Visualizing spatiotemporal dynamics of multicellular cell-cycle progressions with fucci technology

Area of Science:

• Cell Biology
• Molecular Biology
• Developmental Biology

Background:

• Visualizing individual cell-cycle dynamics in multicellular tissues is challenging.
• Monitoring the G1 to S phase transition, crucial for DNA replication, is particularly difficult.

Purpose of the Study:

• To develop genetically encoded fluorescent probes for monitoring cell-cycle progression.
• To utilize ubiquitination oscillators for tracking cell-cycle transitions.

Main Methods:

• Development of Fucci (fluorescent ubiquitination-based cell-cycle indicator) probes.
• Exploitation of cell-cycle-dependent ubiquitination for nuclear labeling (red for G1, green for S/G2/M).
• Generation of cultured cells and transgenic mice expressing Fucci probes.
• Time-lapse imaging of Fucci2-expressing cells for spatiotemporal dynamics.
• High-resolution imaging of fixed Fucci transgenic mouse embryos for proliferation and differentiation mapping.

Main Results:

• Fucci probes successfully label individual nuclei with distinct colors based on cell-cycle phase.
• Time-lapse imaging revealed spatiotemporal patterns of cell-cycle dynamics in cultured cells.
• Imaging of transgenic mouse embryos provided maps of cell proliferation and differentiation in developing organs.

Conclusions:

• Fucci probes offer a powerful tool for visualizing cell-cycle progression in various biological contexts.
• This technology facilitates the study of cell proliferation and differentiation dynamics in development and disease.