Transcriptomes of lineage-specific Drosophila neuroblasts profiled by genetic targeting and robotic sorting

Development. 2016 Feb 1;143(3):411-21. doi: 10.1242/dev.129163. Epub 2015 Dec 23.

Abstract

A brain consists of numerous distinct neurons arising from a limited number of progenitors, called neuroblasts in Drosophila. Each neuroblast produces a specific neuronal lineage. To unravel the transcriptional networks that underlie the development of distinct neuroblast lineages, we marked and isolated lineage-specific neuroblasts for RNA sequencing. We labeled particular neuroblasts throughout neurogenesis by activating a conditional neuroblast driver in specific lineages using various intersection strategies. The targeted neuroblasts were efficiently recovered using a custom-built device for robotic single-cell picking. Transcriptome analysis of mushroom body, antennal lobe and type II neuroblasts compared with non-selective neuroblasts, neurons and glia revealed a rich repertoire of transcription factors expressed among neuroblasts in diverse patterns. Besides transcription factors that are likely to be pan-neuroblast, many transcription factors exist that are selectively enriched or repressed in certain neuroblasts. The unique combinations of transcription factors present in different neuroblasts may govern the diverse lineage-specific neuron fates.

Keywords: Antennal lobe; Mushroom body; Neural stem cell; RNA-seq; Transcription factor; Type II neuroblast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cell Lineage / genetics*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / genetics*
  • Gene Expression Regulation, Developmental
  • Gene Targeting*
  • Neurons / cytology*
  • Robotics*
  • Sequence Analysis, RNA
  • Single-Cell Analysis
  • Transcription Factors / metabolism
  • Transcriptome / genetics*

Substances

  • Drosophila Proteins
  • Transcription Factors