Wtip is required for proepicardial organ specification and cardiac left/right asymmetry in zebrafish

Powell,Rebecca; Bubenshchikova,Ekaterina; Fukuyo,Yayoi; Hsu,Chaonan; Lakiza,Olga; Nomura,Hiroki; Renfrew,Erin; Garrity,Deborah; Obara,Tomoko

doi:10.3892/mmr.2016.5550

Wtip is required for proepicardial organ specification and cardiac left/right asymmetry in zebrafish

Authors:
- Rebecca Powell
- Ekaterina Bubenshchikova
- Yayoi Fukuyo
- Chaonan Hsu
- Olga Lakiza
- Hiroki Nomura
- Erin Renfrew
- Deborah Garrity
- Tomoko Obara
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Affiliations: Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma, OK 73104, USA, Department of Biology, Colorado State University, Fort Collins, CO 80523, USA
Published online on: July 27, 2016 https://doi.org/10.3892/mmr.2016.5550
Pages: 2665-2678
Copyright: © Powell et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Wilm's tumor 1 interacting protein (Wtip) was identified as an interacting partner of Wilm's tumor protein (WT1) in a yeast two-hybrid screen. WT1 is expressed in the proepicardial organ (PE) of the heart, and mouse and zebrafish wt1 knockout models appear to lack the PE. Wtip's role in the heart remains unexplored. In the present study, we demonstrate that wtip expression is identical in wt1a‑, tcf21‑, and tbx18‑positive PE cells, and that Wtip protein localizes to the basal body of PE cells. We present the first genetic evidence that Wtip signaling in conjunction with WT1 is essential for PE specification in the zebrafish heart. By overexpressing wtip mRNA, we observed ectopic expression of PE markers in the cardiac and pharyngeal arch regions. Furthermore, wtip knockdown embryos showed perturbed cardiac looping and lacked the atrioventricular (AV) boundary. However, the chamber‑specific markers amhc and vmhc were unaffected. Interestingly, knockdown of wtip disrupts early left‑right (LR) asymmetry. Our studies uncover new roles for Wtip regulating PE cell specification and early LR asymmetry, and suggest that the PE may exert non‑autonomous effects on heart looping and AV morphogenesis. The presence of cilia in the PE, and localization of Wtip in the basal body of ciliated cells, raises the possibility of cilia-mediated PE signaling in the embryonic heart.

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