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Sensitive detection of lysosomal membrane permeabilization by lysosomal galectin puncta assay

Aits, Sonja LU ; Kricker, Jennifer; Liu, Bin; Ellegaard, Anne Marie; Hämälistö, Saara; Tvingsholm, Siri; Corcelle-Termeau, Elisabeth; Høgh, Søren; Farkas, Thomas and Jonassen, Anna Holm, et al. (2015) In Autophagy 11(8). p.1408-1424
Abstract

Lysosomal membrane permeabilization (LMP) contributes to tissue involution, degenerative diseases, and cancer therapy. Its investigation has, however, been hindered by the lack of sensitive methods. Here, we characterize and validate the detection of galectin puncta at leaky lysosomes as a highly sensitive and easily manageable assay for LMP. LGALS1/galectin-1 and LGALS3/ galectin-3 are best suited for this purpose due to their widespread expression, rapid translocation to leaky lysosomes and availability of high-affinity antibodies. Galectin staining marks individual leaky lysosomes early during lysosomal cell death and is useful when defining whether LMP is a primary or secondary cause of cell death. This sensitive method also reveals... (More)

Lysosomal membrane permeabilization (LMP) contributes to tissue involution, degenerative diseases, and cancer therapy. Its investigation has, however, been hindered by the lack of sensitive methods. Here, we characterize and validate the detection of galectin puncta at leaky lysosomes as a highly sensitive and easily manageable assay for LMP. LGALS1/galectin-1 and LGALS3/ galectin-3 are best suited for this purpose due to their widespread expression, rapid translocation to leaky lysosomes and availability of high-affinity antibodies. Galectin staining marks individual leaky lysosomes early during lysosomal cell death and is useful when defining whether LMP is a primary or secondary cause of cell death. This sensitive method also reveals that cells can survive limited LMP and confirms a rapid formation of autophagic structures at the site of galectin puncta. Importantly, galectin staining detects individual leaky lysosomes also in paraffin-embedded tissues allowing us to demonstrate LMP in tumor xenografts in mice treated with cationic amphiphilic drugs and to identify a subpopulation of lysosomes that initiates LMP in involuting mouse mammary gland. The use of ectopic fluorescent galectins renders the galectin puncta assay suitable for automated screening and visualization of LMP in live cells and animals. Thus, the lysosomal galectin puncta assay opens up new possibilities to study LMP in cell death and its role in other cellular processes such as autophagy, senescence, aging, and inflammation.

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type
Contribution to journal
publication status
published
subject
keywords
Breast involution, C. elegans, Cell death, Galectin, Lysosome
in
Autophagy
volume
11
issue
8
pages
1408 - 1424
publisher
Landes Bioscience
external identifiers
  • scopus:84943747270
ISSN
1554-8627
DOI
10.1080/15548627.2015.1063871
language
English
LU publication?
no
id
03fb0084-6cbd-45fd-b50d-1c740dcec845
date added to LUP
2018-03-05 15:23:28
date last changed
2018-11-18 05:03:28
@article{03fb0084-6cbd-45fd-b50d-1c740dcec845,
  abstract     = {<p>Lysosomal membrane permeabilization (LMP) contributes to tissue involution, degenerative diseases, and cancer therapy. Its investigation has, however, been hindered by the lack of sensitive methods. Here, we characterize and validate the detection of galectin puncta at leaky lysosomes as a highly sensitive and easily manageable assay for LMP. LGALS1/galectin-1 and LGALS3/ galectin-3 are best suited for this purpose due to their widespread expression, rapid translocation to leaky lysosomes and availability of high-affinity antibodies. Galectin staining marks individual leaky lysosomes early during lysosomal cell death and is useful when defining whether LMP is a primary or secondary cause of cell death. This sensitive method also reveals that cells can survive limited LMP and confirms a rapid formation of autophagic structures at the site of galectin puncta. Importantly, galectin staining detects individual leaky lysosomes also in paraffin-embedded tissues allowing us to demonstrate LMP in tumor xenografts in mice treated with cationic amphiphilic drugs and to identify a subpopulation of lysosomes that initiates LMP in involuting mouse mammary gland. The use of ectopic fluorescent galectins renders the galectin puncta assay suitable for automated screening and visualization of LMP in live cells and animals. Thus, the lysosomal galectin puncta assay opens up new possibilities to study LMP in cell death and its role in other cellular processes such as autophagy, senescence, aging, and inflammation.</p>},
  author       = {Aits, Sonja and Kricker, Jennifer and Liu, Bin and Ellegaard, Anne Marie and Hämälistö, Saara and Tvingsholm, Siri and Corcelle-Termeau, Elisabeth and Høgh, Søren and Farkas, Thomas and Jonassen, Anna Holm and Gromova, Irina and Mortensen, Monika and Jäättelä, Marja},
  issn         = {1554-8627},
  keyword      = {Breast involution,C. elegans,Cell death,Galectin,Lysosome},
  language     = {eng},
  month        = {06},
  number       = {8},
  pages        = {1408--1424},
  publisher    = {Landes Bioscience},
  series       = {Autophagy},
  title        = {Sensitive detection of lysosomal membrane permeabilization by lysosomal galectin puncta assay},
  url          = {http://dx.doi.org/10.1080/15548627.2015.1063871},
  volume       = {11},
  year         = {2015},
}