The Fuction And Expression Analysis Of Activating Transcription Factor 4 In Embryo Diapause Termination Of Brine Shrimp,Artemia Sinicain

Artemia sinica is a small crustacean widely living in high salt environment,it is an important aquatic bait with strong resistance to stress and for developmental biology andmolecular biology.Artemia enter diapause stage in response to unfavourable environments and form a special diapause cysts to arrest development temporarily.Activating transcription factor 4(ATF4)is a ubiquitous basic leucine-zipper transcription factor that is a member of the ATF/ cyclic adenosine monophosphate responsive.There are three conserved binding sites in the promoter regions of ATF4 target genes.These include the CRE site,amino acid response element(AARE)site,and nutrient-sensing response element-1(NSRE-1)site.t has been shown that ATF4 is crucial to many physiological activities.Studies have demonstrated that ATF4 is involved in hematopoiesis,lens and skeletal development)learning and memory formation,hypoxia resistance,tumor growth,endoplasmic reticulum(ER)stress,autophagy,glucose metabolism,metabolic responses to amino acid deprivation etc.However,the expression pattern,distribution and functions of As-ATF4 in the post-diapause embryonic,remains unknown.In this study,the complete cDNA of ATF4 from A.sinica(As-ATF4)was cloned.The full-length was 1515 bp,encoding a 306 amino acids,and 34.5 kDa protein.We used quantitative real-time PCR(qPCR)and western blotting to investigate its mRNA expression levels and protein expression levels of As-ATF4,As-CHOP and As-eIF2? during embryonic development.It reduced from 0h to 15 h and increased with the development of Artemia,and increased at important moment with UPR,meanwhile,the expression level of As-ATF4 was also increassed at low and high salinity with UPR.Immunofluorescence(IF)assayed the location of As-ATF4 expression.It was distributed whole body at 0h,15 h and 3days.In this paper,we further analyzed the molecular mechanism of ATF4 of the poor environment of A.sinica,which provided the scientific basis for the study of the molecular mechanism of the embryoregeneration of others.