Deep sequencing of Phox2a nuclei reveals five classes of anterolateral system neurons

TY - JOUR

T1 - Deep sequencing of Phox2a nuclei reveals five classes of anterolateral system neurons

AU - Bell, Andrew M

AU - Utting, Charlotte

AU - Dickie, Allen C

AU - Kucharczyk, Mateusz W

AU - Quillet, Raphaëlle

AU - Gutierrez-Mecinas, Maria

AU - Razlan, Aimi N B

AU - Cooper, Andrew H

AU - Lan, Yuxuan

AU - Hachisuka, Junichi

AU - Weir, Greg A

AU - Bannister, Kirsty

AU - Watanabe, Masahiko

AU - Kania, Artur

AU - Hoon, Mark A

AU - Macaulay, Iain C

AU - Denk, Franziska

AU - Todd, Andrew J

N1 - Publisher Copyright: Copyright © 2024 the Author(s).

PY - 2024/6/4

Y1 - 2024/6/4

N2 - 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.

AB - 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.

KW - Animals

KW - Mice

KW - Homeodomain Proteins/genetics

KW - Spinal Cord/cytology

KW - Neurons/metabolism

KW - High-Throughput Nucleotide Sequencing

KW - Male

KW - Cell Nucleus/metabolism

KW - Transcription Factors/genetics

UR - http://www.scopus.com/inward/record.url?scp=85194866153&partnerID=8YFLogxK

U2 - 10.1073/pnas.2314213121

DO - 10.1073/pnas.2314213121

M3 - Article

C2 - 38805282

SN - 0027-8424

VL - 121

SP - e2314213121

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 23

M1 - e2314213121

ER -