Expression Of V-ATPase Gene During Early Embryo Development Of Chinese Artemia Parthenogenetica

Brine shrimp(Artemia)is widely distributed in inner and salt lake and coasting saltworks,it belongs to Crustacea, Branchiopoda, Anostacea, Artemidae, Artemia. The vacuolar ATPase (V-ATPases) is a large, complex enzyme that can pump H+ outside of the cell or into the lumens of vacuolar organelles to maintain the cytoplasmic pH at a relative stable neutral level. This kind of environment is essential for the survival of cell and can also acidify some intracellular apartments which are contributing to the normal function of endosomes, lysosomes and Golgi apparatus. Following the mRNA sequence of Artemia parthenogenetica V-ATPase gene on GenBank, the article uses total RNA of Artemia franciscana as template and gets 1577bp-long gene fragment by RT-PCR. After the analysis on presumptive amino acid sequence and related protein amino acid sequence supporting by Blastn, software ClustalX 1.83 and MEGA 3.0 software, we demonstrated that this sequence is the V-ATPase gene of Artemia parthenogenetica and revealed that the fragment displayed up to 97% identity with V-ATPase gene in Artemia franciscana available in GenBank Artemia franciscana (AY197611).V-ATPase gene sequences encoded 401 amino acid residues by CLUSTALW (1.83). The deduced Artemia parthenogenetica protein sequence demonstrates a highly degree of amino acid identity with Artemia franciscana—98% and its consanguinity is the closest with A.franciscana. Gene expression analysis using real-time RT-PCR is currently the most sensitive and reproducible method of detecting and quantifying an mRNA transcript.In this paper, the samples of Artemia parthenogenetica were collected at 0 hour, 4 hour, 8 hour, 12 hour, 16 hour and 24 hour. Real-time PCR analysis of the mRNA was used to demonstrate differential expressions through encysted embryo development. The analysis of mRNA expression demonstrated a low level before 12 hour, followed by an increase in 16 hour. The expression of V-ATPase at 24 hour is similar to that in 16 hour. The V-ATPase of brine shrimp embryos may be downregulated under anoxia before 12 hour and the mRNA expression of V-ATPase decreases.The mechanism of V-ATPase for regulated the acidification of the intracellular compartments may represent a critical adaptive trait to anoxia tolerance in this species, Artemia parthenogenetica.