Expression Pattern Of As-ClC Gene In Early Development And Under Salinity Stress Of Artemia Sinica

Artemia sinica live in high salinity water worldwide and are frequently exposed to seasonal salinity changes. Therefore, osmoregulation and the adaption to hypertonic and hypotonicity saline for Artemia sinica are an especially significant aspect of Artemia physiology. The gene of chloride channels protein from Artemia sinica (As-ClC) is a member of the Chloride channels protein family. When cells are exposed to hypotonic or hypotonicity media, An influx or outflux of Cland K+via activation of Cl and K+channels, which leads to movement of water to regulate cell volume reached equilibrium between intracellular osmolarity and extracellular. In addition, GCK-3/ste20(ste20) gene which is a member of serine/threonine kinase superfamily also play a indispensable role in C1C osmoregulation pathway. In yeast, the activation of ste20can trigger a MAPK cascade to regulate cell division, growth, cell cycle and maintain the balance of osmotic pressure in hydrostatic and hypertonic.In this paper, the2201bp complete cDNA sequence of As-ClC was cloned from Artemia sinica which were used for experimental material for the first time using RACE technology. It was predicted to traverse the membrane11times and contained two CBS domain at carboxyterminal cytoplasmic region and a voltage-gated C1C domain.The expression of As-ClC has been researched in diverse stage during embryonic development in Artemia by In situ hybridization (ISH) assay. The result showed that this protein was distributed widely and evenly in the whole body of cell and might be constitutive expression.The expression of As-ClC and GCK-3/ste20have been researched in diverse stage during embryonic development and salinity stress in Artemia by real-time PCR analysis. The result showed that there was a lower expression levels of As-ClC gene after Oh and a higher expression level present to stage of15h and40h when the A.sinica entered to next growth period and also the environment changed, at adult period, As-ClC maintain a higher expression level. The results of Salinity stress assay showed obviously an increasing trend of As-ClC transcripts with increased salinity. The expression of As-ClC was higher in the control group (28‰) than in the experimental group except at salinity for200. Howerver, The expression of GCK-3/ste20revealed an declining trend. This demonstrated that As-CC, a salinity-stress-related gene, probably participated in cell volum-regulation and osmotic regulation, during the early embryonic development of A. sinica. Therefore, researching on salt tolerance genes may have a certain theoretical significance for managing salinization in the future. At the same time, it also has practical value for disaster mitigation and genetically modification on salt-tolerance plants and animals.