Isolation of specific neuron populations from roundworm caenorhabditis elegans

Edward M. Germany, Nataliya Zahayko, Oleh Khalimonchuk

Research output: Contribution to journalArticlepeer-review

Abstract

During the aging process, many cells accumulate high levels of damage leading to cellular dysfunction, which underlies many geriatric and pathological conditions. Post-mitotic neurons represent a major cell type affected by aging. Although multiple mammalian models of neuronal aging exist, they are challenging and expensive to establish. The roundworm Caenorhabditis elegans is a powerful model to study neuronal aging, as these animals have short lifespan, an available robust genetic toolbox, and well-cataloged nervous system. The method presented herein allows for seamless isolation of specific cells based on the expression of a transgenic green fluorescent protein (GFP). Transgenic animal lines expressing GFP under distinct, cell type-specific promoters are digested to remove the outer cuticle and gently mechanically disrupted to produce slurry containing various cell types. The cells of interest are then separated from non-target cells through fluorescence-activated cell sorting, or by anti-GFP-coupled magnetic beads. The isolated cells can then be cultured for a limited time or immediately used for cell-specific ex vivo analyses such as transcriptional analysis by real time quantitative PCR. Thus, this protocol allows for rapid and robust analysis of cell-specific responses within different neuronal populations in C. elegans.

Original languageEnglish (US)
Article numbere60145
JournalJournal of Visualized Experiments
Volume2019
Issue number150
DOIs
StatePublished - Aug 2019

Keywords

  • Aging
  • C
  • Cell isolation
  • Developmental Biology
  • Elegans
  • Green fluorescent protein
  • Issue 150
  • Neuronal markers
  • Neurons

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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