PUBLICATION
Single-copy Loss of Rho Guanine Nucleotide Exchange Factor 10 ( arhgef10) Causes Locomotor Abnormalities in Zebrafish Larvae
- Authors
- Zhang, Y., An, M.X., Gong, C., Li, Y.Y., Wang, Y.T., Lin, M., Li, R., Tian, C.
- ID
- ZDB-PUB-220127-4
- Date
- 2022
- Source
- Biomedical and environmental sciences : BES 35: 35-44 (Journal)
- Registered Authors
- Keywords
- CRISPR/Cas9, Copy loss, Haploinsufficiency, Zebrafish, arhgef10
- MeSH Terms
-
- Spectrophotometry/methods
- Animals
- In Situ Hybridization
- Phenotype
- Blotting, Western
- Real-Time Polymerase Chain Reaction/standards
- CRISPR-Associated Protein 9
- Cells, Cultured
- Annexin A5
- Cell Line
- Sincalide/analysis
- Larva/genetics
- Larva/physiology
- RNA/isolation & purification
- Flow Cytometry
- Cell Proliferation
- Zebrafish/genetics
- Zebrafish/physiology*
- Rho Guanine Nucleotide Exchange Factors/genetics*
- Rho Guanine Nucleotide Exchange Factors/metabolism
- Genotype
- Humans
- Apoptosis
- CRISPR-Cas Systems
- PubMed
- 35078560 Full text @ Biomed. Environ. Sci.
Citation
Zhang, Y., An, M.X., Gong, C., Li, Y.Y., Wang, Y.T., Lin, M., Li, R., Tian, C. (2022) Single-copy Loss of Rho Guanine Nucleotide Exchange Factor 10 ( arhgef10) Causes Locomotor Abnormalities in Zebrafish Larvae. Biomedical and environmental sciences : BES. 35:35-44.
Abstract
Objective To determine if ARHGEF10 has a haploinsufficient effect and provide evidence to evaluate the severity, if any, during prenatal consultation.
Methods Zebrafish was used as a model for generating mutant. The pattern of arhgef10 expression in the early stages of zebrafish development was observed using whole-mount in situ hybridization (WISH). CRISPR/Cas9 was applied to generate a zebrafish model with a single-copy or homozygous arhgef10 deletion. Activity and light/dark tests were performed in arhgef10-/-, arhgef10+/-, and wild-type zebrafish larvae. ARHGEF10 was knocked down using small interferon RNA (siRNA) in the SH-SY5Y cell line, and cell proliferation and apoptosis were determined using the CCK-8 assay and Annexin V/PI staining, respectively.
Results WISH showed that during zebrafish embryonic development arhgef10 was expressed in the midbrain and hindbrain at 36-72 h post-fertilization (hpf) and in the hemopoietic system at 36-48 hpf. The zebrafish larvae with single-copy and homozygous arhgef10 deletions had lower exercise capacity and poorer responses to environmental changes compared to wild-type zebrafish larvae. Moreover, arhgef10 -/- zebrafish had more severe symptoms than arhgef10 +/- zebrafish. Knockdown of ARHGEF10 in human neuroblastoma cells led to decreased cell proliferation and increased cell apoptosis.
Conclusion Based on our findings, ARHGEF10 appeared to have a haploinsufficiency effect.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping