Abstract
Purpose:
MicroRNAs (miRNAs) are a group of small non-coding RNAs, and regulate gene expression at the posttranscriptional level. We generated a corneal endothelial miRNA expression profile of a transgenic Col8a2Q455K/Q455K mouse model of early-onset Fuchs endothelial corneal dystrophy (FECD) to analyze the role of differential miRNA expression in FECD pathogenesis.
Methods:
Clinical confocal microscopy was performed in Col8a2Q455K/Q455K mutant (MUT) and wild-type (WT) mice. Endothelial miRNA expression was analyzed in three respective groups of ten month-old old MUT and WT mice using Taqman Array Rodent MicroRNA A+B cards v3.0 (Applied Biosystems) and the ExpressionSuite version 1.0 software package (Applied Biosystems). Differential expression of individual miRNAs was validated using stem-loop RT qPCR.
Results:
Clinical confocal microscopy confirmed the FECD and normal phenotype of MUT and WT mice. Taqman array analysis demonstrated differential expression of at least 55 miRNAs in MUT compared to WT mice applying 1.5-fold change and p<0.05 as cut-off criteria. Validation experiments using individual stem-loop RT qPCR assays (mmu-miR-362-5p, mmu-miR-10a, mmu-miR-34a, mmu-miR-532-5p, mmu-miR-34c, mmu-miR-322, mmu-miR-320 and mmu-miR-218-1#) showed good consistency with the initial array experiment.
Conclusions:
The present study provides the first miRNA profile in a mouse model of FECD and may serve as a helpful resource for further studies of the disease. Overlap and differences to a previously generated endothelial miRNA expression profile in human FECD are discussed.
Keywords: 481 cornea: endothelium