2. Academic Validation
  3. High-Performance Genetically Encoded Green Fluorescent Biosensors for Intracellular l-Lactate

High-Performance Genetically Encoded Green Fluorescent Biosensors for Intracellular l-Lactate

  • ACS Cent Sci. 2024 Jan 31;10(2):402-416. doi: 10.1021/acscentsci.3c01250.
Saaya Hario 1,  Giang N T Le 1 2,  Hikaru Sugimoto 3,  Kei Takahashi-Yamashiro 4 5,  Suguru Nishinami 6,  Hirofumi Toda 6,  Selene Li 1,  Jonathan S Marvin 7,  Shinya Kuroda 8,  Mikhail Drobizhev 9,  Takuya Terai 1,  Yusuke Nasu 1 10,  Robert E Campbell 1 5 11
Affiliations

Affiliations

  • 1 Department of Chemistry, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
  • 2 Department of Chemistry, University of Toronto, Toronto, Ontario M5S 3H6, Canada.
  • 3 Department of Biochemistry and Molecular Biology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
  • 4 Department of Molecular Pathology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
  • 5 Department of Chemistry, Faculty of Science, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
  • 6 International Institute for Integrative Sleep Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
  • 7 Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, Virginia 20147, United States.
  • 8 Department of Biological Sciences, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
  • 9 Department of Microbiology and Cell Biology, Montana State University, Bozeman, Montana 59717, United States.
  • 10 PRESTO, Japan Science and Technology Agency, Chiyoda-ku, Tokyo 102-0075, Japan.
  • 11 CERVO Brain Research Center and Department of Biochemistry, Microbiology, and Bioinformatics, Université Laval, Québec, Québec G1 V 0A6, Canada.
PMID: 38435524 DOI: 10.1021/acscentsci.3c01250
Abstract

l-Lactate is a monocarboxylate produced during the process of cellular glycolysis and has long generally been considered a waste product. However, studies in recent decades have provided new perspectives on the physiological roles of l-lactate as a major energy substrate and a signaling molecule. To enable further investigations of the physiological roles of l-lactate, we have developed a series of high-performance (ΔF/F = 15 to 30 in vitro), intensiometric, genetically encoded green fluorescent protein (GFP)-based intracellular l-lactate biosensors with a range of affinities. We evaluated these biosensors in cultured cells and demonstrated their application in an ex vivo preparation of Drosophila brain tissue. Using these biosensors, we were able to detect glycolytic oscillations, which we analyzed and mathematically modeled.

Figures
Products