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Comprehensive RNA-Seq Profiling to Evaluate the Sheep Mammary Gland Transcriptome in Response to Experimental
Rohini Chopra-Dewasthaly1, Melanie Korb2, René Brunthaler3
1Division of Clinical Microbiology and Infection Biology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine, Veterinaerplatz 1,Vienna, Austria.
Abstract:
Mycoplasma agalactiae is a worldwide serious pathogen of small ruminants that usually spreads through the mammary route causing acute to subacute mastitis progressing to chronic persistent disease that is hard to eradicate. Knowledge of mechanisms of its pathogenesis and persistence in the mammary gland are still insufficient, especially the host-pathogen interplay that enables it to reside in a chronic subclinical state. This study reports transcriptome profiling of mammary tissue from udders of sheep experimentally infected with M. agalactiae type strain PG2 in comparison with uninfected control animals using Illumina RNA-sequencing (RNA-Seq). Several differentially expressed genes (DEGs) were observed in the infected udders and RT-qPCR analyses of selected DEGs showed their expression profiles to be in agreement with results from RNA-Seq. Gene Ontology (GO) analysis revealed majority of the DEGs to be associated with mycoplasma defense responses that are directly or indirectly involved in host innate and adaptive immune responses. Similar RNA-Seq analyses were also performed with spleen cells of the same sheep to know the specific systemic transcriptome responses. Spleen cells exhibited a comparatively lower number of DEGs suggesting a less prominent host response in this organ. To our knowledge this is the first study that describes host transcriptomics of M. agalactiae infection and the related immune-inflammatory responses. The data provides useful information to further dissect the molecular genetic mechanisms underlying mycoplasma mastitis, which is a prerequisite for designing effective intervention strategies.
Insights
This study reveals host gene expression changes in sheep mammary glands infected with Mycoplasma agalactiae. Understanding these immune responses is key to developing new treatments for mycoplasma mastitis.
Area of Science:
- Veterinary Immunology
- Molecular Biology
- Infectious Diseases
Background:
- Mycoplasma agalactiae causes significant mastitis in small ruminants, leading to persistent infections.
- The host-pathogen interactions enabling chronic, subclinical Mycoplasma agalactiae infections remain poorly understood.
Purpose of the Study:
- To investigate the host transcriptome changes in sheep mammary tissue following experimental Mycoplasma agalactiae infection.
- To characterize the immune and inflammatory responses at the molecular level.
Main Methods:
- Illumina RNA-sequencing (RNA-Seq) was employed for transcriptome profiling of infected and control sheep mammary tissues.
- Quantitative reverse transcription PCR (RT-qPCR) was used to validate the expression of selected differentially expressed genes (DEGs).
- Gene Ontology (GO) analysis was performed to identify biological functions of DEGs.
Main Results:
- Significant differences in gene expression were identified in infected mammary tissues compared to controls.
- RT-qPCR confirmed the RNA-Seq findings for selected DEGs.
- GO analysis indicated that most DEGs are involved in host defense and immune responses (innate and adaptive) against Mycoplasma agalactiae.
- Spleen cells showed fewer DEGs, suggesting a more localized mammary gland response.
Conclusions:
- This is the first study to describe the host transcriptomics of Mycoplasma agalactiae infection in sheep.
- The findings provide crucial insights into the molecular mechanisms of mycoplasma mastitis and host immune responses.
- This data is essential for developing effective intervention strategies against Mycoplasma agalactiae infections.
