TRPV4在Aβ诱导星形胶质细胞活化及介导神经元死亡中的作用

TRPV4 (Transient receptor potential channel vanilloid member 4) is a polymodally gated channel that can be activated by diverse stimuli. It is involved in Aβ-induced hippocampal neuron death. The aim of this study is to uncover the mechanisms of activating TRPV4 in Aβ induced reactive astrocytes and how it contributes to neuronal death. To achieve this aim, we will carry out the studies by using site-directed mutagenesis, protein purification, liposome reconstitution, and single-channel recording of TRPV4 to demonstrate the molecular pathway of Aβ activating TRPV4. We will investigate the expression of TRPV4 in reactive of astrocytes upon Aβ treatment by using RT-PCR, in situ hybridization, and Western blot analysis. Finally, we will demonstrate the role of TRPV4 in astrocytes [Ca2+]I and inflammation, and neuron death induced by Aβ.

瞬时感受器电位离子通道TRPV4受多种理化因素激活，参与Aβ诱导的海马培养神经元死亡。本研究旨在探索TRPV4 在Aβ诱导活化星形胶质细胞中被激活以及引起神经元死亡的机制。本研究将对TRPV4基因定点突变，通过将纯化的TRPV4蛋白或突变蛋白重构入脂质体，用单离子通道记录研究Aβ对TRPV4通道特性的影响，阐明Aβ在激活TRPV4中的作用机制；通过RT-PCR、原位杂交、免疫印迹等方法研究Aβ对星形胶质细胞中TRPV4表达的影响，通过钙成像、siRNA等方法，研究TRPV4在Aβ诱导的星形胶质细胞的钙离子浓度升高、炎症反应及神经元死亡中的作用。