使用放射性标记的2-脱氧-D-葡萄糖测量小鼠组织中葡萄糖利用的实验方案。

Protocol to measure glucose utilization in mouse tissues using radiolabeled 2-deoxy-D-glucose.

作者信息

Wu Cyun-Ming, Li Hao-Yun, Li Wen-Yi, Hsu Chih-Neng, Yang Wei-Shun, Chuang Gwo-Tsang, Che Tony Pan-Hou, Lee Tung-Yuan, Lee Hsiao-Lin, Hee Siow-Wey, Lee Jiin-Horng, Liao Daniel, Liao Karen Chia-Wen, Liu Ya-Wen, Hsuan Chin-Feng, Chang Yi-Cheng, Chu Shao-Lun

机构信息

Department of Medicine, College of Medicine, National Taiwan University, Taipei 100, Taiwan.

Division of Nephrology, Department of Internal Medicine, National Taiwan University Hospital, Yunlin Branch, Yunlin 640, Taiwan.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103478. doi: 10.1016/j.xpro.2024.103478. Epub 2024 Dec 6.

DOI:10.1016/j.xpro.2024.103478

PMID:39644497

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11656071/

Abstract

2-deoxy-D-glucose (2DG) is a glucose analog converted to 2-deoxy-D-glucose-6-phosphate (2DG-6P) by hexokinase in glycolysis. While 2DG commonly measures glucose uptake, 2DG-6P detects glucose utilization. Here, we present a protocol to measure glucose utilization in various tissues after entering a mouse's body using radiolabeled 2DG. We describe steps for preparing mice and chemicals, extracting blood, adding chemicals, and dissolving tissue. We then detail procedures for calculating glucose utilization using the trapezoid rule.

摘要

2-脱氧-D-葡萄糖（2DG）是一种葡萄糖类似物，在糖酵解过程中被己糖激酶转化为2-脱氧-D-葡萄糖-6-磷酸（2DG-6P）。虽然2DG通常用于测量葡萄糖摄取，但2DG-6P可检测葡萄糖利用情况。在此，我们展示了一种使用放射性标记的2DG测量小鼠体内进入各种组织后的葡萄糖利用情况的方案。我们描述了制备小鼠和化学试剂、采血、添加化学试剂以及溶解组织的步骤。然后，我们详细说明了使用梯形法则计算葡萄糖利用情况的程序。