Gonad-inhibiting Hormone(GIH) Of The Macrobrachium Nipponense: Immunolocalization And The Effect Of Atrazine On The Expression Of GIH

Gonad-inhibiting hormone(GIH) belongs to CHH/MIH/GIH neuropeptide family, which includes crustacean hyperglycemic hormone(CHH), molt-inhibiting hormone MIH), gonad-inhibiting hormone(GIH) and mandibular organ-inhibiting hormone(MOIH). Gonad-inhibiting hormone plays a very important role in crustacean. In this study, two types of type II neuropeptide genes encoding gonad inhibiting hormone(GIH-A and GIH-B) were identified. Recombinant peptides related to gonad inhibiting hormone of the Macrobrachium nipponense were expressed in bacterial cells, and then purified. The titer of antibody was detected by double diffusion.The specific antibody titer in the serum of rabbits immunized with the purified protein was 1∶64.Two specific polyclonal antibodies were raised against the mature region of GIH-A and GIH-B. The specificity was detected by Western blotting. Results from Western blot analysis indicated that the two antibodies were specific. Immunofluorescence studies were carried out by using these antibodies to detect the GIH-A and GIH-B in optic ganglia and thoracic ganglion of M. nipponense. GIH was found in both optic ganglia and thoracic ganglion in M. nipponense. This study showed that GIH was expressed not only in the X-organ-sinus gland complex of optic ganglia but also in other tissues in M. nipponense.Atrazine is an endocrine disruptor, which can seriously affect the endocrine system of crustaceans. In this paper, after dealt with the different concentrations of atrazine(0.001 mg / L, 0.01 mg / L, 0.1mg / L, 1mg / L), the expression of GIH-B in the optic ganglia in M. nipponense was detected using fluorescent quantitative RT-PCR. The results show that when atrazine concentration was 0.1mg/L, the expression of GIH-B was affected significantly. There was a clear downward trend.The data will not only facilitate the further research of functions and the mode of actions of GIH, but also can provide a foundation on the research of precocity of M. nipponense in aquiculture. At the same time, the paper can also provide new information on the toxicity of atrazine in crustacean endocrine system.The data will not only facilitate the further research of functions and the mode of actions of GIH, but also can provide a foundation on the research of precocity of M. nipponense in aquiculture.