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Minocycline Recovers MTT-Formazan Exocytosis Impaired by Amyloid Beta Peptide

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Abstract

Minocycline, a tetracycline antibiotic, has been reported to exert beneficial effects in models of Alzheimer’s disease (AD). To characterize the mechanisms underlying the putative minocycline-related neuroprotection, we studied its effect in an in vitro model of AD. Primary hippocampal cultures were treated with β-amyloid peptide (Aβ) and cell viability was assessed by standard MTT-assay. Incubation with 10 μM Aβ for 24 h significantly inhibits cellular MTT-reduction without inducing morphological signs of enhanced cell death or increase in release of lactate dehydrogenase. This indicates that cell viability was not affected. The inhibition of MTT-reduction by Aβ was due to an acceleration of MTT-formazan exocytosis. Intriguingly, the Aβ-triggered increase in MTT-formazan exocytosis was abolished by co-treatment with minocycline. In vehicle-treated cells minocycline had no effect on formazan exocytosis. This hitherto unrecognized property of minocycline has to be noticed in the elucidation of the underlying mechanism of this promising neuroprotectant.

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References

  • Abe K, Saito H (1998) Amyloid beta protein inhibits cellular MTT reduction not by suppression of mitochondrial succinate dehydrogenase but by acceleration of MTT formazan exocytosis in cultured rat cortical astrocytes. Neurosci Res 31:295–305

    Article  CAS  PubMed  Google Scholar 

  • Blum D, Chtarto A, Tenenbaum L, Brotchi J, Levivier M (2004) Clinical potential of minocycline for neurodegenerative disorders. Neurobiol Dis 17:359–366

    Article  CAS  PubMed  Google Scholar 

  • Choi Y, Kim HS, Shin KY, Kim EM, Kim M, Park CH, Jeong YH, Yoo J, Lee JP, Chang KA, Kim S, Suh YH (2007) Minocycline attenuates neuronal cell death and improves cognitive impairment in Alzheimer’s disease models. Neuropsychopharmacology 32:2393–2404

    Article  CAS  PubMed  Google Scholar 

  • Fan R, Xu F, Previti ML, Davis J, Grande AM, Robinson JK, Van Nostrand WE (2007) Minocycline reduces microglial activation and improves behavioral deficits in a transgenic model of cerebral microvascular amyloid. J Neurosci 27:3057–3063

    Article  CAS  PubMed  Google Scholar 

  • Haroon MF, Fatima A, Scholer S, Gieseler A, Horn TF, Kirches E, Wolf G, Kreutzmann P (2007) Minocycline, a possible neuroprotective agent in Leber’s hereditary optic neuropathy (LHON): studies of cybrid cells bearing 11778 mutation. Neurobiol Dis 28:237–250

    Article  CAS  PubMed  Google Scholar 

  • Hong HS, Maezawa I, Yao N, Xu B, Diaz-Avalos R, Rana S, Hua DH, Cheng RH, Lam KS, Jin LW (2007) Combining the rapid MTT formazan exocytosis assay and the MC65 protection assay led to the discovery of carbazole analogs as small molecule inhibitors of Abeta oligomer-induced cytotoxicity. Brain Res 1130:223–234

    Article  CAS  PubMed  Google Scholar 

  • Isobe I, Michikawa M, Yanagisawa K (1999) Enhancement of MTT, a tetrazolium salt, exocytosis by amyloid beta-protein and chloroquine in cultured rat astrocytes. Neurosci Lett 266:129–132

    Article  CAS  PubMed  Google Scholar 

  • Jordan J, Fernandez-Gomez FJ, Ramos M, Ikuta I, Aguirre N, Galindo MF (2007) Minocycline and cytoprotection: shedding new light on a shadowy controversy. Curr Drug Deliv 4:225–231

    Article  CAS  PubMed  Google Scholar 

  • Kerokoski P, Soininen H, Pirttila T (2001) Beta-amyloid (1–42) affects MTT reduction in astrocytes: implications for vesicular trafficking and cell functionality. Neurochem Int 38:127–134

    Article  CAS  PubMed  Google Scholar 

  • Kupsch K, Hertel S, Kreutzmann P, Wolf G, Wallesch CW, Siemen D, Schonfeld P (2009) Impairment of mitochondrial function by minocycline. FEBS J 276:1729–1738

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Schubert D (1997) Cytotoxic amyloid peptides inhibit cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction by enhancing MTT formazan exocytosis. J Neurochem 69:2285–2293

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Schubert D (1998) Steroid hormones block amyloid fibril-induced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) formazan exocytosis: relationship to neurotoxicity. J Neurochem 71:2322–2329

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Peterson DA, Kimura H, Schubert D (1997) Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. J Neurochem 69:581–593

