2025年1月3日
This protocol outlines a method for grafting a magnetic bead into the developing zebrafish heart through microsurgery, enabling the manipulation of mechanical forces in vivo and triggering mechanical stimulus-dependent calcium influx in endocardial cells.
We focus on understanding how mechanical forces shape the zebrafish heart using advanced imaging techniques. By combining our knowledge of biology, physics, microscopy, and computing, we develop tools for optical imaging and image analysis to study how mechanical stimuli influence the way the cardiovascular system develops. A beating heart generates several type of forces, such as pressure for the shear and contractor force.
Although each of these forces has a function in vitro culture systems, it is difficult to separate these parameters in the in vivo heart. By developing our approach, we aim to tackle this challenging task. We have established a novel approach to assess, adapt biological output caused by external force stimulation, and we verified our force spin and constant signal in crosscutting the cardiac cells.
As a result this approach enables us to impetrate other aspects of tissue neurogenesis in the world of mechanical biology. This method enables the study of heart function in zebrafish embryo via targeted mechanical stimulation. Unlike genetic, pharmacological, or optogenetic approaches, pit grafting offers a more direct way to influence heart physiology through mechanical means.
Pit grafting is viable for examining the roles of mechanical forces and calcium influx in cardiac biological processes. It provides means to deepen our understanding of the mechanical transduction pathways involved in heart development and function.
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本研究利用先进的成像技术,探讨机械力如何影响斑马鱼心脏的发育。研究人员采用显微手术将磁性微珠移植到心脏中,从而操控机械刺激,并评估内皮心肌细胞中的钙离子内流情况。
对斑马鱼心脏发育过程中机械力的直接操控,能够精确研究对心脏形态发生至关重要的力传导通路。该技术解决了早期发现阶段的一个关键难题——分离并量化生物力学刺激对瓣膜发育及心内膜细胞信号传导的影响。这一方法可提高心血管药物研发中靶点验证的预测可信度,并为风险调整后的研发组合决策提供依据。
该方法整合了早期发现与临床前研究的界面,将假设驱动的机制性研究与定量的体内验证相结合。