Kotiyal Srishti, Fulbright Alexis, O'Brien Liam K, Evason Kimberley J
Department of Pathology, Department of Oncological Sciences, and Huntsman Cancer Institute, University of Utah School of Medicine.
Department of Pathology, Department of Oncological Sciences, and Huntsman Cancer Institute, University of Utah School of Medicine;
J Vis Exp. 2020 Feb 2(156). doi: 10.3791/60744.
In several transgenic zebrafish models of hepatocellular carcinoma (HCC), hepatomegaly can be observed during early larval stages. Quantifying larval liver size in zebrafish HCC models provides a means to rapidly assess the effects of drugs and other manipulations on an oncogene-related phenotype. Here we show how to fix zebrafish larvae, dissect the tissues surrounding the liver, photograph livers using bright-field microscopy, measure liver area, and analyze results. This protocol enables rapid, precise quantification of liver size. As this method involves measuring liver area, it may underestimate differences in liver volume, and complementary methodologies are required to differentiate between changes in cell size and changes in cell number. The dissection technique described herein is an excellent tool to visualize the liver, gut, and pancreas in their natural positions for myriad downstream applications including immunofluorescence staining and in situ hybridization. The described strategy for quantifying larval liver size is applicable to many aspects of liver development and regeneration.
在几种肝细胞癌（HCC）的转基因斑马鱼模型中，在幼体早期阶段可观察到肝肿大。在斑马鱼HCC模型中量化幼体肝脏大小提供了一种快速评估药物和其他操作对癌基因相关表型影响的方法。在这里，我们展示了如何固定斑马鱼幼体，解剖肝脏周围的组织，使用明场显微镜拍摄肝脏照片，测量肝脏面积，并分析结果。该方案能够快速、精确地量化肝脏大小。由于该方法涉及测量肝脏面积，可能会低估肝脏体积的差异，因此需要补充方法来区分细胞大小变化和细胞数量变化。本文所述的解剖技术是一种出色的工具，可用于在自然位置可视化肝脏、肠道和胰腺，以用于包括免疫荧光染色和原位杂交在内的众多下游应用。所描述的量化幼体肝脏大小的策略适用于肝脏发育和再生的许多方面。
