Optimization of CaMPARI for large-scale, cellular-resolution activity recording in freely-moving mice
CaMPARI 的优化,用于自由移动小鼠的大规模细胞分辨率活动记录
基本信息
- 批准号:10472700
- 负责人:
- 金额:$ 64.81万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-09-01 至 2025-12-31
- 项目状态:未结题
- 来源:
- 关键词:AffinityAreaBehaviorBehavior assessmentBenchmarkingBindingBiological AssayBrainBrain regionCalciumCalcium ionCellsCharacteristicsColorDataDetectionDevelopmentDevicesDorsalEnsureEnvironmentEquipmentFluorescenceFutureGenesGenetic IdentityGoalsHeadHippocampusImmunohistochemistryImplantIn VitroLabelLibrariesLightLightingLinkMapsMeasurementMethodsMicroscopeMicroscopyMonitorMovementMusMutateMutationNeuronsNoiseOpticsPathologyPatternPerformancePopulationPositioning AttributeProcessPropertyProtein EngineeringProteinsResearchResolutionSamplingSignal TransductionStep TestsSurfaceSystemTechniquesTechnologyTestingTissuesVariantViolaViral VectorVisual Cortexadvanced systemage relatedbehavior testbehavioral studycell typecognitive testingcortex mappingdetection sensitivityexperienceexperimental studyflexibilityimaging systemimplantable deviceimprovedin vitro Assayin vitro testingin vivoin vivo evaluationinterestmillimeterminiaturizeneural circuitnovel strategiesoptical imagingoptogeneticspredictive testprotein expressionprotein purificationresponsesample fixationscreeningsensorsingle cell sequencingthree photon microscopytranslational neurosciencetwo-photon
项目摘要
Project Summary
The goal of this proposed research is to optimize a dual-use calcium ion sensor for recording single-cell activity
from the entire dorsal cortex or hippocampus of freely-moving mice. It is widely accepted that optical imaging
with genetically-encoded fluorescent calcium sensors is currently the only method to obtain measurements of
genetically-identified neuronal populations with dense sampling. Over the past several years, the recording
capabilities of two-photon microscopes have been improved to record from millimeter-scale tissue, and new
miniaturized microscopes have been developed to record from moving mice. However, most behaviors arise
from collective interactions between neurons from multiple brain areas, which cannot be simultaneously
monitored with these systems. Therefore, there is a clear need to develop a new approach to directly monitor
the synchronized activity of distributed neural circuits. CaMPARI is a unique calcium sensor that can detect
activity in two calcium-dependent ways: 1) permanent color change (green to red) upon illumination with violet
light, a process known as photoconversion, and 2) dynamic changes in fluorescence intensity. Our data show
that CaMPARI allows recording of brain activity from freely-behaving mice, without using microscope objectives
or implanted devices. Moreover, natural degradation of the red CaMPARI protein enables multiple longitudinal
measurements. However, previous attempts to improve CaMPARI using in vitro assays reduced some of its in
vivo properties, which resulted in low dynamic recording sensitivity and a low photoconversion rate that requires
long illumination times to accumulate a sufficient amount of red protein. Therefore, the project goal is to optimize
CaMPARI to allow sensitive recording of cellular-resolution, cortex-wide activity snapshots in freely-moving mice,
followed by subsequent dynamic recording from the same mouse using two- and three-photon microscopy. To
optimize CaMPARI’s performance, we will combine in vitro testing in purified protein, HEK cells, and neurons,
and the most predictive assay: large-scale in vivo screening of ~30-fold more constructs than previous studies.
Aim 1 will focus on enhancing CaMPARI’s photoconversion efficiency to facilitate large-scale recordings in
freely-moving mice. Aim 2 will focus on improving CaMPARI’s dynamic recording properties and sensitivity. In
Aim 3, we will combine beneficial mutations from Aims 1-2 to generate a new CaMPARI with optimized
photoconversion and dynamic recording capabilities. Our proof-of-concept experiments will demonstrate multi-
regional cortical mapping during a battery of behavioral and cognitive tests to detect cellular-resolution changes
in cortex-wide activity patterns. This optimized CAMPARI is expected to facilitate new hypothesis-driven studies
by providing volumetric, multi-regional brain activity data of genetically-targeted neurons during cognitive and
behavioral testing of freely-moving mice, enabling studies that involve both head-fixation and free movement in
the same mice, and to utilize complementary techniques like optogenetic stimulation and single-cell sequencing
methods to enable studying the properties of active (red-labeled) cells during behavioral studies.
