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Research Articles, Systems/Circuits

Single Nuclei Analyses Reveal Transcriptional Profiles and Marker Genes for Diverse Supraspinal Populations

Zachary Beine, Zimei Wang, Pantelis Tsoulfas and Murray G. Blackmore
Journal of Neuroscience 23 November 2022, 42 (47) 8780-8794; DOI: https://doi.org/10.1523/JNEUROSCI.1197-22.2022
Zachary Beine
1Department of Biomedical Sciences, Marquette University, Milwaukee, Wisconsin 53201
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Zimei Wang
1Department of Biomedical Sciences, Marquette University, Milwaukee, Wisconsin 53201
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Pantelis Tsoulfas
2Department of Neurological Surgery, Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, Florida 33136
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Murray G. Blackmore
1Department of Biomedical Sciences, Marquette University, Milwaukee, Wisconsin 53201
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Abstract

The mammalian brain contains numerous neurons distributed across forebrain, midbrain, and hindbrain that project axons to the lower spinal cord and work in concert to control movement and achieve homeostasis. Extensive work has mapped the anatomic location of supraspinal cell types and continues to establish specific physiological functions. The patterns of gene expression that typify and distinguish these disparate populations, however, are mostly unknown. Here, using adult mice of mixed sex, we combined retrograde labeling of supraspinal cell nuclei with fluorescence-activated nuclei sorting and single-nuclei RNA sequencing analyses to transcriptionally profile neurons that project axons from the brain to lumbar spinal cord. We identified 14 transcriptionally distinct cell types and used a combination of established and newly identified marker genes to assign an anatomic location to each. To validate the putative marker genes, we visualized selected transcripts and confirmed selective expression within lumbar-projecting neurons in discrete supraspinal regions. Finally, we illustrate the potential utility of these data by examining the expression of transcription factors that distinguish different supraspinal cell types and by surveying the expression of receptors for growth and guidance cues that may be present in the spinal cord. Collectively, these data establish transcriptional differences between anatomically defined supraspinal populations, identify a new set of marker genes of use in future experiments, and provide insight into potential differences in cellular and physiological activity across the supraspinal connectome.

SIGNIFICANCE STATEMENT The brain communicates with the body through a wide variety of neuronal populations with distinct functions and differential sensitivity to damage and disease. We have used single-nuclei RNA sequencing technology to distinguish patterns of gene expression within a diverse set of neurons that project axons from the mouse brain to the lumbar spinal cord. The results reveal transcriptional differences between populations previously defined on the basis of anatomy, provide new marker genes to facilitate rapid identification of cell type in future work, and suggest distinct responsiveness of different supraspinal populations to external growth and guidance cues.

  • AAV2-retro
  • growth factor receptor
  • guidance receptor
  • scRNA-seq
  • supraspinal neuron
  • transcription factor

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The Journal of Neuroscience: 42 (47)
Journal of Neuroscience
Vol. 42, Issue 47
23 Nov 2022
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Single Nuclei Analyses Reveal Transcriptional Profiles and Marker Genes for Diverse Supraspinal Populations
Zachary Beine, Zimei Wang, Pantelis Tsoulfas, Murray G. Blackmore
Journal of Neuroscience 23 November 2022, 42 (47) 8780-8794; DOI: 10.1523/JNEUROSCI.1197-22.2022

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Single Nuclei Analyses Reveal Transcriptional Profiles and Marker Genes for Diverse Supraspinal Populations
Zachary Beine, Zimei Wang, Pantelis Tsoulfas, Murray G. Blackmore
Journal of Neuroscience 23 November 2022, 42 (47) 8780-8794; DOI: 10.1523/JNEUROSCI.1197-22.2022
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Keywords

  • AAV2-retro
  • growth factor receptor
  • guidance receptor
  • scRNA-seq
  • supraspinal neuron
  • transcription factor

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