Related Experiment Videos
Rat microglial interleukin-3
P J Gebicke-Haerter1, K Appel, G D Taylor
1Department of Psychiatry, University of Freiburg, Medical School, Germany.
Abstract:
Interleukin-3 (IL-3, multi-CSF) is a growth factor for a variety of hematopoietic progenitor cells. Recently, microglial cells, the resident macrophages of the central nervous system (CNS) have been shown to proliferate in the presence of IL-3 both in vivo and in culture. Data obtained from cultured astrocytes gave rise to the hypothesis that astrocytes synthesize the microglial growth factor. This is the first report identifying rat microglial cells themselves as a source of IL-3. Culture media conditioned by isolated microglia enhanced microglial proliferation above fresh media controls. IL-3 polypeptide was detected in both conditioned media (CM) and in microglial cells by Western blotting and immunoprecipitation. Furthermore, anti-IL-3 antibodies were able to inhibit microglial proliferation induced by conditioned media. mRNAIL-3 was present in single microglial cells as revealed by in situ hybridization. Total RNA prepared from purified microglia yielded a single PCR amplification product. Identity of the PCR product was confirmed by Southern blot hybridization using a cDNAIL-3 probe and by DNA sequencing. Expression of mRNAIL-3 was observed in both absence and presence of lipopolysaccharide, a bacterial endotoxin, that commonly induces expression of inflammatory cytokines and inhibits microglial proliferation. It is concluded that IL-3 expression in ensuring the recruitment of enhanced numbers of immunocompetent cells at sites of lesion. In the light of weak immune reactions in the brain, it is hypothesized that the expression of a characteristic T cell feature in monocyte-derived microglia may be a partial compensation of T cell functions in brain lesions.
Insights
Microglia, the immune cells of the brain, produce Interleukin-3 (IL-3), a growth factor that promotes their own proliferation. This finding suggests IL-3 plays a role in brain immune responses.
Area of Science:
- Neuroimmunology
- Cell Biology
Background:
- Interleukin-3 (IL-3) is a hematopoietic growth factor.
- Microglia, the central nervous system's resident macrophages, proliferate in response to IL-3.
- Astrocytes were previously hypothesized to produce the microglial growth factor.
Purpose of the Study:
- To investigate if microglial cells themselves are a source of IL-3.
- To characterize IL-3 expression and its role in microglial proliferation.
Main Methods:
- Primary rat microglial cell cultures.
- Western blotting and immunoprecipitation to detect IL-3 protein.
- In situ hybridization and RT-PCR to detect IL-3 mRNA.
- Functional assays using anti-IL-3 antibodies.
Main Results:
- Microglial conditioned media enhanced microglial proliferation.
- IL-3 protein and mRNA were detected in isolated microglia.
- Anti-IL-3 antibodies inhibited microglial proliferation induced by conditioned media.
- IL-3 mRNA expression was observed irrespective of lipopolysaccharide stimulation.
Conclusions:
- Rat microglial cells are an endogenous source of IL-3.
- IL-3 produced by microglia contributes to their proliferation.
- Microglial IL-3 may be involved in recruiting immune cells to brain lesions.
- Microglial IL-3 expression might compensate for T cell functions in the brain.