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对表现出对空肠弯曲菌抗性遗传差异的近交系鸡肠道组织的转录组分析。

Transcriptomic analysis of caecal tissue in inbred chicken lines that exhibit heritable differences in resistance to Campylobacter jejuni.

机构信息

The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UK.

The Royal Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire, AL9 7TA, UK.

出版信息

BMC Genomics. 2021 Jun 4;22(1):411. doi: 10.1186/s12864-021-07748-2.

Abstract

BACKGROUND

Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans and the handling or consumption of contaminated poultry meat is a key source of infection. Selective breeding of poultry that exhibit elevated resistance to Campylobacter is an attractive control strategy. Here we studied the global transcriptional response of inbred chicken lines that differ in resistance to C. jejuni colonisation at a key site of bacterial persistence.

RESULTS

Three-week-old chickens of line 6 and N were inoculated orally with C. jejuni strain M1 and caecal contents and tonsils were sampled at 1 and 5 days post-infection. Caecal colonisation was significantly lower in line 6 compared to line N at 1 day post-infection, but not 5 days post-infection. RNA-Seq analysis of caecal tonsils of both lines revealed a limited response to C. jejuni infection compared to age-matched uninfected controls. In line N at days 1 and 5 post-infection, just 8 and 3 differentially expressed genes (DEGs) were detected (fold-change > 2 and false-discovery rate of < 0.05) relative to uninfected controls, respectively. In the relatively resistant line 6, a broader response to C. jejuni was observed, with 69 DEGs relating to immune regulation, cell signalling and metabolism at 1 day post-infection. However, by day 5 post-infection, no DEGs were detected. By far, the greatest number of DEGs were between uninfected birds of the two lines implying that differential resistance to C. jejuni is intrinsic. Of these genes, several Major Histocompatibility Complex class I-related genes (MHCIA1, MHCBL2 and MHCIY) and antimicrobial peptides (MUC2, AvBD10 and GZMA) were expressed to a greater extent in line N. Two genes within quantitative trait loci associated with C. jejuni colonisation were also more highly expressed in line N (ASIC4 and BZFP2). Quantitative reverse-transcriptase PCR analysis of a subset of transcripts confirmed the RNA-Seq results.

CONCLUSIONS

Our data indicate a limited transcriptional response in the caecal tonsils of inbred chickens to intestinal colonisation by Campylobacter but identify a large number of differentially transcribed genes between lines 6 and N that may underlie variation in heritable resistance to C. jejuni.

摘要

背景

空肠弯曲菌是人类细菌性肠胃炎的主要病原体,而接触或食用受污染的禽肉是感染的主要来源。对表现出对空肠弯曲菌更高抗性的家禽进行选择性繁殖是一种有吸引力的控制策略。在这里,我们研究了在细菌持续存在的关键部位对空肠弯曲菌定植具有不同抗性的近交系鸡的全球转录反应。

结果

3 周龄的 6 号线和 N 号线鸡用空肠弯曲菌菌株 M1 经口接种,在感染后 1 天和 5 天分别采集盲肠内容物和扁桃体样本。与感染后 5 天相比,感染后 1 天,6 号线的盲肠定植明显低于 N 号线。对两条线的盲肠扁桃体进行 RNA-Seq 分析表明,与年龄匹配的未感染对照相比,对空肠弯曲菌感染的反应有限。在感染后 1 天和 5 天的 N 号线中,相对于未感染对照,分别仅检测到 8 个和 3 个差异表达基因(DEGs)(倍数变化> 2,错误发现率< 0.05)。在相对抗性的 6 号线中,观察到对空肠弯曲菌的更广泛反应,在感染后 1 天与免疫调节、细胞信号转导和代谢相关的 69 个 DEG。然而,到感染后 5 天,未检测到 DEG。到目前为止,数量最多的 DEG 存在于两条线的未感染鸟类之间,这表明对空肠弯曲菌的不同抗性是内在的。在这些基因中,几种主要组织相容性复合体 I 类相关基因(MHCIA1、MHCBL2 和 MHCIY)和抗菌肽(MUC2、AvBD10 和 GZMA)在 N 线中表达更强烈。与空肠弯曲菌定植相关的数量性状基因座内的两个基因在 N 线中也表达更高(ASIC4 和 BZFP2)。一组转录本的定量逆转录聚合酶链反应分析证实了 RNA-Seq 的结果。

结论

我们的数据表明,近交系鸡的盲肠扁桃体对空肠弯曲菌肠道定植的转录反应有限,但在 6 号线和 N 号线之间确定了大量差异转录的基因,这些基因可能是可遗传抗性的基础对空肠弯曲菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6e41/8176612/3ff1613e7932/12864_2021_7748_Fig1_HTML.jpg

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