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High Throughput Microinjections of Sea Urchin Zygotes
Published on: January 21, 2014
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Spatially mapping gene expression in sea urchin primary mesenchyme cells
Daniel T Zuch1, Cynthia A Bradham2
1Department of Biology, Boston University, Boston, MA, United States; Program in Molecular Biology, Cell Biology and Biochemistry, Boston University, Boston, MA, United States.
Methods in Cell Biology
|April 6, 2019
Summary
Researchers mapped gene expression in sea urchin primary mesenchyme cells (PMCs) during development. This helps understand how these cells position and form the skeleton.
Area of Science:
- Developmental Biology
- Cell Biology
- Molecular Biology
Background:
- Sea urchin embryogenesis involves primary mesenchyme cells (PMCs) migrating and forming a calcium carbonate skeleton.
- Specific genes are known to be expressed in restricted domains within the PMC population.
Purpose of the Study:
- To investigate the molecular mechanisms controlling PMC positioning.
- To characterize the expression profiles of PMC subset-specific genes.
Main Methods:
- Combined RNA fluorescence in situ hybridization and immunolabeling to detect cell-specific mRNA expression in PMCs.
- Confocal microscopy to obtain 3D position and gene expression data for individual PMCs.
- 3D modeling software to analyze PMC positions and relative gene expression levels.
Main Results:
- Detailed spatiotemporal expression patterns of PMC subset-specific genes were mapped.
- 3D models illustrating gene expression within the PMC population were generated.
Conclusions:
- The developed protocol provides insights into the molecular control of PMC positioning.
- This versatile method can be applied to other developmental systems and cell populations.
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