Expression Analysis of CB2-GFP BAC Transgenic Mice

PLoS One. 2015 Sep 25;10(9):e0138986. doi: 10.1371/journal.pone.0138986. eCollection 2015.

Abstract

The endocannabinoid system (ECS) is a retrograde messenger system, consisting of lipid signaling molecules that bind to at least two G-protein-coupled receptors, Cannabinoid receptor 1 and 2 (CB1 and 2). As CB2 is primarily expressed on immune cells such as B cells, T cells, macrophages, dendritic cells, and microglia, it is of great interest how CB2 contributes to immune cell development and function in health and disease. Here, understanding the mechanisms of CB2 involvement in immune-cell function as well as the trafficking and regulation of CB2 expressing cells are crucial issues. Up to now, CB2 antibodies produce unclear results, especially those targeting the murine protein. Therefore, we have generated BAC transgenic GFP reporter mice (CB2-GFPTg) to trace CB2 expression in vitro and in situ. Those mice express GFP under the CB2 promoter and display GFP expression paralleling CB2 expression on the transcript level in spleen, thymus and brain tissue. Furthermore, by using fluorescence techniques we show that the major sources for GFP-CB2 expression are B cells in spleen and blood and microglia in the brain. This novel CB2-GFP transgenic reporter mouse line represents a powerful resource to study CB2 expression in different cell types. Furthermore, it could be used for analyzing CB2-mediated mobilization and trafficking of immune cells as well as studying the fate of recruited immune cells in models of acute and chronic inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / metabolism
  • Brain / metabolism
  • Chromosomes, Artificial, Bacterial / genetics*
  • Green Fluorescent Proteins / blood
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • Mice
  • Mice, Transgenic*
  • Promoter Regions, Genetic
  • Receptor, Cannabinoid, CB2 / blood
  • Receptor, Cannabinoid, CB2 / genetics*
  • Receptor, Cannabinoid, CB2 / metabolism
  • Spleen / metabolism
  • Thymus Gland / metabolism

Substances

  • Cnr2 protein, mouse
  • Receptor, Cannabinoid, CB2
  • Green Fluorescent Proteins

Grants and funding

This work was partly supported by a start-up grant from the Medical Faculty of the University of Bonn "Bonfor" (to ACS and HS) and the Deutsche Forschungsgemeinschaft, FOR 926 (to ACS, RL, BG and AZ). ACS, RL, EB, DMO and AZ are members of the DFG Cluster of Excellence "Immunosensation." The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.