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A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae
Published on: July 18, 2011
Autophagic activity in BC3H1 cells exposed to yessotoxin
Mónica Suárez Korsnes1, Hilde Kolstad2, Charlotte Ramstad Kleiveland3
1Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU) - Campus Ås, P.O. Box 5003, NO-1432 Ås, Norway.
Abstract:
The marine toxin yessotoxin (YTX) can induce programmed cell death through both caspase-dependent and -independent pathways in various cellular systems. It appears to stimulate different forms of cellular stress causing instability among cell death mechanisms and making them overlap and cross-talk. Autophagy is one of the key pathways that can be stimulated by multiple forms of cellular stress which may determine cell survival or death. The present work evaluates a plausible link between ribotoxic stress and autophagic activity in BC3H1 cells treated with YTX. Such treatment produces massive cytoplasmic compartments as well as double-membrane vesicles termed autophagosomes which are typically observed in cells undergoing autophagy. The observed autophagosomes contain a large amount of ribosomes associated with the endoplasmic reticulum (ER). Western blotting analysis of Atg proteins and detection of the autophagic markers LC3-II and SQSTM1/p62 by flow cytometry and immunofluorescence verified autophagic activity during YTX-treatment. The present work supports the idea that autophagic activity upon YTX exposure may represent a response to ribotoxic stress.
Insights
Marine toxin yessotoxin (YTX) induces cell death by activating autophagy, a cellular stress response. This study links YTX-induced ribotoxic stress to significant autophagosome formation in BC3H1 cells.
Area of Science:
- Cell Biology
- Toxicology
- Molecular Biology
Background:
- Marine toxin yessotoxin (YTX) induces programmed cell death via caspase-dependent and -independent pathways.
- YTX exposure causes cellular stress, leading to overlapping and cross-talking cell death mechanisms.
- Autophagy is a critical cellular pathway modulated by stress, influencing cell survival or death.
Purpose of the Study:
- To investigate the potential link between ribotoxic stress and autophagic activity in BC3H1 cells exposed to YTX.
- To elucidate the role of autophagy as a cellular response to YTX-induced stress.
Main Methods:
- BC3H1 cells were treated with yessotoxin (YTX).
- Morphological analysis was performed to identify autophagosomes and ribosomes associated with the endoplasmic reticulum (ER).
- Western blotting, flow cytometry, and immunofluorescence were used to detect autophagy-related (Atg) proteins and markers like LC3-II and SQSTM1/p62.
Main Results:
- YTX treatment resulted in the formation of cytoplasmic compartments and autophagosomes.
- Autophagosomes containing ribosomes attached to the ER were observed.
- Verification of autophagic activity was confirmed through the detection of key autophagic markers and proteins.
Conclusions:
- YTX exposure stimulates autophagic activity in BC3H1 cells.
- Autophagy activation in response to YTX may serve as a protective mechanism against ribotoxic stress.
- This study supports the role of autophagy in mediating cellular responses to marine toxin-induced stress.
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