Related Experiment Video
Updated: Nov 25, 2025
Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry
Published on: June 22, 2017
A Binary Cre Transgenic Approach Dissects Microglia and CNS Border-Associated Macrophages
Jung-Seok Kim1, Masha Kolesnikov1, Shany Peled-Hajaj2
1Department of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel.
Abstract:
The developmental and molecular heterogeneity of tissue macrophages is unravelling, as are their diverse contributions to physiology and pathophysiology. Moreover, also given tissues harbor macrophages in discrete anatomic locations. Functional contributions of specific cell populations can in mice be dissected using Cre recombinase-mediated mutagenesis. However, single promoter-based Cre models show limited specificity for cell types. Focusing on macrophages in the brain, we establish here a binary transgenic system involving complementation-competent NCre and CCre fragments whose expression is driven by distinct promoters: Sall1ncre: Cx3cr1ccre mice specifically target parenchymal microglia and compound transgenic Lyve1ncre: Cx3cr1ccre animals target vasculature-associated macrophages, in the brain, as well as other tissues. We imaged the respective cell populations and retrieved their specific translatomes using the RiboTag in order to define them and analyze their differential responses to a challenge. Collectively, we establish the value of binary transgenesis to dissect tissue macrophage compartments and their functions.
Insights
Researchers developed a new binary transgenic system to precisely target different macrophage populations in the brain, including parenchymal microglia and vasculature-associated macrophages, enabling better study of their functions.
Area of Science:
- Immunology
- Neuroscience
- Cell Biology
Background:
- Tissue macrophages exhibit significant developmental and molecular heterogeneity.
- Macrophages reside in distinct anatomical locations within tissues, performing diverse physiological and pathophysiological roles.
- Existing Cre recombinase-mediated mutagenesis models offer limited specificity for targeting distinct macrophage populations.
Purpose of the Study:
- To develop a novel binary transgenic system for precise targeting of specific tissue macrophage populations.
- To differentiate between parenchymal microglia and vasculature-associated macrophages in the brain.
- To analyze the functional and molecular differences between these distinct macrophage subsets.
Main Methods:
- Generation of Sall1ncre: Cx3cr1ccre mice to target parenchymal microglia.
- Generation of Lyve1ncre: Cx3cr1ccre mice to target vasculature-associated macrophages.
- Utilized in vivo imaging and RiboTag for translatome analysis of targeted cell populations.
Main Results:
- The binary transgenic system successfully achieved specific targeting of distinct macrophage populations.
- Sall1ncre: Cx3cr1ccre mice specifically labeled parenchymal microglia.
- Lyve1ncre: Cx3cr1ccre mice specifically labeled vasculature-associated macrophages in the brain and other tissues.
- Differential translatomes and responses to challenges were identified between the targeted macrophage subsets.
Conclusions:
- Binary transgenesis is a valuable tool for dissecting distinct tissue macrophage compartments.
- This approach enables a deeper understanding of the specific functions of microglia and other tissue macrophages.
- The developed system facilitates the study of macrophage heterogeneity and their roles in health and disease.
