Transcriptional profiling at whole population and single cell levels reveals somatosensory neuron molecular diversity

  1. Isaac M Chiu  Is a corresponding author
  2. Lee B Barrett
  3. Erika K Williams
  4. David E Strochlic
  5. Seungkyu Lee
  6. Andy D Weyer
  7. Shan Lou
  8. Gregory Bryman
  9. David P Roberson
  10. Nader Ghasemlou
  11. Cara Piccoli
  12. Ezgi Ahat
  13. Victor Wang
  14. Enrique J Cobos
  15. Cheryl L Stucky
  16. Qiufu Ma
  17. Stephen D Liberles
  18. Clifford Woolf
  1. Boston Children's Hospital, United States
  2. Harvard Medical School, United States
  3. Medical College of Wisconsin, United States

Abstract

The somatosensory nervous system is critical for the organism's ability to respond to mechanical, thermal, and nociceptive stimuli. Somatosensory neurons are functionally and anatomically diverse but their molecular profiles are not well-defined. Here, we used transcriptional profiling to analyze the detailed molecular signatures of dorsal root ganglion (DRG) sensory neurons. We used two mouse reporter lines and surface IB4 labeling to purify three major non-overlapping classes of neurons: 1)IB4+SNS-Cre/TdTomato+, 2)IB4-SNS-Cre/TdTomato+, and 3)Parv-Cre/TdTomato+ cells, encompassing the majority of nociceptive, pruriceptive, and proprioceptive neurons. These neurons displayed distinct expression patterns of ion channels, transcription factors, and GPCRs. Highly parallel qRT-PCR analysis of 334 single neurons selected by membership of the three populations demonstrated further diversity, with unbiased clustering analysis identifying six distinct subgroups. These data significantly increase our knowledge of the molecular identities of known DRG populations and uncover potentially novel subsets, revealing the complexity and diversity of those neurons underlying somatosensation.

Article and author information

Author details

  1. Isaac M Chiu

    F.M Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    For correspondence
    isaac_chiu@hms.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
  2. Lee B Barrett

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Erika K Williams

    Department of Cell Biology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. David E Strochlic

    Department of Cell Biology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Seungkyu Lee

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Andy D Weyer

    Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Shan Lou

    Dana Farber Cancer Institute, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Gregory Bryman

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. David P Roberson

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Nader Ghasemlou

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Cara Piccoli

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Ezgi Ahat

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Victor Wang

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. Enrique J Cobos

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  15. Cheryl L Stucky

    Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Qiufu Ma

    Dana Farber Cancer Institute, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  17. Stephen D Liberles

    Department of Cell Biology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  18. Clifford Woolf

    F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.

Ethics

Animal experimentation: All animal experiments were conducted according to institutional animal care and safety guidelines at Boston Children's Hospital, in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All animal work was conducted under strict review and guidelines according to the Institutional Animal Care and Use Committee (IACUC) at Boston Children's Hospital, which meets the veterinary standards set by the American Association for Laboratory Animal Science (AALAS). The experiments were reviewed and approved by the IACUC at Boston Children's Hospital under animal protocol number 13-01-2342R.

Reviewing Editor

  1. Jeremy Nathans, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, United States

Publication history

  1. Received: October 2, 2014
  2. Accepted: December 18, 2014
  3. Accepted Manuscript published: December 19, 2014 (version 1)
  4. Version of Record published: January 14, 2015 (version 2)

Copyright

© 2014, Chiu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Isaac M Chiu
  2. Lee B Barrett
  3. Erika K Williams
  4. David E Strochlic
  5. Seungkyu Lee
  6. Andy D Weyer
  7. Shan Lou
  8. Gregory Bryman
  9. David P Roberson
  10. Nader Ghasemlou
  11. Cara Piccoli
  12. Ezgi Ahat
  13. Victor Wang
  14. Enrique J Cobos
  15. Cheryl L Stucky
  16. Qiufu Ma
  17. Stephen D Liberles
  18. Clifford Woolf
(2014)
Transcriptional profiling at whole population and single cell levels reveals somatosensory neuron molecular diversity
eLife 3:e04660.
https://doi.org/10.7554/eLife.04660

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