PUFAs in the Pathways of Cell Fate [Jun 10, 2025] DOJINDO LABORATORIES

Previous Science Note

Polyunsaturated fatty acids (PUFAs) influence cell survival by altering membrane lipid composition, with recent studies showing that changes in PUFA levels and trafficking can either protect neurons from stress or increase cancer cell sensitivity to ferroptosis. This Science Note introduces three studies that uncover how PUFA-containing phospholipids regulate cell death through mitochondrial interactions, lipid remodeling, and tissue-specific lipid depletion.

Neuronal polyunsaturated fatty acids are protective in ALS/FTD (Nature Neuroscience, 2025)

Summary: Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia neurons are depleted of protective polyunsaturated fatty acids (PUFA). Replenishing these fatty acids through diet and by boosting their production inside the cells markedly prolonged the survival of both fly and human neuron models, suggesting a simple protective strategy.

Highlighted technique: This study used iPSC-derived spinal neurons from ALS/FTD patients to test whether increasing lipid unsaturation could protect against neurotoxicity. Overexpressing desaturase enzymes raised PUFA levels and significantly reduced glutamate-induced neuronal death, showing a protective effect in this human disease model.

Related technique Apoptosis Detection

Phospholipids with two polyunsaturated fatty acyl tails promote ferroptosis (Cell, 2024)

Summary: This study found that certain fatty acid treatments cause cells to build up PC-PUFA2, a lipid that binds mitochondria, generates reactive oxygen, and increases ferroptosis sensitivity. PC-PUFA2 was reduced in aging and Huntington disease brains, and mitochondrial antioxidants blocked its damaging effects.

Highlighted technique: The researchers treated cancer cells with PC-PUFA2 lipids and measured ferroptosis sensitivity using lipid peroxidation and viability assays. They further quantified mitochondrial ROS and membrane potential changes, showing that PC-PUFA2 triggers ferroptosis through mitochondrial oxidative stress.

Related technique Lipid Peroxide Detection (used in this article), OCR Assay

Lipid availability influences ferroptosis sensitivity in cancer cells by regulating polyunsaturated fatty acid trafficking (Cell Chemical Biology, 2025)

Summary: When cancer cells are starved of external lipids, they channel PUFAs into membrane phospholipids, which increases their sensitivity to ferroptosis. This suggests that reducing lipid availability around tumors could enhance PUFA driven ferroptosis based cancer therapies.

Highlighted technique: The authors performed LC–MS/MS lipidomics using isotope-labeled PUFA to trace their incorporation into distinct phospholipid classes under lipid-starved conditions. They then assessed ferroptosis sensitivity by combining cell viability assays with lipid peroxidation staining after treatment with ferroptosis inducers.

Related technique Lipid Peroxidation Assay, Cell Proliferation Assay

Related Techniques (click to open/close)

Target	Kit & Probes
Apoptosis detection in multiple samples	Annexin V Apoptosis Plate Assay Kit
Extracellular ATP measurement	Extracellular ATP Assay Kit-Luminescence
Lipid peroxidation detection	Liperfluo(Intracellular), MitoPeDPP(Mitochondrial)
Lipid Peroxidation Assay	Lipid Peroxidation Probe -BDP 581/591 C11-
Mitochondrial superoxide detection	MitoBright ROS Deep Red - Mitochondrial Superoxide Detection
Total ROS detection	Highly sensitive DCFH-DA or Photo-oxidation Resistant DCFH-DA
Ferrous ion (Fe2+) detection	FerroOrange(Intracellular), Mito-FerroGreen(Mitochondrial)
Oxygen consumption rate assay	Extracellular OCR Plate Assay Kit
Mitochondrial membrane potential detection	JC-1 MitoMP Detection Kit, MT-1 MitoMP Detection Kit
Glutathione Quantification	GSSG/GSH Quantification Kit
Cystine Uptake detection	Cystine Uptake Assay Kit
Cell proliferation/ cytotoxicity assay	Cell Counting Kit-8 and Cytotoxicity LDH Assay Kit-WST

Application Note (click to open/close)
> Changes in various indicators of cell death induced by drugs

	HepG2 cells treated with the apoptosis-inducing agent staurosporine or the ferroptosis-inducing agents Erastin and RSL3. After treatment, extracellular LDH, phosphatidylserine, cell viability, intracellular Fe²⁺ and lipid peroxidation were determined. The results showed that apoptosis-induced cells treated with staurosporine showed an increase in phosphatidylserine, a decrease in cell viability and an increase in extracellular LDH, indicating that cell death had occurred. On the other hand, intracellular Fe²⁺, an indicator of ferroptosis, remained unchanged. In cells treated with Erastin, a ferroptosis inducer, intracellular Fe²⁺ increased and cell viability decreased, but extracellular LDH and lipid peroxidation (lipid peroxidation: decrease in red fluorescence and increase in green fluorescence) did not increase. In cells in which ferroptosis was more strongly induced by co-treatment with RSL3 in addition to Erastin, increased intracellular Fe²⁺ and lipid peroxidation were observed. Moreover, decreased cell viability and increased dead cells were detected. Meanwhile, phosphatidylserine showed a lower rate of increase during ferroptosis induction compared to apoptosis-induced cells. These results suggest that cell death can be distinguished by evaluating a combination of cell death indicators.