PUBLICATION
Small GTPase R-Ras participates in neural tube formation in zebrafish embryonic spinal cord
- Authors
- Ohata, S., Uga, H., Okamoto, H., Katada, T.
- ID
- ZDB-PUB-180518-6
- Date
- 2018
- Source
- Biochemical and Biophysical Research Communications 501(3): 786-790 (Journal)
- Registered Authors
- Ohata, Shinya, Okamoto, Hitoshi
- Keywords
- Neural tube defect, R-Ras, Vangl2, Zebrafish
- MeSH Terms
-
- Animals
- Gene Expression Regulation, Developmental
- Membrane Proteins/genetics
- Membrane Proteins/metabolism
- Mutation
- Neural Tube/embryology*
- Neural Tube/metabolism
- Spinal Cord/embryology*
- Spinal Cord/metabolism
- Zebrafish/embryology*
- Zebrafish/genetics
- Zebrafish/metabolism
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 29772239 Full text @ Biochem. Biophys. Res. Commun.
Citation
Ohata, S., Uga, H., Okamoto, H., Katada, T. (2018) Small GTPase R-Ras participates in neural tube formation in zebrafish embryonic spinal cord. Biochemical and Biophysical Research Communications. 501(3):786-790.
Abstract
Ras related (R-Ras), a small GTPase, is involved in the maintenance of apico-basal polarity in neuroepithelial cells of the zebrafish hindbrain, axonal collapse in cultured murine hippocampal neurons, and maturation of blood vessels in adult mice. However, the role of R-Ras in neural tube formation remains unknown. Using antisense morpholino oligonucleotides (AMOs), we found that in the spinal cord of zebrafish embryos, the lumen was formed bilaterally in rras morphants, whereas it was formed at the midline in control embryos. As AMO can cause off-target effects, we generated rras mutant zebrafish lines using CRISPR/Cas9 technology. Although these rras mutant embryos did not have a bilateral lumen in the spinal cord, the following findings suggest that the phenotype is unlikely due to an off-target effect of rras AMO: 1) The rras morphant phenotype was rescued by an injection of AMO-resistant rras mRNA, and 2) a bilaterally segregated spinal cord was not observed in rras mutant embryos injected with rras AMO. The results suggest that the function of other ras family genes may be redundant in rras mutants. Previous research reported a bilaterally formed lumen in the spinal cord of zebrafish embryos with a mutation in a planar cell polarity (PCP) gene, van gogh-like 2 (vangl2). In the present study, in cultured cells, R-Ras was co-immunoprecipitated with Vangl2 but not with another PCP regulator, Pricke1. Interestingly, the interaction between R-Ras and Vangl2 was stronger in guanine-nucleotide free point mutants of R-Ras than in wild-type (WT) or constitutively active forms of R-Ras. R-Ras may regulate neural tube formation in cooperation with Vangl2 in the developing zebrafish spinal cord.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping