{"id":{"repo_id":"missouri","oai_identifier":"oai:mospace.umsystem.edu:10355/5039"},"canonical_url":"https://search.dev.ndltd.org/etd/missouri/oai:mospace.umsystem.edu:10355/5039","repository":{"repo_id":"missouri","name":"University of Missouri","base_url":"https://mospace.umsystem.edu/oai/request"},"display":{"title":"Development of a flexible image-based approach for studying signal transduction in isolated arterioles","abstract":"Myogenic constriction in arterioles occurs when membrane tension leads to smooth muscle depolarization and voltage-gated calcium channel activation. A barrier to understanding the exact signaling events is the limited ability of glass electrodes to follow temporal changes in membrane potential (Em). This study aimed to develop an imaging station to measure Em, [Ca²+]i and diameter in isolated arterioles. A Forster Resonance Enery Transfer (FRET) approach was proposed for Em meaurement due to enhanced sensitivity and temporal responsiveness. FRET indicators used were CC2-DMPE (Coumarin-labelled Phospholipid, Donor, ex: 405nm) and DisBAC₄(3) (Oxonol, acceptor). For measurement of [Ca²+]i Fluo-4 AM (ex 488nm) was used. Initial studies examined toxicity of the multiple fluorophores. Isolated cells remained viable after loading with the fluorophores. Arterioles maintained myogenic, and agonist, responsiveness pre and post dye loading suggesting a lack of toxicity. Spectral characteristics of the fluorophores used in the project required specific optical filter units and development of a. custom built optical system. Using the system, FRET was demonstrated, as was the ability to detect changes in fluorescence signal to known depolarizing stimuli. The developed system is now ready for translation to the study of myogenic signaling in intact and functional arterioles.","abstract_html":"Myogenic constriction in arterioles occurs when membrane tension leads to smooth muscle depolarization and voltage-gated calcium channel activation. A barrier to understanding the exact signaling events is the limited ability of glass electrodes to follow temporal changes in membrane potential (Em). This study aimed to develop an imaging station to measure Em, [Ca²+]i and diameter in isolated arterioles. A Forster Resonance Enery Transfer (FRET) approach was proposed for Em meaurement due to enhanced sensitivity and temporal responsiveness. FRET indicators used were CC2-DMPE (Coumarin-labelled Phospholipid, Donor, ex: 405nm) and DisBAC₄(3) (Oxonol, acceptor). For measurement of [Ca²+]i Fluo-4 AM (ex 488nm) was used. Initial studies examined toxicity of the multiple fluorophores. Isolated cells remained viable after loading with the fluorophores. Arterioles maintained myogenic, and agonist, responsiveness pre and post dye loading suggesting a lack of toxicity. Spectral characteristics of the fluorophores used in the project required specific optical filter units and development of a. custom built optical system. Using the system, FRET was demonstrated, as was the ability to detect changes in fluorescence signal to known depolarizing stimuli. The developed system is now ready for translation to the study of myogenic signaling in intact and functional arterioles.","abstract_has_math":false,"creators":["Ella, Srikanth Reddy"],"institution":"University of Missouri--Columbia","degree_name":"M.S.","degree_level":"Masters","degree_discipline":"Biological engineering (MU)","degree_department":null,"school":null,"contributors":[],"advisors":["Hill, Michael A."],"committee_chairs":[],"committee_members":[],"year":2007,"date_issued":"2007","date_published":"2007","updated_at":"2026-07-24T03:08:43Z","subjects":[],"languages":["eng","English"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier.doi","label":"DOI","values":["https://doi.org/10.32469/10355/5039"],"render_values":[{"text":"https://doi.org/10.32469/10355/5039","href":"https://doi.org/10.32469/10355/5039","code":true}]}]},"links":{"outbound_url":"https://hdl.handle.net/10355/5039","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Hill, Michael A."]},{"key":"dc:creator","label":"Author","values":["Ella, Srikanth Reddy"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2010-01-12T19:08:52Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2010-01-12T19:08:52Z"]},{"key":"dc:date.issued","label":"Date","values":["2007"]},{"key":"dc:publisher","label":"Institution","values":["University of Missouri--Columbia"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biological engineering (MU)"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Masters"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M.S."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Missouri--Columbia"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["English"]},{"key":"dc:language.iso","label":"Language (ISO)","values":["eng"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.doi","label":"DOI","values":["https://doi.org/10.32469/10355/5039"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/10355/5039"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file.","Title from title screen of research.pdf file (viewed on May 11, 2009)","Includes bibliographical references.","Thesis (M.S.) 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Isolated cells remained viable after loading with the fluorophores. Arterioles maintained myogenic, and agonist, responsiveness pre and post dye loading suggesting a lack of toxicity. Spectral characteristics of the fluorophores used in the project required specific optical filter units and development of a. custom built optical system. Using the system, FRET was demonstrated, as was the ability to detect changes in fluorescence signal to known depolarizing stimuli. The developed system is now ready for translation to the study of myogenic signaling in intact and functional arterioles."]},{"key":"dc:source","label":"Dc Source","values":["Submitted by University of Missouri--Columbia Graduate School."]},{"key":"dc:title","label":"Title","values":["Development of a flexible image-based approach for studying signal transduction in isolated arterioles"]}]}],"canonical_facts":{"dc:contributor.advisor":["Hill, Michael A."],"dc:creator":["Ella, Srikanth Reddy"],"dc:date.accessioned":["2010-01-12T19:08:52Z"],"dc:date.available":["2010-01-12T19:08:52Z"],"dc:date.issued":["2007"],"dc:description":["The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file.","Title from title screen of research.pdf file (viewed on May 11, 2009)","Includes bibliographical references.","Thesis (M.S.) 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Isolated cells remained viable after loading with the fluorophores. Arterioles maintained myogenic, and agonist, responsiveness pre and post dye loading suggesting a lack of toxicity. Spectral characteristics of the fluorophores used in the project required specific optical filter units and development of a. custom built optical system. Using the system, FRET was demonstrated, as was the ability to detect changes in fluorescence signal to known depolarizing stimuli. 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