Skip to main content

Main menu

  • HOME
  • CONTENT
    • Early Release
    • Featured
    • Current Issue
    • Issue Archive
    • Collections
    • Podcast
  • ALERTS
  • FOR AUTHORS
    • Information for Authors
    • Fees
    • Journal Clubs
    • eLetters
    • Submit
  • EDITORIAL BOARD
  • ABOUT
    • Overview
    • Advertise
    • For the Media
    • Rights and Permissions
    • Privacy Policy
    • Feedback
  • SUBSCRIBE

User menu

  • Log in
  • My Cart

Search

  • Advanced search
Journal of Neuroscience
  • Log in
  • My Cart
Journal of Neuroscience

Advanced Search

Submit a Manuscript
  • HOME
  • CONTENT
    • Early Release
    • Featured
    • Current Issue
    • Issue Archive
    • Collections
    • Podcast
  • ALERTS
  • FOR AUTHORS
    • Information for Authors
    • Fees
    • Journal Clubs
    • eLetters
    • Submit
  • EDITORIAL BOARD
  • ABOUT
    • Overview
    • Advertise
    • For the Media
    • Rights and Permissions
    • Privacy Policy
    • Feedback
  • SUBSCRIBE
PreviousNext
Brief Communications

Microglial CD33-Related Siglec-E Inhibits Neurotoxicity by Preventing the Phagocytosis-Associated Oxidative Burst

Janine Claude, Bettina Linnartz-Gerlach, Alexei P. Kudin, Wolfram S. Kunz and Harald Neumann
Journal of Neuroscience 13 November 2013, 33 (46) 18270-18276; DOI: https://doi.org/10.1523/JNEUROSCI.2211-13.2013
Janine Claude
1Neural Regeneration Group, Institute of Reconstructive Neurobiology and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Bettina Linnartz-Gerlach
1Neural Regeneration Group, Institute of Reconstructive Neurobiology and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alexei P. Kudin
2Division of Neurochemistry, Department of Epileptology, University of Bonn Medical Center, 53127 Bonn, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Wolfram S. Kunz
2Division of Neurochemistry, Department of Epileptology, University of Bonn Medical Center, 53127 Bonn, Germany
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Harald Neumann
1Neural Regeneration Group, Institute of Reconstructive Neurobiology and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Article Figures & Data

Figures

  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Detection of Siglec-E on microglia. A, Detection of Siglece mRNA in primary microglia and the microglia line. Spleen tissue served as positive control. Siglece transcripts were detected in unstimulated microglia (unstim.) as well as in microglia stimulated with LPS, IFN-γ, IFN-α, and TNF-α. No Siglece transcripts were detected in primary neurons. Gapdh mRNA served as housekeeping standard. Representative data of three independent experiments are shown. Control, Water control. B, Flow cytometry analysis of the microglial line. Siglec-E was detected on unstimulated (unstim.) microglia at low levels. Treatment with interferons (IFN-γ, IFN-α) slightly increased expression of Siglec-E, whereas treatment with LPS or TNF-α had no effect. Representative data of three independent experiments are shown. Isotype, Isotype control antibody. C, Flow cytometry analysis of ex vivo and primary microglia. Low constitutive expression of Siglec-E on CD11b+ and CD45low cells was detected. Representative data of three independent experiments are shown. Isotype, Isotype control antibody. D, Microglia were transduced with lentiviral vectors expressing Siglece (SigE vector) or a control vector. Furthermore, lentiviral knockdown was performed by two lentiviral short-hairpin constructs targeting Siglece (shRNASigE1; shRNASigE2) or a corresponding nontargeting control vector (NTshRNA). qRT-PCR confirmed the successful modification of the microglial line by showing an increased (left graph) or decreased (right graph) Siglece cDNA, respectively. **p ≤ 0.01, ***p ≤ 0.001. E, Flow cytometry analysis confirmed overexpression (left graph) and reduced expression (right graph) of the Siglec-E protein. Representative data of three independent experiments are shown.

  • Figure 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2.

    Siglec-E prevents phagocytosis and the associated reactive oxygen burst after challenge with neural debris. A, Uptake of red fluorescent-labeled neural debris into the microglial line was determined by confocal microscopy and 3D reconstruction. Microglial cells were transduced with the control vector, the Siglece overexpressing vector (SigE vector), the Siglece knockdown vectors (shRNASigE1, shRNASigE2) or the nontargeting vector (NTshRNA). Representative images of three independent experiments are shown. Scale bar, 20 μm. B, Phagocytosis of neural debris was quantified. Overexpression of Siglece mRNA reduced the uptake of neural material, whereas knockdown of Siglece increased the uptake of neural debris. ***p ≤ 0.001. C, Level of superoxide production as determined by DHE staining was quantified in the microglial line. After stimulation with neural debris, DHE intensity was increased after Siglece knockdown compared with the NTshRNA. ***p ≤ 0.001. D, Quantification of superoxide production as determined by DHE staining. After stimulation with neural debris in the presence of either 20 μg/ml SOD1 or 40 nm Trolox, increased DHE intensity after Siglece knockdown was antagonized. ***p ≤ 0.001. E, Quantification of superoxide production of microglial cells using the Amplex Red method. Knockdown of microglial Siglece via shRNASigE1 or shRNASigE2 increased the endogenous production of H2O2 equivalents after stimulation with neural debris compared with cells transduced with a control construct (NTshRNA). Arrow 1, Addition of cells; arrow 2, addition of 12,000 U/ml catalase. Representative data of three independent experiments are shown.

