Live imaging of mitosis in the developing mouse embryonic cortex.

Area of Science:

• Neuroscience
• Developmental Biology
• Cell Biology

Background:

• Mitosis is crucial for organ development, including the brain.
• Aberrant neural progenitor mitosis in the developing cerebral cortex can lead to brain size and function defects.
• Current methods often rely on fixed brain sections, limiting dynamic observation.

Purpose of the Study:

• To detail a protocol for live imaging of mitosis in ex vivo embryonic rodent brain slices.
• To provide a method for analyzing neural progenitor cell division with high temporal and spatial resolution.
• To offer a flexible approach for studying the impact of genetic and anatomical factors on mitosis.

Main Methods:

• Brain extraction, embedding, and vibratome sectioning.
• Staining and culturing of brain slices for live imaging.
• Time-lapse microscopy and post-acquisition analysis of mitosis.

Main Results:

• Demonstration of live imaging of mitosis using vital dyes and transgenic mouse models (histone H2B-EGFP, centrin-EGFP).
• Application of in utero electroporation (mCherry-α-tubulin) for visualizing mitotic events.
• Representative results showcasing the protocol's effectiveness in capturing dynamic mitotic processes.

Conclusions:

• Live imaging of mitosis in brain slices provides a flexible and high-resolution tool for studying neural progenitor cell division.
• This protocol complements existing methods and facilitates the study of genetic regulation of mitosis.
• The approach allows for assessment of age, anatomy, and genetic perturbations in a controlled setting.