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Cells were recorded by perforated-patch clamp technique in whole cell configuration The current-voltage relation for the nilvadipine-sensitive current was bell-shaped and the peak current reached a maximum at -8 mV in the presence and absence of nilvadipine. Nilvadipine block of voltage-gated Ca current was dose-dependent between 1 and 100 μM. The inhibitory actions of nilvadipine on Ca channels could relieve the intracellular Ca increase in GCs and rescue the GCs from death in glaucoma.\n2) Elevation of intracellular Ca^\u003c2+\u003e concentration ([Ca^\u003c2+\u003e]_i) induced by hypoxia in ganglion cells\nApplication of fluo-3 to the cut edge of the optic nerve of 6-week-old rats. The Ca images of sliced retina were captured. A hypoxic condition was created by superfusing the retinal slice with a solution with an oxygen/glucose deprived solution.\nResults : The retrograde staining method stained GC selectively. Fifteen minutes of hypoxic conditions induced [Ca^\u003c2+\u003e]_i increases in GC (Δ0.13±0.03, n=23). Application of 60 μM\nDL-2-amino-5-phosphonovaleric acid counteracted the hypoxia-induced [Ca^\u003c2+\u003e]_i increase in dendrites partially (Δ0.03±0.02, n=4, p\u003c0.005), but not in somata (Δ0.12±0.02, n=9). GC dendrites showed a further increase in [Ca^\u003c2+\u003e]_i after switching back to oxygenated solution (Δ0.14±0.04, n=4). Neither 6-cyano-7-nitroquinoxaline-2,3-dione disodium, DL-threo-β-benzyloxyaspartate, nifedipine nor bepridil inhibited the hypoxia-induced [Ca^\u003c2+\u003e]_i increase. Ca^\u003c2+\u003e-free superfusion prevented the anoxia-induced [Ca^\u003c2+\u003e]_i increase in somata (Δ0.07±0.02, n=5, p\u003c0.005), not in dendrites (Δ0.16±0.005, n=4).\nConclusions : The mechanisms of hypoxia-induced increase in [Ca^\u003c2+\u003e]_i differ between somata and dendrites. 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カルシウムイメージングとパッチクランプによる緑内障治療薬の神経保護作用の検証
https://doi.org/10.24517/00056946
https://doi.org/10.24517/0005694625c66e25-e21a-43a4-8206-5ef63178fac7
名前 / ファイル | ライセンス | アクション |
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ME-PR-SASAKI-T-kaken 2006-3p.pdf (109.3 kB)
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Item type | 報告書 / Research Paper(1) | |||||
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公開日 | 2020-02-21 | |||||
タイトル | ||||||
タイトル | カルシウムイメージングとパッチクランプによる緑内障治療薬の神経保護作用の検証 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | The research in neuroprotection effect of anti-glaucoma drug with Ca imaging and patch clamping | |||||
言語 | ||||||
言語 | jpn | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_18ws | |||||
資源タイプ | research report | |||||
ID登録 | ||||||
ID登録 | 10.24517/00056946 | |||||
ID登録タイプ | JaLC | |||||
著者 |
佐々木, 次壽
× 佐々木, 次壽 |
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著者別表示 |
Sasaki, Tsugihisa
× Sasaki, Tsugihisa |
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書誌情報 |
平成17(2005)年度 科学研究費補助金 基盤研究(C) 研究成果報告書 en : 2005 Fiscal Year Final Research Report 巻 2004-2005, p. 3p., 発行日 2006-03 |
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出版者 | ||||||
