Dataset Information

Effect of dopamine on bone morphogenesis protein-2 expression in human retinal pigment epithelium.

ABSTRACT:

Aim

To investigate the effect of dopamine on bone morphogenesis protein-2 (BMP-2) expression in retinal pigment epithelium (RPE) cells in vitro.

Methods

ARPE-19 cells as a human RPE cell line were cultured with dopamine for different times (2, 4, 6, 8, 12, 16 and 24h) or with different concentrations (0.1, 1, 2, 5, 10, 20, and 100 µg/mL) in vitro. BMP-2 mRNA expression level in ARPE-19 cells was analyzed with real-time polymerase chain reaction (PCR) analysis and BMP-2 protein level was measured with Western blot analysis. The active form of BMP-2 in the culture medium was measured with enzyme-linked immunosorbent assay (ELISA).

Results

The expression level of BMP-2 increased significantly cultured with 20 µg/mL dopamine, at different time points (P<0.05). BMP-2 mRNA level peaked 2h and the protein level peaked at 6 and 8h after treatment. The concentrations of secreted BMP-2 elevated at 12h and peaked at 24h (P<0.05) in a time-dependent manner. Treated with 100 µg/mL dopamine for 6h, the expression levels of BMP-2 mRNA and protein in ARPE-19 cells were enhanced significantly compared to that in the untreated cells (P<0.05). And secreted BMP-2 protein in the cell culture supernatant was also increased (P<0.05).

Conclusion

Dopamine up-regulate BMP-2 expression in RPE cells, and this may be associated with its inhibitive effect on myopia development.

SUBMITTER: Li HH

PROVIDER: S-EPMC5596220 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Publications

Effect of dopamine on bone morphogenesis protein-2 expression in human retinal pigment epithelium.

Li Hong-Hui HH Sun Yan-Li YL Cui Dong-Mei DM Wu Juan J Zeng Jun-Wen JW

International journal of ophthalmology 20170918 9

<h4>Aim</h4>To investigate the effect of dopamine on bone morphogenesis protein-2 (BMP-2) expression in retinal pigment epithelium (RPE) cells <i>in vitro</i>.<h4>Methods</h4>ARPE-19 cells as a human RPE cell line were cultured with dopamine for different times (2, 4, 6, 8, 12, 16 and 24h) or with different concentrations (0.1, 1, 2, 5, 10, 20, and 100 µg/mL) <i>in vitro</i>. BMP-2 mRNA expression level in ARPE-19 cells was analyzed with real-time polymerase chain reaction (PCR) analysis and BMP ...[more]

PMID: 28944194

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Project description:PURPOSE: The molecular mechanisms associated with human retinal pigment epithelium (RPE) development constitute the basis for cell replacement therapy for the treatment of retinal degenerative diseases. In the current study, gene expression analysis of the human fetal RPE during development was performed and was compared with the human native RPE. METHODS: Microdissection of the human RPE at three time points (13 weeks and 16 weeks of gestation and in mature adult eyes) was performed, and total RNA was isolated. Equal amounts of RNA were pooled from two or three independent donor eyes for each time point in each group. Gene expression was analyzed by hybridization to microarray chips. Validation was accomplished by comparing the microarray expression pro?les with quantitative real-time reverse transcriptase-polymerase chain reaction (qRT(2)-PCR) analysis of selected genes and by comparing selected expression pro?les with predicted pro?les based on previous studies. RESULTS: Of the 45,033 probe sets on the microarray, 30,736 were detected. A total of 3,498 differentially expressed genes could be clustered into eight patterns of expression that were statistically significant. Analysis of the expression patterns of genes coding for key functions (pigment synthesis, visual cycle, phagocytosis, adherens and tight junctions, and transcellular transport) indicated that the human RPE achieves a high degree of maturity during early pregnancy. Compared with 154 signature genes in the RPE, 148 candidate genes were identified in this study, including 53 downregulated genes and 5 upregulated genes. The qRT(2)-PCR results showed similar expression trends to those obtained by microarray analysis at the three time points. CONCLUSIONS: This study demonstrated gene expression profiles in the human RPE during normal development. These ?ndings indicate that the human RPE has different expression patterns than those of other animals. The results of this study may be helpful in furthering the understanding of the developmental processes occurring in humans and of the differentiation of RPE cells derived from human embryonic stem cells and from human induced pluripotent stem cells.

Project description:BACKGROUND: To determine level, variability and functional annotation of gene expression of the human retinal pigment epithelium (RPE), the key tissue involved in retinal diseases like age-related macular degeneration and retinitis pigmentosa. Macular RPE cells from six selected healthy human donor eyes (aged 63-78 years) were laser dissected and used for 22k microarray studies (Agilent technologies). Data were analyzed with Rosetta Resolver, the web tool DAVID and Ingenuity software. RESULTS: In total, we identified 19,746 array entries with significant expression in the RPE. Gene expression was analyzed according to expression levels, interindividual variability and functionality. A group of highly (n = 2,194) expressed RPE genes showed an overrepresentation of genes of the oxidative phosphorylation, ATP synthesis and ribosome pathways. In the group of moderately expressed genes (n = 8,776) genes of the phosphatidylinositol signaling system and aminosugars metabolism were overrepresented. As expected, the top 10 percent (n = 2,194) of genes with the highest interindividual differences in expression showed functional overrepresentation of the complement cascade, essential in inflammation in age-related macular degeneration, and other signaling pathways. Surprisingly, this same category also includes the genes involved in Bruch's membrane (BM) composition. Among the top 10 percent of genes with low interindividual differences, there was an overrepresentation of genes involved in local glycosaminoglycan turnover. CONCLUSION: Our study expands current knowledge of the RPE transcriptome by assigning new genes, and adding data about expression level and interindividual variation. Functional annotation suggests that the RPE has high levels of protein synthesis, strong energy demands, and is exposed to high levels of oxidative stress and a variable degree of inflammation. Our data sheds new light on the molecular composition of BM, adjacent to the RPE, and is useful for candidate retinal disease gene identification or gene dose-dependent therapeutic studies.

Dataset Information

Effect of dopamine on bone morphogenesis protein-2 expression in human retinal pigment epithelium.

Aim

Methods

Results

Conclusion

Publications

Effect of dopamine on bone morphogenesis protein-2 expression in human retinal pigment epithelium.

Similar Datasets

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