Expressions Of Trh Gene And APH-1 Gene During Early Embryonic Development Of Artemia Sinica

Artemia sinica belongs to Phylum Arthropoda, Class Crustacea, Subclass Branchiopoda, Order Anostaca, Family Artemiidae and Genus Artemia. It lives in the high-salt salt lakes and salterns diffusely. So, osmoregulation plays an important part in its life. In the different stage of development, osmoregulation is accomplished by different tissues.For example, when Artemia is hatched into nauplius, it is accomplished by salt gland and thoracic epipods show their affects in the sub-adult and adult stages.Diapause eggs suspend in gastrulation , which is significant differences from other embryonic decvelopment.It is improtant to identify Artemia embryo development gene ,and study the function and mechanism of embryo genes suspension and restart. trh gene and APH-1 gene are known as embryonic development genes and transcription factors, encoading a bHLH-PAS domain protein and POU domain protein respectively. In this study, RACE was used to achieve the full 2698bp trh gene cDNA for the first time, then determined the gene sequence for Artemia trh by Bioinformatiic analysi(sGenebank:FJ952937). RT-PCR primer sequences of trh gene were designed according to the full length trh gene, which were used for semi-quantitative RT-PCR of differdnt samples in developmental satages. The rusults showed that the trh expression in pre-nauplius increased till nauplii then decreased in late nauplii. Western blot was used to detect the relationship between enviromental salinity and trh expression, and showed that the relationship positively correlated, indicating that Artemia trh gene had function in osmoregulation. Whole immunohistochemistry staining resuls showed that in the period of pre-naupliu, nauplius, latter-nauplius, trh gene expresses in the salt gland of Artemia sinica. In the same time, lightly expresses in the anlages of thoracic epipod of latter-nauplius. However, in the period of sub-adult and adult, it expresses in the thoracic epipods of Artemia sinica. In this study, the 1987bp full length of APH-1 cDNA was also obtained by RACE(Genebank:FJ952936). Semi-quantitative RT-PCR of APH-1 showed that the APH-1 expression in pre-nauplius increased till nauplii then decreased in late nauplii. Western blot was used to detect the relationship between enviromental salinity and APH-1 expression, and showed that the relationship positively correlated, indicating that Artemia APH-1 gene had function in osmoregulation. Whole immunohistochemistry staining resuls showed that in the period of pre-naupliu, nauplius and latter-nauplius, APH-1 gene expresses in the salt gland of Artemia sinica. The expression can not be detected after the disappearance of salt gland.