A luciferase prosubstrate and a red bioluminescent calcium indicator for imaging neuronal activity in mice

Xiaodong Tian, Yiyu Zhang, Xinyu Li, Ying Xiong, Tianchen Wu and Hui-Wang Ai ()
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Xiaodong Tian: University of Virginia
Yiyu Zhang: University of Virginia
Xinyu Li: University of Virginia
Ying Xiong: University of Virginia
Tianchen Wu: University of Virginia
Hui-Wang Ai: University of Virginia

Nature Communications, 2022, vol. 13, issue 1, 1-13

Abstract: Abstract Although fluorescent indicators have been broadly utilized for monitoring bioactivities, fluorescence imaging, when applied to mammals, is limited to superficial targets or requires invasive surgical procedures. Thus, there is emerging interest in developing bioluminescent indicators for noninvasive mammalian imaging. Bioluminescence imaging (BLI) of neuronal activity is highly desired but hindered by insufficient photons needed to digitalize fast brain activities. In this work, we develop a luciferase prosubstrate deliverable at an increased dose and activated in vivo by nonspecific esterase. We further engineer a bright, bioluminescent indicator with robust responsiveness to calcium ions (Ca2+) and appreciable emission above 600 nm. Integration of these advantageous components enables the imaging of the activity of neuronal ensembles in awake mice minimally invasively with excellent signal-to-background and subsecond temporal resolution. This study thus establishes a paradigm for studying brain function in health and disease.

Date: 2022
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DOI: 10.1038/s41467-022-31673-x

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