Investigation of PRDM10 and PRDM13 Expression in Developing Mouse Embryos by an Optimized PACT-Based Embryo Clearing Method

• Published in: International journal of molecular sciences Vol. 22; no. 6; p. 2892
• Main Authors: Woo, Jiwon, Jin, Byung-Ho, Lee, Mirae, Lee, Eunice Yoojin, Moon, Hyung-Seok, Park, Jeong-Yoon, Cho, Yong-Eun
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 12.03.2021; MDPI
• Subjects: Animals; Cartilage; Clearing; Disintegration; embryo clearing; Embryo, Mammalian; Embryogenesis; Embryonic Development - genetics; Embryos; Gene Expression Regulation, Developmental - genetics; Histone-Lysine N-Methyltransferase - genetics; Imaging, Three-Dimensional - methods; IMPACT; Investigations; Methods; Mice; Microscopy; Nervous system; passive clearing technique; PRDM10; PRDM13; Proteins; Spinal cord; Three dimensional analysis; Transcription Factors - genetics; transparent embryo; IMPACT; PRDM13; embryo clearing; passive clearing technique; transparent embryo; PRDM10
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22062892

Recent developments in tissue clearing methods have significantly advanced the three-dimensional analysis of biological structures in whole, intact tissue, providing a greater understanding of spatial relationships and biological circuits. Nonetheless, studies have reported issues with maintaining structural integrity and preventing tissue disintegration, limiting the wide application of these techniques to fragile tissues such as developing embryos. Here, we present an optimized passive tissue clearing technique (PACT)-based embryo clearing method, initial embedding PACT (IMPACT)-Basic, that improves tissue rigidity without compromising optical transparency. We also present IMPACT-Advance, which is specifically optimized for thin slices of mouse embryos past E13.5. We demonstrate proof-of-concept by investigating the expression of two relatively understudied PR domain (PRDM) proteins, PRDM10 and PRDM13, in intact cleared mouse embryos at various stages of development. We observed strong PRDM10 and PRDM13 expression in the developing nervous system and skeletal cartilage, suggesting a functional role for these proteins in these tissues throughout embryogenesis.