Functional expression of the CXC-chemokine receptor-4/fusin on mouse microglial cells and astrocytes

J Immunol. 1997 Jul 15;159(2):905-11.

Abstract

The mRNA for the seven-transmembrane-spanning G protein-coupled receptor fusin/CXCR-4 is expressed in primary mouse astrocyte cultures and the transformed mouse microglial cell line, N9. Cell surface expression of fusin in these cells was confirmed by staining with a polyclonal anti-fusin Ab. The functional capacity of this chemokine receptor was examined by evaluating the calcium responses following stimulation of glial cells with the CXC-chemokine, stromal-derived cell factor-1alpha (SDF-1alpha). Both astrocytes and microglial cells mobilized calcium following stimulation with chemically synthesized SDF-1alpha. SDF-1alpha- and carbachol-mediated calcium responses of astrocytes were partially inhibited by treatment with pertussis toxin (PTx), suggesting receptor coupling to a combination of G alpha(i) and other G proteins. In contrast, the calcium responses of microglial cells to SDF-1alpha were completely PTx sensitive, while responses to carbachol stimulation were PTx resistant. The ability of SDF-1alpha to induce glial cell migration was also examined. Synthetic SDF-1alpha was a potent chemoattractant for mouse microglial cells at ligand concentrations of 10 to 500 ng/ml; peak responses were noted at 100 ng/ml. In contrast, astrocytes did not migrate toward a gradient of SDF-1alpha. The failure of SDF-1alpha to induce astrocyte migration was specific, as another chemokine, macrophage inflammatory protein-1alpha, triggered astrocyte chemotaxis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Astrocytes / metabolism*
  • Astrocytes / pathology
  • Calcium / metabolism
  • Cell Line, Transformed
  • Cell Movement / drug effects
  • Chemokine CXCL12
  • Chemokines, CXC*
  • Cytokines / pharmacology
  • GTP-Binding Proteins / metabolism
  • Membrane Proteins / biosynthesis*
  • Mice
  • Mice, Inbred BALB C
  • Microglia / metabolism*
  • Microglia / pathology
  • Receptors, CXCR4
  • Receptors, HIV / biosynthesis*

Substances

  • Chemokine CXCL12
  • Chemokines, CXC
  • Cxcl12 protein, mouse
  • Cytokines
  • Membrane Proteins
  • Receptors, CXCR4
  • Receptors, HIV
  • GTP-Binding Proteins
  • Calcium