GEM-IL: A highly responsive fluorescent lactate indicator

Ramsey Bekdash; Jose R Quejada; Shunnosuke Ueno; Fuun Kawano; Kumi Morikawa; Alison D Klein; Kenji Matsumoto; Tetz C Lee; Koki Nakanishi; Amy Chalan; Teresa M Lee; Rui Liu; Shunichi Homma; Chyuan-Sheng Lin; Maria V Yelshanskaya; Alexander I Sobolevsky; Keisuke Goda; Masayuki Yazawa

doi:10.1016/j.crmeth.2021.100092

GEM-IL: A highly responsive fluorescent lactate indicator

Cell Rep Methods. 2021 Oct 21;1(7):100092. doi: 10.1016/j.crmeth.2021.100092. eCollection 2021 Nov 22.

Authors

Ramsey Bekdash^{1

2

3}, Jose R Quejada^{1

2

3}, Shunnosuke Ueno^{1

2

4}, Fuun Kawano^{1

2}, Kumi Morikawa^{1

2}, Alison D Klein^{1

2}, Kenji Matsumoto⁵, Tetz C Lee⁵, Koki Nakanishi⁵, Amy Chalan^{1

2}, Teresa M Lee^{1

6}, Rui Liu⁵, Shunichi Homma⁵, Chyuan-Sheng Lin^{7

8}, Maria V Yelshanskaya⁹, Alexander I Sobolevsky⁹, Keisuke Goda^{4

10

11}, Masayuki Yazawa^{1

2

3}

Affiliations

¹ Columbia Stem Cell Initiative, Columbia University, New York, NY 10032, USA.
² 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.
³ Department of Molecular Pharmacology and Therapeutics, Vagelos College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
⁴ Department of Chemistry, University of Tokyo, Tokyo 113-0033, Japan.
⁵ Division of Cardiology, Department of Medicine, Vagelos College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
⁶ Department of Pediatrics, Vagelos College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
⁷ Department of Pathology and Cell Biology, Vagelos College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
⁸ Transgenic Mouse Shared Resource, Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA.
⁹ Department of Biochemistry and Molecular Biophysics, Vagelos College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
¹⁰ Department of Bioengineering, University of California, Los Angeles, CA 90095, USA.
¹¹ Institute of Technological Sciences, Wuhan University, Hubei 430072, China.

Abstract

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.

Keywords: cancer; cardiac metabolism; cellular metabolism; genetically encoded fluorescent indicators; lactate; metabolite indicator.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Humans
Lactic Acid* / metabolism
Mice
Myocytes, Cardiac / metabolism
Neural Stem Cells* / metabolism
Signal Transduction

Substances

Lactic Acid

Abstract

Publication types

MeSH terms

Substances

Grants and funding