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Extraction of Hemocytes from Drosophila melanogaster Larvae for Microbial Infection and Analysis
Published on: May 24, 2018
Transcriptome-wide analysis of immune responses in Eriocheir sinensis hemocytes after challenge with different
Xingkun Jin1, Weiwei Li2, Minjie Xu3
1Institute of Marine Biology, School of Oceanography, Hohai University, Nanjing, 210098, China; Department of Biology, School of Life Science, East China Normal University, Shanghai, 200241, China.
Abstract:
Knowledge about how Eriocheir sinensis interacts with microorganisms in its ambient environment is still lacking. Using RNA-Seq, we determined the most conserved genes and pathways compared with other animals and detected highly-induced immune genes in E. sinensis hemocytes post-in vivo challenge with different microbial derivatives. In total, 33.2 million high-quality reads were generated and assembled into 177,679 contigs. Completeness assessment and functional annotation were performed. Lipopolysaccharide, peptidoglycan, and β-1, 3-glucan stimulation induced 373, 173, and 108 differentially expressed (DE) transcripts, respectively. GO terms such as 'G-protein-coupled receptor binding', 'negative regulation of mitogen-activated protein kinase activity', and 'positive regulation of blood circulation' were enriched in the DE transcripts. Quantitative real-time PCR validated the data for selected genes. Our data contribute to understanding the immune defense mechanism in E. sinensis and the development of the innate immune system, thereby providing insights into disease control and prevention in aquaculture.
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