Skip to main content

Main menu

  • Home
  • About the Journal
    • General Information
    • Scope
    • Editorial Board
    • Impact & Metrics
    • Benefits of Publishing
    • Advertising/Sponsorship
    • About the Biochemical Society
  • Current Issue
  • For Authors
    • Submit Your Paper
    • Submission Guidelines
    • Editorial Policy
    • Open Access Policy
    • Rights and Permissions
    • Biochemical Society Member Benefits
  • For Librarians
    • Subscriptions and Pricing
    • Check Your Usage
    • Terms and Conditions
      • Biochemical Journal- Terms and Conditions of Usage
    • Open Access Policy
    • FAQs for Librarians
    • Register for Free Trial
  • For Readers
    • Subscribe
    • Rights and Permissions
    • Biochemical Society Member Benefits
    • Journal Access for Biochemical Society Members
    • Request a Free Trial
  • Collections
    • Article Collections
    • Classic Articles
  • Help
    • Technical Support
    • Contact Us
  • Other Publications
    • NEW: Emerging Topics in Life Sciences
    • NEW: Neuronal Signaling
    • Clinical Science
    • Biochemical Journal
    • Biochemical Society Transactions
    • Bioscience Reports
    • Essays in Biochemistry
    • Biochemical Society Symposia
    • Cell Signalling Biology
    • Glossary of Biochemistry and Molecular Biology
    • The Biochemist
    • Biochemical Society

User menu

  • Log-in
  • Subscribe
  • Contact Us

Search

  • Advanced search
  • Other Publications
    • NEW: Emerging Topics in Life Sciences
    • NEW: Neuronal Signaling
    • Clinical Science
    • Biochemical Journal
    • Biochemical Society Transactions
    • Bioscience Reports
    • Essays in Biochemistry
    • Biochemical Society Symposia
    • Cell Signalling Biology
    • Glossary of Biochemistry and Molecular Biology
    • The Biochemist
    • Biochemical Society

Log-in

Sign-up for alerts  
  • My Cart
Biochemical Journal
Browse Archive
Advanced Search
  • Home
  • About the Journal
    • General Information
    • Scope
    • Editorial Board
    • Impact & Metrics
    • Benefits of Publishing
    • Advertising/Sponsorship
    • About the Biochemical Society
  • Current Issue
  • For Authors
    • Submit Your Paper
    • Submission Guidelines
    • Editorial Policy
    • Open Access Policy
    • Rights and Permissions
    • Biochemical Society Member Benefits
  • For Librarians
    • Subscriptions and Pricing
    • Check Your Usage
    • Terms and Conditions
    • Open Access Policy
    • FAQs for Librarians
    • Register for Free Trial
  • For Readers
    • Subscribe
    • Rights and Permissions
    • Biochemical Society Member Benefits
    • Journal Access for Biochemical Society Members
    • Request a Free Trial
  • Collections
    • Article Collections
    • Classic Articles
  • Help
    • Technical Support
    • Contact Us

Research article

Lipid-based DNA/siRNA transfection agents disrupt neuronal bioenergetics and mitophagy

Eleonora Napoli, Siming Liu, Ilaria Marsilio, Konstantinos Zarbalis, Cecilia Giulivi
Biochemical Journal Nov 10, 2017, 474 (23) 3887-3902; DOI: 10.1042/BCJ20170632
Eleonora Napoli
Department of Molecular Biosciences, School of Veterinary Medicine, University of California Davis, Davis, CA 95616, U.S.A.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Siming Liu
Department of Molecular Biosciences, School of Veterinary Medicine, University of California Davis, Davis, CA 95616, U.S.A.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Ilaria Marsilio
Department of Molecular Biosciences, School of Veterinary Medicine, University of California Davis, Davis, CA 95616, U.S.A.Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, Italy
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Konstantinos Zarbalis
Department of Pathology and Laboratory Medicine, UC Davis Medical Center, Sacramento, CA 95817, U.S.A.Institute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Northern California, 2425 Stockton Boulevard, Sacramento, CA 95817, U.S.A.Medical Investigations of Neurodevelopmental Disorders (MIND) Institute, University of California Davis, Sacramento, CA 95817, U.S.A.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
Cecilia Giulivi
Department of Molecular Biosciences, School of Veterinary Medicine, University of California Davis, Davis, CA 95616, U.S.A.Medical Investigations of Neurodevelopmental Disorders (MIND) Institute, University of California Davis, Sacramento, CA 95817, U.S.A.
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • View author's works on this site
  • For correspondence: cgiulivi@ucdavis.edu
  • Article
  • Figures
  • Info & Metrics
  • Supplementary Data
  • PDF
Loading

Abstract

A multitude of natural and artificial compounds have been recognized to modulate autophagy, providing direct or, through associated pathways, indirect entry points to activation and inhibition. While these pharmacological tools are extremely useful in the study of autophagy, their abundance also suggests the potential presence of unidentified autophagic modulators that may interfere with experimental designs if applied unknowingly. Here, we report unanticipated effects on autophagy and bioenergetics in neuronal progenitor cells (NPCs) incubated with the widely used lipid-based transfection reagent lipofectamine (LF), which induced mitochondria depolarization followed by disruption of electron transport. When NPCs were exposed to LF for 5 h followed by 24, 48, and 72 h in LF-free media, an immediate increase in mitochondrial ROS production and nitrotyrosine formation was observed. These events were accompanied by disrupted mitophagy (accumulation of dysfunctional and damaged mitochondria, and of LC3II and p62), in an mTOR- and AMPK-independent manner, and despite the increased mitochondrial PINK1 (PTEN-inducible kinase 1) localization. Evidence supported a role for a p53-mediated abrogation of parkin translocation and/or abrogation of membrane fusion between autophagosome and lysosomes. While most of the outcomes were LF-specific, only two were shared by OptiMEM exposure (with no serum and reduced glucose levels) albeit at lower extents. Taken together, our findings show that the use of transfection reagents requires critical evaluation with respect to consequences for overall cellular health, particularly in experiments designed to address autophagy-inducing effects and/or energy stress.

  • autophagy
  • mitophagy
  • oxidative stress
  • stress
  • transfection agent
  • Abbreviations

    DAPI,
    4′,6-diamidino-2-phenylindole;
    DOPE,
    1,2-dioleoyl-sn-glycero-3-phosphoethanolamine;
    DOTAP,
    N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate;
    G6PD,
    glucose-6-phosphate dehydrogenase;
    HO1,
    heme oxygenase-1;
    LF,
    lipofectamine;
    FI,
    fluorescence intensity;
    nY,
    nitrotyrosine;
    NPCs,
    neuronal progenitor cells;
    RCR,
    respiratory control ratio;
    PINK1,
    PTEN-inducible kinase 1
    • © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society
    View Full Text

    Log in using your username and password

    Enter your Biochemical Journal username.
    Enter the password that accompanies your username.
    Forgot your user name or password?

    Log in through your institution

    You may be able to gain access using your login credentials for your institution. Contact your library if you do not have a username and password.
    If your organization uses OpenAthens, you can log in using your OpenAthens username and password. To check if your institution is supported, please see this list. Contact your library for more details.
    Previous ArticleNext Article
    Back to top

     

     

    December 2017

    Volume: 474 Issue: 23

    Biochemical Journal: 474 (23)
    • Table of Contents
    • About the Cover
    • Index by author

    Actions

    Email

    Thank you for your interest in spreading the word about Biochemical Journal.

    NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

    Enter multiple addresses on separate lines or separate them with commas.
    Lipid-based DNA/siRNA transfection agents disrupt neuronal bioenergetics and mitophagy
    (Your Name) has forwarded a page to you from Biochemical Journal
    (Your Name) thought you would like to see this page from the Biochemical Journal web site.
    Share
    Lipid-based DNA/siRNA transfection agents disrupt neuronal bioenergetics and mitophagy
    Eleonora Napoli, Siming Liu, Ilaria Marsilio, Konstantinos Zarbalis, Cecilia Giulivi
    Biochemical Journal Dec 2017, 474 (23) 3887-3902; DOI: 10.1042/BCJ20170632
    del.icio.us logo Digg logo Reddit logo Technorati logo Twitter logo CiteULike logo Facebook logo Mendeley logo
    Citation Tools
    Lipid-based DNA/siRNA transfection agents disrupt neuronal bioenergetics and mitophagy
    Eleonora Napoli, Siming Liu, Ilaria Marsilio, Konstantinos Zarbalis, Cecilia Giulivi
    Biochemical Journal Dec 2017, 474 (23) 3887-3902; DOI: 10.1042/BCJ20170632

    Citation Manager Formats

    • BibTeX
    • Bookends
    • EasyBib
    • EndNote (tagged)
    • EndNote 8 (xml)
    • Medlars
    • Mendeley
    • Papers
    • RefWorks Tagged
    • Ref Manager
    • RIS
    • Zotero
    Print
    Alerts

    Please log in to add an alert for this article.

    Request Permissions
    Save to my folders

    View Full PDF

     Open in Utopia Docs
    • Tweet Widget
    • Facebook Like

    Jump To

    • Article
      • Abstract
      • Introduction
      • Materials and methods
      • Results
      • Discussion
      • Author Contribution
      • Funding
      • Competing Interests
      • Acknowledgements
      • References
    • Figures
    • Info & Metrics
    • Supplementary Data
    • PDF

    Keywords

    autophagy
    mitophagy
    oxidative stress
    stress
    transfection agent

    Related Articles

    Cited By...

    • Portland Press Homepage
    • Publish With Us
    • Advertising
    • Technical Support
    • Biochemical Journal
    • Clinical Science
    • Essays in Biochemistry
    • Emerging Topics in Life Sciences
    • Biochemical Society Transactions
    • Neuronal Signaling
    • Bioscience Reports
    • Cell Signalling Biology
    • Biochemical Society Symposia

    Portland Press Limited
    Charles Darwin House
    12 Roger Street
    London WC1N 2JU
    Tel: +44(0) 20 7685 2410
    Fax: +44(0) 20 7685 2469
    Email: editorial@portlandpress.com

    The Biochemical Society