磷脂酶Cβ介导的光导蛋白质机器装配与调控的分子机理研究

The phospholipase Cβ(PLCβ) phototransduction protein complexes play vital roles in the phototransduction cascade of vertebrate intrinsic photosensitive retinal ganglion cells (ipRGCs) and invertebrate rhabdomeric photoreceptor cells. In vertebrate, through the PLCβ phototransduction protein complex in the ipGRCs, the light signal is sent to the hypothalamic suprachiasmatic nucleus ( SCN), which is an important circadian pacemaker, to control the circadian rhythem. It has been reported that the components of the PLCβ phototransduction protein complex in the vertebrate ipRGCs are very similar to those of invertebrate rhabdomeric photoreceptor cells. However, the assembly and regulation mechanism of this phototransduction complex still remains unclear. How the PLCβ phototransduction protein complex works in vertebrate ipRGCs is even less understood compared to the invertebrate. In this study, based on our previous studies in Drosophila PLCβ phototransduction protein complex, we combine the molecular biology, biochemistry, structural biology, knock-out mice, viral infection and patch-clamp techniques, to study the assembly and regulation mechanism of the Drosophila and mouse PLCβ phototransduction protein complex, in which the Drosophila INAD/NORPA, INAD/ePKC, TRP/CaM complex and mouse INADL/PLCβ4 complex will be studied with higher priorities. Hopefully, our study will established the biochemical and structural basis for better understanding the biological functions of the PLCβ protein complexes in the phototransduction cascade of Drosophila and mouse.

磷脂酶Cβ介导的光导蛋白质机器，对于非脊椎动物视网膜杆状体感光细胞以及脊椎动物的自主感光神经节细胞的光信号传导都非常重要。在脊椎动物中，自主感光神经节细胞中的磷脂酶Cβ光导蛋白质机器，通过向大脑控制昼夜节律的下丘脑视交叉上核发送光信息，从而对昼夜节律进行调节。研究表明，脊椎动物自主感光神经节细胞的磷脂酶Cβ光导蛋白质机器的组成成分，与非脊椎动物的杆状体感光细胞非常相似，但是目前二者装配与调控的分子机制仍不清楚，对于前者的研究更是处于起步阶段。本项目拟在对果蝇光导机器的研究基础上，以果蝇INAD/NORPA（PLCβ）、INAD/ePKC、TRP/CaM以及小鼠INADL/PLCβ4蛋白复合体为突破口，综合运用分子生物学、生物化学、结构生物学、基因敲除动物模型、病毒侵染及膜片钳等技术，揭示其组装与动态调节的分子机制，探讨该蛋白质机器在模式动物果蝇以及小鼠光信号传导生理过程中的生物学功能。

磷脂酶Cβ介导的光导蛋白质机器，对于非脊椎动物视网膜杆状体感光细胞以及脊椎动物的自主感光神经节细胞的光信号传导都非常重要。在脊椎动物中，自主感光神经节细胞中的磷脂酶Cβ光导蛋白质机器，通过向大脑控制昼夜节律的下丘脑视交叉上核发送光信息，从而对昼夜节律进行调节。研究表明，脊椎动物自主感光神经节细胞的磷脂酶Cβ光导蛋白质机器的组成成分，与非脊椎动物的杆状体感光细胞非常相似，但是目前二者装配与调控的分子机制仍不清楚，对于前者的研究更是处于起步阶段。本项目在既往对果蝇光导机器的研究基础上，以果蝇INAD/NORPA（PLCβ）、TRP/CaM以及小鼠INADL/PLCβ4蛋白复合体为突破口，综合运用了分子生物学、生物化学、结构生物学、基因敲除动物模型、病毒侵染及膜片钳等技术，揭示了其组装与动态调节的分子机制，探讨了该蛋白质机器在模式动物果蝇以及小鼠光信号传导生理过程中的生物学功能。