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Fluorescence Methods to Measure Pexophagy.

Francesco G Barone^1,2, Sylvie Urbé¹, Michael J Clague³

¹Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, UK.

Methods in Molecular Biology (Clifton, N.J.)

|August 8, 2024

Summary

This summary is machine-generated.

We present a method to study pexophagy (selective autophagy of peroxisomes) using fluorescence microscopy and specialized reporters. This approach allows quantitative assessment in various models, aiding research into cellular degradation processes.

Keywords:

Autophagy Fluorescence microscopy Keima-SKL Peroxisomes Pexo-QC Pexophagy

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Monitoring Stub1-Mediated Pexophagy

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Published on: May 12, 2023

Area of Science:

Cell Biology
Molecular Biology
Autophagy Research

Background:

Selective autophagy is a crucial cellular process for maintaining homeostasis.
Pexophagy, the selective degradation of peroxisomes, plays a role in cellular quality control.
Understanding pexophagy mechanisms is vital for various physiological and pathological conditions.

Purpose of the Study:

To describe a quantitative fluorescence microscopy approach for studying pexophagy.
To introduce ratiometric reporters for precise measurement of pexophagy.
To explore inducers of pexophagy and their relationship with mitophagy.

Main Methods:

Utilizing fluorescence microscopy in tissue cell culture models.
Employing ratiometric reporters that specifically localize to peroxisomes.
Assessing pexophagy in fixed and live cells, and whole organisms.

Main Results:

Developed a quantitative method for assessing pexophagy.
Demonstrated the utility of ratiometric reporters for pexophagy studies.
Identified chemical and physiological inducers of pexophagy.

Conclusions:

The described approach provides a robust tool for quantitative pexophagy analysis.
Ratiometric reporters enable precise monitoring of pexophagy across different biological systems.
Further investigation into pexophagy inducers may reveal therapeutic targets.