Chemistry - An Asian Journal
10.1002/asia.201901689
FULL PAPER
5
% CO
2
in air. After 24 h, the medium was changed to fresh DMEM
to K.Hanaoka) of The Ministry of Education, Culture, Sports,
Science, and Technology, Japan.
containing 10% fetal bovine serum and 1% penicillin and 1%
streptomycin, and the cells were maintained at 37˚C under an
atmosphere of 5% CO
removed, the dish was washed with Hanks’ balanced salt solution
HBSS) three times, and then CaPR-1 AM (10 μM) in HBSS containing
.1% DMSO as a cosolvent was added. After incubation at 37˚C for 1 h,
2
in air. Before loading CaPR-1 AM, DMEM was
Keywords: Fluorescent probes • Fluorescence • Calcium •
Neuroscience • Fluorescence spectroscopy
(
0
the medium was removed and the dish was washed with HBSS three
times. The cells were observed in HBSS.
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Imaging in brain slices. All experiments were carried out in accordance
with the regulations and guidelines of the Institutional Animal Care and
Use Committee at The University of Tokyo and were approved by the
institutional review committees of the Graduate School of Medicine, The
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at
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CaPR-1 potassium salt into single PCs through patch pipettes. The
resistance values of patch pipettes were 4–7 MΩ when filled with the
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intracellular solution containing (in mM) 130 K-gluconate, 4 MgCl
2
, 4
Na ATP, 0.4 Na GTP, 10 sodium phosphocreatine, sodium L-
2
2
3
ascorbate, 10 HEPES and 0.1 CaPR-1 (pH 7.3, adjusted with KOH).
EPSPs were recorded with an EPC-9 patch-clamp amplifier and PULSE
software (HEKA, Lambrecht, Germany). For the focal stimulation of CF,
square pulses (0.1 ms, 5–15 V) were applied through stimulation pipettes
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layer adjacent to the soma of the PC of interest. For PF stimulation, five
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NBQX (10 µM) to block Ca2+ influx via VGCC. Bicuculline and NBQX
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Acknowledgements
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[
This work was supported in part by grants by JSPS KAKENHI
Grant Numbers 16H05099 and 18H04609 to K.Hanaoka, and
SENTAN, JST to K.Hanaoka, who was also supported by
Hoansha Foundation, Daiichi Sankyo Foundation of Life Science,
a grant JSPS Core-to-Core program, A. Advanced Research
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Barone, V.; Mennucci, B.; Petersson, G. A.; Nakatsuji, H.; Caricato, M.;
Li, X.; Hratchian, H. P.; Izmaylov, A. F.; Bloino, J.; Zheng, G.;
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Networks, and
a Grant-in-Aid for Scientific Research on
Innovative Areas “Singularity Biology (No.8007)” (JP19H05414
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