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
    • Special Collections
  • EDITORIAL BOARD
    • Editorial Board
    • ECR Advisory Board
    • Journal Staff
  • ABOUT
    • Overview
    • Advertise
    • For the Media
    • Rights and Permissions
    • Privacy Policy
    • Feedback
    • Accessibility
  • SUBSCRIBE

User menu

  • Log out
  • Log in
  • My Cart

Search

  • Advanced search
Journal of Neuroscience
  • Log out
  • 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
    • Special Collections
  • EDITORIAL BOARD
    • Editorial Board
    • ECR Advisory Board
    • Journal Staff
  • ABOUT
    • Overview
    • Advertise
    • For the Media
    • Rights and Permissions
    • Privacy Policy
    • Feedback
    • Accessibility
  • SUBSCRIBE
PreviousNext
Articles, Neurobiology of Disease

Selective Motor Neuron Resistance and Recovery in a New Inducible Mouse Model of TDP-43 Proteinopathy

Krista J. Spiller, Claudia J. Cheung, Clark R. Restrepo, Linda K. Kwong, Anna M. Stieber, John Q. Trojanowski and Virginia M.-Y. Lee
Journal of Neuroscience 20 July 2016, 36 (29) 7707-7717; https://doi.org/10.1523/JNEUROSCI.1457-16.2016
Krista J. Spiller
Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Krista J. Spiller
Claudia J. Cheung
Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Claudia J. Cheung
Clark R. Restrepo
Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Clark R. Restrepo
Linda K. Kwong
Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Anna M. Stieber
Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
John Q. Trojanowski
Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for John Q. Trojanowski
Virginia M.-Y. Lee
Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Virginia M.-Y. Lee
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Article Figures & Data

Figures

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

    Selective MN death is observed in hypoglossal and spinal MNs despite uniformly high levels of hTDP-43 expression in all MN pools. A–C, Immunostaining for human hTDP-43 (green) and VAChT (MNs, red) in rNLS8 mice shows many MNs expressing high levels of hTDP-43 in oculomotor (A) and facial (B) nuclei, as well as in L4 SC (C) at 4 weeks off DOX. D, Almost all MNs of the fast-twitch TA pool (top) or slow-twitch Sol pool (bottom) identified by retrograde labeling with CTB-594 (red) are hTDP-43+ (green). E, There is no difference in hTDP-43 staining intensity (measured in arbitrary fluorescence units or a.u.) between these pools in the same mice at 1 week off DOX, 15–25 MNs per mouse from n = 3. t test, p = 0.97. F, After DOX removal, 65–80% of MNs in lumbar L4–L5 SC, the hypoglossal (XII), trigeminal (V), and facial (VII) nuclei, and ∼1/3 of those in the oculomotor nucleus (III) are hTDP-43+ in rNLS8 mice and these numbers remain stable over the disease course. Data are mean ± SD (n = 4 mice per MN pool). G, Eight weeks of cytoplasmic expression of hTDP-43 induces significant MN loss in lumbar L4–L5 SC and the hypoglossal nucleus (XII), but is not sufficient to induce MN loss in oculomotor, trochlear (III/IV), trigeminal (V), or facial (VII) nuclei in rNLS8 mice. Data are mean ± SD (n = 4 per motor pool), t test, **p < 0.01, ***p < 0.001. Scale bars, 100 μm.

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

    Within the generally vulnerable hypoglossal nucleus, certain MNs are more resistant to degeneration. A, Representative image of the hypoglossal nucleus from an rNLS8 mouse immunostained with VAChT (red) and hTDP-43 (green), with dorsal and ventral subdivisions separated by a dashed line. B, After 8 weeks of transgene expression, MNs are lost from both subdivisions, with a larger percentage lost from the ventral subdivision. Data are mean ± SEM, n = 3 animals, t test. C, D, Muscle denervation is observed in the tongue, but not the masseter muscles, of rNLS8 mice at late disease stages. Representative cryosections of muscles immunostained with VAChT (red) and labeled with α-Btx (green) to identify NMJs shows that, by 8 weeks off DOX, 22 ± 4% of NMJs of the tongue muscle are denervated (C), whereas the masseter muscle NMJs are still intact (D). **p < 0.01, ***p < 0.001. Scale bars, 100 μm.

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

    Fast MNs in the lumbar SC are those that are lost in rNLS8 mice over time. A–C, Immunostaining for VAChT (red) and MMP-9 (green) on SCs of rNLS8 mice 1 week (A), 4 weeks (B), and 8 weeks (C) off DOX shows a loss of fast (MMP-9+) MNs. D, At 1 week off DOX, 49 ± 2% of MNs at L4–L5 are MMP-9+. By 8 weeks off DOX, ∼44% of total MNs have been lost and only ∼11% of those remaining are MMP-9+, with no change in the number of MMP-9− MNs over time. E, MN size distributions of VAChT+ MNs show a decrease in size from 1 week off DOX (gray bars) to 6 weeks off DOX (black bars), consistent with a loss of the largest MNs. Data are mean ± SEM, n = 4 animals. F, Representative cryosection of an rNLS8 lumbar SC 1 week off DOX immunostained for VAChT (green) and calbindin (red). G, Average size of neurons containing calbindin in the superficial dorsal horn did not change between 1 week (gray bar) and 6 weeks off DOX (black bar), but there was a significant decrease in the average soma size of VAChT+ MNs in the ventral horn. This suggests that the reason for the decrease in average size of the MNs is that the largest cells have died, rather than that the tissue has shrunk. Data are mean ± SD (n = 3), ***p < 0.001, **p < 0.01.

