Live-Cell Imaging of Neurotransmitter Release with a Cell-Surface-Anchored DNA-Nanoprism Fluorescent Sensor

Abstract

Neurotransmitters are essential chemical mediators for neuronal communication in variable neuromodulations. However, the progress of neuroscience is hampered by the shortage of suitable sensors to track neurotransmitters with high spatial and temporal resolution. Here, we introduce a self-assembled DNA-nanoprism fluorescent probe capable of nongenetically engineering the cell surface for ultrasensitive imaging of the neurotransmitter release at a single live-cell level. The DNA-nanoprism structure conjugated with three cholesterol tails enables the probe to rapidly and stably anchor on the cell surface within 10 min. The in situ detection of neurotransmitters is achieved by equipping the DNA-nanoprism with an aptamer-based “turn-on” fluorescent sensory module for the transmitter of interest. In a proof-of-concept study, we directly visualized the transient dopamine (DA) release on the cell surface with selective responsivity and high spatiotemporal precision and further explored the dynamic correlation between DA release and calcium influx triggered by high K+. This study provides a robust and sensitive tool for cell-surface-targeted imaging of neuromodulations, which might open up a new avenue to improve the understanding of neurochemistry and advance neuroscience research.

Multiple Origins of Neurons From Secretory Cells

“…Multiplexed nanotools (Farsi et al, 2016 ; Jing et al, 2018 ; Wu et al, 2018 ; Dinarvand et al, 2019 ; Zeng et al, 2020 ) are instrumental in visualizing cell-specific secretion of co-transmitters and the actual balance of neurotransmitters as stereotyped and learned behaviors are generated.…”

Recent Advances on Functional Nucleic-Acid Biosensors

“…For instance, the real-time imaging biosensor can be used to observe the condition of the patient's diseased area at all times during a surgical operation, which can determine the success of the operation [174]. In the clinical research of live animals, real-time imaging can always pay attention to various changes in the animal body, which greatly shortens the time of clinical research [161]. Many technologies support real-time imaging research nowadays.…”

“…The first study was focused on the living cell imaging of neurotransmitter release at the single living cell level. Zeng et al experimentally verified the proposed concept with dopamine (DA) as the target [161]. The researchers designed an aptamer-modified amphiphilic DNA-nanoprism structure.…”

“…By encountering the target protein, FNA binds to the target protein and detaches from the quenching element because of its different affinity for the performance of real-time imaging in living cells.The first study was focused on the living cell imaging of neurotransmitter release at the single living cell level. Zeng et al experimentally verified the proposed concept with dopamine (DA) as the target[161]. The researchers designed an aptamer-modified amphiphilic DNA-nanoprism structure.…”

DNA nanostructure-based nucleic acid probes: construction and biological applications

“…DNA prisms are also used in the sensing of some neurotransmitters. 79 Through a DNAnanoprism with aptamer-based "turn-on" fluorescent sensory modules, it provided a robust and sensitive tool for cell-surfacetargeted imaging of neuromodulations.…”