Effect Of Eyestalk-ablation On Differential Expressed Genes And Profile Of The Important Genes During Ovarian Development Stages Of Shrimp Litopenaeus Vannamei

Litopenaeus vannamei also known as P. vannamei, is a majoy aquaculture speciesin China. In the present study, by using suppression subtractive hybridization (SSH) webuild library of different expression genes between oogonia proliferating (firstphase,before eyestalk-ablation) and primary vitellogenesis (third phase, aftereyestalk-ablation) in ovary and hepatopancreas. The sequencing results were comparedby BLASTx and nucleotide BLAST homology on GenBank, and analysis by NCBIdatabase for differential expressed gene function annotation. Selectting several differentexpression genes validated by the fluorescence quantitative PCR to study the geneexpression abundance during the different stages of the ovarian development. Analyzethe relationship between the expression features and functions of these genes, as well asthe relationship between these genes and ovarian development. Then, by using thefluorescence quantitative PCR, research the expression of hemocyanin and vitellogeninin the ovaries and hepatopancreas, during the ovarian development. And analyze therelationship between hemocyanin and vitellogenesis.The SSH results show that differentially expressed genes in the first stage ofovarian is: polehole-like protein, Thrombospondin, and nuclear autoantigen spermprotein genes. The third stage ovarian differentially expressed genes were: intracellularfatty acid-binding protein; protein disulfide isomerase; and hemocyanin genes. Thedifferentially expressed genes in the first phase of hepatopancreas is: ATP subunit,NADH subunits and ferritin gene. The third stage differentially expressed genes is:vitellogenin and hemocyanin genes. Polehole-like protein and vitellogenin expressed both in the first and third stage. Vitellogenin gene has a strong expression in the thirdstage. Gene expression is related to the status of oocytes division in the first stage andvitellogenic in the third stage.Quantitative PCR results showed that polehole-like protein (PH), nuclearautoantibodies spermatogonial protein (NA), and thrombospondin (THP) geneexpressed primarily in the ovary. The expression of these genes were the highest beforeeyestalk-ablation. With ovarian development, the expression decline gradually, andreached the lowest in the fourth and fifth stages. The main function of PH, NA and othergenes are signal transduction and regulation of cell division. In the early stages, the highexpression and ovarian development are closely related.Real-time quantitative PCR were used to detect the expression of hemocyanin andvitellogenin mRNA in different developmental stages of ovary and hepatopancreas. Theresults showed that hemocyanin mRNA expressed mainly in hepatopancreas, with thedevelopment of the ovaries, the expression had an increasing trend and peaked in thethird stage and subsequently decreased slightly, but remained at a high level, reduced tothe initial level in the sixth stage. The expression of vitellogenin mRNA in ovarianincreased with the development of ovary and highest in the fifth stage and then declinerapidly.The result showed that hemocyanin and ovary development has some correlation.Hemocyanin whether directly or indirectly involved in vitellogenesis and ovariandevelopment should further exploration studies.