Optogenetic control of skeletal muscle excitability

描述(由申请人提供)：几种情况，如疾病或意外和化学创伤，可导致骨骼肌神经控制的丧失。光遗传控制兴奋性是肌肉功能恢复的一种很有前途的选择，它已经彻底改变了神经生物学研究，但从未在成人骨骼肌中尝试过。然而，在体内实际实施光遗传方法之前，需要回答几个基本问题。在这项建议的目标1中，我们将研究激活(例如通道视紫红质2)和沉默激活物(例如视紫红质3)是否在成人骨骼肌纤维中有足够的表达，从而使照明素脉冲可以分别诱发动作电位(AP)或抑制电诱导的AP。这 信息将使我们能够比较地测试(在目标2中)光和电刺激的能力，以实现对表达光学致动器的完整肌肉的机械活动的精细控制。我们将使用两种方法将Opsins编码的质粒特异性地导入肌肉：电穿孔和腺相关病毒。这将使我们能够研究使用无线光电植入设备对完整动物的肌肉进行精细控制的新方法的可行性。这笔赠款中提出的创新研究方法的结果有望为实际使用光遗传学工具铺平道路，以便在由于缺乏神经输入而被认为不活跃时恢复对肌肉输出的控制。

DESCRIPTION (provided by applicant): Several conditions such as diseases or accidental and chirurgical trauma can lead to the loss of nerve control of skeletal muscles. Optogenetic control of excitability, which has revolutionized neurobiological research but has never been attempted in adult skeletal muscles, is a promising alternative for the recovery of muscle function. However, several basic questions need to be answered prior to the practical implementation of optogenetic approaches in vivo. In Aim 1 of this proposal we will investigate whether activating (e.g. channelrhodopsin 2) and silencing actuators (e.g. archeorhodopsin 3) are expressed in sufficient quantities in adult skeletal muscle fibers so that pulses of illuminatin can either elicit action potentials (APs), or inhibit electrically elicited APs, respectively. This information will allow us to comparatively test (in Aim 2) the ability of light and electrical stimulation to attain fine control of the mechanical activity of intact muscles expressing optical actuators. We will use two approaches for the specific transfection of muscle with plasmids encoding for opsins: electroporation and adeno-associated virus. This will enable us to investigate the feasibility of using novel approaches using wireless optoelectronic implantable devices for the fine control of muscles in the intact animal. The outcome of the innovative research approaches proposed in this grant are expected to pave the way for the practical use of optogenetics tools to restore the control of muscle output when they are deemed inactive due to the absence of nerve inputs.