The Mechanism Of Bmp10 And Dusp1 In Apoptosis Of Zebrafish Induced By Low Temperature

Temperature is one of the important factors that limits the distribution of organisms and directly determines the physiological features and behavior of organisms.Fish is the most widespread vertebrates.How the fish adapted to the temperatures changes is an important and meaningful question in the environmental biology.For the tilapia and zebrafish,the gill appears to be the most sensitive tissue to cold-induced damage.Divergent gene expression between zebrafish and tilapia enriched in apoptosis pathway during cold stress at 8?6h,12 h.A transcriptomic comparison followed by experimental verification indicated that the differential expression of dusp1 and bmp10,is likely an important factor contributing to the exacerbated apoptosis in the zebrafish gill.Based on this study,we propose a model to address bmp10 and dusp1 regulate the molecular and cellular processes of zebrafish.This study opens doors for exploring the factors governing the gene of a fish.Bmp10,a cardiac-restricted BMP family member,plays a critical role in regulating development of the heart.Bmp10 is first expressed in the mouse heart at embryonic and subsequently becomes enriched within the trabecular myocardium: a developmental window devoted to cardiac growth and chamber maturation.Bmp10–/– embryos survive,but succumb from heart failure associated with a normally patterned,but severely hypoplastic,heart.However,it is not clear what mechanisms regulate expression of bmp10 during embryonic or postnatal development in zebrafish.Dusp1 is a double-specific phosphatase,a negative regulator of MAPK family,involved in a large number of physiological activities,including cellular signaling,chondrocyte growth,cell metabolism,and so on,dusp1 reduced or induced can lead to apoptosis.Zebrafish as a model organism of vertebrate has many advantages such as its small size,high fecundity,shorter life cycle,larger capacity for generating more offspring.The genome of the zebrafish,Danio rerio,has been fully sequenced.Therefore,this fish is a widely used model for developmental biology and environmental genomics studies.In order to reveal the relationship between bmp10,dusp1 and gill apoptosis,this study firstly uses CRISPR/Cas9 to knock out bmp10,dusp1 during zebrafish embryo development,and detect the apoptosis of gill by TUNEL,pathological detect with HE.Secondly,the dusp1 was knocked down and the apoptotic number was detected in ZF4 cells.The results showed that the bmp10-/-zebrafish is abnormal in the myocardial development,edema at the secondary lamellae were observed,low temperature stress,accelerated myocardial cell necrosis,gill serious edema,resulting in the apoptosis of gill cells.The analysis that the bmp10 target site in the conservative area,so in the zebrafish play an important role in the evolutionary process.The absence of bmp10 will affect the heart development,indicating that bmp10 in the zebrafish plays an important role in regulating heart development process.However,the dusp1-/-zebrafish apoptosis number of gill significantly increased at low temperature,but the gill secondary lamellae did not detect the obvious pathological changes.Dusp1 knock down in ZF4 cells can promote apoptosis at the low temperature.The analysis that the dusp1 sequences did not detect the conservative zone,and the knockout dusp1 would appear to interact with other genes,so the dusp1 zebrafish phenotype was not apparent.In short,this study proves that bmp10,dusp1 participates in the process of low temperature stress in fishes,which plays an important role in regulating apoptosis signaling pathway,knocking down or knocking out bmp10,dusp1,gene function deficiency,inducing cell apoptosis.Bmp10 plays an important role in the heart development of zebrafish.This study lays a foundation for the follow-up study on the molecular mechanism of gene expression and occurrence of endogenous genes in fishes.