All optical control and monitoring of neural activity
All optical control and monitoring of neural activity
批准号:
8638689
负责人:
Samarendra Kumar Mohanty
金额:
$17.61万
依托单位国家:
美国
项目类别:
财政年份:
2013
资助国家:
美国
项目状态:
已结题
起止时间:
2013-09-15 至 2015-08-31
关键词:
Animal ModelCell Culture TechniquesCellsClinical TreatmentControlled StudyDetectionDevelopmentElectric StimulationElectrophysiology (science)ElectroporationEmbryoFrequenciesGanglion Cell LayerGene DeliveryGene TransferGenesGeneticGoalsHuman ActivitiesImageIn VitroInflammatory ResponseInterferometryInterventionLabelLasersLengthLightLocationMeasuresMethodsMolecularMonitorNeuronsOpsinOptical Coherence TomographyOpticsPatientsPatternPeripheralPhasePhotoreceptorsPhysiologic pulsePlaguePlasmidsProteinsRattusReactionResolutionRetinaRetinal Ganglion CellsRetinitis PigmentosaSignal TransductionSourceStagingSystemTechniquesTechnologyTissuesTransfectionViralViral VectorWidthbaseblindcell typehuman diseasein vivokidney cellminimally invasivenanonanometernoveloptogeneticspromoterpublic health relevancerelating to nervous systemsuccess
中文摘要
项目总结:光基因干预神经活动的成功需要优化
英文摘要
Project Summary: Success of optogenetic intervention of neural activity requires optimization of
delivery of genes encoding light sensitive proteins (opsins) to specific cells, and to record the
changes in cells and tissue during optogenetic stimulation. The most-commonly used method for
delivering opsin(s) is use of viral vector, which is prone to cause unexpected inflammatory
responses, immunological reactions, and improper gene integration. Further, the viral methods
limit the size of plasmid that can be packaged and delivered and therefore cannot carry multiple
opsin-encoding genes or large promoters. Further, in several cases of human diseases such as
retinitis pigmentosa (RP) where progressive loss of photoreceptors happens in peripheral retina, it
will be useful to localize the expression of the opsins not only in specific cell types, but in a
restricted spatial region (peripheral in RP). The first aim of the proposal is to optimize a non-viral
delivery method to mitigate the challenges posed by viral delivery. We have recently used focused
near-IR ultrafast laser beam method to deliver opsins (ChR2) into spatially-patterned regions of
neural tissue (retina). However, this technique needs to be optimized so as to minimize the
deleterious effects. Further, it will prove useful to develop a label-free optical technique (in contrast
to electrophysiology) to non-invasively evaluate functional activation of optogenetically-sensitized
neurons with high spatial resolution and large throughput. Recently, we demonstrated use of
Phase-Sensitive Frequency Domain Optical Coherence Tomography (PSFD-OCT) for detection of
fluctuations in optogenetically-stimulated cells. PSFD-OCT is a novel technique based on the
principles of low-coherence interferometry that can detect displacements of the order of tens of
picometers. Because of the low-coherence length of the light, the detected signal has to be within
the coherence length of the light source (~10¿m). This enables PSFD-OCT to investigate sub-
nanometer changes within a very small region of the tissue volume and is suitable for highly
localized detection. The overall aim of this study is to optimize optical delivery of gene encoding
opsins, and develop label-free non-invasive optical readout method based on PSFD-OCT to
monitor the changes in cortical neurons and tissue resulting from the activation.
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依托单位:
海外基金