Membrane Repair and Protectionanatomy2/kadai2011/kadai2011B.pdfDr. Jennifer Lippincott Schwartz...
Transcript of Membrane Repair and Protectionanatomy2/kadai2011/kadai2011B.pdfDr. Jennifer Lippincott Schwartz...
![Page 1: Membrane Repair and Protectionanatomy2/kadai2011/kadai2011B.pdfDr. Jennifer Lippincott Schwartz (NIH) Dr. Tom Kirchhausen (Harvard Medical School) Dr. Kevin Campbell (University of](https://reader036.fdocument.pub/reader036/viewer/2022071421/611a9e2600c4a1639c5f24b8/html5/thumbnails/1.jpg)
二光子レーザー顕微鏡を用いた細胞膜修復機構の観察:二光子レーザー顕微鏡を用いた細胞膜修復機構の観察:
細胞膜修復における細胞膜修復におけるMG53MG53ととdysferlindysferlinの発現についての発現について
市川冬輝市川冬輝 濵田萌濵田萌
香川大学医学部3年香川大学医学部3年
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細胞膜の構造
テキストテキスト
テキストテキスト
テキスト
生物学基礎ホームページより
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FDx (10kD) or HRP(24kD)Scrape or Scratch loading
Bead loadingSyringe loading
How to detect wounded cells in vitro?
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膜プロープFM色素
• FM4−64
• FM1−43
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MG53について
MG53欠損マウス→筋細胞が貧弱
骨格筋の膜修復機構の中核を担う蛋白質
京都大学HPより
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dysferlinについて
• 筋ジストロフィーの原因遺伝子として発見
• 膜修復に関係していることを三宅先生が証明
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マウスの足底筋
DNA
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安静 激しい運動
ラット上腕三頭筋
McNeil et al. (1992) Am. J. Pathol. 140: 1097
激しい運動は膜損傷修復を繰り返す
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Vesicles are important for resealingVesicles are important for resealing
Miyake & McNeil, J Cell Biol, 1995 細胞の分子生物学 第5版
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nature REVIEWS Molecular Cell Biology
コレステロールなど細胞膜構成成分との関わり?
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Kagawa University, Medicine, Miki, Kagawa
LSM710 NLO, Zeiss
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m:Sapphire(Ti:S) Laser SystemChameleon, Coherent
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二光子レーザーアッセイ法
FM1−43
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Plus Ca
Minus Ca
Plus Ca
McNeil & Kirchhausen, nature REVIEWS, Molecular Cell Biology, 2005
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修復成功 修復失敗
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Defective resealing of membrane in DYSF-null
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コレステロール合成阻害剤(スタチン)の副作用
激しい運動後の横紋筋融解症
メカニズム?
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コレステロール合成阻害剤の副作用
横紋筋融解症の発症メカニズム
スタチンが骨格筋線維から細胞膜修復に
必要なコレステロールを減少
⇩
細胞膜修復阻害
⇩
骨格筋線維の死
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シクロデキストリン(MBCD)
⇩
細胞膜のコレステロールを減少
⇩
細胞膜修復は阻害
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Plus Ca2+
Minus Ca2+
MBCD 10mM
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Time (s)
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MBCDを洗浄
⇩
PBSで1時間インキュベート
⇩
細胞内から細胞膜へコレステロールを回復
⇩
細胞膜修復の回復
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MBCD 10mM 10mM MBCD+PBS 60min
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Time (s)
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スタチン(Lovastatin)で5時間⇩
コレステロールの合成を阻害⇩
MBCDで細胞膜のコレステロールを減少⇩
1時間の回復時間⇩
膜修復は回復しない
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水溶性コレステロール
⇩
細胞膜のコレステロールを付与
⇩
細胞膜修復能力は回復
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Statin+ MBCD 10mMStatin
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Statin+ MBCD 10mM+ PBS 60min
Statin+ MBCD 10mM
+ w-Cholesterol 30min
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Time (s)
+MBCD10mM(30min)+PBS(30min)+MBCD10mM(30min)+PBS(60min)
+MBCD10mM(30min)+w-chol 500ug(30min)
PBS(30min)+PBS(30min)
Control
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コレステロール合成阻害剤の副作用
横紋筋融解症の発症メカニズム
スタチンが骨格筋線維から細胞膜修復に
必要なコレステロールを減少
⇩
細胞膜修復阻害
⇩
骨格筋線維の死
![Page 31: Membrane Repair and Protectionanatomy2/kadai2011/kadai2011B.pdfDr. Jennifer Lippincott Schwartz (NIH) Dr. Tom Kirchhausen (Harvard Medical School) Dr. Kevin Campbell (University of](https://reader036.fdocument.pub/reader036/viewer/2022071421/611a9e2600c4a1639c5f24b8/html5/thumbnails/31.jpg)
AcknowledgmentDr. Jennifer Lippincott Schwartz (NIH)Dr. Tom Kirchhausen (Harvard Medical School)Dr. Kevin Campbell (University of Iowa, Antibodies for Dysferlin etc.)Dr. Mark Terasaki (University of Connecticut Health Center)Dr. Susumu Ito (Harvard Medical School)
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THANK YOU
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