ZFIN ID: ZDB-PUB-130710-47
Fluorescent activated cell sorting (FACS) combined with gene expression microarrays for transcription enrichment profiling of zebrafish lateral line cells
Gallardo, V.E., and Behra, M.
Date: 2013
Source: Methods (San Diego, Calif.)   62(3): 226-31 (Journal)
Registered Authors: Behra, Martine, Gallardo, Viviana
Keywords: migrating primordium, neuromast, hair cell progenitor, FACS, zebrafish larva, lateral line
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Cell Differentiation
  • Cell Movement
  • Embryo, Nonmammalian
  • Flow Cytometry
  • Gene Expression Regulation, Developmental*
  • Genes, Reporter
  • Green Fluorescent Proteins/genetics*
  • Green Fluorescent Proteins/metabolism
  • Larva/genetics*
  • Larva/growth & development
  • Larva/metabolism
  • Lateral Line System/cytology
  • Lateral Line System/growth & development
  • Lateral Line System/metabolism*
  • Mechanotransduction, Cellular
  • Oligonucleotide Array Sequence Analysis
  • RNA, Messenger/genetics*
  • RNA, Messenger/metabolism
  • Transcription, Genetic
  • Zebrafish/genetics*
  • Zebrafish/growth & development
  • Zebrafish/metabolism
PubMed: 23791746 Full text @ Methods
ABSTRACT

Transgenic lines carrying fluorescent reporter genes like GFP have been of great value in the elucidation of developmental features and physiological processes in various animal models, including zebrafish. The lateral line (LL), which is a fish specific superficial sensory organ, is an emerging organ model for studying complex cellular processes in the context of the whole living animal. Cell migration, mechanosensory cell development/differentiation and regeneration are some examples. This sensory system is made of superficial and sparse small sensory patches called neuromasts, with less than 50 cells in any given patch. The paucity of cells is a real problem in any effort to characterize those cells at the transcriptional level. We describe here a method which we applied to efficiently separate subpopulation of cells of the LL, using two distinct stable transgenic zebrafish lines, Tg(cldnb:gfp) and Tg(tnks1bp1:EGFP). In both cases, the GFP positive (GFP+) cells were separated from the remainder of the animal by using a Fluorescent Activated Cell Sorter (FACS). The transcripts of the GFP+ cells were subsequently analyzed on gene expression microarrays. The combination of FACS and microarrays is an efficient method to establish a transcriptional signature for discrete cell populations which would otherwise be masked in whole animal preparation.

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