Interfacing Aptamer-Modified Nanopipettes with Neuronal Media and Ex Vivo Brain Tissue

IF 4.6 Q1 CHEMISTRY, ANALYTICAL ACS Measurement Science Au Pub Date : 2023-11-22 DOI:10.1021/acsmeasuresciau.3c00047
Annina Stuber, Anna Cavaccini, Andreea Manole, Anna Burdina, Yassine Massoud, Tommaso Patriarchi, Theofanis Karayannis and Nako Nakatsuka*, 
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Abstract

Aptamer-functionalized biosensors exhibit high selectivity for monitoring neurotransmitters in complex environments. We translated nanoscale aptamer-modified nanopipette sensors to detect endogenous dopamine release in vitro and ex vivo. These sensors employ quartz nanopipettes with nanoscale pores (ca. 10 nm diameter) that are functionalized with aptamers that enable the selective capture of dopamine through target-specific conformational changes. The dynamic behavior of aptamer structures upon dopamine binding leads to the rearrangement of surface charge within the nanopore, resulting in measurable changes in ionic current. To assess sensor performance in real time, we designed a fluidic platform to characterize the temporal dynamics of nanopipette sensors. We then conducted differential biosensing by deploying control sensors modified with nonspecific DNA alongside dopamine-specific sensors in biological milieu. Our results confirm the functionality of aptamer-modified nanopipettes for direct measurements in undiluted complex fluids, specifically in the culture media of human-induced pluripotent stem cell-derived dopaminergic neurons. Moreover, sensor implantation and repeated measurements in acute brain slices was possible, likely owing to the protected sensing area inside nanoscale DNA-filled orifices, minimizing exposure to nonspecific interferents and preventing clogging. Further, differential recordings of endogenous dopamine released through electrical stimulation in the dorsolateral striatum demonstrate the potential of aptamer-modified nanopipettes for ex vivo recordings with unprecedented spatial resolution and reduced tissue damage.

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适配体修饰的纳米吸管与神经元介质和离体脑组织的连接
适配体功能化的生物传感器在复杂环境中监测神经递质具有高选择性。我们翻译了纳米级适配体修饰的纳米吸管传感器来检测内源性多巴胺的体外和体外释放。这些传感器采用带有纳米级孔(直径约10纳米)的石英纳米吸管,这些孔被适体功能化,能够通过靶向特异性构象变化选择性捕获多巴胺。多巴胺结合后适配体结构的动态行为导致纳米孔内表面电荷的重排,导致离子电流的可测量变化。为了实时评估传感器的性能,我们设计了一个流体平台来表征纳米吸管传感器的时间动态。然后,我们通过在生物环境中部署用非特异性DNA修饰的控制传感器和多巴胺特异性传感器来进行差异生物传感。我们的研究结果证实了适配体修饰的纳米吸管在未稀释的复杂液体中直接测量的功能,特别是在人类诱导的多能干细胞衍生的多巴胺能神经元的培养基中。此外,在急性脑切片中植入传感器和重复测量是可能的,这可能是由于纳米级dna填充孔内受保护的传感区域,最大限度地减少了暴露于非特异性干扰物并防止堵塞。此外,通过电刺激在背侧纹状体释放内源性多巴胺的差异记录表明,适体修饰的纳米管具有前所未有的空间分辨率和减少组织损伤的离体记录潜力。
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来源期刊
ACS Measurement Science Au
ACS Measurement Science Au 化学计量学-
CiteScore
5.20
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0.00%
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0
期刊介绍: ACS Measurement Science Au is an open access journal that publishes experimental computational or theoretical research in all areas of chemical measurement science. Short letters comprehensive articles reviews and perspectives are welcome on topics that report on any phase of analytical operations including sampling measurement and data analysis. This includes:Chemical Reactions and SelectivityChemometrics and Data ProcessingElectrochemistryElemental and Molecular CharacterizationImagingInstrumentationMass SpectrometryMicroscale and Nanoscale systemsOmics (Genomics Proteomics Metabonomics Metabolomics and Bioinformatics)Sensors and Sensing (Biosensors Chemical Sensors Gas Sensors Intracellular Sensors Single-Molecule Sensors Cell Chips Arrays Microfluidic Devices)SeparationsSpectroscopySurface analysisPapers dealing with established methods need to offer a significantly improved original application of the method.
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