Phospholipase C (PLC) is a key enzyme that regulates physiological processes via lipid and calcium signaling. Despite advances in protein engineering, no tools are available for direct PLC control. Here, we developed a novel optogenetic tool, light-controlled PLCβ (opto-PLCβ). Opto-PLCβ uses a light-induced dimer module, which directs an engineered PLC to the plasma membrane in a light-dependent manner. Our design includes an autoinhibitory capacity, ensuring stringent control over PLC activity. Opto-PLCβ triggers reversible calcium responses and lipid dynamics in a restricted region, allowing precise spatiotemporal control of PLC signaling. Using our system, we discovered that phospholipase D-mediated phosphatidic acid contributes to diacylglycerol clearance on the plasma membrane. Moreover, we extended its applicability in vivo, demonstrating that opto-PLCβ can enhance amygdala synaptic plasticity and associative fear learning in mice. Thus, opto-PLCβ offers precise spatiotemporal control, enabling comprehensive investigation of PLC-mediated signaling pathways, lipid dynamics, and their physiological consequences in vivo.

磷脂酶 C (PLC) 是一种通过脂质和钙信号传导调节生理过程的关键酶。尽管蛋白质工程取得了进步，但还没有可用于直接 PLC 控制的工具。在这里，我们开发了一种新型光遗传学工具，光控PLCβ（opto-PLCβ）。 Opto-PLCβ 使用光诱导二聚体模块，该模块以光依赖性方式将工程 PLC 引导至质膜。我们的设计包括自动抑制能力，确保对 PLC 活动的严格控制。 Opto-PLCβ 在受限区域触发可逆钙反应和脂质动力学，从而实现 PLC 信号传导的精确时空控制。使用我们的系统，我们发现磷脂酶 D 介导的磷脂酸有助于质膜上二酰甘油的清除。此外，我们扩展了其在体内的适用性，证明 opto-PLCβ 可以增强小鼠杏仁核突触可塑性和联想恐惧学习。因此，opto-PLCβ 提供精确的时空控制，能够全面研究 PLC 介导的信号通路、脂质动力学及其体内生理后果。