Genome-wide scleral micro-and messenger-RNA regulation during myopia development in the mouse

• Type of Material: Article
• Authors: Ravikanth Metlapally, UC Berkeley; Han Na Park, Emory University; Ranjay Chakraborty, Emory University; Kevin K. Wang, UC Berkeley; Christopher C. Tan, Emory University; Jacob G. Light, Emory University; Machelle Pardue, Emory University; Christine F. Wildsoet, UC Berkeley
• Language: English
• Date: 2016-11-01
• Publisher: Association for Research in Vision and Ophthalmology (ARVO)
• Publication Version: Final Published Version
• Copyright Statement: © 2016, Association for Research in Vision and Ophthalmology Inc. All rights reserved.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1167/iovs.16-19563
• Title of Journal or Parent Work: Investigative Ophthalmology & Visual Science
• ISSN: 0146-0404
• Volume: 57
• Issue: 14
• Start Page: 6089
• End Page: 6097
• Grant/Funding Information: Supported by US National Institutes of Health Grants NIH-NEI K12 EY017269 (RM) (Berkeley Clinical Scientist Development Program), NIH-NEI K08 EY022670 (RM), NIH-NEI R01 EY016435 (MTP), NIH-NEI R01 EY012392 (CFW).
• Keywords: mouse; sclera; microRNA; microarray; myopia
• Research Categories: Health Sciences, Opthamology; Engineering, Biomedical

Abstract

PURPOSE. MicroRNA (miRNAs) have been previously implicated in scleral remodeling in normal eye growth. They have the potential to be therapeutic targets for prevention/ retardation of exaggerated eye growth in myopia by modulating scleral matrix remodeling. To explore this potential, genome-wide miRNA and messenger RNA (mRNA) scleral profiles in myopic and control eyes from mice were studied. METHODS. C57BL/6J mice (n = 7; P28) reared under a 12L:12D cycle were form-deprived (FD) unilaterally for 2 weeks. Refractive error and axial length changes were measured using photorefraction and 1310-nm spectral-domain optical coherence tomography, respectively. Scleral RNA samples from FD and fellow control eyes were processed for microarray assay. Statistical analyses were performed using National Institute of Aging array analysis tool; group comparisons were made using ANOVA, and gene ontologies were identified using software available on the Web. Findings were confirmed using quantitative PCR in a separate group of mice (n = 7). RESULTS. Form-deprived eyes showed myopic shifts in refractive error (–2.02 ± 0.47 D; P < 0.01). Comparison of the scleral RNA profiles of test eyes with those of control eyes revealed 54 differentially expressed miRNAs and 261 mRNAs fold-change >1.25 (maximum fold change = 1.63 and 2.7 for miRNAs and mRNAs, respectively) (P < 0.05; minimum, P = 0.0001). Significant ontologies showing gene over-representation (P < 0.05) included intermediate filament organization, scaffold protein binding, detection of stimuli, calcium ion, G protein, and phototransduction. Significant differential expression of Let-7a and miR-16-2, and Smok4a, Prph2, and Gnat1 were confirmed. CONCLUSIONS. Scleral mi- and mRNAs showed differential expression linked to myopia, supporting the involvement of miRNAs in eye growth regulation. The observed general trend of relatively small fold-changes suggests a tightly controlled, regulatory mechanism for scleral gene expression.