- 作者: Gang-Hui Lee, Chia-Lin Chang, Wen-Tai Chiu, Tsun-Hsien Hsiao, Po-Yuan Chen, Kuan-Chieh Wang, Cheng-Hsiang Kuo, Bing-Hung Chen, Guey-Yueh Shi, Hua-Lin Wu and Tzu-Fun Fu
- 作者服務機構: 1.The Institute of Basic Medical Science College of Medicine, National Cheng Kung University, Tainan, Taiwan 2.International Center for Wound Repair and Regeneration, National Cheng Kung University, Tainan, Taiwan 3.Department of Biochemistry and Molecular Biology, National Cheng Kung University College of Medicine, Tainan, Taiwan 4.Cardiovascular Research Center College of Medicine, National Cheng Kung University, Tainan, Taiwan 5.Department of Biomedical Engineering College of Engineering, National Cheng Kung University, Tainan, Taiwan 6.Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University College of Medicine, Tainan, Taiwan 7.Department of Food Safety/ Hygiene and Risk Management College of Medicine, National Cheng Kung University, Tainan, Taiwan 8.Department of Pharmacy College of Pharmacy and Science, Chia Nan University of Pharmacy and Science, Tainan, Taiwan 9.Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan 10.The Institute of Biomedical Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan 11.Cardiovascular Research Center College of Medicine, National Cheng Kung University, Tainan, Taiwan 12.Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University College of Medicine, Tainan, Taiwan
- 中文摘要:
- 英文摘要:
Background
Thrombomodulin (TM), an integral membrane protein, has long been known for its anticoagulant activity. Recent studies showed that TM displays multifaceted activities, including the involvement in cell adhesion and collective cell migration in vitro. However, whether TM contributes similarly to these biological processes in vivo remains elusive.
Methods
We adapted zebrafish, a prominent animal model for studying molecular/cellular activity, embryonic development, diseases mechanism and drug discovery, to examine how TM functions in modulating cell migration during germ layer formation, a normal and crucial physiological process involving massive cell movement in the very early stages of life. In addition, an in vivo assay was developed to examine the anti-hemostatic activity of TM in zebrafish larva.
Results
We found that zebrafish TM-b, a zebrafish TM-like protein, was expressed mainly in vasculatures and displayed anti-hemostatic activity. Knocking-down TM-b led to malformation of multiple organs, including vessels, heart, blood cells and neural tissues. Delayed epiboly and incoherent movement of yolk syncytial layer were also observed in early TM-b morphants. Whole mount immunostaining revealed the co-localization of TM-b with both actin and microtubules in epibolic blastomeres. Single-cell tracking revealed impeded migration of blastomeres during epiboly in TM-b-deficient embryos.
Conclusion
Our results showed that TM-b is crucial to the collective migration of blastomeres during germ layer formation. The structural and functional compatibility and conservation between zebrafish TM-b and mammalian TM support the properness of using zebrafish as an in vivo platform for studying the biological significance and medical use of TM. - 中文關鍵字:
- 英文關鍵字: Thrombomodulin, Collective cell migration, Cytoskeleton, Germ layers formation, Organogenesis, Zebrafish