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Peterson DA, Schubert D (1998) Amyloid beta peptide alters intracellular vesicle trafficking and cholesterol homeostasis. Proc Natl Acad Sci USA 95:13266–13271

    Article  CAS  PubMed  Google Scholar 

  • Lukiw WJ, Pappolla M, Pelaez RP, Bazan NG (2005) Alzheimer’s disease—a dysfunction in cholesterol and lipid metabolism. Cell Mol Neurobiol 25:475–483

    Article  CAS  PubMed  Google Scholar 

  • Mansson R, Morota S, Hansson MJ, Sonoda I, Yasuda Y, Shimazu M, Sugiura A, Yanagi S, Miura H, Uchino H, Elmer E (2010) Minocycline sensitizes rodent and human liver mitochondria to the permeability transition: implications for toxicity in liver transplantation. Hepatology 51:347–348 (author reply 349–350)

    PubMed  Google Scholar 

  • Nikodemova M, Lee J, Fabry Z, Duncan ID (2010) Minocycline attenuates experimental autoimmune encephalomyelitis in rats by reducing T cell infiltration into the spinal cord. J Neuroimmunol 219:33–37

    Article  CAS  PubMed  Google Scholar 

  • Ryu JK, Franciosi S, Sattayaprasert P, Kim SU, McLarnon JG (2004) Minocycline inhibits neuronal death and glial activation induced by beta-amyloid peptide in rat hippocampus. Glia 48:85–90

    Article  PubMed  Google Scholar 

  • Seabrook TJ, Jiang L, Maier M, Lemere CA (2006) Minocycline affects microglia activation, Abeta deposition, and behavior in APP-tg mice. Glia 53:776–782

    Article  PubMed  Google Scholar 

  • Shearman MS, Ragan CI, Iversen LL (1994) Inhibition of PC12 cell redox activity is a specific, early indicator of the mechanism of beta-amyloid-mediated cell death. Proc Natl Acad Sci USA 91:1470–1474

    Article  CAS  PubMed  Google Scholar 

  • Shearman MS, Hawtin SR, Tailor VJ (1995) The intracellular component of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction is specifically inhibited by beta-amyloid peptides. J Neurochem 65:218–227

    Article  CAS  PubMed  Google Scholar 

  • Suzuki T, Araki Y, Yamamoto T, Nakaya T (2006) Trafficking of Alzheimer’s disease-related membrane proteins and its participation in disease pathogenesis. J Biochem 139:949–955

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Zhu S, Drozda M, Zhang W, Stavrovskaya IG, Cattaneo E, Ferrante RJ, Kristal BS, Friedlander RM (2003) Minocycline inhibits caspase-independent and -dependent mitochondrial cell death pathways in models of Huntington’s disease. Proc Natl Acad Sci USA 100:10483–10487

    Article  CAS  PubMed  Google Scholar 

  • Zhang ZY, Zhang Z, Fauser U, Schluesener HJ (2009) Improved outcome of EAN, an animal model of GBS, through amelioration of peripheral and central inflammation by minocycline. J Cell Mol Med 13:341–351

    Article  CAS  PubMed  Google Scholar 

  • Zhu S, Stavrovskaya IG, Drozda M, Kim BY, Ona V, Li M, Sarang S, Liu AS, Hartley DM, Wu DC, Gullans S, Ferrante RJ, Przedborski S, Kristal BS, Friedlander RM (2002) Minocycline inhibits cytochrome c release and delays progression of amyotrophic lateral sclerosis in mice. Nature 417:74–78

    Article  CAS  PubMed  Google Scholar 

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Acknowledgment

This work was supported by funding from Magdeburger Forschungsverbund NBL3 (project number BMBF 01ZZ0407).

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Authors and Affiliations

  1. Institute of Medical Neurobiology, Otto-von-Guericke University Magdeburg, Leipziger Str. 44, 39120, Magdeburg, Germany

    Peter Kreutzmann, Gerald Wolf & Kathleen Kupsch

  2. Institute of Biochemistry and Cell Biology, Otto-von-Guericke University Magdeburg, Leipziger Str. 44, 39120, Magdeburg, Germany

    Peter Kreutzmann

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  1. Peter Kreutzmann
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  2. Gerald Wolf
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  3. Kathleen Kupsch
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Correspondence to Peter Kreutzmann.

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Kreutzmann, P., Wolf, G. & Kupsch, K. Minocycline Recovers MTT-Formazan Exocytosis Impaired by Amyloid Beta Peptide. Cell Mol Neurobiol 30, 979–984 (2010). https://doi.org/10.1007/s10571-010-9528-6

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  • DOI: https://doi.org/10.1007/s10571-010-9528-6

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