Hypoxia/reoxygenation cardiac injury and regeneration in zebrafish adult heart.
Hypoxia/reoxygenation cardiac injury and regeneration in zebrafish adult heart.
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DOI：
10.1371/journal.pone.0053748
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Capogrossi MC
中科院分区：
文献类型：
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作者：
Parente V;Balasso S;Pompilio G;Verduci L;Colombo GI;Milano G;Guerrini U;Squadroni L;Cotelli F;Pozzoli O;Capogrossi MC
成年斑马鱼的心脏在损伤后会自动再生，并已被用于研究心脏修复的机制。然而，目前尚无模拟哺乳动物心脏缺血损伤的斑马鱼模型。我们建立并表征了低氧/复氧(H/R)诱导的斑马鱼心脏损伤以及随后的再生。成年斑马鱼在低氧(H)和常氧对照(C)水中饲养15min，然后将斑马鱼返回C水中。复氧后2-6小时内，二氢乙锭荧光和蛋白质亚硝化显示心脏氧化应激反应，以及炎症反应。我们用转TG(cmlc2：nucDsRed)的斑马鱼来鉴定心肌细胞核。原位末端标记法(TUNEL)显示心肌细胞发生凋亡和坏死，18h后，9.9±2.6%的心肌细胞核为TUNEL+，15.0±2.5%的心肌细胞核为AO+。在30天(D)时间点，心肌细胞死亡恢复到基线(每个时间点n = 为3)。我们通过磷酸组蛋白H3(PHH3)或增殖细胞核抗原(PCNA)的表达来评估心肌细胞的增殖。心肌细胞在H/R后18~24 h开始明显增殖，3~7 d达高峰，30 d基本恢复正常。缺氧再灌注后7d，心肌细胞pH值为17.4±2.3%，增殖细胞核抗原阳性率为7.4±0.6%(各时间点n = 均为3)。用二维超声心动图评价心功能，用左心室舒缩面积计算面积变化分数(FAC)。FAC由常氧时的2 9.3±2.0%降至H/R后18h的16.4±1.8%，1个月后心功能恢复至基础水平(各时间点n = 均为12)。暴露在H/R下的斑马鱼表现出心脏氧化应激和炎症，心肌细胞死亡和增殖的证据。最初的心功能下降之后是完全恢复。与其他心脏损伤模型相比，该模型更接近哺乳动物的再灌注损伤。
the adult zebrafish heart regenerates spontaneously after injury and has been used to study the mechanisms of cardiac repair. However, no zebrafish model is available that mimics ischemic injury in mammalian heart. We developed and characterized zebrafish cardiac injury induced by hypoxia/reoxygenation (H/R) and the regeneration that followed it. adult zebrafish were kept either in hypoxic (H) or normoxic control (C) water for 15 min; thereafter fishes were returned to C water. Within 2–6 hours (h) after reoxygenation there was evidence of cardiac oxidative stress by dihydroethidium fluorescence and protein nitrosylation, as well as of inflammation. We used Tg(cmlc2:nucDsRed) transgenic zebrafish to identify myocardial cell nuclei. Cardiomyocyte apoptosis and necrosis were evidenced by TUNEL and Acridine Orange (AO) staining, respectively; 18 h after H/R, 9.9±2.6% of myocardial cell nuclei were TUNEL+ and 15.0±2.5% were AO+. At the 30-day (d) time point myocardial cell death was back to baseline (n = 3 at each time point). We evaluated cardiomyocyte proliferation by Phospho Histone H3 (pHH3) or Proliferating Cell Nuclear Antigen (PCNA) expression. Cardiomyocyte proliferation was apparent 18–24 h after H/R, it achieved its peak 3–7d later, and was back to baseline at 30d. 7d after H/R 17.4±2.3% of all cardiomyocytes were pHH3+ and 7.4±0.6% were PCNA+ (n = 3 at each time point). Cardiac function was assessed by 2D-echocardiography and Ventricular Diastolic and Systolic Areas were used to compute Fractional Area Change (FAC). FAC decreased from 29.3±2.0% in normoxia to 16.4±1.8% at 18 h after H/R; one month later ventricular function was back to baseline (n = 12 at each time point). zebrafish exposed to H/R exhibit evidence of cardiac oxidative stress and inflammation, myocardial cell death and proliferation. The initial decrease in ventricular function is followed by full recovery. This model more closely mimics reperfusion injury in mammals than other cardiac injury models.
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影响因子：
56.9
作者：
Poss, KD;Wilson, LG;Keating, MT
通讯作者：
Keating, MT
DOI：
10.1007/978-1-61779-328-8_11
发表时间：
2011-01-01
期刊：
NEURODEGENERATION: METHODS AND PROTOCOLS
影响因子：
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作者：
Laird, Angela S.;Robberecht, Wim
通讯作者：
Robberecht, Wim
DOI：
10.1073/pnas.1834204100
发表时间：
2003-09-30
影响因子：
11.1
作者：
Raya, A;Koth, CM;Izpisúa-Belmonte, JC
通讯作者：
Izpisúa-Belmonte, JC
影响因子：
3.7
作者：
Schnabel K;Wu CC;Kurth T;Weidinger G
通讯作者：
Weidinger G
影响因子：
64.8
作者：
通讯作者：
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