Clioquinol induces autophagy in cultured astrocytes and neurons by acting as a zinc ionophore

Mi Ha Park, Sook Jeong Lee, Hyae ran Byun, Yunha Kim, Young Jun Oh, Jae Young Koh, Jung Jin Hwang

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Recent studies have demonstrated that clioquinol, an antibiotic with an anti-amyloid effect, acts as a zinc ionophore under physiological conditions. Because increases in labile zinc may induce autophagy, we examined whether clioquinol induces autophagy in cultured astrocytes in a zinc-dependent manner.Within 1 h of exposure to 0.1-10 μM clioquinol, the levels of microtubule-associated protein 1 light chain 3 (LC3)-II, a marker of autophagy, began to increase in astrocytes. Confocal live-cell imaging of GFP-LC3-transfected astrocytes showed the formation of LC3(+) autophagic vacuoles (AVs), providing a further indication that clioquinol induced autophagy. Addition of 3-methyladenine or small-interfering RNA against autophagy-related gene 6 (ATG6/Beclin-1) blocked clioquinol-induced increases in LC3-II. FluoZin-3 fluorescence microscopy showed that, like the zinc ionophore pyrithione, clioquinol increased intracellular zinc levels in the cytosol and AVs in an extracellular zinc-dependent manner. Zinc chelation with N,N,N',N'-tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) reduced, and addition of zinc increased the levels of LC3-II and LC3(+) puncta, indicating that zinc influx plays a key role therein. Moreover, astrocytes and SH-SY5Y cells expressing mutant huntingtin (mHttQ74) accumulated less aggregates when treated with clioquinol, and this effect was reversed by TPEN. These results indicate that clioquinol-induced autophagy is likely to be physiologically functional.The present study demonstrates that clioquinol induces autophagy in a zinc-dependent manner and contributes to clearance of aggregated proteins in astrocytes and neurons. Hence, in addition to its metal-chelating effect in and around amyloid beta (Aβ) plaques, clioquinol may contribute to the reduction of Aβ loads by activating autophagy by increasing or normalizing intracellular zinc levels in brain cells.

Original languageEnglish
Pages (from-to)242-251
Number of pages10
JournalNeurobiology of Disease
Volume42
Issue number3
DOIs
Publication statusPublished - 2011 Jun 1

Fingerprint

Clioquinol
Ionophores
Autophagy
Astrocytes
Zinc
Neurons
Light
ethylenediamine
Vacuoles
Amyloid
Microtubule-Associated Proteins
Fluorescence Microscopy
Cytosol
Small Interfering RNA

All Science Journal Classification (ASJC) codes

  • Neurology

Cite this

Park, Mi Ha ; Lee, Sook Jeong ; Byun, Hyae ran ; Kim, Yunha ; Oh, Young Jun ; Koh, Jae Young ; Hwang, Jung Jin. / Clioquinol induces autophagy in cultured astrocytes and neurons by acting as a zinc ionophore. In: Neurobiology of Disease. 2011 ; Vol. 42, No. 3. pp. 242-251.
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abstract = "Recent studies have demonstrated that clioquinol, an antibiotic with an anti-amyloid effect, acts as a zinc ionophore under physiological conditions. Because increases in labile zinc may induce autophagy, we examined whether clioquinol induces autophagy in cultured astrocytes in a zinc-dependent manner.Within 1 h of exposure to 0.1-10 μM clioquinol, the levels of microtubule-associated protein 1 light chain 3 (LC3)-II, a marker of autophagy, began to increase in astrocytes. Confocal live-cell imaging of GFP-LC3-transfected astrocytes showed the formation of LC3(+) autophagic vacuoles (AVs), providing a further indication that clioquinol induced autophagy. Addition of 3-methyladenine or small-interfering RNA against autophagy-related gene 6 (ATG6/Beclin-1) blocked clioquinol-induced increases in LC3-II. FluoZin-3 fluorescence microscopy showed that, like the zinc ionophore pyrithione, clioquinol increased intracellular zinc levels in the cytosol and AVs in an extracellular zinc-dependent manner. Zinc chelation with N,N,N',N'-tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) reduced, and addition of zinc increased the levels of LC3-II and LC3(+) puncta, indicating that zinc influx plays a key role therein. Moreover, astrocytes and SH-SY5Y cells expressing mutant huntingtin (mHttQ74) accumulated less aggregates when treated with clioquinol, and this effect was reversed by TPEN. These results indicate that clioquinol-induced autophagy is likely to be physiologically functional.The present study demonstrates that clioquinol induces autophagy in a zinc-dependent manner and contributes to clearance of aggregated proteins in astrocytes and neurons. Hence, in addition to its metal-chelating effect in and around amyloid beta (Aβ) plaques, clioquinol may contribute to the reduction of Aβ loads by activating autophagy by increasing or normalizing intracellular zinc levels in brain cells.",
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Clioquinol induces autophagy in cultured astrocytes and neurons by acting as a zinc ionophore. / Park, Mi Ha; Lee, Sook Jeong; Byun, Hyae ran; Kim, Yunha; Oh, Young Jun; Koh, Jae Young; Hwang, Jung Jin.

In: Neurobiology of Disease, Vol. 42, No. 3, 01.06.2011, p. 242-251.

Research output: Contribution to journalArticle

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