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

Abstract

ObjectiveTo determine if ARHGEF10 has a haploinsufficient effect and provide evidence to evaluate the severity, if any, during prenatal consultation. MethodsZebrafish 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. ResultsWISH 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 aahgef10 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. ConclusionBased on our findings, ARHGEF10 appeared to have a haploinsufficiency effect.