Hunan International Joint Laboratory of Animal Intestinal Ecology and Health, Laboratory of Animal Nutrition and Human Health, College of Life Sciences, Hunan Normal University, Changsha 410081, China.
Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation, Changsha 410081, China.
Int J Mol Sci. 2023 Apr 23;24(9):7740. doi: 10.3390/ijms24097740.
The adhesion G-protein-coupled receptor is a seven-transmembrane receptor protein with a complex structure. Impaired has been found to cause developmental damage to the human brain, resulting in intellectual disability and motor dysfunction. To date, studies on deficiency in zebrafish have been limited to the nervous system, and there have been no reports of its systemic effects on juvenile fish at developmental stages. In order to explore the function of in zebrafish, the CRISPR/Cas9 gene-editing system was used to construct a -knockout zebrafish. Subsequently, the differentially expressed genes (DEGs) at the transcriptional level between the 3 days post fertilization (dpf) homozygotes of the mutation and the wildtype zebrafish were analyzed via RNA-seq. The results of the clustering analysis, quantitative PCR (qPCR), and in situ hybridization demonstrated that the expression of innate immunity-related genes in the mutant was disordered, and multiple genes encoding digestive enzymes of the pancreatic exocrine glands were significantly downregulated in the mutant. Motor ability tests demonstrated that the zebrafish were more active, and this change was more pronounced in the presence of cold and additional stimuli. In conclusion, our results revealed the effect of deletion on the gene expression of juvenile zebrafish and found that the mutant was extremely active, providing an important clue for studying the mechanism of in the development of juvenile zebrafish.
黏附 G 蛋白偶联受体是一种具有复杂结构的七跨膜受体蛋白。研究发现，其功能受损会导致人类大脑发育损伤，从而导致智力残疾和运动功能障碍。迄今为止，关于斑马鱼 缺乏的研究仅限于神经系统，尚未有报道表明其在发育阶段对幼鱼有全身影响。为了研究 在斑马鱼中的功能，我们使用 CRISPR/Cas9 基因编辑系统构建了 - 敲除斑马鱼。随后，通过 RNA-seq 分析了 3 天受精后 (dpf) 突变纯合子与野生型斑马鱼在转录水平上的差异表达基因 (DEGs)。聚类分析、定量 PCR (qPCR) 和原位杂交的结果表明，突变体中先天免疫相关基因的表达紊乱，并且突变体中胰腺外分泌腺的多种消化酶编码基因显著下调。运动能力测试表明， 斑马鱼更活跃，而在存在冷刺激和其他刺激时，这种变化更为明显。总之，我们的结果揭示了 缺失对幼鱼斑马鱼基因表达的影响，并发现 突变体极其活跃，为研究 在幼鱼发育过程中的作用机制提供了重要线索。
