Blockade of chloride intracellular ion channel 1 stimulates Abeta phagocytosis.
In amyloid-beta (Abeta)-stimulated microglial cells, blockade of chloride intracellular ion channel 1 (CLIC1) reverts the increase in tumor necrosis factor-alpha and nitric oxide (NO) production and results in neuroprotection of cocultured neurons. This effect could be of therapeutic efficacy in Alzheimer's disease (AD), where microglial activation may contribute to neurodegeneration, but it could reduce Abeta phagocytosis, which could facilitate amyloid plaque removal. Here, we analyzed the CLIC1 blockade effect on Abeta-stimulated mononuclear phagocytosis. In the microglial cell line BV-2, Abeta25-35 treatment enhanced fluorescent bead phagocytosis, which persisted also in the presence of IAA-94, a CLIC1 channel blocker. The same result was obtained in rat primary microglia and in BV2 cells, where CLIC1 expression had been knocked down with a plasmid producing small interfering RNAs. To address specifically the issue of Abeta phagocytosis, we treated BV-2 cells with biotinylated Abeta1-42 and measured intracellular amyloid by morphometric analysis. IAA-94-treated cells showed an increased Abeta phagocytosis after 24 hr and efficient degradation of ingested material after 72 hr. In addition, we tested Abeta1-42 phagocytosis in adult rat peritoneal macrophages. Also, these cells actively phagocytosed Abeta1-42 in the presence of IAA-94. However, the increased expression of inducible NO synthase (iNOS), stimulated by Abeta, was reverted by IAA-94. In parallel, a decrease in NO release was detected. These results suggest that blockade of CLIC1 stimulates Abeta phagocytosis in mononuclear phagocytes while inhibiting the induction of iNOS and further point to CLIC1 as a possible therapeutic target in AD.
Paradisi
Silvia
S
Department of Cell Biology and Neuroscience, Istituto Superiore di Sanità, Rome, Italy.
Matteucci
Andrea
A
Fabrizi
Cinzia
C
Denti
Michela A
MA
Abeti
Rosella
R
Breit
Samuel N
SN
Malchiodi-Albedi
Fiorella
F
Mazzanti
Michele
M
D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D018118:Chloride Channels / Q000378:metabolism*; D004232:Diuretics / Q000494:pharmacology; D005434:Flow Cytometry; D006016:Glycolates / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D017737:Macrophages, Peritoneal / Q000378:metabolism; D051379:Mice; D017628:Microglia / Q000378:metabolism*; D018613:Microscopy, Confocal; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000187:drug effects; D010587:Phagocytosis / Q000187:drug effects / Q000502:physiology*; D034622:RNA Interference; D051381:Rats
D016229:Amyloid beta-Peptides; D018118:Chloride Channels; C501967:Clic1 protein, mouse; D004232:Diuretics; D006016:Glycolates; C015765:MK 473; D052247:Nitric Oxide Synthase Type II