Novel TIRF microscopy analyzing trafficking & signaling at the cell cortex
新型 TIRF 显微镜分析贩运
基本信息
- 批准号:7432044
- 负责人:
- 金额:$ 248.13万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2007
- 资助国家:美国
- 起止时间:2007-09-30 至 2012-08-31
- 项目状态:已结题
- 来源:
- 关键词:1-Phosphatidylinositol 3-KinaseAbbreviationsAccountingAcousticsAddressAdipocytesAffectAlgorithmsAreaArtsAttenuatedAutomobile DrivingAwardBackBindingBiochemicalBiochemistryBiologicalBiologyBoxingBuffersCaliberCalibrationCell LineCell membraneCell surfaceCellsCellular biologyClathrinCluster AnalysisCollaborationsCollectionCollimatorColorComaCommunitiesComplexComputational BiologyComputer Vision SystemsComputersConflict (Psychology)Confocal MicroscopyCoupledCouplingCuesCytoskeletonDataData SetDefectDepthDevelopmentDiabetes MellitusDiffusionDimensionsDimerizationDisadvantagedDisciplineDockingDown-RegulationDropsDyesEmployee StrikesEndocytosisEngineeringEnsureEnvironmentEventExocytosisEyeFaceFeedbackFiberFigs - dietaryFlareFluorescein-5-isothiocyanateFluorescenceFluorescence MicroscopyFluorescence Recovery After PhotobleachingFluorescent DyesFunctional disorderFundingGenetic ScreeningGermanyGlassGlucose TransporterGlycerolGoalsGrantGreen Fluorescent ProteinsImageImage AnalysisImageryIncidenceInsulinInterdisciplinary StudyInvestmentsJointsKineticsKnowledgeLabelLaboratoriesLasersLearningLegal patentLengthLifeLightLightingLinkLipidsLocalizedLocationMacromolecular ComplexesMalignant NeoplasmsMapsMasksMeasuresMediatingMembraneMembrane MicrodomainsMembrane Protein TrafficMethodologyMethodsMicroscopeMicroscopyMicrotubulesModelingMolecularMonitorMorphologic artifactsMotivationMotorNatureNon-Insulin-Dependent Diabetes MellitusOpticsOrganellesPTEN genePaperParasitesPathway interactionsPenetrationPerformancePersonal SatisfactionPhosphatidylinositolsPhosphoinositide-3-Kinase, Catalytic, Gamma PolypeptidePhosphotransferasesPhotobleachingPhysiologic pulsePlanet MarsPlayPliabilityPositioning AttributePostdoctoral FellowPrivate SectorProbabilityProcessProteinsPublicationsPulse takingPupilQuantum DotsRNA InterferenceRadialRandomizedRangeReagentRecruitment ActivityRefractive IndicesRegulationRelative (related person)ReporterResearchResearch InfrastructureResearch PersonnelResolutionRiskRoleSamplingScanningScienceScientistSeminalSeriesSideSignal PathwaySignal TransductionSilicon DioxideSimulateSiteSmall Interfering RNASolidSolutionsSorting - Cell MovementSourceSpainSpatial DistributionSpecific qualifier valueSpecimenSpeedSpottingsStandards of Weights and MeasuresStructureSupport of ResearchSurfaceSystemTechniquesTechnologyTestingTextThickTimeTotal Internal Reflection FluorescentTouch sensationTrainingTransfectionTubulinVesicleVisualWolvesWorkanalytical methodbasal insulinbaseblood glucose regulationcell cortexcell motilitycell typecellular imagingconceptdaydensitydesigndesireextracellularflotillinfluorescence imagingfluorescence microscopefluorophorehandbookhuman wyatt proteinimage processingimprovedinnovationinsightinstrumentinstrumentationinsulin signalinginterestlensmedical schoolsmicromanipulatormigrationmillisecondmouse wyatt proteinnanometernovelobject shapephotoactivationprototyperadius bone structurereceptorresearch studyresponsescaffoldsimulationsingle moleculesuccesstooltraffickingtrans-Golgi Networktrenduser-friendlyvirtual
项目摘要
My major goal is to advance knowledge about events on or near the plasma membrane. This region
directly controls membrane traffic to and from the cell surface (exo- and endocytosis) and is where
