Single-nucleus and single-cell transcriptomes compared in matched cortical cell types

PLoS One. 2018 Dec 26;13(12):e0209648. doi: 10.1371/journal.pone.0209648. eCollection 2018.

Abstract

Transcriptomic profiling of complex tissues by single-nucleus RNA-sequencing (snRNA-seq) affords some advantages over single-cell RNA-sequencing (scRNA-seq). snRNA-seq provides less biased cellular coverage, does not appear to suffer cell isolation-based transcriptional artifacts, and can be applied to archived frozen specimens. We used well-matched snRNA-seq and scRNA-seq datasets from mouse visual cortex to compare cell type detection. Although more transcripts are detected in individual whole cells (~11,000 genes) than nuclei (~7,000 genes), we demonstrate that closely related neuronal cell types can be similarly discriminated with both methods if intronic sequences are included in snRNA-seq analysis. We estimate that the nuclear proportion of total cellular mRNA varies from 20% to over 50% for large and small pyramidal neurons, respectively. Together, these results illustrate the high information content of nuclear RNA for characterization of cellular diversity in brain tissues.

MeSH terms

  • Animals
  • Cell Lineage / genetics
  • Cell Lineage / physiology
  • Cell Nucleus / genetics*
  • Gene Expression Profiling / methods
  • Mice
  • Neurons / metabolism
  • Sequence Analysis, RNA / methods
  • Single-Cell Analysis*
  • Transcriptome / genetics*
  • Visual Cortex / metabolism*
  • Visual Cortex / physiology