Transcriptional profiling at whole population and single cell levels reveals somatosensory neuron molecular diversity
- Boston Children's Hospital, United States
- Harvard Medical School, United States
- 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.
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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.
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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Transcriptional profiling at whole population and single cell levels reveals somatosensory neuron molecular diversity
eLife 3:e04660.
https://doi.org/10.7554/eLife.04660
