The Role Of Annexin 3 In Embryo Restart Of Artemia Sinica

Artemia sinica is a small crustacean that lives in a high salt water body,It’s also an important bait,extremely resilient,It is also an important laboratory animal in molecular biology and developmental biology,It plays an increasingly important role in embryology and molecular biology.The Annexin family is a multifunctional soluble calcium ion dependent membrane bound protein superfamily,they are essential for intracellular Ca²⁺signaling pathways,It is found in vertebrates,invertebrates,plants and other eukaryotes（except yeasts）and prokaryotes.The animal cell Annexin is regulated by the cytoplasmic free Ca²⁺ratio on the outside of the cell.All in all,a lot of physiological and biochemical processes involve the involvement of animal Annexin,such as regulation of membrane dynamics,participation in Ca²⁺signaling pathway,participation in Ca²⁺concentration regulation and adjustment of cytoskeleton,formation of membrane ion channels,etc.Many physiological processes involve Annexin 3,these include tumor occurrence,drug resistance and metastasis.Therefore,Annexin 3 has a very important role in the process of restarting the embryo of Artemia sinica in China,but its regulatory mechanism is unclear.In this experiment,RACE technology was used for the first time to clone the full length of cDNA of the Chinese Artemia sinica As-anx 3 gene,In present study,the full-length of As-anx 3 cDNA was 1024 bp,encoding a 315 amino acid protein,open reading box containing 948bp,there are four classical ANX domains.The subcellular localization analysis of As-anx was analyzed by immunofluorescence technique,experimental results show that,As-ANX 3 is located on the cell membrane of the Artemia sinica cell.Using real-time PCR and Western blotting,the expression pattern of Annexin is analyzed,As-anx has the highest expression at 0h,with the development of Artemia sinica,the membrane protein that accumulates during diapause is consumed by a little bit,As-anx expression decreased,It is indicated that membrane protein plays an important role in embryo restart.In the experiment of salinity stimulus and temperature stress,As-ANX 3 at temperature 15,when the salinity is28‰,the expression quantity is the highest,and then as the temperature goes up or down,the salinity goes up,the quantity of expression was down-regulated.It can be concluded that As-ANX 3 can be controlled by temperature and salinity,when salinity is too high or too low,As-ANX 3 expression decreased,As-ANX 3 is an important protein in embryo development.This paper analyzes the role of Annexin 3 in the restarting process of Chinese Artemia sinica diapause embryos,the study provides scientific basis for the study of the molecular mechanism of embryo reactivation of crustaceans and other invertebrate animals.