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Cellular and Molecular Atlas of the Macaque Dorsal Horn

Arokiaraj, Cynthia Mary (2022) Cellular and Molecular Atlas of the Macaque Dorsal Horn. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

The spinal cord dorsal horn occupies a key position in the central nervous system, as it is the first site of integration of somatosensory input from the periphery. Local interneurons of the dorsal horn process the incoming peripheral information while projection neurons relay it to supraspinal regions via ascending pathways. However, the exact mechanisms through which this occurs in normal or maladaptive states, and the neuron populations involved, remains to be understood. Identification of the neuronal cell types present in the dorsal horn will enable us to determine their functional organization. Thus, a goal of this thesis is to achieve a consensus in the cell types and their classification.
To do this, we performed single-nucleus RNA-sequencing (snRNA-seq) of the Rhesus macaque dorsal horn. Mapping out the cellular and molecular organization of the non-human primate allows us to develop a reliable species-independent classification scheme for the cell types that can then facilitate translation of rodent dorsal horn studies to primate. From the snRNA-seq, we identified 11 excitatory neuronal clusters (GLUTs) and 5 inhibitory clusters (GABAs). Based on the cytoarchitecture of the spinal cord, layers or laminae have been described along the dorso-ventral axis, and these appear to correspond well with the functional organization of neuron populations. In situ hybridization revealed that a majority of the clusters reside within specific lamina(e). A comparative analysis between clusters of the macaque and mouse meta-analysis showed a strong correspondence between both species as well as several interesting species-specific differences at the gene level.
An additional goal of this thesis is to describe a strategy for the identification of regulatory elements in transcriptionally active regions of individual dorsal horn cell types using single nucleus Assay for Transposase-Accessible Chromatin coupled with sequencing (snATAC-seq). We found that the snATAC-seq derived clusters correspond well with our snRNA-seq clusters i.e., all 16 cell-types identified from the snRNA-seq have good representation in the scATAC-seq data. These data further validate our proposed classification scheme of dorsal horn cell types while also providing candidate regulatory regions that drive expression in specific cell types.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Arokiaraj, Cynthia Marycma83@pitt.educma83
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorSeal, Rebecca Prpseal@pitt.edurpseal
Committee ChairTaylor, Bradley Kbkt@pitt.edubkt
Committee MemberStauffer, Williamwrs@pitt.eduwrs
Committee ChairPfenning, Andreas Rapfenning@cmu.edu
Committee MemberAbraira, Victoriavictoria.abraira@rutgers.edu
Date: 1 January 2022
Date Type: Publication
Defense Date: 15 October 2021
Approval Date: 1 January 2022
Submission Date: 21 November 2021
Access Restriction: 1 year -- Restrict access to University of Pittsburgh for a period of 1 year.
Number of Pages: 185
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Neurobiology
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: Transcriptome, macaque, spinal cord, dorsal horn
Date Deposited: 01 Jan 2022 19:51
Last Modified: 01 Jan 2022 19:51
URI: http://d-scholarship.pitt.edu/id/eprint/41966

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