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

摘要： 在活体大脑中操纵神经元功能的能力对神经元的功能产生了重大影响。 人类健康和基础科学。在过去的半个世纪，通过植入的神经元调制 在特定的大脑区域的微电极已被用于缓解与脑缺血相关的症状。 各种神经元疾病。然而，这种方法需要植入复杂的， 大型和昂贵的电子设备，依赖于外部电源， 神经刺激的实际放置 电极经常产生。最近，分子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， 神经元对光、化学物质或温度的反应。然而，控制这些 蛋白质需要活化剂的侵入性递送。因此，一种非侵入性的、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 未满足的需求 为了应对这一挑战，我们研究了替代方案的潜力， 一种通过非侵入性传输远程控制细胞功能的新方法， 电磁场（EMF）使用表达克隆，我们已经鉴定并克隆了一个 编码对电磁场有反应的蛋白质的基因。这种独特的基因从未被 电磁感知基因（Electromagnetic Perceptive Gene，EPG）是一种新的电磁感知基因。EPG被克隆 并在哺乳动物细胞、神经元培养物和大鼠脑中表达。免疫组 表明EPG的表达局限于哺乳动物细胞膜， 可以在特定的群体中表达，并且在大鼠的特定脑区域中表达。钙 在哺乳动物细胞和表达EPG的培养神经元中的成像表明， EMF激活显著增加细胞内钙浓度，表明 细胞兴奋性此外，无线磁激活大鼠运动皮层的EPG诱导， 运动诱发反应的对侧前肢在体内。我们假设EPG 技术将使无线控制神经元功能成为可能， 时间特异性在这里，我们建议彻底表征EPG在蜂窝， 分子和功能水平。我们还将测试EPG技术的有效性， 无线控制体内神经元的功能 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，并大大有助于理解 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.