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

    Axonal dieback is not uniform from MNs of vulnerable MN pools. A, Representative muscle cryosection from the TA of an end-stage rNLS8 mouse, with overlap of VAChT+ motor terminals (red) and acetylcholine receptors (BTX, green) as an indicator of innervated motor endplates. Denervated NMJs are marked with white asterisks. B, At the same time point, the slow Sol has far fewer denervated NMJs. Scale bar, 100 μm. C, Denervation of the Sol muscle (green) is later than for the fast TA (red) and LGC (black). Data are mean ± SEM, n = 4 per time point. D, At the electron microscopic level, NMJ structural differences are apparent between the Sol and TA muscle from an rNLS8 mouse after 6 weeks of transgene expression. Although the Sol still has a nerve ending (orange arrow) lying in the set of depressions on the muscle fiber surface (pink arrowhead) to form a NMJ, there is no nerve making contact with the motor terminal in the TA and there is evident collagen (marked with #) in the place of a nerve. Scale bar, 500 nm. E, F, Evoked CMAPs in the GC muscle after stimulation of the sciatic nerve in nTg and rNLS8 mice that are still on DOX and off DOX for 4 or 6 weeks. E, Individual traces showing the M-wave, the maximum peak-to-peak value of which is used to calculate the CMAP. F, CMAP measurements significantly decrease with hTDP-43 expression. Data are mean ± SD, n = 3–4 animals per genotype, *p < 0.05, **p < 0.01, ***p < 0.001.

  • Figure 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 5.

    The percentage of MNs in a given MN pool without nuclear TDP-43 expression increases with increasing vulnerability to degenerate. A–D, Immunostaining for total TDP-43 (mouse + human, red) and DAPI on cryosections of the hypoglossal nucleus (A, B) and lumbar SC (C, D) shows the primarily nuclear localization of TDP-43 in the wild-type animals and the primarily cytoplasmic localization and loss of nuclear TDP-43 in rNLS8 mice. Scale bar, 100 μm. E, Percentage of MNs with nuclear TDP-43 at 4 weeks off DOX varies significantly between motor pools, with the two most vulnerable groups, the hypoglossal and L4 MNs, having the fewest MNs with nuclear TDP-43. Data are mean ± SEM, n = 3–4 animals, one-way ANOVA. F, There is no difference in the level of nuclear TDP-43 (red) between vulnerable MMP-9 (green) MNs and resistant MMP-9− MNs at L4. G, Fluorescence intensity (in arbitrary units) of the TDP-43 signal in the nucleus does not differ between MMP-9+ (black bar) and MMP-9− MNs (gray bar). Data are mean ± SEM of 280–300 MNs from 3 animals, p = 0.77.

  • Figure 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 6.

    Resistant FR and S MNs reinnervate muscle and underlie motor recovery after hTDP-43ΔNLS suppression. A, Schematic of experimental timeline. B–D, Transverse cryosections of TA from a nTg mouse (B), an rNLS8 mouse after 6 weeks of transgene expression (C), and an rNLS8 mouse after 12 weeks of transgene suppression (D) immunostained with MHCIIA to label muscle fibers innervated by FR MNs. MHCIIA+ fibers are grouped after recovery, suggesting reinnervation by FR MNs after transgene suppression. Scale bars, 100 μm. E, Average fiber diameter is significantly decreased in the TA muscle after 6 weeks of transgene expression, reflecting muscle atrophy. Atrophy is reversed after 12 weeks of transgene suppression, when the average TA fiber diameter increases beyond nTg controls. Data are mean ± SD from n = 3–4 muscles per treatment, F(2, 8) = 67.8, p < 0.001. F, After 6 weeks, but not 1 week, of hTDP-43 expression, the largest TA-innervating FF MNs are lost and the remaining smaller FR/S MNs sprout to innervate unoccupied motor endplates during recovery from disease. Data are mean ± SEM from n = 3 animals, 20–30 MNs measured from each.

  • Figure 7.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 7.

    Remaining MNs in the lumbar SC change phenotype after hTDP-43 clearance. A, Immunostaining for MMP-9 (green), VAChT (to label MNs, red), and DAPI on cryosections of the lumbar SC of an rNLS8 mouse after only 2 weeks of transgene suppression showing a return of MMP-9 expression, which is lost during the disease course (Fig. 3A–C). Scale bar, 100 μm. B, Average (Avg) MMP-9 perikaryal MN expression at the L4–L5 level did not differ between the crushed side ventral horn and the control (noncrushed) side. Data are mean ± SD, n = 3, p = 0.49. C, Despite previous axonal dieback and lumbar FF MN loss, evoked CMAPs return to predenervation maxima after 8 weeks of transgene suppression, a result of muscle reinnervation and a phenotypic switch of former FR/S MNs. Data are mean ± SD, n = 3–6. D, Timeline of MMP-9 (red) and hTDP-43 (green) expression in rNLS8 mice shown as percentage of the total (ttl) MNs. Data are mean ± SD, n = 2–7 animals per time point, *p < 0.05, **p < 0.01, ***p < 0.001.

Tables

  • Figures
    • View popup
    Table 1.

    Comparative MN vulnerability to degenerate: rNLS8 mouse model versus human

    Motor pool/subtyperNLS8 miceAffected in ALS patients?
    % MNs hTDP-43+ (mean ± SD)a% lost by 8 weeks off DOX (mean ± SD)% MNs w/o nuclear TDP-43 (mean ± SD)b
    Oculomotor34.7 ± 4.7None7.3 ± 4Spared (Okamoto et al., 1993; Kanning et al., 2010)
    Trigeminal64.6 ± 11.1None8.0 ± 2Mild degeneration (Iwanaga et al., 1997)
    Facial71 ± 4.3None18.7 ± 4Mild to moderate degeneration (Iwanaga et al., 1997)
    Hypoglossal76 ± 6.132.2 ± 5.724.7 ± 2∼46% lost in ALS (Kiernan and Hudson, 1991); affected first in bulbar presentation
    Lumbar MNs80.3 ± 3.349.5 ± 0.346 ± 3.2∼50% lost in ALS (Kiernan and Hudson, 1991); affected early in the majority of patients
    TA-innervating MNs100 [48, n = 3]Not measuredNot measuredAffected early in disease (Dengler et al., 1990; Sharma et al., 1995; Kanning et al., 2010)
    Sol-innervating MNs100 [23, n = 3]Not measuredNot measuredSlow motor units are preferentially spared (Patten et al., 1979)
    • ↵aNumbers from rNLS8 mice at 1 week off DOX.

    • ↵bNumbers from rNLS8 mice at 4 weeks off DOX.

Back to top

In this issue

The Journal of Neuroscience: 36 (29)
Journal of Neuroscience
Vol. 36, Issue 29
20 Jul 2016
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by author
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.
Selective Motor Neuron Resistance and Recovery in a New Inducible Mouse Model of TDP-43 Proteinopathy
(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
Selective Motor Neuron Resistance and Recovery in a New Inducible Mouse Model of TDP-43 Proteinopathy
Krista J. Spiller, Claudia J. Cheung, Clark R. Restrepo, Linda K. Kwong, Anna M. Stieber, John Q. Trojanowski, Virginia M.-Y. Lee
Journal of Neuroscience 20 July 2016, 36 (29) 7707-7717; DOI: 10.1523/JNEUROSCI.1457-16.2016

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
Selective Motor Neuron Resistance and Recovery in a New Inducible Mouse Model of TDP-43 Proteinopathy
Krista J. Spiller, Claudia J. Cheung, Clark R. Restrepo, Linda K. Kwong, Anna M. Stieber, John Q. Trojanowski, Virginia M.-Y. Lee
Journal of Neuroscience 20 July 2016, 36 (29) 7707-7717; DOI: 10.1523/JNEUROSCI.1457-16.2016
Twitter logo Facebook 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

Keywords

  • amyotrophic lateral sclerosis
  • motor neuron
  • reinnervation
  • rNLS mice
  • selective vulnerability
  • TDP-43

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

Articles

  • Memory Retrieval Has a Dynamic Influence on the Maintenance Mechanisms That Are Sensitive to ζ-Inhibitory Peptide (ZIP)
  • Neurophysiological Evidence for a Cortical Contribution to the Wakefulness-Related Drive to Breathe Explaining Hypocapnia-Resistant Ventilation in Humans
  • Monomeric Alpha-Synuclein Exerts a Physiological Role on Brain ATP Synthase
Show more Articles

Neurobiology of Disease

  • Targeting Lysine α-Ketoglutarate Reductase to Treat Pyridoxine-Dependent Epilepsy
  • The Role of Neprilysin and Insulin-Degrading Enzyme in the Etiology of Sporadic Alzheimer's Disease
  • Positron emission tomography (PET) neuroimaging of the Pink1-/- rat Parkinson disease model with the norepinephrine transporter (NET) ligand [18F]NS12137
Show more Neurobiology of Disease
  • Home
  • Alerts
  • Follow SFN on BlueSky
  • 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 Notice
  • Contact
  • Accessibility
(JNeurosci logo)
(SfN logo)

Copyright © 2025 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.