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ToolsBell, Andrew M., Utting, Charlotte, Dickie, Allen C., Kucharczyk, Mateusz W., Quillet, Raphaëlle, Gutierrez-Mecinas, Maria, Razlan, Aimi N. B., Cooper, Andrew H., Lan, Yuxuan, Hachisuka, Junichi, Weir, Greg A., Bannister, Kirsty, Watanabe, Masahiko, Kania, Artur, Hoon, Mark A., Macaulay, Iain C., Denk, Franziska and Todd, Andrew J. (2024) Deep sequencing of Phox2a nuclei reveals five classes of anterolateral system neurons. Proceedings of the National Academy of Sciences of the United States of America, 121 (23). ISSN 0027-8424
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Abstract
The anterolateral system (ALS) is a major ascending pathway from the spinal cord that projects to multiple brain areas and underlies the perception of pain, itch, and skin temperature. Despite its importance, our understanding of this system has been hampered by the considerable functional and molecular diversity of its constituent cells. Here, we use fluorescence-activated cell sorting to isolate ALS neurons belonging to the Phox2a-lineage for single-nucleus RNA sequencing. We reveal five distinct clusters of ALS neurons (ALS1-5) and document their laminar distribution in the spinal cord using in situ hybridization. We identify three clusters of neurons located predominantly in laminae I–III of the dorsal horn (ALS1-3) and two clusters with cell bodies located in deeper laminae (ALS4 and ALS5). Our findings reveal the transcriptional logic that underlies ALS neuronal diversity in the adult mouse and uncover the molecular identity of two previously identified classes of projection neurons. We also show that these molecular signatures can be used to target groups of ALS neurons using retrograde viral tracing. Overall, our findings provide a valuable resource for studying somatosensory biology and targeting subclasses of ALS neurons.
| Item Type: | Article |
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| Additional Information: | Data, Materials, and Software Availability: Sequencing data are available to browse via the Broad Institute single-cell portal (63). Raw sequencing data, aligned counts, and the Seurat object are available to download from the Gene Expression Omnibus under accession number GSE240528 (64). This study did not generate new unique reagents or software. Funding Information: This research was funded in whole, or in part, by the Wellcome Trust (Grant numbers 219433/Z/19/Z and 204820/Z/16/Z), the Medical Research Council (Grant numbers MR/T01072X/1, MR/V033638/1, MR/ W004739/1, and MR/W002426/1), the Biotechnology and Biological Sciences Research Council (BB/S017178/1), the Academy of Medical Sciences (Grant number SGL025\\1079), and the Medical Research Foundation (MRF-160-0015-ELP-DENK-C0844). We acknowledge support from the Biotechnology and Biological Sciences Research Council (BBSRC), part of UK Research and Innovation, Core Capability Grant BB/CCG1720/1, and the National Capability Grant BBS/E/ T/000PR9816. This work was supported by the intramural research program of the NIDCR, NIH, project ZIADE000721-22. We acknowledge the assistance of Diane Vaughan, School of Infection and Immunity Flow Cytometry Facility at the University of Glasgow.We are grateful to Brian Roome and Sara Villa-Hernandez for helpful discussion, to Oscar Marin Parra and Eleanor Paul for sharing CckCre and SstCre mice, to Ariel Levine for sharing data, and to Robert Kerr and Iain Plenderleith for expert technical assistance. Rights Retention Statement: For the purpose of Open Access, the authors have applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. |
| Uncontrolled Keywords: | als projection neuron,pain,spinal cord,temperature sensation,general ,/dk/atira/pure/subjectarea/asjc/1000 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 24 Oct 2024 16:30 |
| Last Modified: | 12 Nov 2024 13:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/97188 |
| DOI: | 10.1073/pnas.2314213121 |
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