A novel quantification method for RPE phagocytosis using a VLC-PUFA-based strategy
Gao, F.; Tom, E.; Skowronska-Krawczyk, D.
Show abstract
The vertebrate retinal pigment epithelium (RPE) lies adjacent to the photoreceptors and is responsible for the engulfment and degradation of shed photoreceptor outer segment fragments (POS) through receptor-mediated phagocytosis. Phagocytosis of POS is critical for maintaining photoreceptor function and is a key indicator of RPE functionality. Popular established methods to assess RPE phagocytosis rely mainly on quantifying POS proteins, especially their most abundant protein rhodopsin, or on fluorescent dye conjugation of bulk, unspecified POS components. While these approaches are practical and quantitative, they fail to assess the fate of POS lipids, which make up about 50% of POS by weight and whose processing is essential for life-long functionality of RPE and retina. Here, we have developed a novel VLC-PUFA-based approach for evaluating RPE phagocytic activity by primary bovine and rat RPE and the human ARPE-19 cell line and validated its results using traditional methods. This new approach can be used to detect the dynamic process of phagocytosis at varying POS concentrations and incubation times and offers a robust, unbiased, and reproducible assay that will have utility in studies of POS lipid processing.
Matching journals
The top 5 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Geographic characterization of RPE structure and lipid changes in the PEX1-p.Gly844Asp mouse model for Zellweger spectrum disorder. 95%
- Fatty Acyl Availability Modulates Cardiolipin Composition and Alters Mitochondrial Function in HeLa Cells 94%
- Quantification of esterified oxylipins following HILIC-fractionation of lipid classes 93%
Similar papers in this journal
- Reassessing Retinal Pigment Epithelial Ketogenesis: Enzymatic Assays for Ketone Body Levels Provide Inaccurate Results 95%
- Comparative Proteomic Study of Retinal Ganglion Cells Undergoing Various Types of Cellular Stressors 94%
- Age-related macular degeneration-like phenotypic features develop at the early ages of Cxcr5/Nrf2 double knockout mice: An accelerated AMD model 92%
Similar papers in this journal
- Lysophosphatidylcholine acyltransferase 1 controls the mitochondrial reactive oxygen species generation and survival of the retinal photoreceptor cells 95%
- Proline provides a nitrogen source in the retinal pigment epithelium to synthesize and export amino acids for the neural retina 94%
- Pyruvate dehydrogenase kinase 1 controls triacylglycerol hydrolysis in cardiomyocytes 93%
Similar papers in this journal
- DHA shortage causes the early degeneration of photoreceptors and RPE in mice with peroxisomal β-oxidation deficiency 94%
- Spatially-Resolved Proteomic Analysis of the Lens Extracellular Diffusion Barrier 93%
- Intermittent hypoxia promotes functional neuroprotection from retinal ischemia in untreated first-generation offspring 93%
Similar papers in this journal
- Microsomal triglyceride transfer protein is necessary to maintain lipid homeostasis and retinal function 94%
- Inhibition of hypoxia-inducible factors suppresses subretinal fibrosis 93%
- Lipid storm within the lungs of severe COVID-19 patients: Extensive levels of cyclooxygenase and lipoxygenase-derived inflammatory metabolites 92%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.