Science Note
How Neurons Defend Against Mitochondrial Breakdown [Aug 7, 2025]
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Lipid Peroxidation from Mitochondrial Dysfunction Elicits Neuronal Protective Mechanisms Mitochondrial dysfunction in neurons commonly leads to oxidative damage through lipid peroxidation, threatening membrane integrity and cell viability. Neurons have evolved protective responses to oxidative stress, such as redox-sensing pathways that modulate excitability. A recent study found that mitochondrial fragmentation, increased mitophagy, and upregulation of ATP synthesis genes during wakefulness in sleep-promoting neurons cause lipid peroxidation, which is detected by redox sensors to trigger sleep. |
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Mitochondrial origins of the pressure to sleep (Nature, 2025) Highlighted technique: This study highlights mitophagy as a key mitochondrial response to sleep deprivation in neurons. To detect this, the authors used mito-QC, a genetically encoded fluorescent reporter that shows a red-only signal when mitochondria are delivered to acidic lysosomes, allowing in vivo visualization of mitophagy. Related technique Mitophagy Detection, Glycolysis/OXPHOS Measurement |
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Sleep pressure accumulates in a voltage-gated lipid peroxidation memory (Nature, 2025) Highlighted technique: This study detected lipid peroxidation in the brain using a technique called SMALDI-MSI, which creates high-resolution maps of different lipid types in tissue slices. The researchers found that certain vulnerable lipids, especially those with polyunsaturated fatty acids (PUFAs), were reduced after sleep deprivation, indicating oxidative damage. Related technique Lipid Peroxide Detection, Mitochondrial ROS Detection |
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Related Techniques (click to open/close)
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Application Note I (click to open/close)
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CCCP was added to Parkin-expressing HeLa cells and normal HeLa cells to confirm mitophagy. In the experiment, lysosomes and mitochondria were co-stained, and a strong mitophagic signal was observed on mitochondria and lysosomes in Parkin-expressing HeLa cells.
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Application Note II (click to open/close)
> Induction of Mitophagy in Parkin Expressed HeLa cells
| Induction of mitophagy by carbonyl cyanide m-chlorophenyl hydrazone (CCCP) as a mitochondrial-uncoupling reagent with Parkin expressed HeLa cells. | |
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HeLa cells were seeded on μ-slide 8 well (Ibidi) and cultured at 37oC overnight in a 5%-CO2 incubator. The cells were transfected with Parkin plasmid vector by HilyMax transfection reagent (Code#:H357), and incubated at 37oC overnight. The Parkin expressed HeLa cells were washed with Hanks’ HEPES buffer twice and then incubated at 37oC for 30 minutes with 250 μl of 100 nmol/l Mtphagy Dye working solution containing 100 nmol/l MitoBright LT Deep Red (※). After the washing of the cells with Hanks’ HEPES buffer twice, the culture medium containing 10 μmol/l CCCP was added to the well. After 24 hours incubation, mitophagy was observed by fluorescence microscopy. After removing the supernatant, 250 μl of 1 μmol/l Lyso Dye working solution were added to the cells and incubated at 37oC for 30 minutes. The cells were washed with Hanks’ HEPES buffer twice and then co-localization of Mtphagy, Lyso Dye and MitoBright Deep Red was observed by confocal fluorescence microscopy. Observation of mitophagy using Parkin ※Our sincere apology that we discontinued distributing MitoBright Deep Red.
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