Imaging in vivo acetylcholine release in the peripheral nervous system with a fluorescent nanosensor

Author:

Xia JunfeiORCID,Yang HongrongORCID,Mu MichelleORCID,Micovic Nicholas,Poskanzer Kira E.ORCID,Monaghan James R.,Clark Heather A.ORCID

Abstract

The ability to monitor the release of neurotransmitters during synaptic transmission would significantly impact the diagnosis and treatment of neurological diseases. Here, we present a DNA-based enzymatic nanosensor for quantitative detection of acetylcholine (ACh) in the peripheral nervous system of living mice. ACh nanosensors consist of DNA as a scaffold, acetylcholinesterase as a recognition component, pH-sensitive fluorophores as signal generators, and α-bungarotoxin as a targeting moiety. We demonstrate the utility of the nanosensors in the submandibular ganglia of living mice to sensitively detect ACh ranging from 0.228 to 358 μM. In addition, the sensor response upon electrical stimulation of the efferent nerve is dose dependent, reversible, and we observe a reduction of ∼76% in sensor signal upon pharmacological inhibition of ACh release. Equipped with an advanced imaging processing tool, we further spatially resolve ACh signal propagation on the tissue level. Our platform enables sensitive measurement and mapping of ACh transmission in the peripheral nervous system.

Funder

HHS | NIH | NIH Office of the Director

HHS | NIH | National Institute of Neurological Disorders and Stroke

NSF | ENG | Division of Chemical, Bioengineering, Environmental, and Transport Systems

Publisher

Proceedings of the National Academy of Sciences

Subject

Multidisciplinary

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