Activation of AMPA-type glutamate neuroreceptors in health and disease

人类大脑包含大约860亿个神经元，它们在一个非常复杂的网络中相互通信。这个网络中的动态活动变化构成了我们所有认知过程的基础，例如记忆形成和学习。因此，介导神经元通信的蛋白质形成这些认知过程的分子基础，并且是必需的药理学靶标。该项目的重点是这样一种蛋白质：AMPA型谷氨酸神经受体。这些谷氨酸门控离子通道存在于几乎所有兴奋性突触的突触后膜中，它们的活性对于动作电位的传播至关重要。您将在一个涵盖基础科学和应用科学的项目中研究一种单点突变体（Lurcher），该突变体会导致罕见和复杂的神经发育障碍，包括癫痫发作，以了解疾病的机制和AMPA受体在单分子水平上的激活原理。初步数据显示，该突变体减缓了四聚体AMPA受体内单个亚基的激活，使我们能够研究AMPA受体激活的机制，因为它的亚基一个接一个地激活。本项目的这一部分解决了谷氨酸门控离子通道领域的一个主要开放问题：所有四个亚基对AMPA受体的激活是否有同等的贡献？该项目的应用方面侧重于研究perampanel（一种用于治疗癫痫的市售药物）对突变AMPA受体的影响。该项目的这一部分可以帮助为携带这种突变的患者开发新的治疗方案。主要的实验方法将是膜片钳电生理学，这使我们能够在宏观和单分子水平上记录AMPA受体的活性。单分子记录伴随着广泛的数据分析，因此候选人将发展强大的实验和数据分析技能。候选人将在项目的各个方面（从实验设计，数据收集和分析到手稿撰写）与Baranovic博士（主要主管）密切合作，Baranovic博士还将对他们进行膜片钳电生理学培训。与此同时，候选人将受益于Wyllie教授（第二导师）的丰富经验，与他们合作，他们将有机会将他们的膜片钳经验扩展到神经元培养。该项目还需要与Teuta Pilizota教授（爱丁堡大学）进行跨学科合作，其细菌鞭毛马达分步分析算法将被探索并适用于分析AMPA受体的分步激活。

The human brain contains about 86 billion neurons communicating with each other in a hugely complex network. Dynamic activity changes in this network form the basis of all our cognitive processes, such as memory formation and learning. Consequently, proteins that mediate neuronal communication form the molecular basis of theses cognitive processes and are essential pharmacological targets. This project focuses on one such protein: AMPA-type glutamate neuroreceptors. These glutamate-gated ion channels are found in the postsynaptic membrane of almost every excitatory synapse and their activity is crucial for propagation of the action potential. You will study a single-point mutant (Lurcher) which causes rare and complex neurodevelopmental disorders, including seizures, to understand the mechanism of disease and principles of activation of AMPA receptors at the single-molecule level, in a project encompassing both fundamental and applied science.Preliminary data shows this mutant slows activation of individual subunits within the tetrameric AMPA receptor, allowing us to investigate the mechanism of AMPA receptor activation as its subunits activate one by one. This part of the project addresses one of the main open questions in the field of glutamate-gated ion channels: do all four subunits contribute equally to the activation of an AMPA receptors? The applied aspect of the project focuses on studying the effects of perampanel, a commercially available drug used in treatments against epilepsy, on mutant AMPA receptors. This part of the project could help develop novel treatment options for patients carrying this mutation.The main experimental method will be patch-clamp electrophysiology, which enables us to record the activity of AMPA receptors at the macroscopic and single-molecule level. Single-molecule recordings are accompanied by extensive data analysis, therefore the candidate would develop both strong experimental and data analysis skills. The candidate will be working closely together on all aspects of the project (from experimental design, data collection and analysis to manuscript writing) with Dr. Baranovic (primary supervisor), who would also train them in patch-clamp electrophysiology. At the same time, the candidate would benefit from the extensive experience of Prof. Wyllie (second supervisor), in collaboration with whom they would have a chance to extend their patch-clamp experience to neuronal cultures. The project also entails interdisciplinary collaboration with Prof. Teuta Pilizota (University of Edinburgh), whose algorithm for step analysis of bacterial flagellar motor will be explored and adapted for analysis of stepwise activation of AMPA receptors.