Optogenetic control of skeletal muscle excitability

描述（由申请人提供）：疾病或意外和外科创伤等几种情况可导致骨骼肌神经控制丧失。光遗传学控制兴奋性，已经彻底改变了神经生物学研究，但从未在成人骨骼肌中进行过尝试，是恢复肌肉功能的有希望的替代方法。然而，在体内实际实施光遗传学方法之前，需要回答几个基本问题。在本提案的目的1中，我们将研究激活（如通道视紫红质2）和沉默致动器（如原视紫红质3）是否在成人骨骼肌纤维中表达足够量，从而使照明脉冲分别引发动作电位（ap）或抑制电激发的ap。这

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.