High-resolution optical approaches to study dendritic signaling

研究树突信号传导的高分辨率光学方法

• 批准号: RGPIN-2017-06171
• 负责人: DeKoninck, Paul
• 金额: $ 2.91万
• 依托单位: Université Laval
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=648824
• 关键词: resolution; optical; approaches; study; dendritic

Understanding the mechanisms by which nerve cells transmit information and adapt their communication has progressed through the identification of the molecular components of synapses as well as the electrochemical nature of synaptic signals. However, for the majority of these components and signals, we know little about how they operate, propagate and interact during neurotransmission. To fully understand how synapses work, we must be able to resolve their molecular components and elementary signals within living neural circuits and to monitor them at length and time scales relevant to neurotransmission. Toward this aim, we have developed a Neurophotonics Research Program focussing on the development of advanced optical methods -in some cases beyond the resolution of light- to resolve the spatial and temporal dynamics of proteins and signals during synaptic transmission and to determine how these dynamics affect synaptic function. This Program is at the interface between Optics and Neurobiology. It bridges recent advances in optical methods, fluorescent probes, and cellular and molecular neuroscience. It trains a new generation of HQP in Biophotonics, supported by our Training Program in Biophotonics.***The general goal of our Program is to understand how temporally- and spatially-encoded dendritic electrical signals are decoded into specific information. We wish to understand how neuronal activities are decoded from the initial electrical signals at the membrane into i) transient intracellular signals and then into ii) longer-lasting biochemical signals iii) and synaptic plasticity***To study these problems, our Research Program has progressed in the last term, particularly on the design and improvements of optical methods to resolve and manipulate signaling events in dendrites of cultured neurons with improved spatial and temporal resolutions.***We specifically ask: How is electrical activity propagated from spines to dendrite? How does CaMKII decode the frequencies of calcium oscillations in dendrites? How does CaMKII propagate long-lasting memory? We address these questions by combining optical methods to monitor changes in membrane potentials, calcium imaging, FRET-FLIM to monitor protein interactions and activation, and optical nanoscopy to resolve protein interactions at the nanoscale. ***Our Research Program should help understanding the molecular processes that neurons use to encode their synaptic and electrical activities inside their dendrites. It will continue to advance optical methods to push the limits of resolution of live imaging in neurons. Our Research Program, at the interface between cellular neurobiology and optics, mainly trains physicists, engineers and chemists, who combine their skills to advance our knowledge of neuronal function and to improve our methods for advancing cellular and molecular neuroscience.*****

通过识别突触的分子组成以及突触信号的电化学性质，对神经细胞传递信息和适应其通信的机制的了解取得了进展。然而，对于这些组件和信号中的大多数，我们对它们在神经传递过程中如何运作、传播和相互作用知之甚少。为了充分了解突触是如何工作的，我们必须能够在活的神经回路中解析它们的分子成分和基本信号，并在与神经传递相关的长度和时间尺度上对它们进行监测。为此，我们开发了一个神经光子学研究计划，专注于发展先进的光学方法-在某些情况下超越光的分辨率-以解决突触传递过程中蛋白质和信号的空间和时间动力学，并确定这些动力学如何影响突触功能。该项目处于光学和神经生物学的交界处。它连接了光学方法、荧光探针以及细胞和分子神经科学的最新进展。它在我们的生物光子学培训计划的支持下，在生物光子学方面培训新一代HQP。*我们计划的总体目标是了解时间和空间编码的树枝状电信号是如何解码成特定信息的。我们希望了解神经元活动是如何从膜上的初始电信号解码成I)瞬时细胞内信号，然后变成II)更持久的生化信号III)和突触可塑性*为了研究这些问题，我们的研究计划在最后一学期取得了进展，特别是在设计和改进光学方法来解决和操纵培养神经元树突中的信号事件方面，提高了空间和时间分辨率。*我们特别问：电活动是如何从棘突传播到树突的？CaMKII是如何解码树突中钙振荡的频率的？CaMKII是如何传播持久记忆的？我们通过结合光学方法来监测膜电位的变化，钙成像，FRET-FILM来监测蛋白质的相互作用和激活，以及光学纳米显微镜来解决纳米级的蛋白质相互作用来解决这些问题。*我们的研究计划应该有助于理解神经元用来编码其突触和树突内的电活动的分子过程。它将继续推进光学方法，以突破神经元实时成像的分辨率极限。我们的研究计划位于细胞神经生物学和光学之间，主要培养物理学家、工程师和化学家，他们结合他们的技能来提高我们对神经元功能的知识，并改进我们推进细胞和分子神经科学的方法。