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Related Experiment Video

Updated: May 7, 2025

Lineage Tracing and Clonal Analysis in Developing Cerebral Cortex Using Mosaic Analysis with Double Markers MADM
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In Vivo Clonal Analysis Using MADM with Spatiotemporal Specificity.

Yuwei Peng1,2,3, Efan Wang1,3, Zhongfu Shen1,2,3

  • 1IDG/McGovern Institute of Brain Research, Tsinghua University, Beijing, People's Republic of China.

Methods in Molecular Biology (Clifton, N.J.)
|January 2, 2025
PubMed
Summary

Mosaic analysis with double markers (MADM) enables precise lineage tracing in vivo. This study applies MADM for single-cell resolution analysis of developing mouse neocortex radial glial progenitors.

Keywords:
3D reconstructionClonal analysisInterchromosomal recombinationLineage tracingMADMSerial sectioning

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Genetics

Background:

  • Lineage tracing is crucial for understanding tissue development and cellular dynamics.
  • Mosaic analysis with double markers (MADM) offers a powerful method for in vivo lineage tracing.
  • Radial glial progenitors are key stem cells in the developing mammalian neocortex.

Purpose of the Study:

  • To apply MADM for quantitative lineage analysis of radial glial progenitors.
  • To achieve single-cell resolution in tracing progenitor cell fates.
  • To investigate the developmental dynamics of the mouse neocortex.

Main Methods:

  • Utilizing Cre recombinase-dependent interchromosomal recombination.
  • Restoring stable expression of two fluorescent proteins in individual cells.
  • Applying MADM for sparse labeling of stem and progenitor cells.
  • Performing quantitative lineage analysis at single-cell resolution.

Main Results:

  • MADM successfully enabled sparse labeling of individual radial glial progenitors.
  • Quantitative lineage analysis provided insights into progenitor cell behavior.
  • Single-cell resolution was achieved in tracing cell fates within the developing neocortex.

Conclusions:

  • MADM is a valuable technique for detailed lineage analysis of neural progenitors.
  • This approach facilitates a deeper understanding of neocortical development.
  • Future studies can leverage MADM for complex developmental and disease investigations.