Weizmann Institute of Science, Rehovot 76100, Israel.
Methods. 2012 Jan;56(1):63-8. doi: 10.1016/j.ymeth.2011.09.017. Epub 2011 Sep 22.
The Drosophila heart has become an exciting model for elucidating the molecular basis for cardiac function in higher organisms. To complement the genetic approaches that have recently identified an array of genes essential for cardiac function, we developed a method to obtain optimal semi-thin cross sections of embryonic, larval, and adult fly hearts in a desired orientation. A procedure for fluorescent labeling of these sections with multiple markers has also been developed, allowing the detection of proteins at high subcellular resolution. Sections obtained by our method reveal changes in cell shape between embryonic heart and aorta cardioblasts and elucidate the morphology of the adult heart. Analysis of the adult heart reveals the precise cardiac tube morphology, differential distribution of the extracellular matrix protein Laminin within the cardiac tube, as well as individual hand-positive, and Held Out Wings (HOW)-positive luminal cells that might represent blood cells. In summary, our method enables visualization of cross sections of the embryonic and adult hearts at high resolution while maintaining the ability to co-label the sections with multiple markers, thereby facilitating the analysis of cardiac tube formation and maintenance at different developmental stages.
果蝇心脏已成为阐明高等生物心脏功能分子基础的令人兴奋的模型。为了补充最近确定的一系列对心脏功能至关重要的基因的遗传方法，我们开发了一种方法，以获得所需方向的胚胎、幼虫和成年果蝇心脏的最佳半薄横切片。还开发了一种用多种标记物对这些切片进行荧光标记的程序，允许以高亚细胞分辨率检测蛋白质。通过我们的方法获得的切片显示出胚胎心脏和主动脉心肌细胞之间的细胞形状变化，并阐明了成年心脏的形态。对成年心脏的分析揭示了精确的心脏管形态，细胞外基质蛋白层粘连蛋白在心脏管内的差异分布，以及可能代表血细胞的单个手阳性和伸出翅膀（HOW）阳性腔细胞。总之，我们的方法能够以高分辨率可视化胚胎和成年心脏的横切片，同时保持用多种标记物对切片进行共标记的能力，从而便于在不同发育阶段分析心脏管的形成和维持。
