Identification of miRNomes associated with adult neurogenesis after stroke using Argonaute 2-based RNA sequencing

RNA Biol. 2017 May 4;14(5):488-499. doi: 10.1080/15476286.2016.1196320. Epub 2016 Jun 17.

Abstract

Neurogenesis is associated with functional recovery after stroke. However, the underlying molecular mechanisms have not been fully investigated. Using an Ago2-based RNA immunoprecipitation to immunoprecipated Ago2-RNA complexes followed by RNA sequencing (Ago2 RIP-seq) approach, we profiled the miRNomes in neural progenitor cells (NPCs) harvested from the subventricular zone (SVZ) of the lateral ventricles of young adult rats. We identified more than 7 and 15 million reads in normal and ischemic NPC libraries, respectively. We found that stroke substantially changed Ago2-associated miRNA profiles in NPCs compared to those in non-ischemic NPCs. We also discovered a new complex repertoire of isomiRs and multiple miRNA-miRNA* pairs and numerous novel miRNAs in the non-ischemic and ischemic NPCs. Among them, pc-3p-17172 significantly regulated NPC proliferation and neuronal differentiation. Collectively, the present study reveals profiles of Ago2-associated miRNomes in non-ischemic and ischemic NPCs, which provide a molecular basis to further investigate the role of miRNAs in mediating adult neurogenesis under physiological and ischemic conditions.

Keywords: Ago2; RNA sequencing; microRNA; neural progenitor cells; neurogenesis; stroke.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Adult
  • Analysis of Variance
  • Animals
  • Argonaute Proteins / genetics
  • Argonaute Proteins / metabolism*
  • Cell Proliferation
  • Humans
  • Lateral Ventricles / chemistry
  • Male
  • MicroRNAs / analysis
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Neural Stem Cells / metabolism*
  • Neural Stem Cells / pathology
  • Neurogenesis / physiology*
  • Primary Cell Culture
  • Rats
  • Rats, Wistar
  • Sequence Analysis, RNA
  • Stroke / metabolism*
  • Stroke / pathology
  • Transcriptome

Substances

  • Ago2 protein, rat
  • Argonaute Proteins
  • MicroRNAs