Abstract
Lactate, the product of aerobic glycolysis, plays a dual role as fuel and intercellular signal in inflammation, immune evasion, and tumor progression. The production of lactate by macrophages has been associated with their polarization and function. Here we describe imaging protocols to characterize the metabolism of cultured human macrophages using a genetically encoded fluorescent sensor-specific for lactate. By superfusing cultures with increasing lactate concentrations and pharmacological inhibitors, it is possible to estimate the kinetic parameters of monocarboxylate transporter 4 (MCT4) and lactate production. Practical advice is given regarding sensor expression, imaging, and data analysis. The spatiotemporal resolution of this technique is amenable to the study of fast events at the single-cell level in different immune and other cell types.
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Acknowledgments
We thank Karen Everett for reading the manuscript. The Centro de Estudios Científicos (CECs) is funded by the Chilean Government through the Centers of Excellence Basal Financing Program of CONICYT.
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Baeza-Lehnert, F., Flores, C.A., Guequén, A., Barros, L.F. (2020). Monitoring Lactate Dynamics in Individual Macrophages with a Genetically Encoded Probe. In: Mishra, S. (eds) Immunometabolism. Methods in Molecular Biology, vol 2184. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0802-9_2
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DOI: https://doi.org/10.1007/978-1-0716-0802-9_2
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