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Detection of alternative splicing of fibronectin mRNA in a single cell

T Kumazaki1, Y Mitsui, K Hamada

  • 1Department of Biochemistry and Biophysics, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan. kumazaki@ue.ipc.hiroshima-u.ac.jp

Journal of Cell Science
|April 23, 1999
PubMed

Insights

Single cells exhibit varied fibronectin messenger RNA (mRNA) splicing patterns. Alternative splicing complexity at the IIICS site changes during cellular senescence.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Pre-fibronectin mRNA undergoes alternative splicing at EDA, EDB, and IIICS sites.
  • Understanding single-cell splicing variability is crucial for comprehending gene expression regulation.

Purpose of the Study:

  • To investigate the alternative splicing patterns of fibronectin mRNA within individual cells.
  • To determine the complexity of fibronectin mRNA isoforms generated by alternative splicing.
  • To examine changes in fibronectin mRNA splicing during cellular senescence.

Main Methods:

  • Single-cell reverse transcription-polymerase chain reaction (RT-PCR) analysis.
  • Analysis of alternative splicing at EDA, EDB, and IIICS sites of fibronectin mRNA.

Main Results:

  • Individual cells displayed diverse fibronectin mRNA splicing patterns, producing either single or multiple mRNA forms at splicing sites.
  • Approximately 80% of cells produced both (+) and (-) forms at EDA and EDB sites.
  • Ten distinct combinations of fibronectin mRNA isoforms were observed concerning the IIICS site.
  • Significant alterations in IIICS site alternative splicing were detected during cellular senescence.

Conclusions:

  • Fibronectin mRNA splicing is highly variable at the single-cell level.
  • Cellular senescence is associated with significant changes in fibronectin mRNA alternative splicing patterns at the IIICS site.

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