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Intraperitoneal dye injection method for visualizing the functioning lymphatic vascular system in zebrafish and medaka.
Developmental Dynamics ( IF 2.5 ) Pub Date : 2020-01-10 , DOI: 10.1002/dvdy.143
Erina Saito 1 , Sumio Isogai 2 , Tomonori Deguchi 3 , Kinji Ishida 4 , Takayuki Nozaki 4 , Eri Ishiyama 4 , Marina Wayama 5 , Hiroshi Shimoda 1, 2
Affiliation  

A hierarchically organized lymphatic vascular system extends throughout the vertebrate body for tissue fluid homeostasis, immune trafficking, and the absorption of dietary fats. Intralymphatic dye injection and serial sectioning have been the main tools for visualizing lymphatic vessels. Specific markers for identifying the lymphatic vasculature in zebrafish and medaka have appeared as new tools that enable the study of lymphangiogenesis using genetic and experimental manipulation. Transgenic fishes have become excellent organisms for visualizing the lymphatic vasculature in living embryos, but this method has limited usefulness, especially in later developmental stages. The functional lymphatic endothelium predominantly takes up foreign particles in zebrafish and medaka. We utilized this physiological activity and lymph flow to label lymphatic vessels. Intraperitoneal injection of trypan blue is useful for temporal observations of the lymphatic ducts, which are essential for large-scale genetic screening, while cinnabar (HgS) injection allows identification of the lymphatic endothelium under electron microscopy, avoiding artefactual damage. This injection method, which is not high in cost and does not require high skill or special devices, is applicable to any live fish with functioning lymphatic vessels, even mutants, with high reproducibility for visualizing the entire lymphatic vascular system.

中文翻译:

用于可视化斑马鱼和青鳉功能淋巴血管系统的腹膜内染料注射方法。

一个分层组织的淋巴血管系统延伸到整个脊椎动物体内,用于组织液稳态、免疫运输和膳食脂肪的吸收。淋巴内染料注射和连续切片一直是可视化淋巴管的主要工具。用于识别斑马鱼和青鳉淋巴管系统的特定标记已作为新工具出现,可以使用遗传和实验操作研究淋巴管生成。转基因鱼已成为观察活胚胎淋巴管系统的优秀生物,但这种方法的用处有限,尤其是在发育后期。功能性淋巴内皮主要吸收斑马鱼和青鳉体内的异物。我们利用这种生理活动和淋巴流量来标记淋巴管。腹腔注射台盼蓝可用于淋巴管的时间观察,这对于大规模基因筛查至关重要,而朱砂 (HgS) 注射可在电子显微镜下识别淋巴管内皮,避免人为损伤。这种注射方法成本不高,不需要高技术或特殊设备,适用于任何具有功能性淋巴管的活鱼,甚至是突变体,对整个淋巴管系统的可视化具有很高的重现性。而朱砂 (HgS) 注射可以在电子显微镜下识别淋巴管内皮,避免人为损坏。这种注射方法成本不高,不需要高技术或特殊设备,适用于任何具有功能性淋巴管的活鱼,甚至是突变体,对整个淋巴管系统的可视化具有很高的重现性。而朱砂 (HgS) 注射可以在电子显微镜下识别淋巴管内皮,避免人为损坏。这种注射方法成本不高,不需要高技术或特殊设备,适用于任何具有功能性淋巴管的活鱼,甚至是突变体,对整个淋巴管系统的可视化具有很高的重现性。
更新日期:2020-01-10
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