胞浆
内吞作用
化学
膜
溶酶体
生物物理学
细胞生物学
细胞外
瞬时受体电位通道
细胞内
细胞器
生物化学
生物
受体
酶
作者
Christian Wahl‐Schott,Marc Freichel,Konstantin Hennis,Koenraad Philippaert,Roger Ottenheijm,Volodymyr Tsvilovskyy,Hristo Varbanov
出处
期刊:Handbook of experimental pharmacology
日期:2023-01-01
卷期号:: 277-304
被引量:1
摘要
Endo-lysosomes are membrane-bound acidic organelles that are involved in endocytosis, recycling, and degradation of extracellular and intracellular material. The membranes of endo-lysosomes express several Ca2+-permeable cation ion channels, including two-pore channels (TPC1-3) and transient receptor potential mucolipin channels (TRPML1-3). In this chapter, we will describe four different state-of-the-art Ca2+ imaging approaches, which are well-suited to investigate the function of endo-lysosomal cation channels. These techniques include (1) global cytosolic Ca2+ measurements, (2) peri-endo-lysosomal Ca2+ imaging using genetically encoded Ca2+ sensors that are directed to the cytosolic endo-lysosomal membrane surface, (3) Ca2+ imaging of endo-lysosomal cation channels, which are engineered in order to redirect them to the plasma membrane in combination with approaches 1 and 2, and (4) Ca2+ imaging by directing Ca2+ indicators to the endo-lysosomal lumen. Moreover, we will review useful small molecules, which can be used as valuable tools for endo-lysosomal Ca2+ imaging. Rather than providing complete protocols, we will discuss specific methodological issues related to endo-lysosomal Ca2+ imaging.
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