Abstract
Purpose: :
MiRNAs are small non-coding RNA molecules with regulatory function and marked tissue specificity that can modulate multiple gene targets. They have been detected in body fluids and are associated with various physiologic and pathologic processes. We analyzed aqueous humor (AH) from human subjects undergoing cataract surgery to establish the presence and relative quantities of known miRNAs.
Methods: :
AH was collected from three patients without known ocular diseases other than cataract and a normal systemic history. Quantitative real time PCR was used to detect the different miRNAs present in the pooled AH in a microarray platform containing 257 known human miRNAs. Data was normalized and analyzed using BioRad iQ5 analysis software v2.1. Product melting temperature dissociation curves were used to validate positive calls and any reaction generating multiple peaks was excluded as negative.
Results: :
Ninety-one of 257 tested miRNAs were identified in the pooled AH. The top 5 abundant miRNAs were miR-412, miR-202, miR-486-5p, miR-671-3p, and miR-548a-5p. Four miRNAs, previously shown as unique to saliva, compared with other body fluids, were also detected in AH. In addition, the highly abundant miRNAs in other body fluids, such as miR335 and miR515, were also detected in the AH but at lower expression levels.
Conclusions: :
The presence of miRNAs in AH suggests that they may have functional roles in regulating target genes in tissues lining the anterior chamber. Further analysis of the AH miRNA population may identify potential gene targets and provide insights regarding their role in AH regulation, glaucoma and anterior segment disease processes.
Keywords: aqueous • gene modifiers • detection