Frataxin silencing inactivates mitochondrial complex i in NSC34 motoneuronal cells and alters glutathione homeostasis

Barbara Carletti, Emanuela Piermarini, Giulia Tozzi, Lorena Travaglini, Alessandra Torraco, Anna Pastore, Marco Sparaco, Sara Petrillo, Rosalba Carrozzo, Enrico Bertini, Fiorella Piemonte

Research output: Contribution to journalArticle

Abstract

Friedreich's ataxia (FRDA) is a hereditary neurodegenerative disease characterized by a reduced synthesis of the mitochondrial iron chaperon protein frataxin as a result of a large GAA triplet-repeat expansion within the first intron of the frataxin gene. Despite neurodegeneration being the prominent feature of this pathology involving both the central and the peripheral nervous system, information on the impact of frataxin deficiency in neurons is scant. Here, we describe a neuronal model displaying some major biochemical and morphological features of FRDA. By silencing the mouse NSC34 motor neurons for the frataxin gene with shRNA lentiviral vectors, we generated two cell lines with 40% and 70% residual amounts of frataxin, respectively. Frataxin-deficient cells showed a specific inhibition of mitochondrial Complex I (CI) activity already at 70% OPEN ACCESS residual frataxin levels, whereas the glutathione imbalance progressively increased after silencing. These biochemical defects were associated with the inhibition of cell proliferation and morphological changes at the axonal compartment, both depending on the frataxin amount. Interestingly, at 70% residual frataxin levels, the in vivo treatment with the reduced glutathione revealed a partial rescue of cell proliferation. Thus, NSC34 frataxin silenced cells could be a suitable model to study the effect of frataxin deficiency in neurons and highlight glutathione as a potential beneficial therapeutic target for FRDA.

Original languageEnglish
Pages (from-to)5789-5806
Number of pages18
JournalInternational Journal of Molecular Sciences
Volume15
Issue number4
DOIs
Publication statusPublished - Apr 4 2014

Fingerprint

ataxia
homeostasis
glutathione
neurons
Neurons
Glutathione
Homeostasis
Cell proliferation
genes
peripheral nervous system
Genes
cells
Neurodegenerative diseases
Friedreich Ataxia
pathology
Neurology
Pathology
compartments
cultured cells
mice

Keywords

  • Friedreich's ataxia
  • Glutathione
  • Mitochondrial enzymes
  • Neurodegeneration
  • Oxidative stress

ASJC Scopus subject areas

  • Inorganic Chemistry
  • Catalysis
  • Molecular Biology
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Spectroscopy
  • Medicine(all)

Cite this

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title = "Frataxin silencing inactivates mitochondrial complex i in NSC34 motoneuronal cells and alters glutathione homeostasis",
abstract = "Friedreich's ataxia (FRDA) is a hereditary neurodegenerative disease characterized by a reduced synthesis of the mitochondrial iron chaperon protein frataxin as a result of a large GAA triplet-repeat expansion within the first intron of the frataxin gene. Despite neurodegeneration being the prominent feature of this pathology involving both the central and the peripheral nervous system, information on the impact of frataxin deficiency in neurons is scant. Here, we describe a neuronal model displaying some major biochemical and morphological features of FRDA. By silencing the mouse NSC34 motor neurons for the frataxin gene with shRNA lentiviral vectors, we generated two cell lines with 40{\%} and 70{\%} residual amounts of frataxin, respectively. Frataxin-deficient cells showed a specific inhibition of mitochondrial Complex I (CI) activity already at 70{\%} OPEN ACCESS residual frataxin levels, whereas the glutathione imbalance progressively increased after silencing. These biochemical defects were associated with the inhibition of cell proliferation and morphological changes at the axonal compartment, both depending on the frataxin amount. Interestingly, at 70{\%} residual frataxin levels, the in vivo treatment with the reduced glutathione revealed a partial rescue of cell proliferation. Thus, NSC34 frataxin silenced cells could be a suitable model to study the effect of frataxin deficiency in neurons and highlight glutathione as a potential beneficial therapeutic target for FRDA.",
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author = "Barbara Carletti and Emanuela Piermarini and Giulia Tozzi and Lorena Travaglini and Alessandra Torraco and Anna Pastore and Marco Sparaco and Sara Petrillo and Rosalba Carrozzo and Enrico Bertini and Fiorella Piemonte",
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T1 - Frataxin silencing inactivates mitochondrial complex i in NSC34 motoneuronal cells and alters glutathione homeostasis

AU - Carletti, Barbara

AU - Piermarini, Emanuela

AU - Tozzi, Giulia

AU - Travaglini, Lorena

AU - Torraco, Alessandra

AU - Pastore, Anna

AU - Sparaco, Marco

AU - Petrillo, Sara

AU - Carrozzo, Rosalba

AU - Bertini, Enrico

AU - Piemonte, Fiorella

PY - 2014/4/4

Y1 - 2014/4/4

N2 - Friedreich's ataxia (FRDA) is a hereditary neurodegenerative disease characterized by a reduced synthesis of the mitochondrial iron chaperon protein frataxin as a result of a large GAA triplet-repeat expansion within the first intron of the frataxin gene. Despite neurodegeneration being the prominent feature of this pathology involving both the central and the peripheral nervous system, information on the impact of frataxin deficiency in neurons is scant. Here, we describe a neuronal model displaying some major biochemical and morphological features of FRDA. By silencing the mouse NSC34 motor neurons for the frataxin gene with shRNA lentiviral vectors, we generated two cell lines with 40% and 70% residual amounts of frataxin, respectively. Frataxin-deficient cells showed a specific inhibition of mitochondrial Complex I (CI) activity already at 70% OPEN ACCESS residual frataxin levels, whereas the glutathione imbalance progressively increased after silencing. These biochemical defects were associated with the inhibition of cell proliferation and morphological changes at the axonal compartment, both depending on the frataxin amount. Interestingly, at 70% residual frataxin levels, the in vivo treatment with the reduced glutathione revealed a partial rescue of cell proliferation. Thus, NSC34 frataxin silenced cells could be a suitable model to study the effect of frataxin deficiency in neurons and highlight glutathione as a potential beneficial therapeutic target for FRDA.

AB - Friedreich's ataxia (FRDA) is a hereditary neurodegenerative disease characterized by a reduced synthesis of the mitochondrial iron chaperon protein frataxin as a result of a large GAA triplet-repeat expansion within the first intron of the frataxin gene. Despite neurodegeneration being the prominent feature of this pathology involving both the central and the peripheral nervous system, information on the impact of frataxin deficiency in neurons is scant. Here, we describe a neuronal model displaying some major biochemical and morphological features of FRDA. By silencing the mouse NSC34 motor neurons for the frataxin gene with shRNA lentiviral vectors, we generated two cell lines with 40% and 70% residual amounts of frataxin, respectively. Frataxin-deficient cells showed a specific inhibition of mitochondrial Complex I (CI) activity already at 70% OPEN ACCESS residual frataxin levels, whereas the glutathione imbalance progressively increased after silencing. These biochemical defects were associated with the inhibition of cell proliferation and morphological changes at the axonal compartment, both depending on the frataxin amount. Interestingly, at 70% residual frataxin levels, the in vivo treatment with the reduced glutathione revealed a partial rescue of cell proliferation. Thus, NSC34 frataxin silenced cells could be a suitable model to study the effect of frataxin deficiency in neurons and highlight glutathione as a potential beneficial therapeutic target for FRDA.

KW - Friedreich's ataxia

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KW - Mitochondrial enzymes

KW - Neurodegeneration

KW - Oxidative stress

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