Controlling Spatially Restricted Intracellular Protein-Activity During Embryonic Neuronal Development Using Biomagnetic Nanotechnologies

胚胎神经元发育过程中空间限制性细胞内蛋白活性的调控 生物磁性纳米技术 在哺乳动物胚胎发育过程中，神经元细胞分裂形成不同的细胞区室， 轴突和树突，其细胞质、细胞骨架和血浆的分子组成固有地不同 膜的这些差异是这些区室独特形态和功能的基础， 负责大脑中的定向信息流。而轴突传递化学和电神经元 信号，树突接收并整合它们。这种两极化的建筑源于精确调控的空间结构， 将特定的细胞内蛋白质的活性分离到单个神经元细胞的离散亚细胞区域， 分别决定了轴突和树突的命运。这些蛋白质活性定位的异常导致 有缺陷的神经元极化和严重的人类神经发育病理学，包括智力和 运动障碍、癫痫和自闭症谱系障碍。能够精确地控制时空， 细胞内蛋白质活性将允许神经元极化的定向调节， 用于修复潜在的神经发育病理的方法。迄今为止，没有任何现有技术， 包括领先的分子遗传学、光控蛋白质激活，或使用光遗传学的组合， 可以允许发育中的神经元中细胞内蛋白质活性的持续空间限制。主要目标 这项研究的目的是通过开发基于生物磁的 纳米技术，这将使空间和时间控制细胞内蛋白质的功能， 胚胎神经元具体来说，我们将开发生物磁纳米技术， 激酶LKB 1的活性，以指示培养中胚胎神经元中轴突形成的过程。这样的 该提案需要采用多学科方法，整合神经生物学、材料工程和 生物电子学，用于开发基于蛋白质的神经疗法。许多细胞事件决定了细胞 形态发生，代谢状态，或其独特的生理功能，在所有细胞类型跨越进化遥远的 物种，由特定细胞内蛋白的高度定位和定时活性决定。a的因果作用 在特定细胞事件中的关键细胞内蛋白或控制该事件的能力只能通过以下方式实现： 定向亚细胞定位和蛋白质或其活性的保留。作为当前时空 蛋白质功能的操纵本质上不能允许蛋白质的长期空间限制 功能，我们的研究将适用于许多基本的细胞事件，如极化和迁移，以及 许多控制这些细胞过程的细胞内蛋白质。

Controlling spatially restricted intracellular protein-activity during embryonic neuronal development using biomagnetic nanotechnologies During mammalian embryonic development, neuronal cells polarize to create distinct cellular compartments of the axon and dendrite that inherently differ in the molecular composition of their cytoplasm, cytoskeleton, and plasma membrane. These differences underlie the unique morphology and function of these compartments and are responsible for directed information flow in the brain. Whereas axons transmit the chemical and electrical neuronal signals, the dendrites receive and integrate them. This polarized architecture arises from precisely regulated spatial segregation of specific intracellular proteins’ activities to discrete subcellular regions of a single neuronal cell that respectively dictate the axonal vs. dendritic fate. Aberrations in the localization of these proteins’ activity lead to defective neuron polarization and underlie severe human neurodevelopmental pathologies including intellectual and motor disabilities, epilepsy, and autism spectrum disorders. The ability to exert precise spatio-temporal control on intracellular protein-activity would permit directed regulation of neuronal polarization and may provide new approaches for the repair of the underlying neurodevelopmental pathologies. To date, no existing technologies, including leading molecular-genetics, light-controlled protein activation, or their combination using optogenetics, can allow sustained spatial restriction of intracellular protein-activity in the developing neuron. The main objective of this study is to address this fundamental challenge in neurobiology by developing biomagnetic-based nanotechnologies that will enable the spatial and temporal control of intracellular protein function in developing embryonic neurons. Specifically, we will develop biomagnetic-nanotechnologies to deliver and retain localized activity of the kinase LKB1, to dictate the process of axon formation in embryonic neurons in culture. Such a proposal demands a multi-disciplinary approach that integrates neurobiology, material engineering, and bioelectronics, for the development of protein based neuro-therapeutics. Many cellular events that dictate cell morphogenesis, metabolic state, or its unique physiological functions, in all cell types across evolutionarily distant species, are determined by highly localized and timed activity of specific intracellular proteins. The causative role of a critical intracellular protein in a particular cellular event or the ability to control that event can only be achieved by directed subcellular localization and retention of the protein or its activity. As current methodologies for spatio-temporal manipulation of protein function are inherently incapable of allowing the long-term spatial confinement of protein function, our studies will be applicable to many fundamental cellular events, as polarization and migration, and to the many intracellular proteins that control these cellular processes.