On May 2, 2022, the international academic journalGenesis published “Generation of Piezo1-CreERtransgenic mice for visualization and lineage tracing of mechanical force responsive cells in vivo”, the research results ofthe laboratory of Zhou Bin, Chief Professor of the School of Life Science, Hangzhou Institute for Advanced Study (HIAS), UCAS.

In the process of body growth and tissue repair, cells and tissues are often stimulated by complex mechanical stimulation. Timely sensing of external mechanical signals is of great physiological significance to cells, tissues, and even the whole organism. Mechanosensitive ion channels (MSCs) can help cells sense external mechanical signals and convert them into electrical signals or chemical signals in cells on a millisecond time scale, further activating downstream signaling pathways and regulating the proliferation, differentiation, and migration of cells.

Piezo1 is a mechanosensitive ion channel protein widely found in mammals and is involved in regulating a variety of physiological activities including vascular development, arterial remodeling, and blood pressure regulation. That means Piezo1+ cells play an important role in body growth and tissue homeostasis and repair. The mice of Piezo1-tdTomato (Sanjeev S Ranade, 2014, PNAS, PMID: 24958852) and Piezo1-lacZ (Cahalan, Elife, 2015, PMID: 26001274) are used to map the expression of Piezo1+ cells in their tissues. However, up to now, there has been still a lack of a mouse that can be used to track the fate changes of Piezo1+ cells and their offspring to explore the fate transition of these cells in important physiological processes such as homeostasis development and tissue repair.

Based on the above background, this study constructed and identified a mouse, Piezo1-CreER, that can be used to track Piezo1+ cells in the body, and used this tool to study the dynamic expression changes of Piezo1 in various tissues of mice during their embryo, neonatal and adult stages. In this study, the CRISPR/Cas9 technology was first used to knock the CreER into the starting codon site of the Piezo1 gene by homologous recombination to obtain the Piezo1-CreER mice strain. Piezo1-CreER mice were hybridized with Rosa26-Ai47 mice to obtain Piezo1-CreER; Rosa26-Ai47 mice strains. Induced by tamoxifen, Piezo1⁺ cells in mice expressed green fluorescent proteins, thus enabling lineage tracking of Piezo1⁺ cells and their offspring.

Note: Piezo1-CreER; Schematic diagram of the principle of green fluorescent protein expressed in Piezo1⁺ cells of Rosa26-Ai47 mice after tamoxifen induction

Using this kind of mouse, this study maps the expression of Piezo1 in different stages of mouse development. The study found that Piezo1 was mainly expressed in endothelial cells and hardly in embryonic epithelial cells at day 13.5 of mouse embryonic phase E, indicating that endothelial cells were the main sensor of external mechanical stimulation during mouse embryonic development. On day 4 of the neonatal period of mice, a small number of epithelial cells were also observed to begin to express Piezo1. However, in adult mice aged 10 weeks, Piezo1 was also widely expressed in specific epithelial cells of various tissues and organs, such as ciliary cells of the lung, hepatocytes of the liver, and acinar cells of the pancreas, in addition to its high expression abundance in endothelial cells. In conclusion, this study constructed and identified the mice that can be used to trace Piezo1⁺cells and their offspring in the body, and used these mice to map the dynamic expression of Piezo1 on Day E13.5 of the embryonic phase, Day P4 of the neonatal phase and Week 10 of adult phase.

Note: The schematic diagram of expression abundance of Piezo1 in endothelial cells (ECs) and epithelial cells (EPCs) of mice on Day E13.5 of the embryonic phase, Day P4 of the neonatal phase, and Week 10 of the adult phase.

Li Xufeng, a postgraduate from the research group of Zhou Bin, School of Life Science, Hangzhou Institute for Advanced Study (HIAS), UCAS, is the first author of the paper. Prof. Zhou Bin fromHIAS and Prof. He Lingjuan from Westlake University are the corresponding authors of the paper.The research is strongly supported by the laboratory animal platform and cell analysis technology platform of CEMCS and Westlake University. The work is funded by the Chinese Academy of Sciences, the National Natural Science Foundation of China, the Ministry of Science and Technology of the People's Republic of China, and the Science and Technology Commission of Shanghai Municipality.

Source | School of Life Science

Typesetter | Tong Sihui

Published in Zhejiang

Scan with WeChat

Follow us