Department of Chemistry, School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, 113-0033, Japan.
PRESTO, Japan Science and Technology Agency, Chiyoda-ku, Tokyo, 102-0075, Japan.
Nat Commun. 2023 Oct 27;14(1):6598. doi: 10.1038/s41467-023-42230-5.
L-Lactate is increasingly appreciated as a key metabolite and signaling molecule in mammals. However, investigations of the inter- and intra-cellular dynamics of L-lactate are currently hampered by the limited selection and performance of L-lactate-specific genetically encoded biosensors. Here we now report a spectrally and functionally orthogonal pair of high-performance genetically encoded biosensors: a green fluorescent extracellular L-lactate biosensor, designated eLACCO2.1, and a red fluorescent intracellular L-lactate biosensor, designated R-iLACCO1. eLACCO2.1 exhibits excellent membrane localization and robust fluorescence response. To the best of our knowledge, R-iLACCO1 and its affinity variants exhibit larger fluorescence responses than any previously reported intracellular L-lactate biosensor. We demonstrate spectrally and spatially multiplexed imaging of L-lactate dynamics by coexpression of eLACCO2.1 and R-iLACCO1 in cultured cells, and in vivo imaging of extracellular and intracellular L-lactate dynamics in mice.
L-乳酸在哺乳动物中作为关键代谢物和信号分子的作用日益受到重视。然而，由于 L-乳酸特异性遗传编码生物传感器的选择和性能有限，目前对 L-乳酸的细胞内外动力学的研究受到了阻碍。在这里，我们现在报告了一对光谱和功能上正交的高性能遗传编码生物传感器：一种绿色荧光细胞外 L-乳酸生物传感器，命名为 eLACCO2.1，和一种红色荧光细胞内 L-乳酸生物传感器，命名为 R-iLACCO1。eLACCO2.1 表现出优异的膜定位和强大的荧光响应。据我们所知，R-iLACCO1 及其亲和变体的荧光响应比以前报道的任何细胞内 L-乳酸生物传感器都要大。我们通过在培养细胞中共同表达 eLACCO2.1 和 R-iLACCO1，以及在小鼠体内进行细胞外和细胞内 L-乳酸动力学的成像，实现了 L-乳酸动力学的光谱和空间多重成像。
