An Integrated Nanopipette Platform for Vesicular Neuropeptide Detection and Modulation in Single Living Cells

Objective

Neurodegenerative diseases (NDDs) represent a growing global crisis with profound social and economic implications. As essential signaling molecules, neuropeptides (NPs) packaged within dense-core vesicles (DCVs) play a critical role in modulating neural communication and have been strongly implicated in NDD pathogenesis. The ability to in situ monitor and regulate NPs within individual vesicles is critical for unraveling disease mechanisms and developing early diagnostic and therapeutic strategies. However, intracellular analysis of NPs within DCVs remains a major technical challenge. To address this gap, this project aims to develop a novel double-barrel nanopipette with integrated sensing and regulatory functions. Combined with intracellular vesicle impact electrochemical cytometry, the detection channel modified with stimuli-responsive polymers will enable fast and sensitive in situ quantification of non-electroactive NPs within single vesicles. Taking advantage of dynamic adsorption/desorption of NPs on polymer surfaces, the nanopipette can serve as a direct sampling tool to extract NPs from DCVs, facilitating structural characterization via nanoESI-MS. The delivery channel will enable closed-loop release of therapeutic agents in response to specific stimuli (e.g., ROS or pH), offering a targeted intervention strategy for vesicle function with high controllability. In summary, this project will establish the first nanotool capable of simultaneous in situ quantification, structural analysis and targeted regulation of NPs within DCVs. By combining my expertise in electrochemical sensors based on stimuli-responsive polymers to detect biomolecules with new innovations in materials science, nanotechnology and neural science, this work will provide a transformative paradigm for NDD research. Its successful implementation will not only redefine our understanding and treatment of NDDs, but also generate profound scientific, societal, and economic impacts.