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CX3CR1+ mononuclear phagocytes gene-expression profile based on their location in the epididymis using RNA-sequencing

ABSTRACT: CX3CR1+ mononuclear phagocytes gene-expression profile based on their location in the epididymis using RNA-sequencing

PROVIDER: PRJNA562539 | ENA |

REPOSITORIES: ENA

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CX3CR1+ mononuclear phagocytes gene-expression profile based on their location in the epididymis using RNA-sequencing

Project description:In the epididymis, prevention of autoimmune responses against spermatozoa, while providing protection against pathogens, is important for male fertility. We previously showed that immune cells known as mononuclear phagocytes (MPs) are located either in the epididymal interstitium or in close proximity to the epithelium. In the initial segments (IS), these “intraepithelial” MPs extend slender luminal-reaching projections between epithelial cells. In this study, we performed an in-depth characterization of MPs isolated from IS, caput-corpus, and cauda epididymis of CX3CR1EGFP+/- mice that express EGFP in these cells. Flow cytometry analysis revealed region-specific subsets of MPs that express combinations of markers traditionally described in ‘dendritic cells’ or ‘macrophages’. RNA sequencing identified distinct transcriptomic signatures in MPs from each region, and revealed specific genes involved in inflammatory and anti-inflammatory responses, phagosomal activity, and antigen processing and presentation. Functional fluorescent in vivo labeling assays showed that higher percentages of CX3CR1+ MPs that captured and processed antigens were detected in the IS compared to other regions. Confocal microscopy showed that in the IS and corpus, circulatory antigens were internalized and processed by interstitial and intraepithelial MPs. However, in the cauda only interstitial MPs internalized and processed antigens, while intraepithelial MPs did not take up antigens, indicating that all antigens have been captured before they reached the epithelial lining. Cauda MPs may thus confer a stronger protection against blood-born pathogens compared to proximal regions. By identifying immunoregulatory mechanisms in the epididymis, our study may lead to new therapies for common disorders such as male infertility and epididymitis, and identify potential targets for immuno-contraception.

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Mononuclear phagocytes in muscle after notexin injury in C57Bl/6, Cx3cr1-/-, Ccl2-/-,Cx3cr1-/-Ccl2-/- mice

Project description:Muscle injury triggers inflammation in which infiltrating mononuclear phagocytes are crucial for tissue regeneration. The interaction of the CCL2/CCR2 and CX3CL1/CX3CR1 chemokine axis that guides phagocyte infiltration is incompletely understood. Here, we show that CX3CR1 deficiency promotes muscle repair and rescues Ccl2-/- mice from impaired muscle regeneration as a result of altered macrophage function, not infiltration. Transcriptomic analysis of muscle mononuclear phagocytes reveals that Apolipoprotein E (ApoE) is up-regulated in mice with efficient regeneration. ApoE treatment enhances phagocytosis by mononuclear phagocytes in vitro, and restores phagocytic activity and muscle regeneration in Ccl2-/- mice. Because CX3CR1 deficiency may compensate for defective CCL2-dependant monocyte recruitment by modulating ApoE-dependent macrophage phagocytic activity, targeting CX3CR1 expressed by macrophages might be a powerful therapeutic approach to improve muscle regeneration.

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Transcription profiling of mouse peripheral blood and alveolar recruited mononuclear phagocytes in Pam3CSK4 treated animals

Project description:Aim of this project was to examine the global gene expression profiles of mononuclear phagocytes recruited from peripheral blood to the alveolar space following alveolar deposition of the TLR2-ligand Pam3CSK4 in transgenic CX3CR1+/GFP mice. Experiment Overall Design: Alveolar macrophages (AM) and peripheral blood monocytes (PBM) were isolated from broncho-alveolar lavages and from blood, respectively, using FACS. Expression profiles from AM and PBM of the same mice were compared on the same slides. Each labeled RNA sample contained RNA pooled from six individuals. Four pairs of RNA from corresponding AM and PBM pools were hybridized, giving a total amount of 24 individual mice analyzed. Two hybridizations were performed with AM labeled with Cy3 and PBM with Cy5, two hybridizations were performed with swaped dyes.

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