Chen Chiann-Mun, Miranda António M A, Bub Gil, Srinivas Shankar
Department of Physiology Anatomy and Genetics, University of Oxford Oxford, UK ; Wellcome Trust London, UK.
Department of Physiology Anatomy and Genetics, University of Oxford Oxford, UK.
Front Physiol. 2015 Jan 7;5:508. doi: 10.3389/fphys.2014.00508. eCollection 2014.
The heart is one of the first organs to develop during mammalian embryogenesis. In the mouse, it starts to form shortly after gastrulation, and is derived primarily from embryonic mesoderm. The embryonic heart is unique in having to perform a mechanical contractile function while undergoing complex morphogenetic remodeling. Approaches to imaging the morphogenesis and contractile activity of the developing heart are important in understanding not only how this remodeling is controlled but also the origin of congenital heart defects (CHDs). Here, we describe approaches for visualizing contractile activity in the developing mouse embryo, using brightfield time lapse microscopy and confocal microscopy of calcium transients. We describe an algorithm for enhancing this image data and quantifying contractile activity from it. Finally we describe how atomic force microscopy can be used to record contractile activity prior to it being microscopically visible.
心脏是哺乳动物胚胎发育过程中最早发育的器官之一。在小鼠中,它在原肠胚形成后不久开始形成,主要来源于胚胎中胚层。胚胎心脏的独特之处在于,它必须在进行复杂的形态发生重塑的同时执行机械收缩功能。对发育中心脏的形态发生和收缩活动进行成像的方法不仅对于理解这种重塑如何受到控制很重要,而且对于理解先天性心脏病(CHD)的起源也很重要。在这里,我们描述了使用明场延时显微镜和钙瞬变共聚焦显微镜观察发育中小鼠胚胎收缩活动的方法。我们描述了一种用于增强此图像数据并从中量化收缩活动的算法。最后,我们描述了原子力显微镜如何用于在收缩活动在显微镜下可见之前记录它。