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Related Experiment Video

Updated: Apr 29, 2026

Confocal Microscopy to Measure Three Modes of Fusion Pore Dynamics in Adrenal Chromaffin Cells
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Cell-fusion method to visualize interphase nuclear pore formation.

Kazuhiro Maeshima1, Tomoko Funakoshi2, Naoko Imamoto3

  • 1Cellular Dynamics Laboratory, RIKEN, Wako, Saitama, Japan; Biological Macromolecules Laboratory, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka, Japan; Department of Genetics, School of Life Science, Graduate University for Advanced Studies (Sokendai), Mishima, Shizuoka, Japan.

Methods in Cell Biology
|May 27, 2014
PubMed
Summary

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Researchers developed a novel cell-fusion method to visualize new nuclear pore complex (NPC) formation in living cells. This technique overcomes challenges in observing nascent NPCs during interphase, advancing our understanding of nuclear transport regulation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • The nucleus, a vital organelle in eukaryotic cells, organizes DNA and relies on nuclear pore complexes (NPCs) for molecular transport.
  • NPCs are crucial for cellular function, with their numbers significantly increasing during the cell cycle's interphase.
  • Understanding NPC biogenesis is hindered by the lack of effective visualization methods for newly formed NPCs alongside existing ones.

Purpose of the Study:

  • To develop and detail a novel system for visualizing interphase nuclear pore complex (NPC) formation.
  • To overcome the technical challenges of observing nascent NPCs in the presence of mature NPCs.
  • To provide a validated method for studying NPC biogenesis in cycling human cells.

Main Methods:

  • Utilized a cell-fusion technique (heterokaryon method) to distinguish and visualize newly synthesized NPCs.
Keywords:
Cell cycleCell fusionHeterokaryonsInterphaseNPCNuclear structureNucleocytoplasmic transportNupsPEG

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  • Employed a photobleaching approach to validate the cell-fusion method's efficacy.
  • Applied these methods to investigate the roles of specific proteins and kinases in NPC formation.

    • Main Results:

    • Successfully visualized newly synthesized interphase NPCs using the cell-fusion system.
    • Validated the cell-fusion approach with photobleaching, confirming its reliability.
    • Demonstrated the involvement of cyclin-dependent protein kinases and Pom121 in NPC formation.

    Conclusions:

    • The cell-fusion (heterokaryon) method provides a powerful tool for studying interphase NPC biogenesis.
    • This technique overcomes previous limitations in visualizing nascent NPCs.
    • The findings highlight key regulators of NPC formation in cycling human cells, contributing to nuclear transport research.