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. 2017 Mar 7;18(1):21.
doi: 10.1186/s12863-017-0488-4.

Deciphering alternative splicing and nonsense-mediated decay modulate expression in primary lymphoid tissues of birds infected with avian pathogenic E. coli (APEC)

Affiliations

Deciphering alternative splicing and nonsense-mediated decay modulate expression in primary lymphoid tissues of birds infected with avian pathogenic E. coli (APEC)

Hongyan Sun. BMC Genet. .

Abstract

Background: Avian pathogenic E. coli (APEC) can lead to a loss in millions of dollars in poultry annually because of mortality and produce contamination. Studies have verified that many immune-related genes undergo changes in alternative splicing (AS), along with nonsense mediated decay (NMD), to regulate the immune system under different conditions. Therefore, the splicing profiles of primary lymphoid tissues with systemic APEC infection need to be comprehensively examined.

Results: Gene expression in RNAseq data were obtained for three different immune tissues (bone marrow, thymus, and bursa) from three phenotype birds (non-challenged, resistant, and susceptible birds) at two time points. Alternative 5' splice sites and exon skipping/inclusion were identified as the major alternative splicing events in avian primary immune organs under systemic APEC infection. In this study, we detected hundreds of differentially-expressed-transcript-containing genes (DETs) between different phenotype birds at 5 days post-infection (dpi). DETs, PSAP and STT3A, with NMD have important functions under systemic APEC infection. DETs, CDC45, CDK1, RAG2, POLR1B, PSAP, and DNASE1L3, from the same transcription start sites (TSS) indicate that cell death, cell cycle, cellular function, and maintenance were predominant in host under systemic APEC.

Conclusions: With the use of RNAseq technology and bioinformatics tools, this study provides a portrait of the AS event and NMD in primary lymphoid tissues, which play critical roles in host homeostasis under systemic APEC infection. According to this study, AS plays a pivotal regulatory role in the immune response in chicken under systemic APEC infection via either NMD or alternative TSSs. This study elucidates the regulatory role of AS for the immune complex under systemic APEC infection.

Keywords: Alternative splicing; Bone marrow; Bursa; Nonsense-mediated decay; Thymus.

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Figures

Fig. 1
Fig. 1
The distribution of alternative splicing events in each of chicken three primary lymphoid tissues. a The distribution of alternative splicing events in bone marrow. b The distribution of alternative splicing events in bursa. c The distribution of alternative splicing events in thymus
Fig. 2
Fig. 2
Expression level of PSAP transcript CUFF.22759.5 in the thymus of challenged and non-challenged birds by RT-qPCR. Significance level for differences between groups was calculated by a t-test. **: p < 0.01. a. The gene PSAP and detected transcript CUFF.22759.5 in spliceR. b. PSAP transcript CUFF.22759.5 amplified product in agarose gel electrophoresis. c. RT-qPCR of PSAP transcript CUFF.22759.5 expression level. NC, non-challenged birds; C, challenged birds; M, marker
Fig. 3
Fig. 3
The isoforms containing gene from the same transcription start site in each of three primary lymphoid tissues in contrast susceptible vs. non-challenged birds at 5 days post-infection (dpi) and susceptible vs. resistant birds at 5 dpi. The left of heatmap is gene ID. The right of heatmap is gene name and transcription start site group id in parentheses. The number in rectangle of heatmap is the fold change of different isoforms. +, up-regulated; −, down-regulated; R, ∞. Red color, up-regulated isoforms. Blue color, down-regulated isoforms
Fig. 4
Fig. 4
The isoforms containing gene from the different transcription start site in each of three primary lymphoid tissues in contrast susceptible vs. non-challenged birds at 5 days post-infection (dpi) and susceptible vs. resistant birds at 5 dpi. The left of heatmap is gene ID. The right of heatmap is gene name. The number in rectangle of heatmap is the fold change of different isoforms. +, up-regulated; −, down-regulated; R, ∞. Red color, up-regulated isoforms. Blue color, down-regulated isoforms

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