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Construction of a DNA Methylation Map of Argali Hybrid Sheep During Mo Infection
Qinchuan Zhang1, Shiyi Li1, Guojie Cheng1
1College of Animal Science and Technology, Shihezi University, Shihezi 832003, China.
Abstract:
The DNA methylation landscape in the lungs of argali hybrid sheep infected with Mycoplasma ovipneumoniae (Mo) remains poorly characterized. This study aimed to profile genome-wide DNA methylation using reduced representation bisulfite sequencing (RRBS) and to validate key genes using bisulfite sequencing PCR (BSP), methylation-specific PCR (MSP), and quantitative MSP (QMSP). The results revealed a significant increase in global mCG methylation in -infected lungs. RRBS identified 3691 differentially methylated regions (DMRs), 66.2% of which were hypermethylated. Methylation levels were highest in gene bodies/downstream regions and lowest in promoters/5' untranslated regions. Differentially methylated genes (DMGs) were enriched in immune-inflammatory pathways (e.g., antigen presentation, B-cell receptor signaling, Th17 differentiation) and, to a lesser extent, neural signaling pathways. BSP confirmed the methylation status of hypermethylated (KHDC3L, GILT, OVAR-DRB1, SGK1, ADAM17) and hypomethylated (EFCAB11, AP1B1, TATDN1) DMGs. Independent validation by MSP and QMSP further supported the hypermethylation of SGK1 and GILT in both lung tissue and alveolar macrophages. Quantitative reverse-transcription PCR showed that promoter hypermethylation of KHDC3L, GILT, SGK1, and ADAM17 was associated with transcriptional downregulation, while hypomethylation of AP1B1 correlated with upregulation. In summary, Mo infection induces genome-wide hypermethylation reprogramming that dysregulates key immune-related genes, highlighting potential epigenetic mechanisms in the pathogenesis of mycoplasmal pneumonia.
Insights
Mycoplasma ovipneumoniae infection in sheep lungs causes widespread DNA hypermethylation, altering immune gene expression. This epigenetic reprogramming highlights novel mechanisms in pneumonia pathogenesis.
Area of Science:
- Epigenetics
- Genomics
- Veterinary Medicine
Background:
- The DNA methylation patterns in argali hybrid sheep lungs infected with Mycoplasma ovipneumoniae (Mo) are largely unknown.
- Understanding these epigenetic changes is crucial for elucidating the pathogenesis of Mo-induced pneumonia.
Purpose of the Study:
- To profile the genome-wide DNA methylation landscape in Mo-infected sheep lungs.
- To identify differentially methylated regions and genes associated with Mo infection.
- To validate key findings using molecular techniques and assess their impact on gene expression.
Main Methods:
- Reduced representation bisulfite sequencing (RRBS) for genome-wide DNA methylation profiling.
- Bisulfite sequencing PCR (BSP), methylation-specific PCR (MSP), and quantitative MSP (QMSP) for validation.
- Quantitative reverse-transcription PCR (qRT-PCR) to assess gene expression.
Main Results:
- Mo infection significantly increased global mCG methylation in sheep lungs, with 66.2% of identified differentially methylated regions (DMRs) being hypermethylated.
- Differentially methylated genes (DMGs) were enriched in immune-inflammatory and neural signaling pathways.
- Promoter hypermethylation correlated with transcriptional downregulation of key immune genes (e.g., KHDC3L, GILT, SGK1, ADAM17), while hypomethylation of AP1B1 correlated with upregulation.
Conclusions:
- Mycoplasma ovipneumoniae infection induces significant genome-wide DNA hypermethylation reprogramming in sheep lungs.
- This epigenetic dysregulation affects crucial immune-related genes, suggesting a role in mycoplasmal pneumonia pathogenesis.
- The study identifies potential epigenetic targets for understanding and managing ovine respiratory diseases.