extracellular signals are amplified and modulated by assembly of signaling scaffolds. The introduction
of total internal reflection fluorescence (TIRF) microscopy, a technique that allows unprecedented
axial resolution, has revolutionized studies of dynamic processes at the cell cortex. I propose 1) to
develop two highly innovative multi-angle TIRF microscopes and 2) to apply these instruments
towards the elucidation of mechanisms that regulate exo- and endocytosis. These microscopes will
allow the penetration depth of the light beam to be varied rapidly and avoid traditional imaging
artifacts. Together with new analytical methods, they will permit high-resolution 3D imaging of a ~50-
1000 nanometer cortical region of living cells. Additionally, a highly innovative FRAP implementation
will allow us to `pulse' photoactivate single vesicles and track their fate. I will use this novel
instrumentation to expand our ongoing studies on exo- and endocytic traffic. A main new goal will be
to elucidate mechanisms in the vesicular trafficking pathways that regulate levels of glucose
transporters (Glut4) at the cell surface, a process whose dysfunction leads to type 2 diabetes. I will
test the hypothesis that the exocyst complex participates in the spatial regulation of the insulin
responsiveness of Glut4 vesicle exocytosis. Using photoactivatable Glut4-Dendra I will determine
whether insulin signaling triggers a switch from lipid raft to clathrin-mediated endocytic pathways. To
address where PI3K signaling acts, I will implement inducible dimerization technology to rapidly turn
on/off PI(3,4,5)P3 at the plasma membrane. The innovative approaches of this proposal capitalize on
my unique expertise in interdisciplinary research spanning instrumentation, cell biology, and
quantitative biology and will fundamentally impact biology and a medically important field.
我的主要目标是推进有关质膜上或附近事件的知识。这一地区
直接控制进出细胞表面的膜运输(胞吞和胞吞),
细胞外信号通过信号支架的组装而被放大和调节。引入
全内反射荧光(TIRF)显微镜,一种允许前所未有的技术,
轴向分辨率,彻底改变了在细胞皮层的动态过程的研究。我建议1)
开发两种高度创新的多角度TIRF显微镜,2)应用这些仪器
对阐明机制,调节外吞和内吞。这些显微镜将
允许光束的穿透深度快速变化
藏物再加上新的分析方法,它们将允许对约50- 500微米的物体进行高分辨率3D成像。
1000纳米的活细胞皮层区域。此外,高度创新的FRAP实施
将允许我们“脉冲”光激活单个囊泡并跟踪它们的命运。我会用这本小说
仪器,以扩大我们正在进行的研究外吞和内吞交通。一个主要的新目标是
阐明调节葡萄糖水平的囊泡运输途径的机制
2型糖尿病是由细胞表面的谷氨酸转运蛋白(Glut 4)引起的,这一过程的功能障碍导致2型糖尿病。我会
检验外囊复合物参与胰岛素空间调节的假设
Glut 4囊泡胞吐反应。使用可光活化的Glut 4-Dendra I将确定
胰岛素信号传导是否触发从脂筏到网格蛋白介导的内吞途径的转换。到
为了解决PI 3 K信号传导作用的问题,我将实施诱导型二聚化技术,
在质膜上开/关PI(3,4,5)P3。本提案的创新方法利用了
我在跨学科研究方面的独特专业知识,涵盖仪器、细胞生物学和
定量生物学,并将从根本上影响生物学和医学重要领域。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(2)
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Derek K. Toomre其他文献
Mechanisms of vesicle docking and fusion pore expansion by calcium-sensor synaptotagmin-1
- DOI:
10.1016/j.bpj.2023.11.2328 - 发表时间:
2024-02-08 - 期刊:
- 影响因子:
- 作者:
Sudheer Kumar Cheppali;Maria Tsemperouli;David Chetrit;Felix Rivera-Molina;Derek K. Toomre;Erdem Karatekin - 通讯作者:
Erdem Karatekin
Derek K. Toomre的其他文献
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{{ truncateString('Derek K. Toomre', 18)}}的其他基金
New Toolkit for Imaging and Controlling Early Ciliogenesis
用于成像和控制早期纤毛发生的新工具包
- 批准号:
8621325 - 财政年份:2014
- 资助金额:
$ 248.13万 - 项目类别:
Super-Resolution Structured Illumination Microscope (SIM)
超分辨率结构照明显微镜 (SIM)
- 批准号:
7793345 - 财政年份:2010
- 资助金额:
$ 248.13万 - 项目类别:
Novel TIRF microscopy analyzing trafficking & signaling at the cell cortex
新型 TIRF 显微镜分析贩运
- 批准号:
7892704 - 财政年份:2009
- 资助金额:
$ 248.13万 - 项目类别:
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