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Exp Eye Res. 2015 Mar;132:73-7. doi: 10.1016/j.exer.2015.01.019. Epub 2015 Jan 22.

Human aqueous humor exosomes.

Author information

1
Department of Ophthalmology, Duke University Medical Center, Durham, NC, USA.
2
Department of Ophthalmology, Duke University Medical Center, Durham, NC, USA; Department of Biomedical Engineering, Duke University Medical Center, Durham, NC, USA.
3
Department of Cellular Biology and Anatomy, Medical College of Georgia, Georgia Regents University, Augusta, GA, USA; James and Jean Culver Vision Discovery Institute, Georgia Regents University, Augusta, GA, USA. Electronic address: yutliu@gru.edu.

Abstract

Aqueous humor (AH) is a dynamic intraocular fluid that supports the vitality of tissues that regulate intraocular pressure. We recently discovered that extracellular nanovesicles called exosomes are a major constituent of AH. Exosomes function in extracellular communication and contain proteins and small RNA. Our goal was to characterize the physical properties of AH exosomes and their exosomal RNA (esRNA) content. We isolated exosomes from human AH collected during cataract surgery from five patients using serial ultracentrifugation. We measured the size and concentration of AH exosomes in solution using nanoparticle tracking analysis. We found a single population of vesicles having a mean size of 121 ± 11 nm in the unprocessed AH. Data show that centrifugation does not significantly affect the mean particle size (121 ± 11 nm versus 124 ± 21 nm), but does impact the final number of exosomes in solution (87% loss from the unprocessed AH; n = 5). We extracted esRNA from the pooled human AH samples using miRCURY RNA isolation kit from Exiqon. The quality of extracted esRNA was evaluated using Agilent Bioanalyzer 2100 and was used to generate a sequencing library for small RNA sequencing with Illumina MiSeq sequencer. More than 10 different miRNAs were identified; abundant species included miR-486-5p, miR-204, and miR-184. We found that the majority of extracellular vesicles in the AH were in the exosome size range, suggesting that miRNAs housed within exosomes may function in communication between AH inflow and outflow tissues.

KEYWORDS:

Aqueous humor; Exosomal RNA; Exosome; Glaucoma; miRNA

PMID:
25619138
PMCID:
PMC4352394
DOI:
10.1016/j.exer.2015.01.019
[Indexed for MEDLINE]
Free PMC Article

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