that the PCP components Daam1, WGEF, Prickle and Strabismus are required for primary cilia formation, which is in turn required for nephric morphogenesis. Xenopus offers experimental advantages including the facile introduction of exogenous constructs to block or activate signaling pathways, rapid development and easy visualization of the forming kidney under the surface ectoderm. Second, using transgenic approaches, we will address in living animals the role of PCP/ non-canonical Wnt signals in shaping and moving cells that are undergoing/ contributing to kidney tubule morphogenesis. Finally, an over-expression screen will be performed to discover new regulators of tubule formation. Overall, this work is novel in assessing the roles of the PCP components in kidney development;in utilizing transgenic approaches in Xenopus laevis to visualize and/ or target kidney development in living embryos;and in performing an unbiased over- expression screen to reveal additional and likely novel components involved in kidney tubulogenesis. In sum, we will enhance our understanding of kidney morphogenesis, which is likely to be relevant to obtaining insights on later pathologies that arise in humans.
Wnt signaling plays a crucial role in kidney development, and disruption of Wnt signaling results in kidney ailments including congenital cystic kidney diseases (polycystic kidney disease and nephronopthisis) and cancer (Wilms tumor and renal cell carcinoma). My research project focuses on the role the planar cell polarity (PCP) pathway, a Wnt trajectory involved in shaping tissues, in the formation and shaping of kidney tubules. Using the Xenopus laevis (frog) experimental model, this proposal aims both to determine if disruption of candidate PCP components leads to alterations in kidney tube formation, and to discover novel components affecting tubulogenesis, taking advantage of innovative methods such as live imaging of transgenic animals.
|Valls, Gabriela; Codina, Montserrat; Miller, Rachel K et al. (2016) Upon Wnt stimulation, Rac1 activation requires Rac1 and Vav2 binding to p120-catenin. J Cell Sci 129:2120-3|
|DeLay, Bridget D; Krneta-Stankic, Vanja; Miller, Rachel K (2016) Technique to Target Microinjection to the Developing Xenopus Kidney. J Vis Exp :|
|Krneta-Stankic, Vanja; DeLay, Bridget D; Miller, Rachel K (2016) Xenopus: leaping forward in kidney organogenesis. Pediatr Nephrol :|
|Munoz, William A; Lee, Moonsup; Miller, Rachel K et al. (2014) Plakophilin-3 catenin associates with the ETV1/ER81 transcription factor to positively modulate gene activity. PLoS One 9:e86784|
|Markham, Nicholas O; Doll, Caleb A; Dohn, Michael R et al. (2014) DIPA-family coiled-coils bind conserved isoform-specific head domain of p120-catenin family: potential roles in hydrocephalus and heterotopia. Mol Biol Cell 25:2592-603|
|Chang, Daniel R; Martinez Alanis, Denise; Miller, Rachel K et al. (2013) Lung epithelial branching program antagonizes alveolar differentiation. Proc Natl Acad Sci U S A 110:18042-51|
|Miller, Rachel K; Hong, Ji Yeon; MuÃ±oz, William A et al. (2013) Beta-catenin versus the other armadillo catenins: assessing our current view of canonical Wnt signaling. Prog Mol Biol Transl Sci 116:387-407|