Methods for the quantification of lysosomal membrane permeabilization : A hallmark of lysosomal cell death
Aits, Sonja LU
; Jäättelä, Marja
and Nylandsted, Jesper
(2015)
In Methods in Cell Biology
126.
p.261-285
- Abstract
- Lysosomal cell death is triggered by lysosomal membrane permeabilization (LMP) and subsequent release of lysosomal hydrolases from the lysosomal lumen into the cytosol. Once released into the cytosol, the lysosomal cathepsin proteases act as executioner proteases for the subsequent cell death-either autonomously without caspase activation or in concert with the classical apoptotic machinery. Lysosomal cell death usually remains functional in apoptosis-resistant cancer cells and thus holds great potential as a therapeutic strategy for circumventing apoptosis deficiency in cancers. Notably, lysosomal cell death also plays an important role in normal physiology, e.g., during the regression of the mammary gland. Here we present four... (More)
- Lysosomal cell death is triggered by lysosomal membrane permeabilization (LMP) and subsequent release of lysosomal hydrolases from the lysosomal lumen into the cytosol. Once released into the cytosol, the lysosomal cathepsin proteases act as executioner proteases for the subsequent cell death-either autonomously without caspase activation or in concert with the classical apoptotic machinery. Lysosomal cell death usually remains functional in apoptosis-resistant cancer cells and thus holds great potential as a therapeutic strategy for circumventing apoptosis deficiency in cancers. Notably, lysosomal cell death also plays an important role in normal physiology, e.g., during the regression of the mammary gland. Here we present four complementary methods for the quantification and visualization of LMP during the onset of death: (1) enzymatic activity measurements of released lysosomal hydrolases in the cytosol after digitonin extraction, (2) direct visualization of LMP by monitoring the release of fluorescent dextran from lysosomes into the cytosol, (3) immunocytochemistry to detect cathepsins released into the cytosol, and (4) detection of the translocation of galectins to damaged lysosomes. The methods presented here can ideally be combined as needed to provide solid evidence for LMP after a given cytotoxic stimuli. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/9bfcb808-d742-42ea-a6f8-a1baa35d0d5e
- author
- Aits, Sonja
LU
; Jäättelä, Marja
and Nylandsted, Jesper
- publishing date
- 2015
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- subject
- keywords
- Cathepsins, Death pathway, Galectins, LMP, Lysosomal cell death, Lysosomal membrane permeabilization, Method
- host publication
- Lysosomes and Lysosomal Diseases
- series title
- Methods in Cell Biology
- editor
- Platt, F. and Platt, N.
- volume
- 126
- pages
- 261 - 285
- external identifiers
-
- scopus:84922947461
- pmid:25665450
- ISSN
- 0091-679X
- ISBN
- 978-0-12-800079-3
- DOI
- 10.1016/bs.mcb.2014.10.032
- language
- English
- LU publication?
- no
- id
- 9bfcb808-d742-42ea-a6f8-a1baa35d0d5e
- date added to LUP
- 2018-03-05 15:15:41
- date last changed
- 2024-05-27 08:03:31
@inbook{9bfcb808-d742-42ea-a6f8-a1baa35d0d5e,
abstract = {{Lysosomal cell death is triggered by lysosomal membrane permeabilization (LMP) and subsequent release of lysosomal hydrolases from the lysosomal lumen into the cytosol. Once released into the cytosol, the lysosomal cathepsin proteases act as executioner proteases for the subsequent cell death-either autonomously without caspase activation or in concert with the classical apoptotic machinery. Lysosomal cell death usually remains functional in apoptosis-resistant cancer cells and thus holds great potential as a therapeutic strategy for circumventing apoptosis deficiency in cancers. Notably, lysosomal cell death also plays an important role in normal physiology, e.g., during the regression of the mammary gland. Here we present four complementary methods for the quantification and visualization of LMP during the onset of death: (1) enzymatic activity measurements of released lysosomal hydrolases in the cytosol after digitonin extraction, (2) direct visualization of LMP by monitoring the release of fluorescent dextran from lysosomes into the cytosol, (3) immunocytochemistry to detect cathepsins released into the cytosol, and (4) detection of the translocation of galectins to damaged lysosomes. The methods presented here can ideally be combined as needed to provide solid evidence for LMP after a given cytotoxic stimuli.}},
author = {{Aits, Sonja and Jäättelä, Marja and Nylandsted, Jesper}},
booktitle = {{Lysosomes and Lysosomal Diseases}},
editor = {{Platt, F. and Platt, N.}},
isbn = {{978-0-12-800079-3}},
issn = {{0091-679X}},
keywords = {{Cathepsins; Death pathway; Galectins; LMP; Lysosomal cell death; Lysosomal membrane permeabilization; Method}},
language = {{eng}},
pages = {{261--285}},
series = {{Methods in Cell Biology}},
title = {{Methods for the quantification of lysosomal membrane permeabilization : A hallmark of lysosomal cell death}},
url = {{http://dx.doi.org/10.1016/bs.mcb.2014.10.032}},
doi = {{10.1016/bs.mcb.2014.10.032}},
volume = {{126}},
year = {{2015}},
}