PDF(Pubmed)
Abstract：
For decades, the expression of immediate early genes (IEGs) such as FOS has been the most widely used molecular marker representing neuronal activation. However, to date, there is no equivalent surrogate available for the decrease of neuronal activity. Here, we developed an optogenetic-based biochemical screen in which population neural activities can be controlled by light with single action potential precision, followed by unbiased phosphoproteomic profiling. We identified that the phosphorylation of pyruvate dehydrogenase (pPDH) inversely correlated with the intensity of action potential firing in primary neurons. In in vivo mouse models, monoclonal antibody-based pPDH immunostaining detected activity decreases across the brain, which were induced by a wide range of factors including general anesthesia, chemogenetic inhibition, sensory experiences, and natural behaviors. Thus, as an inverse activity marker (IAM) in vivo, pPDH can be used together with IEGs or other cell-type markers to profile and identify bi-directional neural dynamics induced by experiences or behaviors.
摘要：
几十年来,例如FOS的立即早期基因(IEGs)的表达是最广泛使用的代表神经元激活的分子标记。然而,到目前为止,没有等效的替代可用于减少神经元活动。这里,我们开发了一种基于光遗传学的生化筛选，其中群体神经活动可以通过具有单动作电位精度的光控制，然后是无偏的磷酸化蛋白质组分析。我们确定丙酮酸脱氢酶（pPDH）的磷酸化与原代神经元中动作电位放电的强度成反比。在体内小鼠模型中，基于单克隆抗体的pPDH免疫染色检测到整个大脑的活性降低，这是由包括全身麻醉在内的多种因素引起的，化学发生抑制，感官体验，和自然行为。因此，作为体内的反向活动标记（IAM），pPDH可以与IEG或其他细胞类型标记物一起使用以分析和鉴定由经验或行为诱导的双向神经动力学。
