Transcriptomics and Phenotypic Analysis of gpr56 Knockout in Zebrafish

Abstract

The adhesion G-protein-coupled receptor is a seven-transmembrane receptor protein with a complex structure. Impaired GPR56 has been found to cause developmental damage to the human brain, resulting in intellectual disability and motor dysfunction. To date, studies on gpr56 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 gpr56 in zebrafish, the CRISPR/Cas9 gene-editing system was used to construct a gpr56-knockout zebrafish. Subsequently, the differentially expressed genes (DEGs) at the transcriptional level between the 3 days post fertilization (dpf) homozygotes of the gpr56 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 gpr56−/− 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 gpr56 deletion on the gene expression of juvenile zebrafish and found that the gpr56 mutant was extremely active, providing an important clue for studying the mechanism of gpr56 in the development of juvenile zebrafish.

In Search of a Target Gene for a Desirable Phenotype in Aquaculture: Genome Editing of Cyprinidae and Salmonidae Species

“…The targets of studies include lysosomes [ 170 , 171 ], endoplasmic reticulum [ 172 ], membranes [ 173 ], membrane channels [ 174 ], cytoskeleton [ 175 ], recombination processes [ 176 ], aneuploidy [ 177 ], expression regulation [ 178 ], and signaling [ 179 ]. There is research on the genetic basis of the organism’s response to oxidative stress [ 180 , 181 , 182 , 183 , 184 ], hypoxia [ 185 , 186 , 187 ], sodium ion uptake from the surrounding aquatic environment and ammonia excretion [ 188 , 189 , 190 ], inflammation processes [ 191 , 192 ], immunity [ 193 , 194 , 195 , 196 ] and disease resistance [ 197 , 198 , 199 , 200 , 201 ].…”

“…A series of works is devoted to the study the functions of genes involved in the immune response reaction. The gpr56 gene coordinates the expression of other immune response genes and digestive enzyme genes [ 193 ]. The socs3b , mitfa , CD18 , and lcp1 genes control the morphology, number, or migration of neutrophils and macrophages to the site of inflammation [ 154 , 159 , 194 , 195 ].…”