The local transcriptome in the synaptic neuropil revealed by deep sequencing and high-resolution imaging

Neuron. 2012 May 10;74(3):453-66. doi: 10.1016/j.neuron.2012.02.036.

Abstract

In neurons, dendritic protein synthesis is required for many forms of long-term synaptic plasticity. The population of mRNAs that are localized to dendrites, however, remains sparsely identified. Here, we use deep sequencing to identify the mRNAs resident in the synaptic neuropil in the hippocampus. Analysis of a neuropil data set yielded a list of 8,379 transcripts of which 2,550 are localized in dendrites and/or axons. Using a fluorescent barcode strategy to label individual mRNAs, we show that their relative abundance in the neuropil varies over 3 orders of magnitude. High-resolution in situ hybridization validated the presence of mRNAs in both cultured neurons and hippocampal slices. Among the many mRNAs identified, we observed a large fraction of known synaptic proteins including signaling molecules, scaffolds and receptors. These results reveal a previously unappreciated enormous potential for the local protein synthesis machinery to supply, maintain and modify the dendritic and synaptic proteome.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Animals
  • Animals, Newborn
  • CA1 Region, Hippocampal / cytology
  • Cells, Cultured
  • Cluster Analysis
  • Computational Biology
  • Dendrites / metabolism
  • Gene Expression Profiling
  • Gene Library
  • High-Throughput Nucleotide Sequencing / methods*
  • In Vitro Techniques
  • Male
  • Microdissection
  • Neuroglia / physiology
  • Neurons / cytology
  • Neurons / metabolism*
  • Neuropil / cytology
  • Neuropil / metabolism*
  • Oligonucleotide Array Sequence Analysis
  • Protein Biosynthesis
  • RNA, Messenger / metabolism
  • Rats
  • Synapses / genetics
  • Synapses / metabolism*
  • Transcriptome / physiology*

Substances

  • RNA, Messenger