Characterization of the mechanisms underlying Alzheimer's disease and multiple sclerosis
department or school: Medicine, Faculty of
data provider: DSpace
degree campus: UBCV
creator: Herculano, Bruno de Araujo
issued: 2019-04-30T15:32:43Z
issued: 2019
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: Deposition of the amyloid β protein (Aβ) into neuritic plaques is the neuropathological hallmark of Alzheimer's Disease (AD). Aβ is generated through the cleavage of the Amyloid Precursor Protein (APP) by β-secretase and γ-secretase. Currently, the evaluation of APP cleavage by β-secretase in experimental settings has largely depended on models that do not replicate the physiological conditions of this process. We have developed a chimeric protein construct, ASGβ, incorporating the β-site cleavage sequence of APP targeted by β-secretase and its intracellular trafficking signal into a Phosphatase-eGFP secreted reporter system. Upon cleavage by β-secretase, ASGβ releases a phosphatase-containing portion that can be measured in the culture medium, and an intracellular fraction that can be detected through western blot. Subsequently, we have generated a cell line stably expressing ASGβ that can be utilized to assay β-secretase in real time. Our findings suggest this system could be a high-throughput tool to screen compounds that aim to modulate β-secretase activity and Aβ production under physiological conditions, as well as evaluating factors that regulate this cleavage. Multiple sclerosis (MS) is an inflammatory disease of the central nervous system characterized by myelin loss and neuronal dysfunction. Although the majority of patients do not present familial aggregation, Mendelian forms have been described and more recently we have identified novel mutations in 2 genes, NLRP12 and NCOA3 in familial forms of the disease. Our findings show that one of the discovered mutations on NLRP12 affects the maturation of Caspase-1, and that the mutation discovered on NCOA3 leads to a higher basal level of inducible Nitric Oxide Synthase (iNOS). These findings show novel mechanisms causing the disruption of immune and inflammatory responses that could in turn lead to the symptoms observed in MS patients. Our study suggests that further characterization of the pathways affected b…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/70000
note: CHARACTERIZATION OF THE MECHANISMS UNDERLYING ALZHEIMER’S DISEASE AND MULTIPLE SCLEROSIS by Bruno de Araujo Herculano B.Sc. Biological Sciences, Universidade Federal de Santa Catarina, 2009 M.Sc. Integrated Biosciences, The University of Tokyo, 2011 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Neuroscience) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) April 2019 © Bruno de Araujo Herculano, 2019 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the dissertation entitled: Characterization of the mechanisms underlying Alzheimer's Disease and Multiple Sclerosis submitted by Bruno de Araujo Herculano in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Neuroscience Examining Committee: Dr. Weihong Song Supervisor Dr. Jason Snyder Supervisory Committee Member Supervisory Committee Member Dr. Yu Tian Wang University Examiner Dr. Honglin Luo University Examiner Additional Supervisory Committee Members: Dr. Lynn Raymond Supervisory Committee Member Dr. Shernaz Bamji Supervisory Committee Member iii Abstract Deposition of the amyloid β protein (Aβ) into neuritic plaques is the neuropathological hallmark of Alzheimer's Disease (AD). Aβ is generated through the cleavage of the Amyloid Precursor Protein (APP) by β-secretase and γ-secretase. Currently, the evaluation of APP cleavage by β-secretase in experimental settings has largely depended on models that do not replicate the physiological conditions of this process. We have developed a chimeric protein construct, ASGβ, incorporating the β-site cleavage sequence of APP targeted by β-secretase and its intracellular trafficking signal into a Phosphatase-eGFP secreted reporter system. Upon cleavage by β-secretase, ASGβ releases a phosphatase-containing portion that can be measured in the culture mediu…
has type: Thesis/Dissertation
date issued: 2019-09
is shown at: 10.14288/1.0378524
language: eng
degree discipline: Neuroscience
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
type: Text
identifier URI: http://hdl.handle.net/2429/70000