Single-copy Loss of Rho Guanine Nucleotide Exchange Factor 10 (arhgef10) Causes Locomotor Abnormalities in Zebrafish Larvae

• 作者: ZHANG Yi AN Ming Xing GONG Chen LI Yang Yang WANG Yu Tong LIN Meng LI Rong TIAN Chan
• 来源期刊: 生物医学与环境科学（英文版）
• 年，卷（期）: 2022,（1）
• 所属期刊栏目: Original Articles
• 页码范围: 35-44
• 页数: 10页
• 语种: 英文
• DOI: 10.3967/bes2022.005

摘要

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.