The role of a calcium permeable TRP channel in endocytosis

钙渗透性 TRP 通道在内吞作用中的作用

• 批准号: 9325812
• 负责人: Liang-Wei Gong
• 金额: $ 39.34万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9325812
• 关键词: Alzheimer' s Disease; Animal Model; Animals; Biological; Biological Assay; Brain; Brain Diseases; Calcium; Cations; Cells; Chromaffin Cells; Complement; Data; Disease; Electric Capacitance; Endocytic Vesicle; Endocytosis; Epilepsy; Event; Exocytosis; Extracellular Space; Family; Genes; Glutamate Transporter; Health; Human; Image; Ions; Kinetics; Knock-out; Knockout Mice; Knowledge; Link; Measurement; Measures; Mediating; Mission; Molecular; Mutagenesis; Mutate; Neurons; Outcome Measure; PHluorin; Parkinson Disease; Pathogenesis; Play; Process; Public Health; Research; Risk; Role; Route; Synaptic Transmission; Synaptic Vesicles; Synaptophysin; Testing; Transgenic Animals; base; disability; human disease; inhibitor/antagonist; knock-down; knockout animal; live cell imaging; member; mouse model; mutant; novel therapeutics; receptor; research study; small hairpin RNA; synaptotagmin I; voltage

 DESCRIPTION (provided by applicant): Synaptic vesicle exocytosis is a fundamental process mediating brain function; subsequent synaptic vesicle endocytosis is important to maintain synaptic transmission. Since malfunctions in synaptic vesicle endocytosis have been linked to human diseases, such as Parkinson's disease, epilepsy, and Alzheimer's disease, elucidating the molecular mechanisms for synaptic vesicle endocytosis will have implications not only for understanding neuronal function but also will provide potential targets for developing novel therapies to treat brain diseases. While it is well established that Ca2+ is critical for exocytosi, increasing evidence suggests that Ca2+ may also be important for endocytosis. Although the importance for endocytosis of Ca2+ influx from extracellular space is accepted, the identity of the Ca2+ influx route for endocytosis remains unclear, as Ca2+-permeable channels other than voltage-gated Ca2+ channels may be involved. Transient receptor potential (TRP) channels are a family of non-selective cation channels with high expression levels in the brain. TRP melastatin 7 (TRPM7), a member of the TRP superfamily, is a Ca2+- permeable channel. We hypothesize that the Ca2+-permeable TRPM7, located on endocytic vesicles, acts as the Ca2+ influx route to regulate endocytosis. Our hypothesis is well supported by the preliminary data described in this proposal. We will test this hypothesis using biophysical and live-cell imaging assays in chromaffin cells and neurons to measure the outcome of molecular biological manipulations such as transgenetic TRPM7 knockout animal models and shRNA-mediated gene knockdown.

 描述（由申请人提供）：突触小泡胞吐作用是介导大脑功能的基本过程；随后的突触小泡胞吞作用对于维持突触传递非常重要，因为突触小泡胞吞作用的功能障碍与人类疾病有关，例如帕金森病、癫痫和癫痫。阿尔茨海默病，阐明突触小泡内吞作用的分子机制将具有重要意义虽然 Ca2+ 对胞吐作用至关重要，但越来越多的证据表明 Ca2+ 可能对内吞作用也很重要。细胞外空间的 Ca2+ 流入已被接受，但内吞作用的 Ca2+ 流入途径的身份仍不清楚，因为电压门控 Ca2+ 通道以外的 Ca2+ 通透通道可能是瞬时受体电位 (TRP) 通道是大脑中高表达的非选择性阳离子通道家族，TRP 褪黑素 7 (TRPM7) 是 TRP 超家族的成员，是一种 Ca2+- 通透性通道。 Ca2+ 渗透性 TRPM7 位于内吞囊泡上，充当调节内吞作用的 Ca2+ 流入途径。我们的假设得到了所描述的初步数据的充分支持。在本提案中，我们将使用嗜铬细胞和神经元中的生物物理和活细胞成像测定来测试这一假设，以测量转基因 TRPM7 敲除动物模型和 shRNA 介导的基因敲除等分子生物学操作的结果。