2. Academic Validation
  3. A high-performance fluorescent sensor spatiotemporally reveals cell-type specific regulation of intracellular adenosine in vivo

A high-performance fluorescent sensor spatiotemporally reveals cell-type specific regulation of intracellular adenosine in vivo

  • Nat Commun. 2025 May 7;16(1):4245. doi: 10.1038/s41467-025-59530-7.
Qingpeng Wei # 1,  Zexiao Bai # 2 3,  Lei Wang # 4 5,  Jing Wang 2 6,  Yipan Wang 4 5,  Yufei Hu 1,  Shiyi Ding 3 7 8,  Zhixiong Ma 2 3,  Chun Li 2 3,  Yumo Li 2 3,  Yizhou Zhuo 4 5,  Wenzhe Li 1,  Fei Deng 4 5,  Bingjie Liu 1,  Pengcheng Zhou 7 8,  Yulong Li 9 10 11,  Zhaofa Wu 12 13,  Jing Wang 14 15
Affiliations

Affiliations

  • 1 State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, China.
  • 2 State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
  • 3 University of Chinese Academy of Sciences, Beijing, China.
  • 4 State Key Laboratory of Membrane Biology, School of Life Sciences, Peking University, Beijing, China.
  • 5 PKU-IDG/McGovern Institute for Brain Research, Beijing, China.
  • 6 Department of Anesthesiology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.
  • 7 Interdisciplinary Center for Brain Information, the Brain Cognition and Brain Disease Institute, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
  • 8 Faculty of Life and Health Sciences, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
  • 9 State Key Laboratory of Membrane Biology, School of Life Sciences, Peking University, Beijing, China. yulongli@pku.edu.cn.
  • 10 PKU-IDG/McGovern Institute for Brain Research, Beijing, China. yulongli@pku.edu.cn.
  • 11 National Biomedical Imaging Center, Peking University, Beijing, China. yulongli@pku.edu.cn.
  • 12 State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China. wuzhaofa@genetics.ac.cn.
  • 13 University of Chinese Academy of Sciences, Beijing, China. wuzhaofa@genetics.ac.cn.
  • 14 State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, China. wangjingsioc@pku.edu.cn.
  • 15 National Biomedical Imaging Center, Peking University, Beijing, China. wangjingsioc@pku.edu.cn.
  • # Contributed equally.
PMID: 40335490 DOI: 10.1038/s41467-025-59530-7
Abstract

Adenosine (Ado), a nucleoside bridging intracellular metabolism with intercellular communication, plays an essential role in regulating processes such as sleep and seizure. While the functions of extracellular Ado ("eAdo") are well documented, our knowledge about the distribution and regulatory functions of intracellular Ado ("iAdo") is limited by a lack of methods for detecting iAdo in vivo. Here, we develop HypnoS, a genetically encoded fluorescent sensor for iAdo characterized by its high sensitivity, specificity, spatiotemporal resolution, and rapid response (sub-seconds). HypnoS enables real-time visualization of iAdo dynamics in live cultures, acute brain slices, flies, and freely moving mice. Using HypnoS for dual-color mesoscopic imaging in mice, we show that seizure-induced iAdo waves propagated across the cortex, following calcium signals. Additionally, two-photon imaging reveals that iAdo decays more rapidly in astrocytes than in neurons during seizures. Moreover, by recording iAdo dynamics in the basal forebrain during the sleep-wake cycle, we observe that iAdo signals are present during wakefulness and rapid eye movement (REM) sleep, regulated by equilibrative Nucleoside Transporters (ENT1/2). Thus, HypnoS is a versatile and powerful tool for investigating the biological functions of iAdo across a range of physiological and pathological states.

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