Resources
Authors & Affiliations
Zacharoula Kagiampaki-Baimpaki, Valentin Rohner, Cedric Kiss, Sebastiano Curreli, Alexander Dieter, Maria Wilhelm, Masaya Harada, Sian N. Duss, Jan Dernic, Musadiq A. Bhat, Xuehan Zhou, Luca Ravotto, Tim Ziebarth, Laura Moreno Wasielewski, Latife Sönmez, Dietmar Benke, Bruno Weber, Johannes Bohacek, Andreas Reiner, J. Simon Wiegert, Tommaso Fellin, Tommaso Patriarchi
Abstract
Norepinephrine (NE) is one of the main neuromodulators in the brain influencing a plethora of behavioral outputs including both autonomic and cognitive functions, such as emotional learning and stress responses, through a brain-wide network of projections emanating from the Locus Coeruleus (LC). Genetically encoded indicators engineered from G-protein-coupled receptors (GPCRs) have been proven essential tools for neuroscience enabling in vivo imaging of neuromodulators with high spatiotemporal resolution. Current state-of-the-art NE sensors allow for in vivo optical detection of NE, however, they exhibit slow kinetics and they rely exclusively on an Alpha-2a adrenergic receptor (AR) scaffold limiting their compatibility with drugs targeting this AR subtype. Here, we introduce a family of sensitive multicolor NE indicators, which includes nLightG (green) and nLightR (red), based on an Alpha-1 AR subtype, that exhibit improved sensitivity, ligand selectivity, and kinetics compared to the existing sensors. Through a combination of in vitro, ex vivo, and in vivo experimental approaches, we validate the properties and distinct pharmacological profiles of these new indicators, benchmarking them against the established ones. Leveraging in vivo multisite fiber photometry recordings, we successfully monitor optogenetically induced NE release in both the mouse LC and hippocampus simultaneously. In addition, two-photon imaging of nLightG revealed NE transients linked to locomotion and reward in the dorsal CA1 region of the hippocampus. Thus, the introduction of these sensitive NE indicators expands the toolkit for investigating the NE system comprehensively.