CRISPR/Cas9‐induced nos2b mutant zebrafish display behavioral abnormalities

Abstract

The immunomodulatory function of nitric oxide synthase (NOS2) has been extensively studied. However, some behavioral abnormalities caused by its mutations have been found in a few rodent studies, of which the molecular mechanism remains elusive. In this research, we generated nos2b gene knockout zebrafish (nos2bsou2/sou2) using CRISPR/Cas9 approach and investigated their behavioral and molecular changes by doing a series of behavioral detections, morphological measurements, and molecular analyses. We found that, compared with nos2b+/+ zebrafish, nos2bsou2/sou2 zebrafish exhibited enhanced motor activity; additionally, nos2bsou2/sou2 zebrafish were characterized by smaller brain size, abnormal structure of optic tectum, reduced mRNA level of presynaptic synaptophysin and postsynaptic homer1, and altered response to sodium nitroprusside/methylphenidate hydrochloride treatment. These findings will likely contribute to future studies of behavioral regulation.

Effects of Inducible Nitric Oxide Synthase (iNOS) Gene Knockout on the Diversity, Composition, and Function of Gut Microbiota in Adult Zebrafish

“…In zebrafish genome, there are two iNOS genes, nos2a and nos2b . The construction of zebrafish with nos2a and nos2b single knockouts at the early stage has already been completed [ 32 , 33 ]. Crossbreeding of homozygous iNOS-deficient zebrafish gave rise to the F0 progeny that were heterozygous double mutants.…”

Transcriptome analysis revealed gene expression feminization of testis after exogenous tetrodotoxin administration in pufferfish Takifugu flavidus

“…After sex is determined by histological observation, 3 or 4 gonad samples are mixed into one pool for each FTG, MTG, FCG and MCG group. Total RNAs of each group were extracted using RNAiso Plus (Takara, Japan) [ 89 ]. The agarose electrophoresis was used to detect the integrity of total RNA, and the Agilent 2100 Bioanalyzer (Agilent Technologies, USA) was used to detect the concentration and purity of total RNA.…”

“…The gene names and the primers are listed in Table S 9 . Gadph was used as an internal control [ 89 , 109 , 110 ]. The relative abundances of mRNA transcripts were evaluated using the formula: R = 2 −ΔΔCt [ 111 ].…”