PUBLICATION
Live imaging and conditional disruption of native PCP activity using endogenously tagged zebrafish sfGFP-Vangl2
- Authors
- Jussila, M., Boswell, C.W., Griffiths, N.W., Pumputis, P.G., Ciruna, B.
- ID
- ZDB-PUB-220925-2
- Date
- 2022
- Source
- Nature communications 13: 5598 (Journal)
- Registered Authors
- Ciruna, Brian
- Keywords
- none
- MeSH Terms
-
- Animals
- Cell Polarity*/genetics
- Embryonic Development/genetics
- Membrane Proteins/genetics
- Membrane Proteins/metabolism
- Zebrafish*
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 36151137 Full text @ Nat. Commun.
Citation
Jussila, M., Boswell, C.W., Griffiths, N.W., Pumputis, P.G., Ciruna, B. (2022) Live imaging and conditional disruption of native PCP activity using endogenously tagged zebrafish sfGFP-Vangl2. Nature communications. 13:5598.
Abstract
Tissue-wide coordination of polarized cytoskeletal organization and cell behaviour, critical for normal development, is controlled by asymmetric membrane localization of non-canonical Wnt/planar cell polarity (PCP) signalling components. Understanding the dynamic regulation of PCP thus requires visualization of these polarity proteins in vivo. Here we utilize CRISPR/Cas9 genome editing to introduce a fluorescent reporter onto the core PCP component, Vangl2, in zebrafish. Through live imaging of endogenous sfGFP-Vangl2 expression, we report on the authentic regulation of vertebrate PCP during embryogenesis. Furthermore, we couple sfGFP-Vangl2 with conditional zGrad GFP-nanobody degradation methodologies to interrogate tissue-specific functions for PCP. Remarkably, loss of Vangl2 in foxj1a-positive cell lineages causes ependymal cell cilia and Reissner fiber formation defects as well as idiopathic-like scoliosis. Together, our studies provide crucial insights into the establishment and maintenance of vertebrate PCP and create a powerful experimental paradigm for investigating post-embryonic and tissue-specific functions for Vangl2 in development and disease.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping