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, Development/Plasticity/Repair

Characterization of Long Descending Premotor Propriospinal Neurons in the Spinal Cord

Yingchun Ni, Homaira Nawabi, Xuefeng Liu, Liu Yang, Kazunari Miyamichi, Andrea Tedeschi, Bengang Xu, Nicholas R. Wall, Edward M. Callaway and Zhigang He
Journal of Neuroscience 9 July 2014, 34 (28) 9404-9417; https://doi.org/10.1523/JNEUROSCI.1771-14.2014
Yingchun Ni
1F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Homaira Nawabi
1F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Xuefeng Liu
1F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Liu Yang
1F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kazunari Miyamichi
2Howard Hughes Medical Institute, Department of Biology, Stanford University, Stanford, California 94305, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Andrea Tedeschi
1F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Bengang Xu
1F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nicholas R. Wall
3Systems Neurobiology Laboratory, The Salk Institute for Biological Studies and Neurosciences Graduate Program, University of California, San Diego, La Jolla, California 92037
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Edward M. Callaway
3Systems Neurobiology Laboratory, The Salk Institute for Biological Studies and Neurosciences Graduate Program, University of California, San Diego, La Jolla, California 92037
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Edward M. Callaway
Zhigang He
1F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115,
  • 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.

    Trans-synaptic labeling of long descending premotor PPNs at the cervical and thoracic spinal levels. A, A schematic diagram of the experimental paradigm. A mixture of Rabies-GFP and AAV-G is injected into the hindlimb muscle TA of a P3 wild-type mouse. Coinfected motor neurons will produce rabies viral particles to be transported across synapses to their premotor interneurons in the lumbar spinal cord and PPNs in the upper spinal cord. B, An image of a transverse lumbar spinal section after injection of TA muscle with rabies virus and AAV-G shows infected motor neurons and local premotor interneurons. A coinfected motor neuron is highlighted in the boxed area. C–G, High-magnification images show the marked motor neuron in B-expressing rabies protein (rabies-GFP), ChAT, and rabies glycoprotein mRNA. The section is counterstained with nuclei marker DAPI. H, I, Transverse sections from thoracic (H) and cervical (I) levels show rabies-GFP expressing long descending PPNs. The inset images in H and I are close-up views of the PPNs marked in the boxed area. CC, Central canal; Ipsi, ipsilateral side; Contra, contralateral side. Scale bars: B, 100 μm; C–G, 10 μm; H, I, 200 μm.

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

    Distribution and morphology of the PPNs in the upper spinal cord for TA muscle. A, A photograph shows a spinal cord before (top) and after (bottom) clarification treatment. B, A projection image of stitched stacks of confocal images of intact spinal cord C1–T13 stained with GFP antibody shows the distribution of PPNs. Two areas in the spinal cord are highlighted by asterisks. C, D, High-magnification images of the two areas indicated by asterisks in B. E, A dot map illustrates the longitudinal distribution of PPNs. Each PPN from an individual sample is represented by a single colored dot. The middle line of the spinal cord is marked by a horizontal dotted line in the spinal cord diagram. Ipsi, Ipsilateral side. F, G, Dot maps show transverse distribution patterns of cervical (F) and thoracic (G) PPNs. Each color represents one sample. H, Correlation analysis of the four samples with Pearson's coefficient shows the degree of consistency in the distribution patterns among the four samples: 0 represents zero degree of correlation, and 1 represents full degree of correlation. T1–T4 represent the four distribution patterns at the thoracic level, and C1–C4 represents the four distribution patterns at the cervical level. I, Bar graphs show the percentage of PPNs on ipsilateral (Ipsi) and contralateral (Contra) sides of the spinal cord (Ipsi, 75 ± 1%; Contra, 25 ± 1%; n = 3). J, Bar graphs show the percentage of PPNs at the cervical and thoracic spinal levels (cervical, 3 ± 0.8%; thoracic = 97 ± 0.8%; n = 3). K, Bar graphs compare the average diameters of PPN cell bodies at the cervical and thoracic levels (cervical, 28 ± 0.75 μm, n = 36; thoracic, 20 ± 0.17 μm, n = 337). Scale bars: B, 500 μm; C, D, 30 μm. *p < 0.05, Student's t test.

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

    Sensory synaptic inputs to PPNs. A, An image of a wild-type mouse spinal section immunostained with antibodies against parvalbumin and VGluT1 labeling sensory fibers and their axonal terminals. The section is counterstained with nuclei marker DAPI. B, A diagram of sensory axons and their terminals in the spinal cord. The large and small boxed areas mark the location of the regions shown in A and C, respectively. C, Immunostaining of a thoracic spinal section with antibodies against GFP (rabies-GFP), parvalbumin, and VGluT1 together with DAPI labeling. Sensory axonal terminals (purple) are colabeled with both parvalbumin and VGluT1. The PPN marked with an arrowhead does not have any synaptic connection with sensory fibers, while the other PPN marked with an arrow is contacted by several sensory boutons, which is further highlighted in the inset. D, The dot map represents the distribution pattern of thoracic PPNs contacted (red squares) and not contacted (black squares) by sensory axons. Each square represents one neuronal cell body. E–G, Distribution of sensory axonal projections originating from the lumbar level in the thoracic spinal cord. E, A thoracic spinal section immunostained with antibodies against GFP and parvalbumin to colabel lumbar sensory axons. The section is counterstained with DAPI. F, High-magnification image of the boxed area in E with GFP, parvalbumin, and DAPI staining shows no GFP-positive sensory axons. G, High-magnification image of the same area in F with only GFP and DAPI channels further identifies the lack of GFP-positive sensory projections. H–J, Distribution of sensory axonal projections originating from the lumbar level in the lumbar spinal cord. H, A lumbar spinal section shows sensory axonal projections. I, High-magnification image of the boxed area in H with GFP, parvalbumin, and DAPI staining shows GFP-positive and parvalbumin-positive sensory axons. J, High-magnification image of the same area in I with only GFP and DAPI channels further identifies the GFP-positive sensory projections in the area. CC, Central canal; Ipsi, ipsilateral side. Scale bars: A, 100 μm; C, 20 μm; E, 100 μm; F, G, I, J, 10 μm.

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

    CST presynaptic inputs to PPNs. A, A thoracic transverse section from an Emx-Cre × Rosa-STOP-tdTomato mouse immunostained with an antibody against tdTomato shows the CST main bundles and axonal ramifications. The section is counterstained with DAPI. B, A diagram of CST axonal bundles and axonal ramifications in the spinal cord. The boxed area represents the location of the region shown in A. C, Immunostaining of a thoracic spinal section from Emx-Cre × Rosa-STOP-tdTomato mouse with antibodies against GFP (rabies-GFP), tdTomato, and VGluT1 together with DAPI labeling. CST axonal terminals (purple) are colabeled with both tdTomato and VGluT1. The boxed area in the image highlights the CST synaptic contact with the PPN. The inset is a close-up view of the synaptic contact. D, A dot map represents the distribution of thoracic PPNs contacted (red squares) and not contacted (black squares) by CST axons. E, A cervical section from an Emx-Cre × Rosa-STOP-tdTomato mouse immunostained with antibodies against GFP (rabies-GFP), tdTomato, and VGluT1 shows a rabies-infected cervical PPN contacted by a CST bouton (tdTomato+ and VGluT1+). The section is counterstained with DAPI. A high-magnification image of the bouton is shown in the inset. F, The distribution of cervical PPNs with (red squares) or without (black squares) CST boutons in the cervical spinal cord. CC, Central canal; Ipsi, ipsilateral side. Scale bars: A, 50 μm; C, 20 μm; E, 15 μm.

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

    5-HT presynaptic inputs to PPNs. A, An immunostaining image of wild-type mouse spinal cord section with 5-HT antibody. B, A diagram of 5-HT axons in the spinal cord. C, Immunostaining of a thoracic spinal section of a wild-type mouse with antibodies against GFP (rabies-GFP), 5-HT, and NeuN together with DAPI labeling. The boxed area highlights the 5-HT contact with the PPN, and the inset is the close-up view of the bouton. D, A dot map represents the distribution patterns of thoracic PPNs contacted (red square) and not contacted (black square) by 5-HT axons. E, An image of a wild-type mouse cervical spinal section immunostained with antibodies against GFP (rabies-GFP) and 5-HT showing a rabies-infected cervical PPN contacted by two 5-HT boutons. A high-magnification image of the boutons is shown in the inset. The spinal section is counterstained with NeuN and DAPI. F, The distribution of cervical PPNs with (red squares) or without (black squares) 5-HT boutons in the cervical spinal cord. Scale bars: A, 100 μm; C, 20 μm; E, 15 μm.

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

    Summary of sensory, CST, and 5-HT synaptic contacts with PPNs. A, Bar graphs represent the average numbers of sensory, CST, or 5-HT synaptic boutons that each PPN receives. B, The spinal cord is divided into four regions based on the anatomy of a transverse section, as follows: (1) ipsilateral dorsal area; (2) contralateral dorsal area; (3) contralateral ventral area; and (4) ipsilateral ventral area. C, D, Bar graphs show the percentage of thoracic (C) and cervical (D) PPNs with CST (white bar) or 5-HT (gray bar) synaptic contacts in the four regions of the spinal cord. **p < 0.01, n = 10, one-way ANOVA followed by Fisher's LSD test.

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

    Excitatory and inhibitory synaptic inputs to thoracic and cervical PPNs. A, B, Images of thoracic (A) and cervical (B) spinal transverse sections show rabies-infected PPNs receiving VGluT2-positive excitatory synaptic boutons. NeuN and DAPI costaining are used to label neurons and nuclei. C, The distribution of thoracic (black squares) and cervical (white squares) PPNs with excitatory synaptic boutons in the spinal cord. D, E, Images of thoracic (D) and cervical (E) spinal transverse sections show rabies-infected PPNs receiving VGAT-positive inhibitory synapses. F, The distribution of thoracic (black squares) and cervical (white squares) PPNs with inhibitory synaptic boutons in the spinal cord. G, Bar graphs show the average number of excitatory and inhibitory synaptic boutons that each PPN receives. Scale bars: A, B, D, E, 20 μm.

  • Figure 8.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 8.

    Molecular characterizations of PPNs. A–D, A PPN with in situ hybridization detected VGluT2 mRNA in the cell body, and immunostained with antibodies against GFP (rabies-GFP) and NeuN. E–H, A PPN with in situ hybridization detected VIAAT mRNA in the cell body, and immunostained with antibodies against GFP (rabies-GFP) and NeuN. I, An image of a thoracic spinal section immunostained with rabies-GFP and ChAT antibodies. The central canal region is highlighted in the boxed area. J–L, High-magnification images of the area show two rabies-infected PPNs without ChAT expression. M, A calbindin-expressing PPN in the spinal cord section immunostained with rabies-GFP and calbindin antibodies. N–P, High-magnification images show the colocalization of rabies-GFP with calbindin in the identified PPN. Q, Bar graphs show the percentages of excitatory and inhibitory PPNs. R, A table illustrates the results of PPNs tested with individual neuronal markers. Scale bars: A–H, 20 μm; I, 100 μm; J–L, 20 μm; M, 150 μm; N–P, 10 μm.

  • Figure 9.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 9.

    Chx10-expressing neurons in the spinal cord. A, An image of a thoracic transverse section from a wild-type mouse immunostained with antibodies against Chx10 and NeuN together with DAPI labeling. The two Chx10-positive neurons highlighted in the boxed area are shown in the inset images. B, A diagram shows the location of Chx10-expressing neurons in the spinal cord. The boxed area shows the location of the region in A. C–E, Images of immunostaining of a thoracic spinal section from wild-type mice with antibodies against GFP (rabies-GFP) and Chx10 show a rabies-infected PPN expressing Chx10 (arrow). F, Dot maps represent the distribution patterns of PPNs with (green squares) or without (black squares) Chx10 expression at the thoracic spinal level. G, Bar graphs show the percentage of Chx10-positive thoracic PPNs among the total population of PPNs in their respective area. CC, Central canal; Ipsi, ipsilateral side. Scale bars: A, 100 μm; C–E, 20 μm.

Back to top

In this issue

The Journal of Neuroscience: 34 (28)
Journal of Neuroscience
Vol. 34, Issue 28
9 Jul 2014
  • 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.
Characterization of Long Descending Premotor Propriospinal Neurons in the Spinal Cord
(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
Characterization of Long Descending Premotor Propriospinal Neurons in the Spinal Cord
Yingchun Ni, Homaira Nawabi, Xuefeng Liu, Liu Yang, Kazunari Miyamichi, Andrea Tedeschi, Bengang Xu, Nicholas R. Wall, Edward M. Callaway, Zhigang He
Journal of Neuroscience 9 July 2014, 34 (28) 9404-9417; DOI: 10.1523/JNEUROSCI.1771-14.2014

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
Characterization of Long Descending Premotor Propriospinal Neurons in the Spinal Cord
Yingchun Ni, Homaira Nawabi, Xuefeng Liu, Liu Yang, Kazunari Miyamichi, Andrea Tedeschi, Bengang Xu, Nicholas R. Wall, Edward M. Callaway, Zhigang He
Journal of Neuroscience 9 July 2014, 34 (28) 9404-9417; DOI: 10.1523/JNEUROSCI.1771-14.2014
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

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

Development/Plasticity/Repair

  • Change of spiny neuron structure in the basal ganglia song circuit and its regulation by miR-9 during song development
  • Stereotyped Spatiotemporal Dynamics of Spontaneous Activity in Visual Cortex Prior to Eye Opening
  • The epigenetic reader PHF23 is required for embryonic neurogenesis
Show more Development/Plasticity/Repair
  • 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.