Increased expression of M1 and M2 phenotypic markers in isolated microglia after four-day binge alcohol exposure in male rats

Hui Peng, Chelsea R. Geil Nickell, Kevin Y. Chen, Justin A. McClain, Kimberly Nixon

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78 Citations (SciVal)

Abstract

Microglia activation and neuroinflammation are common features of neurodegenerative conditions, including alcohol use disorders (AUDs). When activated, microglia span a continuum of diverse phenotypes ranging from classically activated, pro-inflammatory (M1) microglia/macrophages to alternatively activated, growth-promoting (M2) microglia/macrophages. Identifying microglia phenotypes is critical for understanding the role of microglia in the pathogenesis of AUDs. Therefore, male rats were gavaged with 25% (w/v) ethanol or isocaloric control diet every 8 h for 4 days and sacrificed at 0, 2, 4, and 7 days after alcohol exposure (e.g., T0, T2, etc.). Microglia were isolated from hippocampus and entorhinal cortices by Percoll density gradient centrifugation. Cells were labeled with microglia surface antigens and analyzed by flow cytometry. Consistent with prior studies, isolated cells yielded a highly enriched population of brain macrophages/microglia (>95% pure), evidenced by staining for the macrophage/microglia antigen CD11b. Polarization states of CD11b+CD45low microglia were evaluated by expression of M1 surface markers, major histocompatibility complex (MHC) II, CD32, CD86, and M2 surface marker, CD206 (mannose receptor). Ethanol-treated animals begin to show increased expression of M1 and M2 markers at T0 (p = n.s.), with significant changes at the T2 time point. At T2, expression of M1 markers, MHC-II, CD86, and CD32 were increased (p < 0.05) in hippocampus and entorhinal cortices, while M2 marker, CD206, was increased significantly only in entorhinal cortices (p < 0.05). All effects resolved to control levels by T4. In summary, four-day binge alcohol exposure produces a transient increase in both M1 (MHC-II, CD32, and CD86) and M2 (CD206) populations of microglia isolated from the entorhinal cortex and hippocampus. Thus, these findings that both pro-inflammatory and potentially beneficial, recovery-promoting microglia phenotypes can be observed after a damaging exposure of alcohol are critically important to our understanding of the role of microglia in the pathogenesis of AUDs.

Original languageEnglish
Pages (from-to)29-40
Number of pages12
JournalAlcohol
Volume62
DOIs
StatePublished - Aug 2017

Bibliographical note

Funding Information:
This work was funded by National Institutes of Health grants R01AA016959 (KN), F31AA023459 (CRGN) and R03NS089433 (HP), T32 DA016176 (CRGN, JAM), University of Kentucky Center for Drug & Alcohol Research (pilot project to JAM) and the University of Kentucky Department of Pharmaceutical Sciences.

Publisher Copyright:
© 2017 Elsevier Inc.

Keywords

  • Alcoholism
  • Ethanol
  • Flow cytometry
  • Microglia
  • Neurodegeneration
  • Neuroinflammation

ASJC Scopus subject areas

  • Health(social science)
  • Biochemistry
  • Toxicology
  • Neurology
  • Behavioral Neuroscience

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