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Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
Published on: January 30, 2014
Microglia derived from aging mice exhibit an altered inflammatory profile
Amanda Sierra1, Andres C Gottfried-Blackmore, Bruce S McEwen
1Laboratory of Neuroendocrinology, Rockefeller University, New York, NY, USA.
Abstract:
Microglia play a critical role in neurodegenerative diseases and in the brain aging process. Yet, little is known about the functional dynamics of microglia during aging. Thus, using young and aging transgenic mice expressing enhanced-green fluorescent protein (EGFP) under the promoter of the c-fms gene for macrophage-colony stimulating factor receptor, we evaluated in vivo-induced inflammatory responses of EGFP-expressing microglia sorted by flow cytometry. Aging microglia were characterized by the presence of lipofuscin granules, decreased processes complexity, altered granularity, and increased mRNA expression of both pro-inflammatory (TNFalpha, IL-1beta, IL-6) and anti-inflammatory (IL-10, TGFbeta1) cytokines. Following lipopolysaccharide (LPS) challenge (1 mg/kg, 3 h), aging microglia exhibit increased basal expression of TNFalpha, IL-1beta, IL-6, and IL-10. Yet, the fold-over-basal LPS response remained constant across age, implying that the inflammatory machinery in aging microglia is functional and adjusted to the basal state. Gender differences were not overall observed across the treatments (age, LPS). The low but sustained production of pro-inflammatory cytokines by aging microglia may have a profound impact in the brain aging process.
Insights
Aging microglia exhibit altered inflammatory responses and cytokine production, potentially impacting brain aging. Their inflammatory machinery remains functional but adjusted to a higher basal state.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are crucial in neurodegeneration and brain aging.
- Functional dynamics of aging microglia remain poorly understood.
Purpose of the Study:
- To investigate in vivo inflammatory responses of aging microglia.
- To characterize age-related changes in microglial inflammatory profiles.
Main Methods:
- Utilized young and aging transgenic mice expressing EGFP under the c-fms promoter.
- Sorted EGFP-expressing microglia via flow cytometry for analysis.
- Assessed inflammatory responses following lipopolysaccharide (LPS) challenge.
Main Results:
- Aging microglia showed lipofuscin, reduced complexity, altered granularity, and increased pro- and anti-inflammatory cytokine mRNA.
- Aging microglia had higher basal TNFα, IL-1β, IL-6, and IL-10 expression.
- LPS-induced inflammatory response magnitude was consistent across ages, indicating functional adjustment.
Conclusions:
- Aging microglia display a distinct inflammatory profile with elevated basal cytokine levels.
- The inflammatory machinery in aging microglia is functional but adapted to a heightened basal state.
- Sustained low-level pro-inflammatory cytokine production by aging microglia may influence brain aging.
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