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. 2012 Sep 6;13:459.
doi: 10.1186/1471-2164-13-459.

Profiling microRNAs in Lung Tissue From Pigs Infected With Actinobacillus Pleuropneumoniae

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Free PMC article

Profiling microRNAs in Lung Tissue From Pigs Infected With Actinobacillus Pleuropneumoniae

Agnieszka Podolska et al. BMC Genomics. .
Free PMC article

Abstract

Background: MicroRNAs (miRNAs) are a class of non-protein-coding genes that play a crucial regulatory role in mammalian development and disease. Whereas a large number of miRNAs have been annotated at the structural level during the latest years, functional annotation is sparse. Actinobacillus pleuropneumoniae (APP) causes serious lung infections in pigs. Severe damage to the lungs, in many cases deadly, is caused by toxins released by the bacterium and to some degree by host mediated tissue damage. However, understanding of the role of microRNAs in the course of this infectious disease in porcine is still very limited.

Results: In this study, the RNA extracted from visually unaffected and necrotic tissue from pigs infected with Actinobacillus pleuropneumoniae was subjected to small RNA deep sequencing. We identified 169 conserved and 11 candidate novel microRNAs in the pig. Of these, 17 were significantly up-regulated in the necrotic sample and 12 were down-regulated. The expression analysis of a number of candidates revealed microRNAs of potential importance in the innate immune response. MiR-155, a known key player in inflammation, was found expressed in both samples. Moreover, miR-664-5p, miR-451 and miR-15a appear as very promising candidates for microRNAs involved in response to pathogen infection.

Conclusions: This is the first study revealing significant differences in composition and expression profiles of miRNAs in lungs infected with a bacterial pathogen. Our results extend annotation of microRNA in pig and provide insight into the role of a number of microRNAs in regulation of bacteria induced immune and inflammatory response in porcine lung.

Figures

Figure 1
Figure 1
The distribution of raw reads versus read lengths in both libraries. X axis shows the insert length while Y axis represents raw reads. Necrotic area sample is marked in blue whereas visually unaffected area sample is marked in red.
Figure 2
Figure 2
The distribution of raw reads between various RNA classes in A) visually unaffected sample and B) necrotic sample.
Figure 3
Figure 3
Profile of reads distribution for read cluster annotated as both miR-664-5p and SNORA36. S1-necrotic, S2-visually unaffected sample.
Figure 4
Figure 4
RT-qPCR analysis of expression of 13 selected unique miRNAs and one snoRNA. miR-d5 represents a novel unannotated microRNA. Data presented are normalized to the reference genes, LOG2-transformed relative quantities. The level of expression for control sample is set to zero.
Figure 5
Figure 5
Sampling areas of the porcine lungs infected with Actinnobacillus pleuropneumoniae. 1 – Necrotic area, within pulmonary lesion. 2 – Demarcation zone, border area between visually unaffected and pulmonary lesions. 3 – Visually unaffected area, which was sampled either from unaffected lung lobe (3 to the left) or from site as distant from pulmonary lesion as possible (3 to the right).

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