title: Sequencing of the first ayu (Plecoglossus altivelis) macrophage transcriptome and microarray development for investigation the effect of LECT2 on macrophages.
abstract: Macrophages play an important role in first-line host defense of innate immune in fishes. However, it is difficult to investigate cellular mechanism of immune response in fish species with little genomic information available. Here we present the first use of RNA-Sequencing to study the macrophage transcriptome of ayu, Plecoglossus altivelis, which is an economically important fish in East Asia. De novo assembly generated 49,808 non-redundant consensus sequences, among which 23,490 transcripts found respective coding sequences. 15,707 transcripts are predicted to be involved in known metabolic or signaling pathways. The sequences were then used to develop a microarray for measurement the effect of recombinant LECT2 on ayu macrophages. LECT2 altered expression of a variety of genes mainly implicated in actin cytoskeleton, pattern recognition receptors and cytokines. Meanwhile, LECT2 enhanced phagocytosis, bacterial killing, and respiratory burst in ayu macrophages, which supported the thought derived from the microarray data that LECT2 activates macrophages. In conclusion, our results contribute to understanding the specific regulation mechanism of LECT2 in macrophage activation, and the combination of transcriptome analysis and microarray assay is a good method for screening a special tissue or cell response to a stimulus or pathogen in non-model fish species.
authors.affiliation: School of Marine Sciences, Ningbo University, Ningbo City, Zhejiang Province 315211, China.
authors.forename: Xin-Jiang
authors.initials: XJ
authors.lastname: Lu
authors.forename: Xing-Yi
authors.initials: XY
authors.lastname: Hang
authors.forename: Ling
authors.initials: L
authors.lastname: Yin
authors.forename: Yu-Qing
authors.initials: YQ
authors.lastname: He
authors.forename: Jiong
authors.initials: J
authors.lastname: Chen
authors.forename: Yu-Hong
authors.initials: YH
authors.lastname: Shi
authors.forename: Chang-Hong
authors.initials: CH
authors.lastname: Li
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001483:Base Sequence; D015723:Gene Library; D060226:Head Kidney / Q000166:cytology; D059014:High-Throughput Nucleotide Sequencing / Q000662:veterinary; D008264:Macrophages / Q000378:metabolism*; D046228:Microarray Analysis / Q000379:methods / Q000662:veterinary*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D036441:Osmeriformes / Q000235:genetics*; D010587:Phagocytosis / Q000276:immunology; D060888:Real-Time Polymerase Chain Reaction; D016897:Respiratory Burst; D013045:Species Specificity; D059467:Transcriptome / Q000235:genetics*