A Novel Acetylcholine Nanosensor for Single Vesicle Storage and Sub-Quantal Exocytosis in Living Neurons and Organoids.

Journal: Angewandte Chemie (International ed. in English)
Published Date: Jan 10, 2026

Abstract

Acetylcholine (ACh) is a critical neurotransmitter that regulates diverse physiological functions, such as cognition and muscle contraction, through synaptic transmission. However, in situ quantitative chemical analysis of single-vesicle storage and release dynamics at single-cell level remains a major technical challenge, hindering mechanistic understanding of cholinergic synaptic plasticity across physiological and pathological states. To address this, we developed an Ti3C2Tx MXene/enzyme-functionalized (M@E@CF) nanosensor, enabling real-time monitoring of vesicular ACh storage and exocytotic release in primary cholinergic neurons and human spinal cord organoids. Our findings reveal a sub-quantal release mode in both mouse and human-derived neurons. To further elucidate the regulatory principles of exocytosis kinetics, we classified single-vesicle exocytosis patterns based on signal peak shapes using a 1D convolutional neural network (1D-CNN) deep learning model, uncovering significant differences in the number of released molecules and kinetic parameters across modes. Critically, in Down syndrome models, we observed significantly reduced single vesicle ACh storage and release alongside an elevated release fraction, concurrent with shortened fusion pore durations during exocytosis. Thus, the M@E@CF nanosensor platform establishes a versatile tool for spatiotemporal investigation of neurotransmitter storage and release dynamics, providing a critical technical foundation for exploring physiological functions and pathological mechanisms of the cholinergic system.

Authors

  • Wanying Zhu
    School of Pharmacy, Nanjing Medical University, Nanjing, 211166, China.
  • Yufan Zhang
    School of Pharmacy, Nanjing Medical University, Nanjing, 211166, China.
  • Hanwen Yu
    School of Mechanical and Automotive Engineering, Liaocheng University, Liaocheng, China.
  • Da Wang
    Department of Colorectal Surgery, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China; Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education, Key Laboratory of Molecular Biology in Medical Sciences, Zhejiang Province, China; The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
  • Xinyue Zhang
    Department of Radiology, Changhai Hospital.
  • Xiaowen Du
    Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, Beijing 100081, China.
  • Shanshan Wu
    Ningbo Key Laboratory of Genomic Medicine and Birth Defects Prevention, The Affiliated Women and Children's Hospital of Ningbo University, Ningbo, 315021, China.
  • Bingzhi Li
    College of Food Science and Engineering, Northwest A&F University, Yangling 712100, Shaanxi, China.
  • Yizhou Zhang
    Institute of Advanced Materials and Flexible Electronics (IAMFE), School of Chemistry and Materials Science, Nanjing University of Information Science & Technology, Nanjing 210044, China.
  • Hongliang Xin
    Department of Pharmaceutics, School of Pharmacy, Nanjing Medical University, Nanjing, China.
  • Xing Guo
    Shanxi Key Laboratory of Micro Nano Sensors & Artificial Intelligence Perception, College of Information and Computer, Taiyuan University of Technology, Taiyuan 030024, China.
  • Yan Liu
    Department of Clinical Microbiology, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai, 200072, People's Republic of China.

Keywords

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