Imaging neuropeptide release at synapses with a genetically engineered reporter
Research on neuropeptide function has advanced rapidly, yet there is still no spatio-temporally resolved method to measure the release of neuropeptides in vivo. Here we introduce Neuropeptide Release Reporters (NPRRs): novel genetically-encoded sensors with high temporal resolution and genetic specificity. Using the Drosophila larval neuromuscular junction (NMJ) as a model, we provide evidence that NPRRs recapitulate the trafficking and packaging of native neuropeptides, and report stimulation-evoked neuropeptide release events as real-time changes in fluorescence intensity, with sub-second temporal resolution.
© 2019, Ding et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Received: February 27, 2019. Accepted: June 25, 2019. Accepted Manuscript published: June 26, 2019 (version 1). Version of Record published: July 4, 2019 (version 2). We thank T Südhof for advice and encouragement at the inception of this project, T Littleton for suggesting GCaMP as a potential reporter for DCV release, B Pfeiffer for advice on construct design, members of the Anderson laboratory for critical feedback, K Zinn for helpful comments on the manuscript, P Goel for the information on antibodies, B Kiragasi for Master-9 programming, K Chen and D Ma for advice on imaging analysis and programming, G Mancuso and C Chiu for administrative assistance and lab management, and A Sanchez for maintenance of fly stocks. This work was supported by NIH BRAIN Initiative grant R21EY026432 and R01 DA031389 (to DJA) and NS091546 (to DD). DJA is an Investigator of the Howard Hughes Medical Institute.
Published - elife-46421-v3.pdf