项目总结
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(0)
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Robert E. Campbell其他文献
Chemi-genetic sensors for metal ions using green fluorescent protein
使用绿色荧光蛋白的金属离子化学遗传传感器
- DOI:
- 发表时间:
2022 - 期刊:
- 影响因子:0
- 作者:
Takuya Terai;Robert E. Campbell - 通讯作者:
Robert E. Campbell
細胞内イメージングを目指したケミジェネティックNa+センサーの開発と評価
用于细胞内成像的化学基因Na+传感器的开发和评估
- DOI:
- 发表时间:
2022 - 期刊:
- 影响因子:0
- 作者:
竹内志織;朱文超;寺井琢也;Robert E. Campbell - 通讯作者:
Robert E. Campbell
Pushing the wavelength frontier for genetically encoded biosensors of neural activity
推动神经活动基因编码生物传感器的波长前沿
- DOI:
- 发表时间:
2022 - 期刊:
- 影响因子:0
- 作者:
Issei Yamaguchi;Yusuke Nasu;Takuya Terai;Robert E Campbell;Robert E. Campbell - 通讯作者:
Robert E. Campbell
High-Performance Intensiometric Direct- and Inverse-Response Genetically Encoded Biosensors for Citrate
用于柠檬酸盐的高性能强度直接和反向响应基因编码生物传感器
- DOI:
10.1021/acscentsci.0c00518 - 发表时间:
2020-07 - 期刊:
- 影响因子:18.2
- 作者:
Yufeng Zhao;Yi Shen;Yurong Wen;Robert E. Campbell - 通讯作者:
Robert E. Campbell
An Experimental Infection Model in Sheep and Goats to Evaluate Salmonella Colonization in Deep Tissue Lymph Nodes and after Carcass Vascular Rinsing with Bacteriophages in Goats.
绵羊和山羊的实验感染模型,用于评估山羊深部组织淋巴结和胴体血管用噬菌体冲洗后沙门氏菌的定植。
- DOI:
10.1016/j.jfp.2024.100312 - 发表时间:
2024 - 期刊:
- 影响因子:2
- 作者:
Koeun Hwang;Serhat Al;Robert E. Campbell;Kathleen Glass;Kurt D. Vogel;J. R. Claus - 通讯作者:
J. R. Claus
Robert E. Campbell的其他文献
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{{ truncateString('Robert E. Campbell', 18)}}的其他基金
High-Resolution Bidirectional Optical-Acoustic Mesoscopic Neural Interface for Image-Guided Neuromodulation in Behaving Animals - RF1 Admin Supplement
用于行为动物图像引导神经调节的高分辨率双向光声介观神经接口 - RF1 管理补充
- 批准号:
10712937 - 财政年份:2023
- 资助金额:
$ 64.81万 - 项目类别:
High-resolution bidirectional optical-acoustic mesoscopic neural interface for image-guided neuromodulation in behaving animals
用于行为动物图像引导神经调节的高分辨率双向光声介观神经接口
- 批准号:
10407380 - 财政年份:2022
- 资助金额:
$ 64.81万 - 项目类别:
Optimization of CaMPARI for large-scale, cellular-resolution activity recording in freely-moving mice
CaMPARI 的优化,用于自由移动小鼠的大规模细胞分辨率活动记录
- 批准号:
10293936 - 财政年份:2021
- 资助金额:
$ 64.81万 - 项目类别:
Northern Lights collaboration for better 2-photon probes
北极光合作打造更好的 2 光子探测器
- 批准号:
9336980 - 财政年份:2015
- 资助金额:
$ 64.81万 - 项目类别:
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