The multiple roles of mitochondria in synaptic transmission

描述（由申请人提供）：我们的总体目标是确定影响神经递质释放的线粒体机制以及这些机制对不同突触类型的影响。神经末梢中的线粒体很好地影响神经递质的释放，但它们的影响方式一直难以澄清。线粒体功能的许多方面直接涉及突触可塑性，但由于这些活动的交织性质（ATP产生; Ca 2+和Na+处理;质子的挤出;活性氧的释放），很难确定产生主要影响的那些。第二个需要澄清的领域是线粒体在不同形式的短期突触可塑性中的作用。虽然线粒体在强直后增强突触强度中具有既定的作用，但对其对其他形式的短期突触可塑性的影响知之甚少。最后，虽然我们知道线粒体影响神经递质的释放和突触可塑性在大的神经末梢很少知道他们的影响，在小的终端，典型的哺乳动物中枢神经系统。这些是我们知识中的明显空白，特别是突触可塑性允许突触强度的变化，这是学习和记忆的基础现象。也许更令人不安的是，线粒体功能障碍被发现在许多神经退行性疾病的中心，其发病机制和进展知之甚少。核心假设是线粒体通过多种机制影响神经递质的释放，神经末梢的结构及其放电历史决定了哪种机制是有影响的。我们带来了一个综合的电生理，成像和遗传学的方法来解决这个假设在果蝇神经末梢在体内，我们引入了一种新的外周突触与一个单一的释放网站作为模型的中央突触具有相同的架构。我们将测试线粒体Ca 2+摄取限制短序列动作电位期间Ca 2+瞬变和神经递质释放幅度的能力-这是中枢神经元中常见的放电模式（目的1）。重点将放在单一的释放网站的神经末梢，我们观察线粒体有一个贪婪的胃口Ca 2+。我们将确定这些末端中的线粒体是否更有效地吸收Ca 2+，因为它们能够直接从Ca 2+微区吸收Ca 2+（Aim 2）。我们将确定线粒体ATP的产生，而不是Ca 2+的摄取，是否是维持持续神经放电过程中同步释放的主要机制（目的3）。最后，我们将测试在强直后增强递质释放中线粒体Ca 2+释放的要求，并检查线粒体和内质网之间的Ca 2+转移（目的4）。了解线粒体功能如何影响突触传递在非病理条件下将提供所需的基础，了解线粒体在病理条件下的作用。线粒体是人体所有细胞内的细胞器，产生我们大部分的能量。它们集中在神经末梢内，在那里它们为神经之间的通信提供动力，这是大脑的基本活动。然而，人们对它们促进神经系统功能的方式知之甚少，这是令人不安的，因为线粒体功能障碍与许多神经系统疾病有关。我们目前正在研究线粒体如何影响健康神经之间的交流，以便我们可以了解它们可能参与神经退行性疾病的方式。

DESCRIPTION (provided by applicant): Our overall goal is to determine the mitochondrial mechanisms that influence neurotransmitter release and the impact of these mechanisms across different synapse types. Mitochondria in nerve terminals are well placed to influence neurotransmitter release but their means of influence have resisted clarification. Many facets of mitochondrial function have been directly implicated in synaptic plasticity but due to the interwoven nature of these activities (ATP production; Ca2+ and Na+ handling; extrusion of protons; release of reactive oxygen species) it has been difficult to identify those that make the primary impact. A second area that requires clarification is the role of mitochondria in different forms of short-term synaptic plasticity. Although mitochondria have an established role in the post-tetanic potentiation of synaptic strength, little is known about their impact on other forms of short-term synaptic plasticity. Lastly, while we know that mitochondria influence neurotransmitter release and synaptic plasticity in large nerve terminals very little is known about their influence in small terminals, typical of the mammalian CNS. These are glaring gaps in our knowledge, particularly as synaptic plasticity allows for changes in synaptic strength, a phenomenon underlying learning and memory. More troubling perhaps, is that mitochondrial dysfunction is found at the epicenter of many neurodegenerative conditions for which the pathogenesis and progression are poorly understood. The central hypothesis is that mitochondria influence neurotransmitter release through multiple mechanisms, and the architecture of the nerve terminal and its firing history determines which mechanism is influential. We bring a combined electrophysiological, imaging and genetic approach to address this hypothesis at Drosophila nerve terminals in vivo, and we introduce a novel peripheral synapse with a single release-site as a model for central synapses with the same architecture. We will test the ability of mitochondrial Ca2+ uptake to limit the amplitude of Ca2+ transients and neurotransmitter release during short trains of action potentials - a firing pattern common in central neurons (Aim 1). Emphasis will be placed on single release-site nerve terminals where we observe mitochondria to have a voracious appetite for Ca2+. We will determine if mitochondria in these terminals are more effective at taking up Ca2+ because they are able to take up Ca2+ directly from Ca2+ microdomains (Aim 2). We will determine whether mitochondrial ATP production, rather than Ca2+ uptake, is the principle mechanism that maintains synchronous release during sustained nerve firing (Aim 3). Finally we will test the requirement for mitochondrial Ca2+ release in the post-tetanic potentiation of transmitter release, and examine the transfer of Ca2+ between mitochondria and the endoplasmic reticulum (Aim 4). An understanding of how mitochondrial function influences synaptic transmission under non-pathological conditions will provide the foundation required to understand the role of mitochondria in pathological conditions. PUBLIC HEALTH RELEVANCE: Mitochondria are organelles within all cells of the human body that generate most of our energy. They concentrate within nerve endings where they power communication between nerves, a fundamental activity of the brain. However, little is known about the way in which they contribute to the function of the nervous system and this is troubling, as mitochondrial malfunction is implicated in many diseases of the nervous system. We are currently examining how mitochondria influence the communication between healthy nerves so that we may understand the ways in which they may become involved in neurodegenerative conditions.