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Multiplexed visualization of dynamic signaling networks using genetically encoded fluorescent protein-based biosensors

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Abstract

Cells rely on a complex, interconnected network of signaling pathways to sense and interpret changes in their extracellular environment. The development of genetically encoded fluorescent protein (FP)-based biosensors has made it possible for researchers to directly observe and characterize the spatiotemporal dynamics of these intracellular signaling pathways in living cells. However, detailed information regarding the precise temporal and spatial relationships between intersecting pathways is often lost when individual signaling events are monitored in isolation. As the development of biosensor technology continues to advance, it is becoming increasingly feasible to image multiple FP-based biosensors concurrently, permitting greater insights into the intricate coordination of intracellular signaling networks by enabling parallel monitoring of distinct signaling events within the same cell. In this review, we discuss several strategies for multiplexed imaging of FP-based biosensors, while also underscoring some of the challenges associated with these techniques and highlighting additional avenues that could lead to further improvements in parallel monitoring of intracellular signaling events.

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Acknowledgments

We wish to thank members of the Zhang lab for helpful discussions, in particular Fabian Hertel for comments on the manuscript. This work is supported by the National Institutes of Health (R01DK073368 and DP1CA174423 to J.Z.).

Conflict of interest

The authors declare they have no competing financial interests.

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Authors and Affiliations

  1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Charlene Depry, Sohum Mehta & Jin Zhang

  2. The Solomon H. Snyder Department of Neuroscience and Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Jin Zhang

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  1. Charlene Depry
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  2. Sohum Mehta
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  3. Jin Zhang
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Correspondence to Jin Zhang.

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This article is published as part of the Special Issue on “Measuring and manipulating biochemical signals, mechanical forces and metabolites in living cells, tissues and organisms.”

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Depry, C., Mehta, S. & Zhang, J. Multiplexed visualization of dynamic signaling networks using genetically encoded fluorescent protein-based biosensors. Pflugers Arch - Eur J Physiol 465, 373–381 (2013). https://doi.org/10.1007/s00424-012-1175-y

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