Chapter title |
Characterization of Endo-Lysosomal Cation Channels Using Calcium Imaging.
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---|---|
Chapter number | 637 |
Book title |
Endolysosomal Voltage-Dependent Cation Channels
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Published in |
Handbook of experimental pharmacology, March 2023
|
DOI | 10.1007/164_2023_637 |
Pubmed ID | |
Book ISBNs |
978-3-03-131522-0, 978-3-03-131523-7
|
Authors |
Wahl-Schott, Christian, Freichel, Marc, Hennis, Konstantin, Philippaert, Koenraad, Ottenheijm, Roger, Tsvilovskyy, Volodymyr, Varbanov, Hristo |
Abstract |
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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