Two-photon imaging of Zn[superscript 2+] dynamics in mossy fiber boutons of adult hippocampal slices
Author(s)Khan, Mustafa; Goldsmith, Christian R.; Huang, Zhen; Georgiou, John; Luyben, Thomas T.; Roder, John C.; Lippard, Stephen J.; Okamoto, Kenichi; ... Show more Show less
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Mossy fiber termini in the hippocampus accumulate Zn[superscript 2+], which is released with glutamate from synaptic vesicles upon neural excitation. Understanding the spatiotemporal regulation of mobile Zn[superscript 2+] at the synaptic level is challenging owing to the difficulty of visualizing Zn[superscript 2+] at individual synapses. Here we describe the use of zinc-responsive fluorescent probes together with two-photon microscopy to image Zn[superscript 2+] dynamics mediated by NMDA receptor-dependent long-term potentiation induction at single mossy fiber termini of dentate gyrus neurons in adult mouse hippocampal slices. The membrane-impermeant fluorescent Zn[superscript 2+] probe, 6-CO2H-ZAP4, was loaded into presynaptic vesicles in hippocampal mossy fiber termini upon KCl-induced depolarization, which triggers subsequent endocytosis and vesicular restoration. Local tetanic stimulation decreased the Zn[superscript 2+] signal observed at individual presynaptic sites, indicating release of the Zn[superscript 2+] from vesicles in synaptic potentiation. This synapse-level two-photon Zn[superscript 2+] imaging method enables monitoring of presynaptic Zn[superscript 2+] dynamics for improving the understanding of physiological roles of mobile Zn[superscript 2+] in regular and aberrant neurologic function.
DepartmentMassachusetts Institute of Technology. Department of Chemistry; Massachusetts Institute of Technology. Department of Physics
Proceedings of the National Academy of Sciences of the United States of America
National Academy of Sciences (U.S.)
Khan, M., C. R. Goldsmith, Z. Huang, J. Georgiou, T. T. Luyben, J. C. Roder, S. J. Lippard, and K. Okamoto. “Two-Photon Imaging of Zn2+ Dynamics in Mossy Fiber Boutons of Adult Hippocampal Slices.” Proceedings of the National Academy of Sciences 111, no. 18 (April 22, 2014): 6786–6791.
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