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Imaging Flow Cytometry Quantifies Neural Genome Dynamics.

Nadine Michel1, Usnish B Majumdar2, Joanne Lannigan3

  • 1Department of Biochemistry & Molecular Genetics, University of Virginia School of Medicine, Neuroscience Graduate Program, Charlottesville, Virginia, 22908.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|May 8, 2019
PubMed
Summary
This summary is machine-generated.

DNA damage is common during development. This study uses imaging flow cytometry to quantify DNA double-strand breaks in neural progenitor cells, revealing their dynamics during neurogenesis.

Keywords:
NPCsTuj1double strand breakshiPSCsimaging flow cytometrynestinneurogenesisneuronal differentiationstem cellsγH2AX

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Area of Science:

  • Neuroscience
  • Genetics
  • Cell Biology

Background:

  • Somatic mosaicism arises from imperfect DNA repair, leading to genomic variations within an individual.
  • Brain somatic mosaicism is significant due to its lifelong impact on neuronal diversity and behavior.
  • Understanding DNA damage during early neurodevelopment is key to explaining brain mosaicism.

Purpose of the Study:

  • To develop and apply a novel single-cell method for quantifying DNA double-strand break (DNA DSB) dynamics in early human neurodevelopment.
  • To investigate DNA DSB accumulation in neural progenitor cells (NPCs) during neurogenesis.

Main Methods:

  • Utilized imaging flow cytometry (IFC) on human-induced pluripotent stem cell-derived NPCs.
  • Quantified DNA DSBs by measuring γH2AX foci.
  • Employed complementary single-cell techniques: fluorescent microscopy, conventional flow cytometry, and comet assay.

Main Results:

  • Established an increase in DNA DSBs during early neurodevelopment in NPCs.
  • Demonstrated dose-dependent and sensitive detection of γH2AX foci using IFC.
  • Revealed the dynamics of DNA DSBs in both proliferating and differentiating neural cells.

Conclusions:

  • Imaging flow cytometry provides a powerful single-cell approach to study DNA DSB dynamics during neurogenesis.
  • The findings offer insights into the origins of brain somatic mosaicism and neuronal diversity.