Implication of the small GTPase Rac1 in the generation of reactive oxygen species in response to β-amyloid in C6 astroglioma cells

Mina Lee, Hye Jin You, Sung Hoon Cho, Chang Hoon Woo, Min Hyuk Yoo, Eun Hye Joe, Jae Hong Kim

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Exogenous application of β-amyloid (Aβ25-35, a fragment of Aβ1-42) significantly elevated levels of reactive oxygen species (ROS) in C6 astroglioma cells, as measured by confocal microscopic analysis of H2O2-sensitive 2′,7′-dichlorofluorescin fluorescence. Subsequent characterization of the signalling path-way revealed that expression of RacN17, a dominant-negative Rac1 mutant, completely blocked Aβ25-35-induced generation of ROS, which is indicative of the crucial role played by Rac GTPase in this process. To better understand the downstream mediators affected by Rac, we assessed the degree to which inhibition of cytosolic phospholipase A2 (cPLA2) and 5-lipoxygenase (5-LO) contributed to the response and found that inhibition of either enzyme completely blocked Aβ25-35-induced ROS generation, indicating its dependence on arachidonic acid synthesis and metabolism to leukotrienes (e.g. leukotriene B4). Consistent with those findings, Aβ25-35 Rac-dependently stimulated translocation of 5-LO to the nuclear envelope and increased intracellular levels of leukotriene B4, while exogenous application of leukotriene B4 increased intracellular H2O2 via BLT, its cell-surface receptor. In addition to the aforementioned downstream mediators, inhibition of phosphoinositide 3-kinase (PI 3-kinase), an enzyme situated upstream of Rac, also completely blocked Aβ25-35-induced H2O2 generation. Our findings thus demonstrate that PI 3-kinase, Rac, cPLA2 and 5-LO are all essential components of the β-amyloid signaling cascade leading to generation of ROS.

Original languageEnglish
Pages (from-to)937-943
Number of pages7
JournalBiochemical Journal
Volume366
Issue number3
DOIs
Publication statusPublished - 2002 Sep 15

Keywords

  • Cytosolic phospholipase A2
  • Leukotriene B
  • Signal transduction

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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