PUBLICATION
Loss of Wnt16 Leads to Skeletal Deformities and Downregulation of Bone Developmental Pathway in Zebrafish
- Authors
- Qu, X., Liao, M., Liu, W., Cai, Y., Yi, Q., Long, J., Tan, L., Deng, Y., Deng, H., Chen, X.
- ID
- ZDB-PUB-210703-25
- Date
- 2021
- Source
- International Journal of Molecular Sciences 22(13): (Journal)
- Registered Authors
- Keywords
- CRISPR-Cas9, RNA sequencing, skeletal development, wnt16, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Bone and Bones/abnormalities*
- Computational Biology/methods
- Disease Models, Animal
- Gene Expression Profiling
- Gene Knockout Techniques
- Gene Ontology
- Molecular Sequence Annotation
- Musculoskeletal Abnormalities/diagnosis
- Musculoskeletal Abnormalities/genetics*
- Musculoskeletal Abnormalities/metabolism*
- Osteogenesis/genetics*
- Phenotype
- Transcriptome
- Wnt Proteins/chemistry
- Wnt Proteins/deficiency*
- Wnt Proteins/metabolism
- Zebrafish/genetics*
- Zebrafish Proteins/chemistry
- Zebrafish Proteins/deficiency*
- Zebrafish Proteins/metabolism
- PubMed
- 34206401 Full text @ Int. J. Mol. Sci.
Citation
Qu, X., Liao, M., Liu, W., Cai, Y., Yi, Q., Long, J., Tan, L., Deng, Y., Deng, H., Chen, X. (2021) Loss of Wnt16 Leads to Skeletal Deformities and Downregulation of Bone Developmental Pathway in Zebrafish. International Journal of Molecular Sciences. 22(13):.
Abstract
Wingless-type MMTV integration site family, member 16 (wnt16), is a wnt ligand that participates in the regulation of vertebrate skeletal development. Studies have shown that wnt16 can regulate bone metabolism, but its molecular mechanism remains largely undefined. We obtained the wnt16-/- zebrafish model using the CRISPR-Cas9-mediated gene knockout screen with 11 bp deletion in wnt16, which led to the premature termination of amino acid translation and significantly reduced wnt16 expression, thus obtaining the wnt16-/- zebrafish model. The expression of wnt16 in bone-related parts was detected via in situ hybridization. The head, spine, and tail exhibited significant deformities, and the bone mineral density and trabecular bone decreased in wnt16-/- using light microscopy and micro-CT analysis. RNA sequencing was performed to explore the differentially expressed genes (DEGs). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis found that the down-regulated DEGs are mainly concentrated in mTOR, FoxO, and VEGF pathways. Protein-protein interaction (PPI) network analysis was performed with the detected DEGs. Eight down-regulated DEGs including akt1, bnip4, ptena, vegfaa, twsg1b, prkab1a, prkab1b, and pla2g4f.2 were validated by qRT-PCR and the results were consistent with the RNA-seq data. Overall, our work provides key insights into the influence of wnt16 gene on skeletal development.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping