中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2026, Vol. 21 ›› Issue (7): 3083-3091.doi: 10.4103/NRR.NRR-D-24-01607

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

用于选择性稀疏标记星形胶质细胞的双腺病毒系统

  

  • 出版日期:2026-07-15 发布日期:2026-03-31
  • 基金资助:
    本研究获得国家自然科学基金、广东省自然科学基金等项目资助。

Dual adeno-associated virus system for selective and sparse labeling of astrocytes

Mei Li1, #, Zhuang Liu2, 3, 4, #, Ruixi Chen5, Ziyue Zhao2, 3, 4, Qingqing Zhou5, Ning Zheng6, Jie Wang4, 7, *, Hanbing Wang1, *   

  1. 1Department of Anesthesiology, The First People’s Hospital of Foshan, Foshan, Guangdong Province, China; 
    2National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, Hubei Province, China; 
    3University of Chinese Academy of Sciences, Beijing, China; 
    4Department of Radiology, Songjiang Research Institute, Shanghai Key Laboratory of Emotions and Affective Disorders, Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China; 
    5Department of Anesthesiology; Brain Research Center; Zhongnan Hospital, Wuhan University, Wuhan, Hubei Province, China; 
    6Clinical & Technical Support, Philips Healthcare, Wuhan, Hubei Province, China; 
    7Institute of Neuroscience and Brain Diseases; Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, Hubei Province, China
  • Online:2026-07-15 Published:2026-03-31
  • Contact: Jie Wang, PhD, jie.wang@shsmu.edu.cn; Hanbing Wang, PhD, fswhbing@126.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 32271148 (to JW); the National Key Research and the Development Program of China, No. 2023M740625 (to ML); the Natural Science Foundation of Guangdong Province, Nos. 2021B1515120050 (to HW) and 2023A1515110782 (to ML); and Key R&D Program of Ningxia Hui Autonomous Region, No. 2024BEG02027 (to JW).

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摘要：

星形胶质细胞是中枢神经系统中数量最多的胶质细胞，具有多种功能，在结构完整性、突触形成和神经传导方面发挥着关键作用。这些细胞具有明显的区域异质性，对不同的神经系统疾病表现出不同的反应。星形胶质细胞形态和功能的这种多样性对于了解正常大脑功能和神经系统疾病的潜在机制至关重要。针对这一问题，实验开发了一种双腺病毒系统（dual-AAV, DAS），用于小鼠大脑中星形胶质细胞的选择性和稀疏标记，并在多个脑区验证了其高效性。通过GfaABC1D启动子驱动Cre重组酶和增强型绿色荧光蛋白表达，利用尾静脉注射PHP.eB病毒或立体定位注射，在C57BL/6J小鼠的12个脑区实现特异性标记。结合共聚焦显微镜三维重建，实验揭示了星形胶质细胞在分支复杂度、面积大小等方面的形态异质性。此外，通过坐骨神经分支选择性损伤模型和化学遗传学操作，双腺病毒系统不仅能清晰呈现细胞形态，还可调控细胞功能，显示出其在形态与功能研究中的双重潜力。进一步分析表明，不同脑区的形态差异可能与其分区功能相关。这种创新方法不仅能让人们更全面地了解星形胶质细胞在神经损伤中的变化，还能让人们更全面地了解星形胶质细胞在多种神经疾病中的变化，因此对未来的研究大有可为。选择性标记和研究不同脑区星形胶质细胞的能力为揭示这些重要细胞的复杂性及其在生理和病理条件下的作用提供了强有力的工具。

https://orcid.org/0000-0002-2997-3450 (Jie Wang);

https://orcid.org/0000-0003-0145-0081 (Hanbing Wang)

关键词: 星形胶质细胞, 化学基因调控, 双腺病毒系统, 胶质纤维酸性蛋白（GfaABC1D）启动子, 分层聚类方法, 形态参数分析, PHP.eB, Sholl 分析, 神经损伤, 稀疏标记

Abstract:

Astrocytes are the most abundant glial cells in the central nervous system. They perform a diverse array of functions, with a critical role in structural integrity, synapse formation, and neurotransmission. These cells exhibit substantial regional heterogeneity and display variable responses to different neurological diseases. Such diversity in astrocyte morphology and function is essential for understanding both normal brain function and the underlying mechanisms of neurological disorders. To investigate this heterogeneity, we developed a novel method for the selective and sparse labeling of astrocytes in various brain regions. This technique utilizes a dual adeno-associated virus system that allows for the expression of Cre recombinase and enhanced green fluorescent protein under the control of the glial fibrillary acidic protein (GfaABC1D) promoter. The system was tested in C57BL/6J mice and successfully labeled astrocytes across multiple brain regions. The method enabled the detailed visualization of individual astrocytes—including their intricate peripheral processes—through three-dimensional reconstructions from confocal microscopy images. Furthermore, the labeling efficiency of this dual adeno-associated virus technology was validated by examining astrocyte function in a spared nerve injury model and through chemogenetic modulation. This innovative approach holds great promise for future research because it enables a more comprehensive understanding of astrocyte variation not only in spared nerve injury but also in a broad spectrum of neurological diseases. The ability to selectively label and study astrocytes in different brain regions provides a powerful tool for exploring the complexities of these essential cells and their roles in physiological and pathological conditions.

Key words: astrocytes, chemogenetic modulation, dual-adeno-associated virus system, glial fibrillary acidic protein (GfaABC1D) promoter, hierarchical clustering approach, morphological parameter analysis, PHP.eB, Sholl analysis, spared nerve injury, sparse labeling