Project description:The transcriptome profiling five tissues of juvenile Eriocheir sinensis, including gill, muscle, thoracic ganglion, eyestalk and hepatopancreas, were sequenced to get the basic dataset for constructed a genome-scale metabolic network model for E. sinensis. The model was used to predict the optimal nutrient requirements of E. sinensis in feed and suggestions for feed improvement were put forward based on the simulation results.
Project description:We sequenced the mRNA in the Y-organ of normal and eyestalk-ablated Eriocheir Sinensis and obtained the transcriptome profiling (RNA-seq) of the two samples with Next-generation sequencing (NGS) technique. The unigene expression of the two samples were compared and the differetially expressed unigenes were listed.
Project description:Spiroplasma eriocheiris, has been identified as a novel lethal pathogen of Eriocheir sinensis tremor disease (TD), one neurological disease with typically paroxysmal tremors of the pereiopod. This pathogen infected and multiplied in the hemocytes of E. sinensis as the first target cells, and then follow the blood circulation to infect the crab other tissues. S. eriocheiris infected the nerves tissue is the directly reason of TD. But the pathogenic mechanism of TD was still few known. Firstly, in the current study, the phosphoproteomic changes of E. sinensis thoracic ganglion after S. eriocheiris infection were obtained. KEGG pathway analysis show Wnt signaling pathways was restrained, corresponding many nervous system development and signal transmission pathway also destroyed. Base on the identified modified sequence, several peptides (GSK3β, SYN, VAMP and SNAMP-25) were selected to synthesize chemically and prepare phosphorylated antibodies. The differentially expressed phosphorylated proteins GSK3β-pSer9, VAMP-pSer72, SNAP25-pSer102, and SYN-pSer134 in thoracic ganglion were significantly down-regulated verified by immunohistochemistry and western blot, this results are similar with the phosphoproteomic. By qRT-PCR, western blot, RNA interference and inhibitor experiments, when the S. eriocheiris infected the hemocytes of crab, the GSK-3β and β-Catenin in Wnt-β-Catenin pathway were restrained similar in the thoracic ganglion. The S. eriocheiris can restricts the hemocytes Wnt-β-Catenin pathway to help itself infection no matter in vivo or in vitro. Neurotransmitter metabolite analysis showed that four kinds of neurotransmitters (5-Hydroxy-L-tryptophan, Serotonin, Acetylcholine and γ-Amino-butyric acid) in thoracic ganglion were metabolic disorders in E. sinensis thoracic ganglion after S. eriocheiris infection. The present work could serve as a basis for understanding the role of Wnt signaling pathway in the process of S. eriocheiris invasion E. sinensis hemocytes and causing paroxysmal tremors of E. sinensis the pereiopod.
