Human aqueous humor exosomes

W. Michael Dismuke, Pratap Challa, Iris Navarro, W. Daniel Stamer, Yutao Liu

Research output: Contribution to journalArticle

41 Scopus citations

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±11nm in the unprocessed AH. Data show that centrifugation does not significantly affect the mean particle size (121±11nm versus 124±21nm), 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.

Original languageEnglish (US)
Pages (from-to)73-77
Number of pages5
JournalExperimental eye research
Volume132
DOIs
StatePublished - Mar 1 2015

Keywords

  • Aqueous humor
  • Exosomal RNA
  • Exosome
  • Glaucoma
  • MiRNA

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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  • Cite this

    Dismuke, W. M., Challa, P., Navarro, I., Stamer, W. D., & Liu, Y. (2015). Human aqueous humor exosomes. Experimental eye research, 132, 73-77. https://doi.org/10.1016/j.exer.2015.01.019