Understanding the role of Basigin in regulating Integrin activation at glial focal adhesion complexes in the peripheral nerves of Drosophila melanogaster
department or school: Science, Faculty of
department or school: Zoology, Department of
data provider: DSpace
degree campus: UBCV
creator: Roth, Sophie Faith
issued: 2024-03-19T08:05:50Z
issued: 2024
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: In the nervous system, glial cells play an important role in ensheathing and protecting nerves from damage, however, the employed mechanisms to properly ensheath nerves are still unknown. In Drosophila melanogaster, the perineurial glia (PG) are a conserved class of glia that surround peripheral nerves and ensure proper ensheathment through interactions with the extracellular matrix (ECM). Glia-ECM interactions occur through focal adhesions (FAs) comprised of integrins and intracellular proteins including talin and vinculin. Loss of FAs leads to the loss of the perineurial glial sheath and disruption of nervous system function. Integrins also associate with the transmembrane protein Basigin (aka Neuroplastin) in the PG. Loss of Basigin leads to constriction across nerves causing deformation of the glial membrane, cytoskeleton and the overlying ECM. The loss of Basigin phenotype can be rescued by a loss of integrin, leading to the hypothesis that Basigin knockdown in the PG leads to increased integrin activation. We performed FLIM-FRET (Fluorescence Lifetime Imaging Microscopy - Förster Resonance Energy Transfer) using paired donor-acceptor sensors inserted in talin that vary in sensitivity to force to quantify changes in integrin activation. Compared to controls, we observed a significant decrease in the FLIM-FRET efficiency in Basigin knockdown animals. This result implies that the sensor is being stretched due to the lengthening of talin, likely because of increased integrin activation. However, the excitation lasers for both the donor and acceptor were turned on when collecting FLIM-FRET images, which led to a false positive FRET signal with donor-only controls and prompted the reinterpretation of results. We also observed a significant increase in the integrated density of vinculin in the PG when Basigin is reduced. Since vinculin binds and stabilizes talin in an extended state, an increase in vinculin recruitment suggests an increase in talin lengthening and th…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/87597
note: UNDERSTANDING THE ROLE OF BASIGIN IN REGULATING INTEGRIN ACTIVATION AT GLIAL FOCAL ADHESION COMPLEXES IN THE PERIPHERAL NERVES OF DROSOPHILA MELANOGASTER by Sophie Faith Roth B.Sc., Wilfrid Laurier University, 2021 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Zoology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) March 2024 © Sophie Faith Roth, 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: Understanding the role of Basigin in regulating integrin activation at glial focal adhesion complexes in the peripheral nerves of Drosophila melanogaster submitted by Sophie Faith Roth in partial fulfilment of the requirements for the degree of Master of Science in Zoology Examining Committee: Dr. Vanessa Auld, Professor, Zoology, UBC Supervisor Dr. Calvin Roskelley, Professor, Cellular & Physiological Sciences, UBC Supervisory Committee Member Dr. Ninan Abraham, Professor, Zoology, UBC Additional examiner Additional Supervisory Committee Members: Dr. Kenji Sugioka, Assistant Professor, Zoology, UBC Supervisory Committee Member iii Abstract In the nervous system, glial cells play an important role in ensheathing and protecting nerves from damage, however, the employed mechanisms to properly ensheath nerves are still unknown. In Drosophila melanogaster, the perineurial glia (PG) are a conserved class of glia that surround peripheral nerves and ensure proper ensheathment through interactions with the extracellular matrix (ECM). Glia-ECM interactions occur through focal adhesions (FAs) comprised of integrins and intracellular proteins including talin and vinculin. Loss of FAs leads to the loss of the perineurial glial sheath and disruption of nervous system function. Integrins also associate with the transmembrane protein Basigin (aka Neuroplastin) in the PG. L…
has type: Thesis/Dissertation
date issued: 2024-05
is shown at: 10.14288/1.0440705
language: eng
degree discipline: Zoology
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: Auld, Vanessa
type: Text
identifier URI: http://hdl.handle.net/2429/87597