sDarken: Next generation genetically encoded fluorescent sensors for serotonin

Martin Kubitschke; Monika C. M. Müller; Lutz Wallhorn; Mauro Pulin; Manuel Mittag; Stefan Pollok; Tim Ziebarth; Svenja Bremshey; Jill Gerdey; Kristin Carolin Claussen; Kim Renken; Juliana Groß; Pascal Gneiße; Niklas Meyer; J. Simon Wiegert; Andreas Reiner; Martin Fuhrmann; Olivia Andrea Masseck

doi:10.1038/s41467-022-35200-w

NeuroscienceNature CommunicationsDec 6, 2022

Peer-reviewed journal article

sDarken: Next generation genetically encoded fluorescent sensors for serotonin

Martin Kubitschke iD, Monika C. M. Müller iD, Lutz Wallhorn iD, Mauro Pulin iD, Manuel Mittag iD, Stefan Pollok iD, Tim Ziebarth iD, Svenja Bremshey iD, Jill Gerdey iD, Kristin Carolin Claussen, Kim Renken iD, Juliana Groß iD, Pascal Gneiße iD, Niklas Meyer iD, J. Simon Wiegert iD, Andreas Reiner iD, Martin Fuhrmann iD, Olivia Andrea Masseck iD

Abstract

We developed a new family of genetically encoded serotonin (5-HT) sensors (sDarken) on the basis of the native 5-HT1A receptor and circularly permuted GFP. sDarken 5-HT sensors are bright in the unbound state and diminish their fluorescence upon binding of 5-HT. Sensor variants with different affinities for serotonin were engineered to increase the versatility in imaging of serotonin dynamics. Experiments in vitro and in vivo showed the feasibility of imaging serotonin dynamics with high temporal and spatial resolution. As demonstrated here, the designed sensors showed excellent membrane expression, have high specificity, a superior signal-to-noise ratio, detect the endogenous release of serotonin and are suitable for two-photon in vivo imaging.

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