Columbia Stem Cell Initiative, Columbia University, New York, NY 10032, USA.
Department of Rehabilitation and Regenerative Medicine, Vagelos College of Physicians and Surgeons, Columbia University, 650 West 168th Street, BB1108/BB1109D, New York, NY 10032, USA.
Cell Rep Methods. 2021 Oct 21;1(7):100092. doi: 10.1016/j.crmeth.2021.100092. eCollection 2021 Nov 22.
Lactate metabolism has been shown to have increasingly important implications in cellular functions as well as in the development and pathophysiology of disease. The various roles as a signaling molecule and metabolite have led to interest in establishing a new method to detect lactate changes in live cells. Here we report our development of a genetically encoded metabolic indicator specifically for probing lactate (GEM-IL) based on superfolder fluorescent proteins and mutagenesis. With improvements in its design, specificity, and sensitivity, GEM-IL allows new applications compared with the previous lactate indicators, Laconic and Green Lindoblum. We demonstrate the functionality of GEM-IL to detect differences in lactate changes in human oncogenic neural progenitor cells and mouse primary ventricular myocytes. The development and application of GEM-IL show promise for enhancing our understanding of lactate dynamics and roles.
乳酸代谢在细胞功能以及疾病的发生和病理生理学方面的重要性日益增加。作为一种信号分子和代谢物,乳酸具有多种作用,这促使人们产生了用一种新方法来检测活细胞中乳酸变化的兴趣。在这里,我们报告了基于超折叠荧光蛋白和诱变技术开发的一种专门用于探测乳酸的遗传编码代谢指示剂(GEM-IL)。通过改进其设计、特异性和灵敏度,与之前的乳酸指示剂 Laconic 和 Green Lindoblum 相比,GEM-IL 具有新的应用前景。我们证明了 GEM-IL 检测人类致癌神经祖细胞和小鼠原代心室肌细胞中乳酸变化差异的功能。GEM-IL 的开发和应用有望增强我们对乳酸动力学和作用的理解。