Optical Tools to Study Purinergic Signaling - Administrative Supplement

研究嘌呤能信号传导的光学工具 - 行政补充

• 批准号: 10580281
• 负责人: Mathew Tantama
• 金额: $ 24.1万
• 依托单位: WELLESLEY COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580281
• 关键词: Address; Administrative Supplement; Affect; Affinity; Biological; Brain; Cell Communication; Cell Volumes; Cell physiology; Cell surface; Cells; Chemotaxis; Communication; Complex; Coupling; Disease; Elements; Energy Metabolism; Event; Immune; Immune response; Infection; Injury; Learning; Long-Term Effects; Measurement; Measures; Mediating; Mediator of activation protein; Metabolic; Methods; Microglia; Molecular; Neuroglia; Neurons; Nucleotides; Optics; Organ; Pain; Pathology; Protein Engineering; Purinoceptor; Receptor Activation; Regulation; Sampling; Sepsis; Series; Signal Transduction; Specimen; System; Tissues; autocrine; cell type; extracellular; imaging study; insight; multiplexed imaging; novel; paracrine; sensor; tool; virtual

Project Summary/Abstract Extracellular ATP and ADP are mediators of purinergic signaling in cell-to-cell communication in virtually every tissue. Autocrine and paracrine purinergic signaling contributes to normal cellular functions such as immune cell chemotaxis and cell volume regulation, and purinergic signaling also contributes to pathology in injury, infection, sepsis, and a number of diseases. Notably, extracellular ATP and ADP mediate purinergic signaling between cells with both short-term and long-term effects. However, it has been a challenge to study the cellular and molecular mechanisms by which ATP and ADP control distinct signaling events because their levels are spatially heterogeneous and purinergic signaling can vary significantly from cell-to-cell. Clearly, direct measurements of extracellular nucleotides are needed, but current methods are not well adapted to measuring fluctuations in the low extracellular nucleotide concentrations expected in live specimens. To overcome this barrier, we will use protein engineering to develop high affinity cell-surface sensors that can be used to resolve the complex spatial and temporal dynamics of ATP and ADP release, clearance, and purinergic receptor activation. To demonstrate the application of our sensors and also guide their optimization, we will study neuron-glia purinergic signaling in the brain. In particular, we will use our sensors to learn how metabolic state affects the function of microglia, the resident immune cells in the brain, in regulating the regional purinergic dynamics that impact local neuronal activity. The proposed series of multiplexed imaging studies that pair our purinergic sensors with sensors of downstream signaling should directly visualize core elements of neuron-glia purinergic communication in an integrated manner. This should provide valuable insight into novel mechanisms responsible for the coupling between brain energy metabolism and excitability. Furthermore, the genetically-encoded tools developed through this proposal should be broadly applicable to deeper study of any purinergic system.

项目总结/文摘