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Neuro Imaging

開催日 2014/9/11
時間 10:00 - 11:00
会場 Room H(304)
Chairperson(s) 喜多村 和郎 / Kazuo Kitamura (東京大学大学院医学系研究科 神経生理学 / Department of Neurophysiology, Graduate School of Medicine, The University of Tokyo, Japan)
今井 猛 / Takeshi Imai (理化学研究所 発生・再生科学総合研究センター(理研CDB) / RIKEN Center for Developmental Biology, Japan)

新規の高感度緑色カルシウム指示薬Cal-520を用いたin vitro およびin vivoにおける神経活動のイメージング
Highly sensitive calcium imaging of neural activity in vitro and in vivo with a new fluorescent calcium indicator, Cal-520

  • O1-H-2-3
  • 竹内 敦也 / Atsuya Takeuchi:1 多田 真弓 / Mayumi Tada:1 橋爪 幹 / Miki Hashizume:2 喜多村 和郎 / Kazuo Kitamura:1 狩野 方伸 / Masanobu Kano:1 
  • 1:東京大院・医・神経生理 / Dept Neurophysiol, Univ of Tokyo, Tokyo, Japan 2:埼玉医科大学医学部生化学 / Dept of Biochemistry, Faculty of Medicine, Saitama Medical University, Saitama, Japan 

In neurons, action potentials (APs) induce transient increase in intracellular calcium concentration. Synthetic calcium indicators are widely used for monitoring neuronal activity in vitro and in vivo. However, calcium signals produced by synthetic fluorescent calcium indicators are sometimes suffered from a low signal-to-noise ratio (SNR), which precludes the detection of APs particularly in neurons with low firing rates in vivo.

Here, we present the data from comparative in vitro and in vivo analysis of two different synthetic calcium indicators for monitoring the activity of individual neurons: 1) Cal-520 : a recently developed calcium indicator dye and 2) Oregon Green BAPTA-1(OGB-1) : the most commonly used calcium indicator dye.

We demonstrate that Cal-520 is sufficiently sensitive to reliably detect single APs both in vitro and in vivo. In neocortical neurons, calcium signals were linearly correlated with the number of APs, and the SNR was > 6 for in vitro slice preparations and > 1.6 for in vivo anesthetized mice. In cerebellar Purkinje cells, dendritic calcium transients evoked by climbing fiber inputs were clearly observed in anesthetized mice with a high SNR and fast decay time. These characteristics of Cal-520 are a great advantage over those of OGB-1 for monitoring neural activity both in vitro and in vivo.

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