{"id":{"repo_id":"utswmed","oai_identifier":"oai:utswmed-ir.tdl.org:2152.5/10606"},"canonical_url":"https://search.dev.ndltd.org/etd/utswmed/oai:utswmed-ir.tdl.org:2152.5/10606","repository":{"repo_id":"utswmed","name":"University of Texas Southwestern Medical Center","base_url":"https://utswmed-ir.tdl.org/server/oai/request"},"display":{"title":"Single Molecule-Based Computer Vision Analysis of Cortical Actin-Plasma Membrane Protein Interplay","abstract":"The file named "NGO-PRIMARY-2025.pdf" is the primary dissertation file. The file named "1745621082149-Supplementary Video.zip" is a zipped file folder that contains 10 movie files (MP4 format), and a description of each video is included in Appendix B of the primary file (pages 148-149).","abstract_html":"The file named "NGO-PRIMARY-2025.pdf" is the primary dissertation file. The file named "1745621082149-Supplementary Video.zip" is a zipped file folder that contains 10 movie files (MP4 format), and a description of each video is included in Appendix B of the primary file (pages 148-149).","abstract_has_math":false,"creators":["Ngo, Huong Tra"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Chen, Elizabeth","Jaqaman, Khuloud","Rajaram, Satwik","Cleaver, Ondine"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-06-03T19:50:05Z","date_published":"2025-06-03T19:50:05Z","updated_at":"2026-07-24T05:52:17Z","subjects":["Actin Cytoskeleton","Actins","Cell Membrane","Membrane Proteins"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["1522122299"],"render_values":[{"text":"1522122299","href":null,"code":true}]}]},"links":{"outbound_url":"https://hdl.handle.net/2152.5/10606","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Chen, Elizabeth","Jaqaman, Khuloud","Rajaram, Satwik","Cleaver, Ondine"]},{"key":"dc:creator","label":"Author","values":["Ngo, Huong Tra"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2025-06-03T19:50:05Z","2025-05","May 2025"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","text"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Actin Cytoskeleton","Actins","Cell Membrane","Membrane Proteins"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://hdl.handle.net/2152.5/10606","1522122299"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The file named "NGO-PRIMARY-2025.pdf" is the primary dissertation file. The file named "1745621082149-Supplementary Video.zip" is a zipped file folder that contains 10 movie files (MP4 format), and a description of each video is included in Appendix B of the primary file (pages 148-149).","The spatiotemporal organization of plasma membrane proteins is regulated by cortical actin, yet quantitatively resolving the interplay between plasma membrane proteins and the cortical actin dynamics remains challenging. To address this, I developed a multiscale imaging and quantitative computer vision tool that integrate Single-Molecule Imaging and Fluorescence Speckle Microscopy (SMI-FSM) to quantitatively analyze how cortical actin dynamics can influence plasma membrane organization in live endothelial cells. Application of this tool, complemented with other image-based analysis - including conditional colocalization analysis, showed that the dynamic organization of cell surface receptor CD36 varies across different membrane subregions and is modulated by cortical actin dynamics. Cortical actin dynamics were predictive of differences in plasma membrane mobility near cell edges versus closer to the cell center. In addition, we found that global changes in actin properties caused by perturbation were not necessarily reflected in the cortical actin properties near plasma membrane proteins, and that the changes in plasma membrane protein properties upon perturbation varied based on the local cortical actin environment. Because CD36 has short intracellular domains and no known molecular link to cortical actin, we also identified the β1-integrin subunit and several tetraspanins - CD9, CD81, and CD151, as part of the previously unknown link between CD36 and cortical actin. We found that CD36 is recruited into complexes/nanodomains containing these proteins. The point mutation G12V altered CD36's interactions with integrins and tetraspanins, changing its membrane organization, weakening its relationship to actin dynamics, and abolishing CD36 downstream signaling in response to thrombospondin-1. These findings suggest that integrins and tetraspanins may also mediate the coupling between many other plasma membrane constituents and cortical actin. Here, we highlight the strength of imaging-based studies in that they reveal subcellular spatial information that is difficult to obtain otherwise. Given the widespread use of single molecular imaging as a method to study the spatiotemporal organization of plasma membrane proteins and the versatility of SMI-FSM, we expect it to be widely applicable to enable future investigation of the influence of cortical actin architecture and dynamics on different plasma membrane proteins, especially in the context of actin-dependent cellular processes."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf","application/zip"]},{"key":"dc:title","label":"Title","values":["Single Molecule-Based Computer Vision Analysis of Cortical Actin-Plasma Membrane Protein Interplay"]}]}],"canonical_facts":{"dc:contributor":["Chen, Elizabeth","Jaqaman, Khuloud","Rajaram, Satwik","Cleaver, Ondine"],"dc:creator":["Ngo, Huong Tra"],"dc:date":["2025-06-03T19:50:05Z","2025-05","May 2025"],"dc:description":["The file named "NGO-PRIMARY-2025.pdf" is the primary dissertation file. The file named "1745621082149-Supplementary Video.zip" is a zipped file folder that contains 10 movie files (MP4 format), and a description of each video is included in Appendix B of the primary file (pages 148-149).","The spatiotemporal organization of plasma membrane proteins is regulated by cortical actin, yet quantitatively resolving the interplay between plasma membrane proteins and the cortical actin dynamics remains challenging. To address this, I developed a multiscale imaging and quantitative computer vision tool that integrate Single-Molecule Imaging and Fluorescence Speckle Microscopy (SMI-FSM) to quantitatively analyze how cortical actin dynamics can influence plasma membrane organization in live endothelial cells. Application of this tool, complemented with other image-based analysis - including conditional colocalization analysis, showed that the dynamic organization of cell surface receptor CD36 varies across different membrane subregions and is modulated by cortical actin dynamics. Cortical actin dynamics were predictive of differences in plasma membrane mobility near cell edges versus closer to the cell center. In addition, we found that global changes in actin properties caused by perturbation were not necessarily reflected in the cortical actin properties near plasma membrane proteins, and that the changes in plasma membrane protein properties upon perturbation varied based on the local cortical actin environment. Because CD36 has short intracellular domains and no known molecular link to cortical actin, we also identified the β1-integrin subunit and several tetraspanins - CD9, CD81, and CD151, as part of the previously unknown link between CD36 and cortical actin. We found that CD36 is recruited into complexes/nanodomains containing these proteins. The point mutation G12V altered CD36's interactions with integrins and tetraspanins, changing its membrane organization, weakening its relationship to actin dynamics, and abolishing CD36 downstream signaling in response to thrombospondin-1. These findings suggest that integrins and tetraspanins may also mediate the coupling between many other plasma membrane constituents and cortical actin. Here, we highlight the strength of imaging-based studies in that they reveal subcellular spatial information that is difficult to obtain otherwise. Given the widespread use of single molecular imaging as a method to study the spatiotemporal organization of plasma membrane proteins and the versatility of SMI-FSM, we expect it to be widely applicable to enable future investigation of the influence of cortical actin architecture and dynamics on different plasma membrane proteins, especially in the context of actin-dependent cellular processes."],"dc:format":["application/pdf","application/zip"],"dc:identifier":["https://hdl.handle.net/2152.5/10606","1522122299"],"dc:language":["en"],"dc:subject":["Actin Cytoskeleton","Actins","Cell Membrane","Membrane Proteins"],"dc:title":["Single Molecule-Based Computer Vision Analysis of Cortical Actin-Plasma Membrane Protein Interplay"],"dc:type":["Thesis","text"]},"updated_at":"2026-07-24T05:52:17Z"}