Author response: Imaging neuropeptide release at synapses with a genetically engineered reporter

Ding, Keke; Han, Yifu; Seid, Taylor W; Buser, Christopher; Karigo, Tomomi; Zhang, Shishuo; Dickman, Dion; Anderson, David J.

doi:10.7554/elife.46421.025

Cited by 1 publication

(1 citation statement)

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“…Fast-scan cyclic voltammetry offers higher temporal resolution but has so far only been used successfully in detecting one neuropeptide methionine enkephalin (Calhoun et al, 2019). Peptides tagged with large fluorescent proteins or reporters such as EGFP, pHluorin, and GCaMP provide a relatively fast readout of peptide release and describe knockout phenotypes related to peptide release in vitro, but this approach is limited to modified peptides, not the endogenous peptides, and is hardly possible to apply in vivo (Burke et al, 1997;de Wit et al, 2009;Ding et al, 2019;Lang et al, 1997;van den Pol, 2012;Xia et al, 2009). Finally cellbased neurotransmitter fluorescent engineered reporters (CNiFERs) have been used to detect neuropeptides; however, the need to implant exogenous cells in specific brain regions limits the utility of this approach (Jones et al, 2018;Lacin et al, 2016;Xiong et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

A toolkit of highly selective and sensitive genetically encoded neuropeptide sensors

Wang

Qian

Zhao

et al. 2022

Preprint

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Neuropeptides are key signaling molecules in the endocrine and nervous systems that regulate many critical physiological processes, including energy balance, sleep and circadian rhythms, stress, and social behaviors. Understanding the functions of neuropeptides in vivo requires the ability to monitor their dynamics with high specificity, sensitivity, and spatiotemporal resolution; however, this has been hindered by the lack of direct, sensitive and non-invasive tools. Here, we developed a series of GRAB (G protein-coupled receptor activation‒based) sensors for detecting somatostatin (SST), cholecystokinin (CCK), corticotropin-releasing factor (CRF), neuropeptide Y (NPY), neurotensin (NTS), and vasoactive intestinal peptide (VIP). These fluorescent sensors utilize the corresponding GPCRs as the neuropeptide-sensing module with the insertion of a circular-permutated GFP as the optical reporter. This design detects the binding of specific neuropeptides at nanomolar concentration with a robust increase in fluorescence. We used these GRAB neuropeptide sensors to measure the spatiotemporal dynamics of endogenous SST release in isolated pancreatic islets and to detect the release of both CCK and CRF in acute brain slices. Moreover, we detect endogenous CRF release induced by stressful experiences in vivo using fiber photometry and 2-photon imaging in mice. Together, these new sensors establish a robust toolkit for studying the release, function, and regulation of neuropeptides under both physiological and pathophysiological conditions.

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