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Synthetic Condensed 1,4-naphthoquinone Derivative Shifts Neural Stem Cell Differentiation by Regulating Redox State

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Abstract

Naphthoquinones are bioactive compounds widespread in nature that impact on several cellular pathways, including cell proliferation and survival, by acting as prooxidants and electrophiles. We have previously described the role of the synthetic isoxazole condensed 1,4-naphthoquinone derivative 1a in preventing apoptosis induced by distinct stimuli in several cell models. In addition, apoptosis regulators and executioners may control neural stem cell (NSC) fate, without involving cell death per se. Here, we hypothesize that 1a might also play a role in NSC fate decision. We found that exposure to 1a shifts NSC differentiation potential from neurogenic to gliogenic lineage and involves the generation of reactive oxygen species, without increasing cell death. Modulation of caspases and calpains, using cysteine protease inhibitors, failed to mimic 1a effects. In addition, incubation with the naphthoquinone derivative resulted in upregulation and nuclear translocation of antioxidant responsive proteins, Nrf2 and Sirt1, which in turn may mediate 1a-directed shift in NSC differentiation. In fact, antioxidants halted the shift in NSC differentiation potential from neurogenic to gliogenic lineage, while strongly reducing reactive oxygen species generation and Nrf2 and Sirt1 nuclear translocation in NSC exposed to 1a. Collectively, these data support a new role for a specific naphthoquinone derivative in NSC fate decision and underline the importance of redox environment control.

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Abbreviations

DCF:

2′,7′-Dichlorofluorescein

FBS:

Fetal bovine serum

GFAP:

Glial fibrillary acidic protein

H2DCFDA:

2′,7′-Dichlorodihydrofluorescein diacetate

LA:

Lipoic acid

NAC:

N-Acetylcysteine

Nrf2:

Nuclear factor erythroid 2-related factor 2

NSC:

Neural stem cells

NS-TGFP:

Adherent mouse Tau-GFP NSC

PBS:

Phosphate buffer saline

ROS:

Reactive oxygen species

Sirt1:

Sirtuin 1

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Acknowledgments

We are grateful to Elsa Abranches and Evguenia Bekman (Instituto de Medicina Molecular, University of Lisbon, Lisbon, Portugal) for skillful technical assistance in establishing NS-TGFP cell line culture conditions. The authors also thank Carlos Ribeiro (iMed.UL, University of Lisbon, Lisbon, Portugal) for providing part of the freshly synthesized 1a. We also thank Maria João Gama, Elsa Rodrigues and Andreia Carvalho iMed.UL, University of Lisbon, Lisbon, Portugal) for technical assistance and reagents for ROS assays and Nrf2 detection. This work was supported by grants PTDC/SAU-NMC/117877/2010 and Pest-OE/SAU/UI4013/2011, and fellowship SFRH/BD/42008/2007 (DMS) from Fundação para a Ciência e a Tecnologia, Portugal.

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Authors and Affiliations

  1. Research Institute for Medicines and Pharmaceutical Sciences (iMed.UL), Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal

    Daniela M. Santos, Maria M. M. Santos, Rui Moreira, Susana Solá & Cecília M. P. Rodrigues

  2. Department of Biochemistry and Human Biology, Faculty of Pharmacy, University of Lisbon, Av. Prof. Gama Pinto, 1649-003, Lisbon, Portugal

    Susana Solá & Cecília M. P. Rodrigues

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  1. Daniela M. Santos
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  2. Maria M. M. Santos
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  4. Susana Solá
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Correspondence to Susana Solá.

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Supplemental Fig. 1

Chemical formula of naphtho[2,3-d]isoxazole-4,9-dione-3-carboxylate 1a. (TIFF 5965 kb)

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Santos, D.M., Santos, M.M.M., Moreira, R. et al. Synthetic Condensed 1,4-naphthoquinone Derivative Shifts Neural Stem Cell Differentiation by Regulating Redox State. Mol Neurobiol 47, 313–324 (2013). https://doi.org/10.1007/s12035-012-8353-y

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