出版者 | 金沢大学附属病院 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | 1)パッチクランプ法による神経節細胞Ca電流への緑内障治療薬の効果: 金魚網膜を酵素的に処理し神経節細胞を単離,顕微鏡下に同定した.穿孔パッチクランプをかけwhole cell configurationにてCa電流を測定した. 網膜神経節細胞L-型Caチャネルへのニルバジピンの最小有効濃度が1μMであること,保持電位が-28〜+50mVでCa電流の抑制効果を認めることを認めた.これらの結果と内服投与後の眼内濃度よりニルバジピンは神経節細胞に直接作用して保護しうると推測された.これらの結果はJournal of ocular pharmacology and therapeuticsに平成17年12月に投稿した. 2)Caイメージング法による神経節細胞の低酸素負荷による細胞内Ca^<2+>濃度([Ca^<2+>]_1)上昇の解析.ラットを用いて,新たに開発した網膜スライス内神経節細胞(GC)選択的Caメージング法を用いて神経節細胞内Ca濃度測定を行った.スライス標本を低O_2条件にするとGCの[Ca^<2+>]_1上昇が認められた,GC(0.14±0.04, Mean±SD)。この[Ca^<2+>]_i上昇はNMDA型glu受容体阻害薬APV(60μM)でGCにおいて樹上突起部で抑制された,神経節細胞層(0.12±0.01,), IPL(0.04±0.04, p<0.005).CNQXはGCの[Ca^<2+>]_i上昇を抑制しなかった.L型Caチャネル阻害薬ニフェジピンはGCでは抑制しなかった(0.13±0.01). 結論:低酸素条件下でのGC樹上突起部での[Ca^<2+>]_I)上昇はNMDA受容体を介する. これらの結果はJapanese Journal of Ophthalmologyに平成18年3月に投稿予定である. |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | 1) The effect of nilvadipine on the voltage-gated Ca channel of retinal ganglion cells (GC). GC were enzymatically dissociated from isolated retina. Cells were recorded by perforated-patch clamp technique in whole cell configuration The current-voltage relation for the nilvadipine-sensitive current was bell-shaped and the peak current reached a maximum at -8 mV in the presence and absence of nilvadipine. Nilvadipine block of voltage-gated Ca current was dose-dependent between 1 and 100 μM. The inhibitory actions of nilvadipine on Ca channels could relieve the intracellular Ca increase in GCs and rescue the GCs from death in glaucoma. 2) Elevation of intracellular Ca^<2+> concentration ([Ca^<2+>]_i) induced by hypoxia in ganglion cells Application of fluo-3 to the cut edge of the optic nerve of 6-week-old rats. The Ca images of sliced retina were captured. A hypoxic condition was created by superfusing the retinal slice with a solution with an oxygen/glucose deprived solution. Results : The retrograde staining method stained GC selectively. Fifteen minutes of hypoxic conditions induced [Ca^<2+>]_i increases in GC (Δ0.13±0.03, n=23). Application of 60 μM DL-2-amino-5-phosphonovaleric acid counteracted the hypoxia-induced [Ca^<2+>]_i increase in dendrites partially (Δ0.03±0.02, n=4, p<0.005), but not in somata (Δ0.12±0.02, n=9). GC dendrites showed a further increase in [Ca^<2+>]_i after switching back to oxygenated solution (Δ0.14±0.04, n=4). Neither 6-cyano-7-nitroquinoxaline-2,3-dione disodium, DL-threo-β-benzyloxyaspartate, nifedipine nor bepridil inhibited the hypoxia-induced [Ca^<2+>]_i increase. Ca^<2+>-free superfusion prevented the anoxia-induced [Ca^<2+>]_i increase in somata (Δ0.07±0.02, n=5, p<0.005), not in dendrites (Δ0.16±0.005, n=4). Conclusions : The mechanisms of hypoxia-induced increase in [Ca^<2+>]_i differ between somata and dendrites. The NMDA channel of dendrites seems to be the main route of Ca^<2+> influx. |
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内容記述 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 研究課題/領域番号:16591741, 研究期間(年度):2004-2005 | |||||
内容記述 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 出典:「カルシウムイメージングとパッチクランプによる緑内障治療薬の神経保護作用の検証」研究成果報告書 課題番号16591741 (KAKEN:科学研究費助成事業データベース(国立情報学研究所)) 本文データは著者版報告書より作成 |
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著者版フラグ | ||||||
出版タイプ | AM | |||||
出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/search/?qm=00338188 | |||||
関連名称 | https://kaken.nii.ac.jp/search/?qm=00338188 | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16591741/ | |||||
関連名称 | https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16591741/ | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-16591741/165917412005kenkyu_seika_hokoku_gaiyo/ | |||||
関連名称 | https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-16591741/165917412005kenkyu_seika_hokoku_gaiyo/ |