基于转录组测序筛选肉鸡腹水综合征相关候选基因

doi:10.11843/j.issn.0366-6964.2025.02.008

摘要/Abstract

摘要：

旨在从心和肺组织中筛选介导肉鸡腹水综合征发生的关键候选基因与通路。本研究以33日龄快速型白羽肉鸡父系为素材，选取患腹水综合征公鸡5只，正常公鸡3只，通过剖检与血液生化指标测定确定腹水综合征公鸡发病情况，分为腹水组与正常组，腹水组5只公鸡，正常组3只公鸡。对腹水组与正常组进行血液生化指标测定与比较分析，解剖取左心室心肌组织制作HE切片，取心、肺组织样品进行转录组测序。血液生化结果显示腹水组红细胞压积(HCT)、血红蛋白(HGB)和粒细胞百分比(GRA)非常显著高于正常组(P＜0.01)。腹水组淋巴细胞百分比(LYM)非常显著低于正常组(P＜0.01)。腹水组二氧化碳分压(PCO2)、总钙(Ca)含量非常显著高于正常组(P＜0.01)，腹水组剩余碱(Beb)含量显著低于正常组(P＜0.05)。HE切片结果显示，正常组心肌组织肌原纤维排列整齐，细胞间未见血细胞沉积；腹水组心肌细胞中肌原纤维结构紊乱，大量血细胞沉积在肌原纤维之间。转录组分析筛选到肺组织差异表达基因969个，包括417个上调基因和552个下调基因，心组织差异表达基因809个，包括397个上调基因和412个下调基因。通路富集显著富集到了细胞周期(mitotic cell cycle)、着丝粒复合物组装(centromere complex assembly)、氧气结合(oxygen binding)、神经活性配体-受体相互作用(neuroactive ligand-receptor interaction)、细胞黏附分子(cell adhesion molecules)、细胞衰老(cellular senescence)、细胞外基质-受体相互作用(ECM-receptor interaction), 运动蛋白(motor proteins)。从富集通路中筛选与肉鸡腹水综合征相关的基因进行实时荧光定量PCR验证，结果KIF20A、NUSAP1、HBAD、TFRC、HBBA基因在腹水组和正常组间差异表达趋势与转录组测序结果一致，可作为肉鸡腹水综合征重要候选基因。本研究利用转录组技术筛选到与肉鸡腹水综合征的发生相关的重要通路和候选基因，对研究肉鸡腹水综合征的发病机制和预防具有十分重要的意义。

关键词: 肉鸡, 腹水综合征, 转录组, 差异表达基因

Abstract:

This study aimed to identify key candidate genes and pathways in cardiac and pulmonary tissues that mediate the development of ascites syndrome in broiler chickens.Taking the sires of fast-growing white-feathered broilers at 33 days of age as the research materials, 5 roosters with ascites syndrome and 3 normal roosters were selected. The occurrence of ascites syndrome in the roosters was determined through necropsy and the measurement of blood biochemical indicators. The subjects were categorized into an ascites group (5 roosters) and a control group (3 roosters).The results of blood biochemical analysis indicated that hematocrit (HCT), hemoglobin (HGB), and granulocyte percentage (GRA) in the ascites group were significantly higher than those in the normal group (P < 0.01).The lymphocyte percentage (LYM) in the ascites group was significantly lower than that in the normal group (P < 0.01).Additionally, the partial pressure of carbon dioxide (PCO₂) and total calcium (Ca) levels were significantly elevated in the ascites group compared to the normal group (P < 0.01), while the base excess (Beb) content was significantly lower in the ascites group (P < 0.05).The results of HE staining showed that in the normal group, the myofibrils in the myocardial tissue were arranged neatly, with no evidence of blood cell deposition between cells. In the ascites group, myocardial fibers exhibited structural disarray, and a large number of blood cells were deposited between the myofibrils. Transcriptome analysis identified 969 differentially expressed genes (DEGs) in lung tissue, including 417 upregulated genes and 552 downregulated genes. In cardiac tissue, 809 DEGs were identified, comprising 397 upregulated genes and 412 downregulated genes. Pathway enrichment analysis revealed significant enrichment in processes such as the mitotic cell cycle, centromere complex assembly, oxygen binding, neuroactive ligand-receptor interaction, cell adhesion molecules, cellular senescence, extracellular matrix (ECM)-receptor interaction, and motor proteins. Genes associated with ascites syndrome in broiler chickens were selected from enriched pathways and validated using real-time quantitative PCR. The results showed that the expression trends of KIF20A, NUSAP1, HBAD, TFRC, and HBBA genes differed between the ascites and normal groups, consistent with transcriptome sequencing findings, and these genes could be regarded as important candidate genes for ascites syndrome in broilers.The study, utilizing transcriptome technology, has identified significant pathways and candidate genes related to the development of ascites syndrome in broiler chickens, providing important insights into the disease mechanism and prevention strategies.

