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Expression Profile of Rat Hippocampal Neurons Treated with the Neuroprotective Compound 2,4-Dinitrophenol: Up-Regulation of cAMP Signaling Genes

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Abstract

2,4-Dinitrophenol (DNP) is classically known as a mitochondrial uncoupler and, at high concentrations, is toxic to a variety of cells. However, it has recently been shown that, at subtoxic concentrations, DNP protects neurons against a variety of insults and promotes neuronal differentiation and neuritogenesis. The molecular and cellular mechanisms underlying the beneficial neuroactive properties of DNP are still largely unknown. We have now used DNA microarray analysis to investigate changes in gene expression in rat hippocampal neurons in culture treated with low micromolar concentrations of DNP. Under conditions that did not affect neuronal viability, high-energy phosphate levels or mitochondrial oxygen consumption, DNP induced up-regulation of 275 genes and down-regulation of 231 genes. Significantly, several up-regulated genes were linked to intracellular cAMP signaling, known to be involved in neurite outgrowth, synaptic plasticity, and neuronal survival. Differential expression of specific genes was validated by quantitative RT-PCR using independent samples. Results shed light on molecular mechanisms underlying neuroprotection by DNP and point to possible targets for development of novel therapeutics for neurodegenerative disorders.

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Acknowledgments

This article was supported by grants from Howard Hughes Medical Institute, Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/Brazil), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ/Brazil) and Instituto Nacional de Neurociência Translacional (INNT/Brazil) (to STF). We thank Lorena Chávez González, Simón Guzmán León, José Luis Santillán Torres and Jorge Ramírez for expert assistance with microarray analysis, and Gerardo Coello, Gustavo Corral and Ana Patricia Gómez for genArise software assistance. CTM and EDN acknowledge the support of Associação Beneficente Alzira Denise Hertzog Silva (ABADHS) and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP).

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Authors and Affiliations

  1. Instituto de Bioquímica Médica, Programa de Bioquímica e Biofísica Celular, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, 21944-590, Brazil

    Adriano Sebollela, Léo Freitas-Corrêa, Fábio F. Oliveira, Ana Paula Wasilewska-Sampaio, Juliana Camacho-Pereira, Antonio Galina, Fernanda G. De Felice & Sérgio T. Ferreira

  2. Laboratório de Neurociências (LIM27), Instituto de Psiquiatria, Faculdade de Medicina da Universidade de São Paulo, São Paulo, SP, 05403-010, Brazil

    Camila T. Mendes & Emmanuel Dias-Neto

  3. Laboratório de Bioinformática, Hospital do Câncer AC Camargo, São Paulo, SP, Brazil

    Helena Brentani

  4. Laboratório de Bioinformática e Biologia Computacional, Serviço de Pesquisa Clínica, Coordenação de Pesquisa (CPQ), Instituto Nacional do Câncer (INCA), Rio de Janeiro, RJ, 20231-050, Brazil

    Fabio Passetti

  5. Centro de Pesquisas do Hospital do Cancer, Sao Paulo, SP, 01509-900, Brazil

    Emmanuel Dias-Neto

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  1. Adriano Sebollela
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  2. Léo Freitas-Corrêa
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  3. Fábio F. Oliveira
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  4. Camila T. Mendes
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  5. Ana Paula Wasilewska-Sampaio
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  6. Juliana Camacho-Pereira
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  7. Antonio Galina
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  9. Fabio Passetti
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  10. Fernanda G. De Felice
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  11. Emmanuel Dias-Neto
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  12. Sérgio T. Ferreira
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Corresponding author

Correspondence to Sérgio T. Ferreira.

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Supplemental Figure I

Representative DCF fluorescence images for detection of ROS levels (TIFF 79 kb)

Supplemental Table I

Genes up-regulated by DNP treatment (DOC 330 kb)

Supplemental Table II

Genes down-regulated by DNP treatment (DOC 298 kb)

Supplemental Table III

Kegg pathways with low representation in the up-regulated dataset (DOC 81 kb)

Supplemental Table IV

Kegg pathways with low representation in the down-regulated dataset (DOC 33 kb)

Supplemental Table V

Sequences of oligonucleotide primers used for real-time PCR (DOC 68 kb)

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Sebollela, A., Freitas-Corrêa, L., Oliveira, F.F. et al. Expression Profile of Rat Hippocampal Neurons Treated with the Neuroprotective Compound 2,4-Dinitrophenol: Up-Regulation of cAMP Signaling Genes. Neurotox Res 18, 112–123 (2010). https://doi.org/10.1007/s12640-009-9133-y

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