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A genetically encoded far-red fluorescent indicator for imaging synaptically released Zn 2+

2023; American Association for the Advancement of Science; Volume: 9; Issue: 9 Linguagem: Inglês

10.1126/sciadv.add2058

2375-2548

Tianchen Wu, Manoj Kumar, Jing Zhang, Shengyu Zhao, Mikhail Drobizhev, Mason McCollum, Charles T. Anderson, Ying Wang, Antje Pokorny, Xiaodong Tian, Yiyu Zhang, Thanos Tzounopoulos, Hui‐wang Ai,

Advanced biosensing and bioanalysis techniques

Synaptic zinc ion (Zn2+) has emerged as a key neuromodulator in the brain. However, the lack of research tools for directly tracking synaptic Zn2+ in the brain of awake animals hinders our rigorous understanding of the physiological and pathological roles of synaptic Zn2+. In this study, we developed a genetically encoded far-red fluorescent indicator for monitoring synaptic Zn2+ dynamics in the nervous system. Our engineered far-red fluorescent indicator for synaptic Zn2+ (FRISZ) displayed a substantial Zn2+-specific turn-on response and low-micromolar affinity. We genetically anchored FRISZ to the mammalian extracellular membrane via a transmembrane (TM) ⍺ helix and characterized the resultant FRISZ-TM construct at the mammalian cell surface. We used FRISZ-TM to image synaptic Zn2+ in the auditory cortex in acute brain slices and awake mice in response to electric and sound stimuli, respectively. Thus, this study establishes a technology for studying the roles of synaptic Zn2+ in the nervous system.