Next-generation multicolor indicators for in vivo imaging of norepinephrine

Year: 2026

Type: article

Abstract: Norepinephrine (NE) is a critical neuromodulator that dynamically shapes brain states and behavior. Genetically encoded fluorescent NE indicators have enabled in vivo visualization of NE release, yet their limited sensitivity and spectral flexibility constrain widespread use in complex experimental paradigms. Here we developed next-generation green and red fluorescent NE indicators, named nLightG2 and nLightR2, respectively. We systematically compared them to other state-of-the-art indicators and found that both indicators improve the detection of endogenous NE release across preparations. Using dual-color photometry, both nLightG2 and nLightR2 could reliably track physiologically relevant NE release along with neuronal activity in different brain areas. Using two-photon imaging, nLightR2 permitted simultaneous dual-color imaging of NE release and astrocytic activity in the hippocampus, while nLightG2 enabled the detection of spatiotemporally discrete NE release events in the visual cortex of awake mice. This improved toolkit will prove useful for dissecting the spatiotemporal complexity of norepinephrine signaling in the brain.

Source: Nature Methods

DOI: 10.1038/s41592-026-03006-z

Institutions: University of Zurich, Italian Institute of Technology, University of Colorado Anschutz, Heidelberg University, Medizinische Fakultät Mannheim, University of Lausanne, University College London, Ruhr University Bochum, University of Cologne, ETH Zurich

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $12,690

Funders: Wellcome Trust, Deutsche Forschungsgemeinschaft, Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung, National Institutes of Health, Horizon 2020 Framework Programme, National Institute of Neurological Disorders and Stroke

Awards: G2138814873, G2237738014, G3244279416, Readout and control of spatiotemporal neuronal codes for behavior, Engineering biosensors for drug profiling across systems, G657093227, Metabolic Imaging of Subiculum in Alzheimer's Disease, Regulation of Striatal Acetylcholine Transmission by Choinergic Interneurons, When the locus coeruleus speaks up in sleep: monoaminergic signaling as integral part of mammalian sleep regulation and function

OpenAlex ID: W7131633380