Effect of trpa1b knockdown on high temperature tolerance in zebrafish(Danio rerio)
To explore transient receptor potential ankyrin 1b(trpa1b)gene on zebrafish(Danio rerio)under high temperature conditions,zebrafish model with trpa1b gene deletion was constructed by CRISPR/Cas9 gene knockout technology.Survival,tissue apoptosis and other related gene expression changes were investigated in Trpa1b knock-out(trpa1b-/-)zebrafish and wild-type(WT)zebrafish exposed to water temperature from 28 ℃ to 40 ℃ at a warming rate of 1 ℃/h per hour by histological and transcriptome methods.Zebrafish tissues of different genotypes were sampled at 40℃.It was found that the overall survival time of trpa1b-/-homozygotes was shorter under high temperature.TUNEL staining showed that there was significantly higher apoptosis in gill and brain of trpa1b-/-ho-mozygotes than that in wild-type zebrafish(P<0.05).Transcriptome sequencing analysis for gill tissues of different genotypes under normal temperature(28℃)and high temperature(38℃and 40℃)revealed that the expression level of hspa1b and hsp90aa1.1in trpa1b-/-homozygotes was significantly down regulated under high temperature(P<0.001),and that the expression level of intracellular calcium ions,ubiquitin protein transferase,lysosome and ribosome related genes were changed,which impaired their viability under heat stimulation.The findings indicate that Trpa1b deletion will lead to more obvious high-temperature intolerance in zebrafish due to the fact that trpa1b gene is an important gene required to maintain high temperature tolerance in zebrafish,which provides reference for further research on the mechanism of high temperature tolerance in fish.
Danio reriotrpa1bgene knockouthigh temperature tolerance
万方数据
维普
