Expression Pattern Of BAG-4in The Embryonic Developing Processes Of Artemia Sinica

Artemia，brine shrimp，is a sort of tiny crustaceans and widely distributed in NorthEurope, Asia, America, central America and Africa.Resources of Artemia sinica are abundantin China and concentratively distributed both around Xinjiang, Qinghai-Tibet and salterns byBohai Bay, Liaodong Bay, Laizhou Bay, salt lakes in Inner Mongolia etc. Artemia sinica isthe insect with characters of possessing highly unsaturated fatty acid and high proteins.Artemia sinica became fodders for producing frys and shrimps and a suitable material forstudying embryonic developing processes with its delayed development.The family of BAG protein, also called Bcl-2-associated athanogene, is a family ofantiapoptotic proteins which is found in recent years. Its secretion of proteins can regulatevarious physiological processes when combined with ATP enzyme of heat-shock protein70.One member named BAG-4in the family of BAG protein,also called the silence of deathdomain can integrate with the death domain of TNFR-I to keep embracer of death fromcombining with cellular apoptosis signaling proteins.In this way cellular apoptosis isprevented because the access is blocked.Artemia sinica occurs diapause when in the menacedconditions which lead to cellular apoptosis and embryonic demise when diapaused muchlonger.However,BAG-4belongs to the related proteins with regulations of cellular apoptosisin the embryonic developing processes of Artemia sinica.Up until now,there is no researchabout BAG-4of Artemia sinica reported. This research is great importance and significancefor it is about to study the structure, function of BAG-4and the expressing pattern in theembryo of Artemia sinica for the first time in the world.This research adopted experimental methods of integral immune tissues to study theexpressing locations of BAG-4in different developing stages of Artemia sinica.Resultsshowed that, one end of the embryo of Artemia sinica expressed positive vividly whendeveloping to0hours,5hours and10hours; the front end of the nauplius expressed a littlewhen developed to15hours. BAG-4protein of Artemia sinica expressed mainly in the chestand abdomen,and most concentrated in the head at later stages of development of20hours,40hours,3days and5days.Expression in the head became more and more obvious with theearly development of the embryo of Artemia sinica.It showed basically the same experimentaldata of positive expressing location compared to integral immune tissues when using analysis of tissue immunohistochemistry to analyse the expressing location at different stages ofdevelopment of BAG-4.This research also used the method of western blot to study expressing amounts ofBAG-4protein at different stages of development of Artemia sinica.The study was detectedby western blot test.The test results showed that the expressing amounts of BAG-4protein atdifferent stages of development trend to increase first and then decrease under the sameconditions of temperature, salinity and light intensity.The expressing amount of BAG-4protein culminated at the point of10hours and15hours.BAG-4expressed higher amountsmainly at the eraly stages of the development of the embryo of Artemia sinica.The expressingamounts of BAG-4protein of Artemia sinica developing to10hours increased with the stressof salinity condition.In another word,the amounts increased with the growth of thesalinity.The amounts increaed until the salinity reached200‰.This is obvious that thegrowth of salinity will enhance the expressing amounts of BAG-4protein.To sum up what has been reported, BAG-4, which can widely distribute at parts ofArtemia sinica, possesses the ability to be involved in the early development of the embryosof Artemia sinica and expressed higher level under the condition of salinity stress. Thismight be related to the participation of BAG-4in the TNFR-I signaling pathway to inhibitcellular apoptosis.