Imaging in vivo acetylcholine release in the peripheral nervous system with a fluorescent nanosensor.
Imaging in vivo acetylcholine release in the peripheral nervous system with a fluorescent nanosensor.
复制标题
使用荧光纳米传感器对周围神经系统中乙酰胆碱的释放进行体内成像。
DOI：
10.1073/pnas.2023807118
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发表时间：
2021
影响因子：
11.1
通讯作者：
Clark,HeatherA
中科院分区：
文献类型：
--
作者：
Xia,Junfei;Yang,Hongrong;Mu,Michelle;Micovic,Nicholas;Poskanzer,KiraE;Monaghan,JamesR;Clark,HeatherA
监测突触传递过程中神经递质释放的能力将对神经系统疾病的诊断和治疗产生重大影响。在这里，我们提出了一种基于DNA的酶纳米传感器的定量检测乙酰胆碱（ACh）在周围神经系统的活小鼠。乙酰胆碱纳米传感器由DNA作为支架，乙酰胆碱酯酶作为识别组分，pH敏感荧光团作为信号发生器，α-银环蛇毒素作为靶向部分组成。我们证明了纳米传感器在活体小鼠下颌下神经节中的实用性，以灵敏地检测范围为0.228至358 μM的ACh。此外，传出神经的电刺激后的传感器响应是剂量依赖性的，可逆的，并且我们观察到ACh释放的药理学抑制后传感器信号减少了1.76%。配备了先进的成像处理工具，我们进一步在组织水平上空间解析ACh信号传播。我们的平台可以灵敏地测量和绘制周围神经系统中的ACh传输。
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.