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.
Project description:As a novel lethal pathogen of Eriocheirsinensistremor disease,Spiroplasma eriocheiris, has led into catastrophic economic losses in aquaculture.The hemocytes of E. sinensis is the first target cells of S. eriocheiris. Our study is the first time designed to understanding of the phosphoproteome and N-glycoproteome dynamics of E. sinensis hemocytesunderS. eriocheiris infected at the early phage. In the current study, the phosphoproteome and N-glycoproteome changes of E. sinensishemocytesafter S. eriocheiris infection were obtained usingtandem mass tags(TMT) labeling and affinity enrichment followed by high-resolution Liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) analysis. Using a 1.2-fold change in expression as a physiologically significant benchmark.In total, 549 differentially expressed phosphoproteins were reliably quantified, including 278 up-regulated phosphoproteinsand 271 down-regulated phosphoproteins; 167 differentially expressed N-glycosylated proteins were reliably quantified, including 79 up-regulated glycosylated proteins and 88 down-regulated glycosylated proteins subsequented to S. eriocheiris infection.Gene ontology (GO) annotation, protein domain annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation, subcellular localization annotation and protein-protein interaction network were used to analysis those significantly differentlyexpression proteins shown that many biological process and pathway are participate inS. eriocheiris infected host cell, such as phagocytosis, ECM-receptor interaction, lysosome, prophenoloxidase system, and so on. Our study could serve as a basis to understand the relationship between E. sinensisand the pathogen S. eriocheiris, and alsoprovide reference to study protein phosphorylation and N-glycosylationin other crustaceans.
2026-04-27 | PXD014116 | Pride
Project description:Eriocheir sinensis infected with Spiroplasma eriocheiris Raw sequence reads
Project description:Endosymbiotic bacteria associated with eukaryotic hosts are omnipresent in nature, particularly in insects. Studying the bacterial side of host-symbiont interactions is, however, often limited by the unculturability and genetic intractability of the symbionts. Spiroplasma poulsonii is a maternally transmitted bacterial endosymbiont that is naturally associated with several Drosophila species. S. poulsonii strongly affects its host’s physiology, for example by causing male killing or by protecting it against various parasites. Despite intense work on this model since the 1950s, attempts to cultivate endosymbiotic Spiroplasma in vitro have failed so far. Here, we developed a method to sustain the in vitro culture of S. poulsonii by optimizing a commercially accessible medium. We also provide a complete genome assembly, including the first sequence of a natural plasmid of an endosymbiotic Spiroplasma species. Last, by comparing the transcriptome of the in vitro culture to the transcriptome of bacteria extracted from the host, we identified genes putatively involved in host-symbiont interactions. This work provides new opportunities to study the physiology of endosymbiotic Spiroplasma and paves the way to dissect insect-endosymbiont interactions with two genetically tractable partners.