Bioengineering a novel electromagnetic perspective gene as a tool for wireless control of excitable cells

摘要： 在活着的大脑中操纵神经元功能的能力对 人类健康和基础科学。在过去的半个世纪里，通过植入性的神经元调制 特定大脑区域的微电极已被用来缓解与脑部疾病相关的症状 各种神经元性疾病。然而，这种方法的缺点是需要植入复合体， 依赖外部电源的大型扩展电子设备，以及 神经刺激的实际放置会产生意想不到的不良副作用 电极通常会产生。近年来，分子生物学和合成生物学的主要进展 促进了受体和通道的克隆和优化，从而允许调节 神经元对光、化学物质或温度作出反应。然而，对这些的控制 蛋白质需要侵入性地传递激活剂。因此，一种非侵入式、远程控制的 能够以非侵入性方式操纵特定神经元群体的神经调节剂是一种 未得到满足的需求。 为了迎接这一挑战，我们调查了一种替代方案的可能性，并 一种新的通过传输非侵入性的、 电磁场(EMF)。使用表达克隆，我们已经识别并克隆了单个 编码成一种对电动势有反应的蛋白质的基因。这种独特的基因从未有过 以前的特征被称为电磁感知基因(EPG)。EPG基因被克隆 并在哺乳动物细胞、神经元培养和大鼠脑中表达。免疫组织化学 表明EPG的表达仅限于哺乳动物细胞膜，而且它 可以在特定的人群中表达，也可以在大鼠的特定大脑区域表达。钙 哺乳动物细胞和培养的神经元表达EPG的成像显示远程 电磁场的激活显著增加了细胞内钙浓度，表明 细胞兴奋性。此外，大鼠运动皮质EPG的无线磁激活可诱导 在体对侧前肢的运动诱发反应。我们假设EPG 技术将使无线控制神经元功能与细胞、区域和 时间特定性。在这里，我们建议彻底描述细胞内的EPG， 分子水平和功能水平。我们还将测试EPG技术的有效性，以 无线控制活体内的神经元功能。 我们预计这项新技术将改变神经调节的未来， 补充现有的神经调节工具，并极大地有助于理解 复杂的神经回路。

Abstract: The ability to manipulate neuronal function in the living brain has a major impact both on human health and basic sciences. In the past half century, neuronal modulation via implantable microelectrodes in specific brain regions has been used to relief symptoms associated with a variety of neuronal disorders. However, this approach suffers from the need to implant complex, large and expansive electronic devices that depend on an external power supply, and the unexpected and undesirable side effects that the actual placement of the neurostimulation electrodes often produces. Recently, major advances in molecular and synthetic biology facilitated the cloning and optimization of receptors and channels that allow modulation of neuronal function in response to light, chemicals or temperature. However, the control of these proteins requires invasive delivery of the activator. Therefore, a non-invasive, remote controlled neuromodulator that can manipulate specific neuronal population in a non-invasive manner is an unmet need. To embark upon this challenge, we have investigated the potential of an alternative and novel method to remotely control cellular function through the transmission of non-invasive, electromagnetic fields (EMF). Using expression cloning, we have identified and cloned a single gene that encodes to a protein that responds to EMF. This unique gene has never been characterized before and was termed electromagnetic perceptive gene (EPG). EPG was cloned and expressed in mammalian cells, neuronal cultures and in rat’s brain. Immunohistochemistry showed that the expression of EPG is confined to the mammalian cell membrane, and that it can be expressed in a specific population, and in specific brain regions of the rat. Calcium imaging in mammalian cells and cultured neurons expressing EPG demonstrated that remote activation by EMF significantly increases intracellular calcium concentrations, indicative of cellular excitability. Moreover, wireless magnetic activation of EPG in rat motor cortex induced motor evoked responses of the contralateral forelimb in vivo. We hypothesize that the EPG technology will enable wireless control of neuronal function with cell, region and temporal specificity. Here we propose to thoroughly characterize the EPG in the cellular, molecular and functional levels. We will also test the effectiveness of the EPG technology to wirelessly control neuronal function in vivo. We anticipate that this new technology would transform the future of neuromodulation, complement existing neuromodulation tools, and considerably contribute to the understanding of complex neural circuits.