Effects of IAG Recombinant Plasmid Injection on the Expression of IAG, mhc, gasp and Secondary Sexual Characteristics of Procambarus clarkii
CHEN Yi-Jun, XIE Ying-Lin, ZHAO Tong, YING Ke, LI Yan-He*
Key Laboratory of Freshwater Biological Breeding, Ministry of Agriculture and Rural Affairs/National Experimental Teaching Demonstration Center for Aquaculture, Huazhong Agricultural University, Wuhan 430070, China
Abstract：The insulin-like androgenic gland hormone (IAG) gene plays an important role in sexual differentiation and growth in decapod crustaceans. To investigate the effects of recombinant plasmids containing IAG on the expression of endogenous genes (IAG) and growth-related genes myosin heavy chain(mhc) and growth arrest-specific protein (gasp) as well as secondary sexual characteristics in Procambarus clarkii. This study involved injecting different doses (0.5, 1.0 and 3.0 μg/g) of IAG recombinant plasmids into the muscle of healthy minor female P. clarkii weighing between 6.5~16.5 g. The weight, abdominal development, and expression of genes IAG, mhc, and gasp in the tissues of the experimental shrimp were monitored at 1, 7, 14, and 28 d, respectively. The results showed that compared with the control group, the copy number of plasmids in the experimental group was positively correlated with the concentration of injection. The copy number in each tissue showed a distribution pattern of muscle>gonads>appendages>hepatopancreas. The degree of plasmid digestion was the highest between 3 and 7 d, and at 28 d, the plasmid was almost completely digested. After 1 d of injection, the expression of mhc in muscles significantly increased at injection concentrations of 1.0 and 3.0 μg/g (P<0.05), and body weight gain was also significantly higher than that of the control group (P<0.05). Within 28 d after injection, female crayfish with an injection concentration of 3.0 μg/g had the fastest ovarian development, while the control group had the slowest development. No masculinization was observed in the abdomen of all female shrimps. Before the mass digestion of plasmids, the expression levels of IAG in the gonads and gasp in the muscles of the experimental group were lower than those in the control group, and their expression was inhibited. The results showed that the injection of IAG recombinant plasmid had certain effects on ovarian development and expression of IAG, mhc, gasp. This study provides reference for research on IAG function and gender control.
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