Characterizing the neuronal expression of neuronal PAS-domain containing protein 4 and aryl hydrocarbon receptor nuclear translocator 2 in multiple sclerosis related inflammatory settings in vitro
department or school: Medicine, Faculty of
department or school: Pathology and Laboratory Medicine, Department of
data provider: DSpace
degree campus: UBCV
creator: Abdul-Rahman, Raneen
issued: 2024-11-20T23:28:30Z
issued: 2024
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Multiple sclerosis (MS) is characterized by inflammatory demyelination and axonal loss, which contribute to irreversible clinical disability. Both hallmarks affect calcium homeostasis, partly due to mediators released by inflammatory cells, like glutamate and hydrogen peroxide (H₂O₂). Nuclear calcium transients can activate neuroprotective factors, including neuronal PAS-domain containing protein 4 (NPAS4), a neuronal activity-dependent gene. NPAS4 and aryl hydrocarbon receptor nuclear translocator 2 (ARNT2) work independently or in combination to regulate genes responsible for neuronal survival. We sought to determine the effects of soluble inflammatory mediators and inflammatory cells relevant to MS pathogenesis on NPAS4 and ARNT2 expression. Murine neuron-enriched cortical cultures were treated with increasing concentrations of glutamate (1-30µM) and H₂O₂ (12.5-100µM) or co-cultured in pilot experiments with resting or activated CD4⁺ T-cells. NPAS4 and ARNT2 protein expression were characterized via immunocytochemistry by examining the neuronal nuclear mean fluorescent intensity (MFI), the percentage of positive cells, and staining intensity changes using fluorescence microscopy across time and concentrations. In untreated conditions, NPAS4 was detected in less than 10% of neurons, while greater than 70% of neurons expressed low-medium ARNT2 levels. Glutamate increased the NPAS4 MFI and percentage of NPAS4-positive neurons, with increased staining intensity observed early at all doses. ARNT2 staining levels were similar across the glutamate doses and times. Prolonged glutamate exposure (10-30µM) was associated with morphological changes and cell loss. H₂O₂ (50-100µM) increased the NPAS4 MFI and percentage of NPAS4-expressing cells at later time points (5hrs and greater), with a simultaneous decrease and loss of ARNT2 expression. Exposure to H₂O₂ at 50-100µM tended to impact neuronal cell viability with significant cell loss by 9 hours compared to controls. Lastly…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/89692
note: Characterizing the Neuronal Expression of Neuronal PAS-Domain Containing Protein 4 and Aryl Hydrocarbon Receptor Nuclear Translocator 2 in Multiple Sclerosis Related Inflammatory Settings In Vitro by Raneen Abdul-Rahman A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in The Faculty of Graduate and Postdoctoral Studies (Pathology and Laboratory Medicine) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) November 2024 © Raneen Abdul-Rahman, 2024 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the thesis entitled: Characterizing the Neuronal Expression of Neuronal PAS-Domain Containing Protein 4 and Aryl Hydrocarbon Receptor Nuclear Translocator 2 in Multiple Sclerosis Related Inflammatory Settings In Vitro Submitted by Raneen Abdul-Rahman in partial fulfilment of the requirements for the degree of Master of Science in Pathology and Laboratory Medicine Examining Committee: Jacqueline Quandt, Assistant Professor, Pathology and Laboratory Medicine, UBC Supervisor Wayne Moore, Clinical Professor, Pathology and Laboratory Medicine, UBC Supervisory Committee Member Kevin Bennewith, Professor, Pathology and Laboratory Medicine, UBC Additional Examiner Clare Beasley, Associate Professor, Psychiatry, UBC Additional Examiner Additional Supervisory Committee Members: Shernaz Bamji, Professor, Cellular and Physiological Sciences, UBC Supervisory Committee Member Yu Tian Wang, Professor, Medicine, UBC Supervisory Committee Member iii Abstract Multiple sclerosis (MS) is characterized by inflammatory demyelination and axonal loss, which contribute to irreversible clinical disability. Both hallmarks affect calcium homeostasis, partly due to mediators released by inflammatory cells, like glutamate and hydrogen peroxide (H2O2). Nuclear calcium transients can activate neuroprotective factors, including neuronal PAS-domain containing protein 4 (NPAS4)…
has type: Thesis/Dissertation
date issued: 2025-05
is shown at: 10.14288/1.0447298
language: eng
degree discipline: Pathology and Laboratory Medicine
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
contributor: Quandt, Jacqueline
type: Text
identifier URI: http://hdl.handle.net/2429/89692