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Zeljka Smit-McBride, Sharon L. Oltjen, Leonard M. Hjelmeland; MicroRNA Expression In The Aging Mouse RPE/choroid. Invest. Ophthalmol. Vis. Sci. 2011;52(14):16.
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© ARVO (1962-2015); The Authors (2016-present)
MicroRNAs are important regulators of many cellular functions due to their ability to target mRNAs for translational inhibition and/or degradation. In studies of human fetal RPE cells miR-204/211 were identified as crucial regulators of epithelial barrier function and cell physiology, while in ARPE-19 cells it was demonstrated that miR-9 was regulated by retinoic acid and reactive oxygen species, indicating that miR-9 could be important in maintaining RPE function. To investigate potential age-related regulation of miRNAs in the RPE, we conducted a survey of 375 mouse miRNAs using real-time PCR analysis.
Eighteen month old C57BL/6J males were obtained from the NIA, and two month old males from The Jackson Laboratory. Research was conducted in compliance with the ARVO guide to the care and use of lab animals. Total RNA from the RPE/choroid was extracted and purified using standard methods. Samples of total RNA were then analyzed by Exiqon, Inc. using the miRCURY LNA Universal RT miRNA Mouse and Rat PCR panel I.
Out of the 375 miRNA assays 268 were successfully assessed with sufficient signal (Cp < 37, or 5 Cp less than negative control) in all RPE/choroid samples. MiR-204, miR-211, and miR-9 were all detected (Cp=20-23). Using a ddCp threshold of 1.5, 19 miRNAs were upregulated with age and 20 miRNAs were downregulated using the same criterion. The expression of previously identified miR-204/211 and miR-9 was not altered with age. MiRNAs related to apoptosis (miR-96, miR-34a, and miR-29b), inflammation (miR-20b), APP (miR-298), atherosclerosis (miR-27b) and aging (miR-106a) showed the biggest expression changes.
Our study validated previous observations in the literature of the expression of miR-204, miR-211, and miR-9 in the RPE. In addition, we demonstrated age-related regulation of miRNAs which are important mediators of apoptosis, inflammation and aging.
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