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Detection of apoptotic cells based on in situ hybridization chain reaction using specific hairpins

Mei Yang1, Ruihua Ji1, Zhengqing Zhao2

  • 1Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China.

Apoptosis : an International Journal on Programmed Cell Death
|November 2, 2022
PubMed

Insights

A new method, In Situ Hybridization Chain Reaction (in situ HCR), detects apoptotic DNA fragmentation. This technique identifies DNA fragments in the cytoplasm, potentially indicating early apoptosis, and offers a cost-effective alternative to TUNEL assays.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Programmed cell death (apoptosis) is characterized by DNA fragmentation.
  • Terminal deoxynucleotidyl transferase mediated-dUTP Nick End Labeling (TUNEL) is the current standard for in situ DNA fragmentation detection.
  • There is a need for alternative methods to study apoptosis and DNA fragmentation.

Purpose of the Study:

  • To establish a novel method for in situ detection of apoptotic DNA fragments.
  • To compare the new method with the established TUNEL assay.
  • To investigate the potential of the new method for early apoptosis detection.

Main Methods:

  • Development of In Situ Hybridization Chain Reaction (in situ HCR) for detecting DNA fragments.
  • Comparative analysis of in situ HCR and TUNEL staining on apoptotic cells.
  • Investigation of DNA fragment localization during apoptosis.

Main Results:

  • The in situ HCR method successfully detected apoptotic DNA fragments in situ.
  • A majority of in situ HCR-positive cells were also TUNEL-positive.
  • In situ HCR identified DNA fragments in the cytoplasm, suggesting early apoptosis stages and cytoplasmic transfer.
  • The in situ HCR method is cost-effective and suitable for large-scale tissue analysis.

Conclusions:

  • In situ HCR is a viable and potentially superior method for detecting DNA fragmentation in apoptosis.
  • The technique offers advantages over TUNEL, including cost-effectiveness and detection of cytoplasmic DNA fragments.
  • In situ HCR may provide insights into the early stages and mechanisms of apoptosis.
Keywords:
ApoptosisHybridization chain reactionIn situ detection

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