DT-Diaphorase Prevents Aminochrome-Induced Lysosome Dysfunction in SH-SY5Y Cells
- 38 Downloads
Aminochrome has been reported to induce lysosomal dysfunction by inhibiting the vacuolar H-type ATPase localized in lysosome membrane. DT-diaphorase has been proposed to prevent aminochrome neurotoxicity but it is unknown whether this enzyme prevents aminochrome-induced lysosomal dysfunction. In the present study, we tested the protective role of DT-diaphorase in lysosomal dysfunction by generating a cell line (SH-SY5YsiNQ7) with a stable expression of a siRNA against DT-diaphorase with only 10% expression of mRNA enzyme. The cells differentiated with retinoic acid and 12-o-tetradecanoylphorbol-13-acetate show a significant increase in the expression of tyrosine hydroxylase, vesicular monoamine transporter-2, and dopamine transporter. The incubation of SH-SY5YsiNQ7 cells with 10 μM aminochrome resulted in a significant decrease of lysosome pH determined by using acridine orange, while aminochrome has no effect on SH-SY5Y cells. These results support the proposed protective role of DT-diaphorase against aminochrome-induced lysosomal dysfunction.
KeywordsDT-Diaphorase Lysosomal dysfunction Aminochrome Dopamine Neurotoxicity Neuroprotection
This research was supported by FONDECYT (1170033).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that there is no conflict of interest.
- Aguirre P, Urrutia P, Tapia V, Villa M, Paris I, Segura-Aguilar J, Núñez MT (2012) The dopamine metabolite aminochrome inhibits mitochondrial complex I and modifies the expression of iron transporters DMT1 and FPN1. Biometals 25:795–803. https://doi.org/10.1007/s10534-012-9525-y CrossRefPubMedGoogle Scholar
- Arriagada A, Paris I, Sanchez de las Matas MJ et al (2004) On the neurotoxicity of leukoaminochrome o-semiquinone radical derived of dopamine oxidation: mitochondria damage, necrosis and hydroxyl radical formation. Neurobiol Dis 16:468–477. https://doi.org/10.1016/j.nbd.2004.03.014 CrossRefPubMedGoogle Scholar
- Cuevas C, Huenchuguala S, Muñoz P, Villa M, Paris I, Mannervik B, Segura-Aguilar J (2015) Glutathione transferase-M2-2 secreted from glioblastoma cell protects SH-SY5Y cells from aminochrome neurotoxicity. Neurotox Res 27:217–228. https://doi.org/10.1007/s12640-014-9500-1 CrossRefPubMedGoogle Scholar
- Huenchuguala S, Muñoz P, Zavala P, Villa M, Cuevas C, Ahumada U, Graumann R, Nore BF, Couve E, Mannervik B, Paris I, Segura-Aguilar J (2014) Glutathione transferase M2 protects glioblastoma cells against aminochrome toxicity by preventing autophagy and lysosome dysfunction. Autophagy 10:618–630. https://doi.org/10.4161/auto.27720 CrossRefPubMedPubMedCentralGoogle Scholar
- Huenchuguala S, Muñoz P, Segura-Aguilar J (2017) The importance of mitophagy in maintaining mitochondrial function in U373MG cells. Bafilomycin A1 restores aminochrome-induced mitochondrial damage. ACS Chem Neurosci 8:2247–2253. https://doi.org/10.1021/acschemneuro.7b00152 CrossRefPubMedGoogle Scholar
- Paris I, Perez-Pastene C, Cardenas S, Iturriaga-Vasquez P, Muñoz P, Couve E, Caviedes P, Segura-Aguilar J (2010) Aminochrome induces disruption of actin, alpha-, and beta-tubulin cytoskeleton networks in substantia-nigra-derived cell line. Neurotox Res 18:82–92. https://doi.org/10.1007/s12640-009-9148-4 CrossRefPubMedGoogle Scholar
- Zecca L, Fariello R, Riederer P, Sulzer D, Gatti A, Tampellini D (2002) The absolute concentration of nigral neuromelanin, assayed by a new sensitive method, increases throughout the life and is dramatically decreased in Parkinson’s disease. FEBS Lett 510:216–220. https://doi.org/10.1016/S0014-5793(01)03269-0 CrossRefPubMedGoogle Scholar