Transcriptome Analysis and Identification of Differentially Expressed Transcripts of Immune-Related Genes in Spleen of Gosling and Adult Goose.
Transcriptome Analysis and Identification of Differentially Expressed Transcripts of Immune-Related Genes in Spleen of Gosling and Adult Goose.
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DOI：
10.3390/ijms160922904
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发表时间：
2015-09-22
影响因子：
5.6
通讯作者：
Cheng A
中科院分区：
文献类型：
--
作者：
Wang A;Liu F;Chen S;Wang M;Jia R;Zhu D;Liu M;Sun K;Wu Y;Chen X;Cheng A
鹅（Anser cygnoides）是一种重要的经济家禽品种，具有营养价值高、羽毛品质好、经济效益高等特点。然而，小鹅比成年鹅对病原体更高的易感性的分子机制仍然知之甚少。本研究对1只2周龄雏鹅和1只成年鹅的脾脏组织切片进行了比较分析。雏鹅的脾脏主要由间充质组成，伴有散在的淋巴细胞，而成年鹅的脾脏实质发育良好。为了研究鹅免疫相关基因，我们使用配对末端测序技术（Illumina）对脾脏样品进行了深度转录组和基因表达分析。使用Trinity软件和TGICL软件总共组装了50，390个unigenes。此外，这些组装的unigenes与基因描述和基因本体论（GO）分析进行了注释。通过京都基因和基因组百科全书（KEGG）分析，我们研究了558个重要的免疫相关的unigenes和23个预测的细胞因子。此外，还鉴定了22个在雏鹅和成年鹅之间存在差异表达的免疫相关基因，其中表达差异最大的3个基因分别为免疫球蛋白α重链（IgH）、甘露聚糖结合凝集素丝氨酸蛋白酶1亚型X1（MASP 1）和C-X-C趋化因子受体4型（CXCR 4）。最后，在这22个差异表达的免疫相关基因中，包括肿瘤坏死因子受体超家族成员13 B（TNFRSF 13 B）、C-C基序趋化因子4样（CCL 4）、CXCR 4、白细胞介素2受体α（IL 2 RA）、MHC I类重链（MHCIα）、抗原加工转运蛋白2（TAP 2）IgH的7个基因通过定量实时PCR（qRT-PCR）得到确认。除TNFRSF 13 B外，其他候选基因在成年鹅中的表达水平均上调。对雏鹅和成年鹅脾脏转录组的比较分析有助于更好地了解鹅免疫分子发育。
The goose (Anser cygnoides), having high nutritional value, high-quality feathers and high economic benefit, is an economically important poultry species. However, the molecular mechanisms underlying the higher susceptibility to pathogens in goslings than in adult geese remains poorly understood. In this study, the histological sections of spleen tissue from a two-week-old gosling and an adult goose, respectively, were subjected to comparative analysis. The spleen of gosling was mainly composed of mesenchyma, accompanied by scattered lymphocytes, whereas the spleen parenchyma was well developed in the adult goose. To investigate goose immune-related genes, we performed deep transcriptome and gene expression analyses of the spleen samples using paired-end sequencing technology (Illumina). In total, 50,390 unigenes were assembled using Trinity software and TGICL software. Moreover, these assembled unigenes were annotated with gene descriptions and gene ontology (GO) analysis was performed. Through Kyoto encyclopedia of genes and genomes (KEGG) analysis, we investigated 558 important immune-relevant unigenes and 23 predicted cytokines. In addition, 22 immune-related genes with differential expression between gosling and adult goose were identified, among which the three genes showing largest differences in expression were immunoglobulin alpha heavy chain (IgH), mannan-binding lectin serine protease 1 isoform X1 (MASP1) and C–X–C chemokine receptor type 4 (CXCR4). Finally, of these 22 differentially expressed immune-related genes, seven genes, including tumor necrosis factor receptor superfamily member 13B (TNFRSF13B), C-C motif chemokine 4-like (CCL4), CXCR4, interleukin 2 receptor alpha (IL2RA), MHC class I heavy chain (MHCIα), transporter of antigen processing 2 (TAP2) IgH, were confirmed by quantitative real-time PCR (qRT-PCR). The expression levels of all the candidate unigenes were up-regulated in adult geese other than that of TNFRSF13B. The comparative analysis of the spleen transcriptomes of gosling and adult goose may promote better understanding of immune molecular development in goose.