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Day-night rhythm in murine retinal pigment epithelium

ABSTRACT: Day-night rhythm in murine retinal pigment epithelium

PROVIDER: PRJNA564263 | ENA |

REPOSITORIES: ENA

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Day-night rhythm in murine retinal pigment epithelium

Project description:Murine RPE transcriptomes were generated for 6 time points during a 24 hour period. Our data demonstrates that genes involved in ATP generation have a high expression at night whereas genes involved in β-oxidation and cholesterol synthesis show a high expression in the morning. Additionally, genes were identified in the glycerophospholipid metabolism pathway, suggesting the generation of glycerophospholipids at night. Finally, genes involved in intracellular trafficking with different expression patterns during the day-night cycle were identified and we identified genes involved in actin cytoskeleton building, remodelling and crosslinking with a high expression in the morning.

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The circadian clock in the retinal pigment epithelium controls the diurnal rhythm of phagocytic activity

Project description:The circadian clock in the retinal pigment epithelium controls the diurnal rhythm of phagocytic activity

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Functional Retinal Pigment Epithelium-like Cells from Human Fibroblasts

Project description:The retinal pigment epithelium (RPE) provides vital support to photoreceptor cells and its dysfunction is associated with the onset and progression of age-related macular degeneration (AMD). Surgical provision of RPE cells may ameliorate AMD and thus it would be valuable to develop sources of patient-matched RPE cells for this application of regenerative medicine. We describe here the generation of functional RPE-like cells from fibroblasts that represent an important step toward that goal. We identified candidate master transcriptional regulators of RPEs using a novel computational method and then used these regulators to guide exploration of the transcriptional regulatory circuitry of RPE cells and to reprogram human fibroblasts into RPE-like cells. The RPE-like cells share key features with RPEs derived from healthy individuals, including morphology, gene expression and function, and thus represent a step toward the goal of generating patient-matched RPE cells for treatment of macular degeneration. Expression analysis was performed on induced retinal pigment epithelium-like cells.

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Project description:The diurnal peak of phagocytosis by the retinal pigment epithelium (RPE) of photoreceptor outer segments is under circadian control, and it is believed that this process involves interactions from both the retina and RPE. Previous studies have demonstrated that a functional circadian clock exists within multiple retinal cell types and RPE cells. Thereby, the aim of the current study was to determine whether the circadian clock in the retina and or RPE controls the diurnal phagocytic peak of photoreceptor outer segments and whether selective disruption of the circadian clock in the RPE would affect RPE cells function and the viability during aging. To that aim, we first generated and validated an RPE tissue-specific KO of the essential clock gene, Bmal1, and then we determined the daily rhythm in phagocytic activity by the RPE in mice lacking a functional circadian clock in the retina or RPE. Then using electroretinography, spectral domain-optical coherence tomography, and optomotor response measurements of visual function we determined the effect of Bmal1 removal in young (6-month-old) and old (18-months old) mice. RPE morphology and lipofuscin accumulation was also determined in young and old mice. Our data show that the circadian clock in the RPE controls the daily diurnal phagocytic peak of POS. Surprisingly, the lack of a functional RPE circadian clock or the diurnal phagocytic peak does not result in any detectable age-related degenerative phenotype in the retina or RPE. Thus, our results demonstrate that the circadian clock in the RPE controls the daily peak in the phagocytic activity. However, the loss of the circadian clock in the RPE does not result in deterioration of photoreceptors or the RPE during aging.

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Oxidative stress alters transcript localization of disease-causing genes in the retinal pigment epithelium

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