The Role Of As-HIFh-1? In The Early Embryonic Development Of Artemia Sinica

Hypoxia-inducible factor-1?(HIF-1?)plays an important role in the process of biological hypoxia stress and is essential for the survival of organisms.Under normoxic conditions,HIF-1? cannot exist stably in cells,and it will be degraded by 26 S ubiquitinated proteasome with the participation of oxygen;While under hypoxic conditions,HIF-1? can escape degradation and interact with HIF-1? binds to form a heterodimer,which regulates the expression of downstream target genes.In higher animals,it mainly increases the body's oxygen supply by promoting angiogenesis and increasing the number of red blood cells.In addition,HIF-1? can also upregulate glucose transporter and the expression of glycolytic enzymes accelerates the delivery of nutrients to cells to meet the metabolic needs of cell growth under hypoxic conditions.Artemia sinica undergoes hypoxia during early embryonic development,diapause,and release of diapause.Therefore,this experiment analyzed the role of HIF-1? in the early embryonic development of Artemia sinica,as well as the response of HIF-1? under hypoxic stress and the mutual regulation relationship with other related genes in the process of maintaining the stability of the body's physiological functions.In this study,the HIF-1? gene sequence with a length of 1459 bp was obtained by RACE technology,and the length of the open reading frame was 1275 bp,encoding a total of 424 amino acids.The bioinformatics analysis of the sequence showed that the HIF-1? protein is a hydrophilic protein with a molecular weight of about 47.5k Da,and the protein does not contain signal peptides and transmembrane regions.Detecting the distribution of HIF-1? in Artemia sinica by immunofluorescence experiments found that HIF-1? is expressed at all stages of Artemia sinica embryonic development and throughout the body.It has no tissue specificity and is mainly located in the nucleus and cytoplasm.Real-time q PCR and Western blotting were used to analyze the changes of HIF-1? expression in each stage of Artemia sinica early embryonic development and hypoxia stress at the gene and protein levels,respectively.The results showed that in the early stages of Artemia sinica embryonic development,the expression of HIF-1? showed a trend of first increasing and then decreasing.When the embryo developed to 15 h,the expression reached the maximum;In the hypoxia stress experiment,as the duration of hypoxia increased,the expression of Artemia sinica HIF-1? increased significantly.The expression level of HIF-1? protein is roughly consistent with the change trend of gene expression.In addition,this experiment explored the regulatory relationship between HIF-1? and p300/CBP,REDD1,Hsp26 and Hsp70 at the protein level through Western blotting in the early embryonic development and cell hypoxic stress response of Artemia sinica.The results showed that the expression of p300/CBP,REDD1 and Hsp70 were positively correlated with HIF-1?,while the expression of Hsp26 was not significantly correlated with HIF-1?.As an important transcriptional regulatory factor in cells,HIF-1? can maintain the constant function and physiological state of Artemia diapause embryos,and at the same time play an important role in the process of Artemia diapause and reopening embryo development.This study provides reference materials for further analysis of Artemia sinica early embryonic development,release of diapause and response to hypoxia stress.It has certain theoretical significance and has potential research and application value for the development and utilization of Artemia protein sources.