The miRNA pathway controls rapid changes in activity-dependent synaptic structure at the Drosophila melanogaster neuromuscular junction

PLoS One. 2013 Jul 2;8(7):e68385. doi: 10.1371/journal.pone.0068385. Print 2013.

Abstract

It is widely accepted that long-term changes in synapse structure and function are mediated by rapid activity-dependent gene transcription and new protein synthesis. A growing amount of evidence suggests that the microRNA (miRNA) pathway plays an important role in coordinating these processes. Despite recent advances in this field, there remains a critical need to identify specific activity-regulated miRNAs as well as their key messenger RNA (mRNA) targets. To address these questions, we used the larval Drosophila melanogaster neuromuscular junction (NMJ) as a model synapse in which to identify novel miRNA-mediated mechanisms that control activity-dependent synaptic growth. First, we developed a screen to identify miRNAs differentially regulated in the larval CNS following spaced synaptic stimulation. Surprisingly, we identified five miRNAs (miRs-1, -8, -289, -314, and -958) that were significantly downregulated by activity. Neuronal misexpression of three miRNAs (miRs-8, -289, and -958) suppressed activity-dependent synaptic growth suggesting that these miRNAs control the translation of biologically relevant target mRNAs. Functional annotation cluster analysis revealed that putative targets of miRs-8 and -289 are significantly enriched in clusters involved in the control of neuronal processes including axon development, pathfinding, and growth. In support of this, miR-8 regulated the expression of a wingless 3'UTR (wg 3' untranslated region) reporter in vitro. Wg is an important presynaptic regulatory protein required for activity-dependent axon terminal growth at the fly NMJ. In conclusion, our results are consistent with a model where key activity-regulated miRNAs are required to coordinate the expression of genes involved in activity-dependent synaptogenesis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / metabolism
  • Drosophila melanogaster / physiology
  • Gene Expression Profiling
  • Gene Expression Regulation*
  • Gene Ontology
  • Larva / genetics
  • Larva / metabolism
  • Larva / physiology
  • MicroRNAs / genetics*
  • Motor Neurons / metabolism
  • Neuromuscular Junction / genetics*
  • Neuromuscular Junction / metabolism
  • Neuromuscular Junction / physiology
  • Oligonucleotide Array Sequence Analysis
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics
  • Synapses / genetics*
  • Synapses / metabolism
  • Synapses / physiology

Substances

  • MIRN1 microRNA, Drosophila
  • MIRN289 microRNA, Drosophila
  • MIRN314 microRNA, Drosophila
  • MIRN958 microRNA, human
  • MicroRNAs
  • RNA, Messenger
  • mir-8 microRNA, Drosophila

Associated data

  • GEO/GSE43945