Optogenetics & Synthetic Biology Interdisciplinary Research Center, State Key Laboratory of Bioreactor Engineering, Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China; Research Unit of New Techniques for Live-cell Metabolic Imaging, Chinese Academy of Medical Sciences, Beijing 100730, China; CAS Center for Excellence in Brain Science, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Center for Translational Medicine, The Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, 600 Yishan Road, Shanghai 200233, China.
Cell Metab. 2023 Jan 3;35(1):200-211.e9. doi: 10.1016/j.cmet.2022.10.002. Epub 2022 Oct 28.
Despite its central importance in cellular metabolism, many details remain to be determined regarding subcellular lactate metabolism and its regulation in physiology and disease, as there is sensitive spatiotemporal resolution of lactate distribution, and dynamics remains a technical challenge. Here, we develop and characterize an ultrasensitive, highly responsive, ratiometric lactate sensor, named FiLa, enabling the monitoring of subtle lactate fluctuations in living cells and animals. Utilizing FiLa, we demonstrate that lactate is highly enriched in mammalian mitochondria and compile an atlas of subcellular lactate metabolism that reveals lactate as a key hub sensing various metabolic activities. In addition, FiLa sensors also enable direct imaging of elevated lactate levels in diabetic mice and facilitate the establishment of a simple, rapid, and sensitive lactate assay for point-of-care clinical screening. Thus, FiLa sensors provide powerful, broadly applicable tools for defining the spatiotemporal landscape of lactate metabolism in health and disease.
尽管乳酸在细胞代谢中具有核心重要性，但关于其亚细胞代谢及其在生理和疾病中的调节，仍有许多细节需要确定，因为乳酸的分布具有敏感的时空分辨率，其动态变化仍然是一项技术挑战。在这里，我们开发并表征了一种超灵敏、高响应性的比率型乳酸传感器，命名为 FiLa，能够监测活细胞和动物中微妙的乳酸波动。利用 FiLa，我们证明了乳酸在哺乳动物线粒体中高度富集，并编制了一个亚细胞乳酸代谢图谱，揭示了乳酸作为一种关键的枢纽，能够感知各种代谢活动。此外，FiLa 传感器还能够直接成像糖尿病小鼠中升高的乳酸水平，并促进建立一种简单、快速和敏感的即时检验（point-of-care clinical screening）乳酸分析方法。因此，FiLa 传感器为定义健康和疾病状态下乳酸代谢的时空景观提供了强大的、广泛适用的工具。
