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Biolistic expression of the macrophage colony stimulating factor receptor in organotypic cultures induces an inflammatory response
Author(s) -
Mitrasinovic Olivera M.,
Robinson Christopher C.,
Tenen Daniel G.,
Lee Yuen Ling,
Poon Clara,
Murphy Greer M.
Publication year - 2004
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.20168
Subject(s) - microglia , integrin alpha m , proinflammatory cytokine , biology , microbiology and biotechnology , receptor , transfection , neuroglia , immunology , cell culture , inflammation , neuroscience , central nervous system , biochemistry , genetics
The receptor for macrophage colony‐stimulating factor (M‐CSFR; c‐fms ) is expressed at increased levels by microglia in Alzheimer's disease (AD) and in mouse models for AD. Increased expression of M‐CSFR on cultured microglia results in a strong proinflammatory response, but the relevance of this cell culture finding to intact brain is unknown. To determine the effects of increased microglial expression of M‐CSFR in a complex organotypic environment, we developed a system for biolistic transfection of microglia in hippocampal slice cultures. The promoter for the Mac‐1 integrin α subunit CD11b is active in cells of myeloid origin. In the brain, CD11b expression is restricted to microglia. Constructs consisting of the promoter for CD11b and a c‐fms cDNA or an enhanced green fluorescent protein (EGFP) cDNA were introduced into monotypic cultures of microglia, neurons, and astrocytes. Strong CD11b promoter activity was observed in microglia, whereas little activity was observed in other cell types. Biolistic transfection of organotypic hippocampal cultures with the CD11b/ c‐fms construct resulted in expression of the c‐fms mRNA and protein that was localized to microglia. Furthermore, biolistic overexpression of M‐CSFR on microglia resulted in significantly increased production by the hippocampal cultures of the proinflammatory cytokines interleukin (IL)‐1α macrophage inflammatory protein (MIP‐1α), and trends toward increased production of IL‐6 and M‐CSF. These findings demonstrate that microglial overexpression of M‐CSFR in an organotypic environment induces an inflammatory response, and suggest that increased microglial expression of M‐CSFR could contribute to the inflammatory response observed in AD brain. © 2004 Wiley‐Liss, Inc.