All three subtypes of inositol 1,4,5-trisphosphate receptor (IP3R) are intracellular Ca2+ channels that are co-regulated by IP3 and Ca2+. This allows IP3Rs to evoke regenerative Ca2+ signals, the smallest of which are Ca2+ puffs that reflect the coordinated opening of a few clustered IP(3)Rs. We use total internal reflection microscopy (TIRF) microscopy to record Ca2+ signals in HEK cells expressing all three IP3R subtypes or a single native subtype. Ca2+ puffs are less frequent in cells expressing one IP3R subtype, commensurate with them expressing fewer IP(3)Rs than wild-type cells. However, all three IP3R subtypes generate broadly similar Ca2+ puffs with similar numbers of IP(3)Rs contributing to each. This suggests that IP3R clusters may be assembled by conserved mechanisms that generate similarly sized clusters across different IP3R expression levels. The Ca2+ puffs evoked by IP(3)R2 had slower kinetics and more prolonged durations, which may be due to IP3 binding with greater affinity to IP(3)R2. We conclude that Ca2+ puffs are the building blocks for the Ca2+ signals evoked by all IP(3)Rs.
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Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, JapanJapan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
Futatsugi, A
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Nakamura, T
Yamada, MK
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Yamada, MK
Ebisui, E
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Ebisui, E
Nakamura, K
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Nakamura, K
Uchida, K
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Uchida, K
Kitaguchi, T
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Kitaguchi, T
Takahashi-Iwanaga, H
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Takahashi-Iwanaga, H
Noda, T
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Noda, T
Aruga, J
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Aruga, J
Mikoshiba, K
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机构:Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
机构:Univ Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, Japan
Miyakawa, T
Maeda, A
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Maeda, A
Yamazawa, T
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Yamazawa, T
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Hirose, K
Kurosaki, T
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Kurosaki, T
Iino, M
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Univ Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, JapanUniv Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, Japan
机构:
Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, JapanJapan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
Futatsugi, A
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Nakamura, T
Yamada, MK
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Yamada, MK
Ebisui, E
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Ebisui, E
Nakamura, K
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机构:Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
Nakamura, K
Uchida, K
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Uchida, K
Kitaguchi, T
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Kitaguchi, T
Takahashi-Iwanaga, H
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机构:Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
Takahashi-Iwanaga, H
Noda, T
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机构:Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
Noda, T
Aruga, J
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机构:Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
Aruga, J
Mikoshiba, K
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机构:Japan Sci & Technol Agcy, Calcium Oscillat, Int Cooperat Res Project, Tokyo 1080071, Japan
机构:Univ Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, Japan
Miyakawa, T
Maeda, A
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机构:Univ Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, Japan
Maeda, A
Yamazawa, T
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机构:Univ Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, Japan
Yamazawa, T
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Hirose, K
Kurosaki, T
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机构:Univ Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, Japan
Kurosaki, T
Iino, M
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Univ Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, JapanUniv Tokyo, Fac Med, Dept Pharmacol, CREST,Japan Sci & Technol,Bunkyo Ku, Tokyo 113, Japan