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Prog Brain Res. 2012;196:235-63. doi: 10.1016/B978-0-444-59426-6.00012-4.

Optogenetic reporters: Fluorescent protein-based genetically encoded indicators of signaling and metabolism in the brain.

Author information

  • 1Department of Neurobiology, Harvard Medical School, Boston, MA, USA.

Abstract

Fluorescent protein technology has evolved to include genetically encoded biosensors that can monitor levels of ions, metabolites, and enzyme activities as well as protein conformation and even membrane voltage. They are well suited to live-cell microscopy and quantitative analysis, and they can be used in multiple imaging modes, including one- or two-photon fluorescence intensity or lifetime microscopy. Although not nearly complete, there now exists a substantial set of genetically encoded reporters that can be used to monitor many aspects of neuronal and glial biology, and these biosensors can be used to visualize synaptic transmission and activity-dependent signaling in vitro and in vivo. In this review, we present an overview of design strategies for engineering biosensors, including sensor designs using circularly permuted fluorescent proteins and using fluorescence resonance energy transfer between fluorescent proteins. We also provide examples of indicators that sense small ions (e.g., pH, chloride, zinc), metabolites (e.g., glutamate, glucose, ATP, cAMP, lipid metabolites), signaling pathways (e.g., G protein-coupled receptors, Rho GTPases), enzyme activities (e.g., protein kinase A, caspases), and reactive species. We focus on examples where these genetically encoded indicators have been applied to brain-related studies and used with live-cell fluorescence microscopy.

Copyright © 2012 Elsevier B.V. All rights reserved.

PMID:
22341329
[PubMed - indexed for MEDLINE]
PMCID:
PMC3494096
Free PMC Article

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