Activation of AMPA-type glutamate neuroreceptors in health and disease

人类大脑包含大约860亿个神经元，在一个极其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，如记忆的形成和学习。因此，介导神经元通讯的蛋白质构成了这些认知过程的分子基础，也是重要的药理靶点。这个项目的重点是一种这样的蛋白质：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上，它们的活动对动作电位的传播至关重要。你将研究一个单点突变(Lurcher)，它会导致罕见和复杂的神经发育障碍，包括癫痫发作，以在单分子水平上了解疾病的机制和AMPA受体激活的原理，这个项目既包括基础科学，也包括应用科学。初步数据显示，该突变减缓了四聚体AMPA受体内个别亚基的激活，使我们能够研究AMPA受体激活的机制，因为它的亚基逐个激活。该项目的这一部分解决了谷氨酸门控离子通道领域中的一个主要悬而未决的问题：所有四个亚基对AMPA受体的激活是否都有同等的贡献？该项目的应用方面侧重于研究潘生丁对突变的AMPA受体的影响，潘生丁是一种用于治疗癫痫的商业药物。该项目的这一部分将有助于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，它使我们能够在宏观和单分子水平上记录AMPA受体的活动。单分子记录伴随着广泛的数据分析，因此应聘者将发展出很强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写作)与巴拉诺维奇博士(首席主管)密切合作，后者还将对他们进行膜片钳电生理学方面的培训。同时，候选人将从Wyllie教授(第二主管)的丰富经验中受益，与Wyllie教授合作，他们将有机会将他们的膜片钳经验扩展到神经元培养。该项目还需要与爱丁堡大学的Teuta Pilizota教授进行跨学科合作，他的细菌鞭毛马达阶跃分析算法将被探索并应用于AMPA受体的阶跃激活分析。

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.