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Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos
Published on: February 22, 2016
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Investigation of PRDM10 and PRDM13 Expression in Developing Mouse Embryos by an Optimized PACT-Based Embryo Clearing
Jiwon Woo1,2,3,4, Byung-Ho Jin1,5,6, Mirae Lee1,2,4
1Department of Neurosurgery, The Spine and Spinal Cord Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul 06273, Korea.
International Journal of Molecular Sciences
|April 3, 2021
Summary
We developed new tissue clearing methods for fragile embryos, improving structural integrity and transparency. This technique revealed new insights into PRDM10 and PRDM13 protein roles in developing nervous and skeletal systems.
Area of Science:
- Developmental Biology
- Biotechnology
- Molecular Biology
Background:
- Tissue clearing advances 3D analysis of biological structures.
- Fragile tissues like embryos pose challenges due to structural disintegration.
- Existing methods struggle with maintaining integrity in delicate samples.
Purpose of the Study:
- To optimize passive tissue clearing technique (PACT)-based methods for embryo clearing.
- To enhance tissue rigidity without compromising optical transparency.
- To investigate the expression of PRDM10 and PRDM13 in cleared mouse embryos.
Main Methods:
- Developed initial embedding PACT (IMPACT)-Basic for general embryo clearing.
- Developed IMPACT-Advance for thin slices of mouse embryos (post-E13.5).
- Utilized cleared embryos to examine PRDM10 and PRDM13 expression patterns.
Main Results:
- IMPACT-Basic and IMPACT-Advance improve tissue rigidity and optical transparency.
- Successfully visualized PRDM10 and PRDM13 expression in intact, cleared mouse embryos.
- Observed strong PRDM10 and PRDM13 expression in the developing nervous system and skeletal cartilage.
Conclusions:
- Optimized PACT-based methods (IMPACT) enable robust clearing of fragile embryos.
- PRDM10 and PRDM13 are expressed in key embryonic tissues, suggesting crucial roles.
- This technique facilitates further investigation of gene function in 3D embryonic development.
