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SPARC_A DNA‐based optical nanosensor for in vivo imaging of acetylcholine in the peripheral nervous system
Author(s) -
Clark Heather,
Monaghan James,
Yang Hongrong,
Xia Junfei,
Mu Michelle,
Lovely Alexander,
Micovic Nicholas
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.05947
Subject(s) - acetylcholine , nanosensor , neuroscience , in vivo , neurotransmitter , cholinergic , acetylcholine receptor , chemistry , biology , microbiology and biotechnology , pharmacology , receptor , central nervous system , nanotechnology , biochemistry , materials science
The study of neurotransmitter signaling helps to understand fundamental neuronal circuits and develop strategies against neurodegenerative diseases. Acetylcholine (ACh) plays a pivotal role in modulating neuron functions. Monitoring its physiological level in real‐time and probing its biological distribution is of crucial importance. However, the current techniques are subject to temporal and spatial limitations which impede their further in vivo applications. Herein we report the development of a ACh nanosensor by using DNA as a scaffold, acetylcholinesterase as a recognition component, pH‐sensitive fluorophores as signal generator, and α‐bungarotoxin as a targeting ligand. The nanosensor was delivered to the submandibular ganglion in living mice by microinjection for in vivo imaging of endogenous ACh release. The ACh nanosensor selectively binds to acetylcholine receptors on the post‐synaptic neurons, and display a sensitive response to physiological ACh range from 0.228 μM to 358 μM in ex vivo calibration. The ACh nanosensors also respond to endogenous ACh release triggered by electrical stimulation in a reversible and dose‐dependent manner. We also found that the treatment of cholinergic inhibitor, Vesamicol, significant suppresses the response of the ACh nanosensor. This work provides a tool for in vivo imaging of ACh dynamics with unprecedented capabilities to enhance the spatial and temporal resolution of detection. We envision such a sensor platform could be applied to imaging other neurotransmitters in mammalian system as well as extended to other organ systems in the peripheral nervous system Support or Funding Information NIH common fund_SPARC_Project number: 1OT2OD024909‐01DNA‐based acetylcholine (ACh) nanosensors detect the ACh in the submandibular ganglion (SMG) of living mice. (A) ACh sensors were microinjected into the SMG of a living mouse. With the targeting ability of bungarotoxin, the ACh nanosensor can immobilize in the synaptic cleft of the mouse. (B) The mechanism of ACh nanosensors. (D) ΔF/F from pHAb channel in response to electrical stimulation (indicated by red arrow).