Phospho-regulation as a driver of inhibitory synapse function

磷酸化作为一种塑造翻译后蛋白质功能的方式已经被研究了很长时间。在大脑中，磷酸化已成为将活动依赖信息转导到突触功能改变的最有效途径之一。大量文献支持磷酸化是中枢神经系统内谷氨酸能兴奋性突触后信号转导的重要组成部分。然而，新出现的证据表明，gaba能抑制性突触后也经历活动依赖的可塑性。基于有限的候选研究，少数gaba能蛋白最近被证明在抑制性突触磷酸化。虽然这支持磷酸化可能是调节抑制性突触的机制，但尚未尝试深入分析磷酸化如何与抑制性突触功能耦合。此外，目前尚不清楚哪些激酶存在并因此作用于抑制性突触。因此，抑制性突触磷酸化蛋白质组目前是一个“黑盒子”，这是理解活动如何改变抑制对形成经验依赖的大脑功能很重要的一个重要障碍。在这里，我建议使用一种前沿的体内化学遗传学方法，结合CRISPR-depletion、光遗传学、电生理学和成像技术，鉴定抑制突触中的磷酸化蛋白，以了解激酶如何协调抑制突触复杂而基本的工作。

Phosphorylation has long been studied as a way to shape protein function after translation. In the brain, phosphorylation has emerged as one of the most efficient ways to transduce activity-dependent information to altered synaptic function. Extensive literature supports phosphorylation as an essential component of signal transduction at glutamatergic excitatory postsynapses within the CNS. Emerging evidence, however, demonstrates that GABAergic inhibitory postsynapses also undergo activity dependent plasticity. Based on limited candidate studies, a handful of GABAergic proteins have recently been shown to be phosphorylated at inhibitory synapses. While this supports phosphorylation as a likely mechanism to modulate inhibitory synapses, an in-depth analysis of how phosphorylation is coupled to inhibitory synapses function has not been attempted. Moreover, it is still unclear as to which kinases are present and therefore act at inhibitory synapses. Thus, the inhibitory synaptic phospho-proteome is currently a “black box”, which is a significant barrier to understanding how activity modifies inhibition important for shaping experience-dependent brain function. Here, I propose to use a cutting edge in vivo chemico-genetic approach to identify phosphorylated proteins at inhibitory synapses in combination with CRISPR-depletion, optogenetics, electrophysiology and imaging to understand how kinases orchestrate the complex yet fundamental workings of the inhibitory synapse.