Pedersen Stine Falsig, Nilius Bernd
Department of Molecular Biology, University of Copenhagen, Copenhagen, Denmark.
Methods Enzymol. 2007;428:183-207. doi: 10.1016/S0076-6879(07)28010-3.
Transient receptor potential (TRP) channels are unique cellular sensors responding to a wide variety of extra- and intracellular signals, including mechanical and osmotic stress. In recent years, TRP channels from multiple subfamilies have been added to the list of mechano- and/or osmosensitive channels, and it is becoming increasingly apparent that Ca(2+) influx via TRP channels plays a crucial role in the response to mechanical and osmotic perturbations in a wide range of cell types. Although the events translating mechanical and osmotic stimuli into regulation of TRP channels are still incompletely understood, the specific mechanisms employed vary between different TRP isoforms, and probably include changes in the tension and/or curvature of the lipid bilayer, changes in the cortical cytoskeleton, and signaling events such as lipid metabolism and protein phosphorylation/dephosphorylation. This chapter describes candidate mechanosensitive channels from mammalian TRP subfamilies, discusses inherent and technical issues potentially confounding evaluation of mechano- and/or osmosensitivity, and presents methods relevant to the study of TRP channel regulation by mechanical and osmotic stimuli and involvement in cell volume regulation.
瞬时受体电位(TRP)通道是独特的细胞传感器,可响应多种细胞外和细胞内信号,包括机械和渗透压应激。近年来,多个亚家族的TRP通道已被列入机械敏感和/或渗透压敏感通道名单,并且越来越明显的是,通过TRP通道的Ca(2+)内流在多种细胞类型对机械和渗透压扰动的反应中起关键作用。尽管将机械和渗透压刺激转化为TRP通道调节的事件仍未完全了解,但不同TRP同工型采用的具体机制有所不同,可能包括脂质双层张力和/或曲率的变化、皮质细胞骨架的变化以及脂质代谢和蛋白质磷酸化/去磷酸化等信号事件。本章描述了来自哺乳动物TRP亚家族的候选机械敏感通道,讨论了可能混淆机械敏感性和/或渗透压敏感性评估的内在和技术问题,并介绍了与研究TRP通道受机械和渗透压刺激调节以及参与细胞体积调节相关的方法。