A novel reporter mouse line for studying alveolar macrophages

Alveolar macrophages (AMs) are self-maintained immune cells that play vital roles in lung homeostasis and immunity. Although reporter mice and culture systems have been established for studying macrophages, an accurate and specific reporter line for alveolar macrophage study is still not available. Here we reported a novel Rspo1-tdTomato gene reporter mouse line that could specifically label mouse AMs in a cell-intrinsic manner. Using this reporter system, we visualized the dynamics of alveolar macrophages intravitally under steady state and characterized the alveolar macrophage differentiation under in vitro condition. By performing ATAC-seq, we found that insertion of the tdTomato cassette in the Rspo1 locus increased the accessibility of a PPARE motif within the Rspo1 locus and revealed a potential regulation by key transcription factor PPAR-? for alveolar macrophage differentiation in vitro and in vivo. Consistently, perturbation of PPAR-? by its agonist rosiglitazone or inhibitor GW9662 resulted in corresponding alteration of tdTomato expression in alveolar macrophages together with the transcription of PPAR-? downstream target genes. Furthermore, global transcriptomic analyses of AMs from the wild type mice and the Rspo1-tdTomato mice showed comparable gene expression profiles, especially those AM-specific genes, confirming that the insertion of the tdTomato cassette in the Rspo1 locus does not impact the cell identity and biological function of AMs under normal condition. Taken together, our study provides an alternative tool for in vivo and in vitro labeling of alveolar macrophages with high specificity which could also be utilized as an indicator of PPAR-? activity for future development of PPAR-? specific targeting drugs.

Automated summary

We reported a novel Rspo1-tdTomato gene reporter mouse line that specifically labels mouse alveolar macrophages (AMs) and visualizes their dynamics and differentiation processes. Insertion of the tdTomato cassette in the Rspo1 locus increased the accessibility of a PPARE motif, potentially regulating AM differentiation via the transcription factor PPAR-γ. Comparative transcriptomic analysis showed similar gene expression profiles between AMs from Rspo1-tdTomato mice and wild type mice, confirming the unaffected cell identity and function of AMs. This study provides an alternative tool for specific labeling of AMs in vivo and in vitro and can serve as an indicator for PPAR-γ activity in drug development.