Key words: broiler, ascites syndrome, transcriptome, differentially expressed genes

中图分类号:

S831.2

苏蒙, 刘莎, 宋丹丽, 高倩梅, 郑麦青, 文杰, 赵桂苹, 李庆贺. 基于转录组测序筛选肉鸡腹水综合征相关候选基因[J]. 畜牧兽医学报, 2025, 56(2): 559-570.

SU Meng, LIU Sha, SONG Danli, GAO Qianmei, ZHENG Maiqing, WEN Jie, ZHAO Guiping, LI Qinghe. Identification of Candidate Genes Associated with Ascites Syndrome in Broilers Based on Transcriptome Sequencing[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(2): 559-570.

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基因 Gene	引物序列(5′→3′) Primer sequence	产物长度/bp Product length
NUSAP1	F: AGATTCGAAGAGTGGCGGTT	145
NUSAP1	R: AACAGTTTATCGGCCCGGAG	145
HBAD	F: AACGCTGTGAAGAACGTGGA	197
HBAD	R: GACACGGCAGACAGGAACTT	197
KIF20A	F: GAAACCCCAACAGCAGCG	401
KIF20A	R: ACCTATGTGCTGCTTGTCCC	401
TFRC	F: TTATCGTGGACGAATCGAGC	239
TFRC	R: CCTCACCAGCCTCAAAGGAG	239
HBBA	F: GAGCAAGAGCCCAGACCTC	131
HBBA	R: ACATTCGGCCACATTGACCT	131
β-actin	F: GAGAAATTGTGCGTGACATCA	152
β-actin	R: CCTGAACCTCTCATTGCCA	152

富集类型 Category	编号 ID	条目/通路 Term/Pathway	P值 P-value	基因 Gene
GO 条目 GO term	GO: 0000278	细胞周期	8.28×10^-6	NUSAP1/CCNA2/CDK1
GO 条目 GO term	GO: 0034508	着丝粒复合物组装	3.23×10^-5	CENPO/CENPW/CENPN
GO 条目 GO term	GO: 0019825	氧气结合	0.001 3	HBBA/HBAD/HBA1
KEGG 通路 KEGG pathway	gga04080	神经活性配体-受体相互作用	8.51×10^-5	TSPO2/P2RY10/ADRA2BL2/EDN1/GABBR2
KEGG 通路 KEGG pathway	gga04514	细胞黏附分子	0.001 3	ITGA4/MHCY11/MHCY4/
KEGG 通路 KEGG pathway	gga04218	细胞衰老	5.23×10^-5	MHCY4/MHCY14/CCNB3/
KEGG 通路 KEGG pathway	gga04512	细胞外基质-受体相互作用	0.001 3	LAMB3/COL6A6/ITGA2B
KEGG 通路 KEGG pathway	gga04814	运动蛋白	0.000 433	KIF11/CENPE/KIF20A

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