Systematic Analysis of Smooth Muscle and Cartilage Ring Formation during Mouse Tracheal Tubulogenesis
Haoyu Wu,
Ping Wang,
Ziying Liu,
Chunyan Lu,
Wenguang Yin
Affiliations
Haoyu Wu
State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, GMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou Medical University, Guangzhou, Guangdong, ChinaGuangzhou National Laboratory, No. 9 XingDaoHuanBei Road, Guangzhou International Bio Island, Guangzhou, Guangdong Province, China
Ping Wang
Guangzhou Women and Children's Medical Center, Guangzhou, Guangdong, China
Ziying Liu
State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, GMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou Medical University, Guangzhou, Guangdong, China
Chunyan Lu
State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, GMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou Medical University, Guangzhou, Guangdong, China
Wenguang Yin
State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, GMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou Medical University, Guangzhou, Guangdong, ChinaGuangzhou National Laboratory, No. 9 XingDaoHuanBei Road, Guangzhou International Bio Island, Guangzhou, Guangdong Province, China, GMU-GIBH Joint School of Life Sciences, Gu
The trachea tube is the exclusive route to allow gas exchange between the external environment and the lungs. Recent studies have shown the critical role of mesenchymal cells in tracheal tubulogenesis. Improved methods for studying the dynamics of the tracheal mesenchyme development are needed to investigate the cellular and molecular mechanisms during tracheal tubulogenesis. Here, we describe a detailed protocol for a systematic analysis of tracheal tube development to enable observing tracheal smooth muscle (SM) and cartilage ring formation. We describe immunostaining, confocal and stereomicroscopy imaging, and quantitative methods to study the process of tracheal SM and cartilage ring development, including SM cell alignment, polarization, and changes in cell shape as well as mesenchymal condensation. The technologies and approaches described here not only improve analysis of the patterning of the developing trachea but also help uncover the mechanisms underlying airway disease. This protocol also provides a useful technique to analyze cell organization, polarity, and nuclear shape in other organ systems.