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3.

    Siglec-E has anti-inflammatory effects and binds to neurons and astrocytes. A, qRT-PCR to detect Il-1β, Tnfsf2 (TNF-α), and Nos2 cDNA after 16 h incubation of microglia with neural debris. Microglia with knockdown of Siglece (shRNASigE1, shRNASigE2) showed a significant increase in gene transcription of Il-1β and Tnfsf2 (TNF-α) in the presence of neural debris compared with the control vector (NTshRNA). *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001; n.s., not significant. B, C, Binding of Siglec-E to sialic acid residues of neural cells. Neurons/astrocytes were either untreated or treated with sialidase and then incubated with the Siglec-E:Fc fusion protein. Removal of sialic acids led to a decreased binding of Siglec-E:Fc to neurons (B) and astrocytes (C). Representative images of three independent experiments are shown. Scale bar, 30 μm.

  • Figure 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4.

    Neurite protective effect of Siglec-E in neuron–microglia cocultures. A, Loss of neurites after knockdown of Siglece in microglia. Neurons were cocultured for 48 h with microglia. Siglece overexpression increased whereas knockdown of Siglece decreased the relative neurite length. Representative images without sialidase treatment of three independent experiments are shown. Scale bar, 30 μm. B, Relative neurite length and neuronal cell body density were quantified. Although the neuronal cell body density was unchanged (right graph), the relative neurite length was dependent on the expression level of Siglec-E on the microglial surface (left graph). After sialidase treatment, the relative neurite length was significantly reduced. **p ≤ 0.01, ***p ≤ 0.001. C, Neurons were cocultured with microglia in the presence of Trolox. Reduced neurite length after knockdown of Siglec-E was restored after treatment with 40 nm Trolox. ***p ≤ 0.001.

Back to top

In this issue

The Journal of Neuroscience: 33 (46)
Journal of Neuroscience
Vol. 33, Issue 46
13 Nov 2013
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by author
  • Advertising (PDF)
  • Ed Board (PDF)
Email

Thank you for sharing this Journal of Neuroscience article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Microglial CD33-Related Siglec-E Inhibits Neurotoxicity by Preventing the Phagocytosis-Associated Oxidative Burst
(Your Name) has forwarded a page to you from Journal of Neuroscience
(Your Name) thought you would be interested in this article in Journal of Neuroscience.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Print
View Full Page PDF
Citation Tools
Microglial CD33-Related Siglec-E Inhibits Neurotoxicity by Preventing the Phagocytosis-Associated Oxidative Burst
Janine Claude, Bettina Linnartz-Gerlach, Alexei P. Kudin, Wolfram S. Kunz, Harald Neumann
Journal of Neuroscience 13 November 2013, 33 (46) 18270-18276; DOI: 10.1523/JNEUROSCI.2211-13.2013

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Respond to this article
Request Permissions
Share
Microglial CD33-Related Siglec-E Inhibits Neurotoxicity by Preventing the Phagocytosis-Associated Oxidative Burst
Janine Claude, Bettina Linnartz-Gerlach, Alexei P. Kudin, Wolfram S. Kunz, Harald Neumann
Journal of Neuroscience 13 November 2013, 33 (46) 18270-18276; DOI: 10.1523/JNEUROSCI.2211-13.2013
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Responses to this article

Respond to this article

Jump to comment:

No eLetters have been published for this article.

Related Articles

Cited By...

More in this TOC Section

  • Heteromodal Cortical Areas Encode Sensory-Motor Features of Word Meaning
  • Pharmacologically Counteracting a Phenotypic Difference in Cerebellar GABAA Receptor Response to Alcohol Prevents Excessive Alcohol Consumption in a High Alcohol-Consuming Rodent Genotype
  • Neuromuscular NMDA Receptors Modulate Developmental Synapse Elimination
Show more Brief Communications
  • Home
  • Alerts
  • Visit Society for Neuroscience on Facebook
  • Follow Society for Neuroscience on Twitter
  • Follow Society for Neuroscience on LinkedIn
  • Visit Society for Neuroscience on Youtube
  • Follow our RSS feeds

Content

  • Early Release
  • Current Issue
  • Issue Archive
  • Collections

Information

  • For Authors
  • For Advertisers
  • For the Media
  • For Subscribers

About

  • About the Journal
  • Editorial Board
  • Privacy Policy
  • Contact
(JNeurosci logo)
(SfN logo)

Copyright © 2023 by the Society for Neuroscience.
JNeurosci Online ISSN: 1529-2401

The ideas and opinions expressed in JNeurosci do not necessarily reflect those of SfN or the JNeurosci Editorial Board. Publication of an advertisement or other product mention in JNeurosci should not be construed as an endorsement of the manufacturer’s claims. SfN does not assume any responsibility for any injury and/or damage to persons or property arising from or related to any use of any material contained in JNeurosci.