Analysis of Cardiac Chamber Development During Mouse Embryogenesis Using Whole Mount Epifluorescence.
Analysis of Cardiac Chamber Development During Mouse Embryogenesis Using Whole Mount Epifluorescence.
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DOI：
10.3791/59413
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发表时间：
2019-04-17
期刊：
影响因子：
--
通讯作者：
Nam YJ
中科院分区：
文献类型：
--
作者：
Zhang Z;Nam YJ
该协议的目标是描述使用心室特异性荧光记者敲入小鼠（MLC-2v-tdTomato 小鼠）在心脏发育过程中解剖小鼠胚胎和可视化胚胎小鼠心室的方法。心脏发育涉及线性心管形成、心管成环和四腔分隔。这些复杂的过程在所有脊椎动物中都高度保守。小鼠胚胎心脏已广泛用于心脏发育研究。然而，由于其尺寸极小，解剖小鼠胚胎心脏在技术上具有挑战性。此外，心室形成的可视化通常需要原位杂交、使用 LacZ 报告小鼠的 β-半乳糖苷酶染色或切片胚胎心脏的免疫染色。在这里，我们描述了如何解剖小鼠胚胎心脏并使用整体落射荧光显微镜直接可视化 MLC-2v-tdTomato 小鼠的心室形成。通过这种方法，可以直接检查心管形成和循环以及四室形成，而无需对小鼠胚胎进行进一步的实验操作。虽然本协议中使用 MLC-2v-tdTomato 报告基因敲入小鼠系作为示例，但该协议可以应用于其他心脏特异性荧光报告基因转基因小鼠系。我们提出了使用整体落射荧光显微镜检查从心室特异性 MLC-2v-tdTomato 报告基因敲入小鼠解剖的小鼠胚胎上检查小鼠心脏发育的方案。这种方法使我们能够直接可视化小鼠心脏发育过程中心室形成的每个阶段，而无需劳动密集型的组织化学方法。
The goal of this protocol is to describe a method for dissection of mouse embryos and visualization of embryonic mouse ventricular chambers during heart development using ventricular specific fluorescent reporter knock-in mice (MLC-2v-tdTomato mice). Heart development involves a linear heart tube formation, the heart tube looping, and four chamber septation. These complex processes are highly conserved in all vertebrates. Mouse embryonic heart has been widely used for heart developmental studies. However, due to their extremely small size, dissecting mouse embryonic hearts is technically challenging. In addition, visualization of cardiac chamber formation often needs in situ hybridization, beta-galactosidase staining using LacZ reporter mice or immunostaining of sectioned embryonic hearts. Here, we describe how to dissect mouse embryonic hearts and directly visualize ventricular chamber formation of MLC-2v-tdTomato mice using whole mount epifluorescent microscopy. With this method, it is possible to directly examine heart tube formation and looping, and four chamber formation without further experimental manipulation of mouse embryos. Although MLC-2v-tdTomato reporter knock-in mouse line is used in this protocol as an example, this protocol can be applied to other heart-specific fluorescent reporter transgenic mouse lines. We present the protocols to examine mouse heart development using whole mount epifluorescent microscopy on mouse embryos dissected from ventricular specific MLC-2v-tdTomato reporter knock-in mice. This method allows us to directly visualize each stage of the ventricular formation during mouse heart development without labor-intensive histochemical methods.
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影响因子：
11.8
作者：
Wu, San-pin;Cheng, Chiang-Min;Lanz, Rainer B.;Wang, Tiannan;Respress, Jonathan L.;Ather, Sameer;Chen, Wen;Tsai, Shaw-Jenq;Wehrens, Xander H. T.;Tsai, Ming-Jer;Tsai, Sophia Y.
通讯作者：
Tsai, Sophia Y.
DOI：
10.1073/pnas.90.11.5157
发表时间：
1993-06-01
影响因子：
11.1
作者：
OBRIEN, TX;LEE, KJ;CHIEN, KR
通讯作者：
CHIEN, KR
影响因子：
4.6
作者：
Nam, Young-Jae;Lubczyk, Christina;Munshi, Nikhil V.
通讯作者：
Munshi, Nikhil V.
影响因子：
56.9
作者：
Bao, ZZ;Bruneau, BG;Cepko, CL
通讯作者：
Cepko, CL
影响因子：
4.8
作者：
Chen, J;Kubalak, SW;Chien, KR
通讯作者：
Chien, KR