Deletion of the murine cytochrome P450 Cyp2j locus by fused BAC-mediated recombination identifies a role for Cyp2j in the pulmonary vascular response to hypoxia.
Deletion of the murine cytochrome P450 Cyp2j locus by fused BAC-mediated recombination identifies a role for Cyp2j in the pulmonary vascular response to hypoxia.
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DOI：
10.1371/journal.pgen.1003950
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发表时间：
2013-11
期刊：
影响因子：
4.5
通讯作者：
Bloch KD
中科院分区：
文献类型：
--
作者：
Zhou GL;Beloiartsev A;Yu B;Baron DM;Zhou W;Niedra R;Lu N;Tainsh LT;Zapol WM;Seed B;Bloch KD
环氧二十碳三烯酸（Epoxyeicosatrienoic Acids，EAE）具有血管活性和心脏保护功能。迄今为止，这些短寿命介质对病理生理学的贡献的遗传分析已被啮齿类动物中与人CYP 2 J2同线的基因组部分的等位基因扩增所混淆，CYP 2 J2是编码负责人类中雌二醇形成的主要细胞色素P450环氧合酶之一的基因。小鼠有八个潜在的功能基因，可以指导合成具有与CYP 2 J2相似性质的环氧合酶。作为理解小鼠Cyp 2 j基因座作用的第一步，我们使用同源和定点重组策略的组合，创建了携带跨越与CYP 2 J2同线的整个区域的626-kb缺失的小鼠。还创建了其中基因座缺失通过转基因递送编码人CYP 2 J2的BAC序列来补充的小鼠品系。在基线时，野生型、无效和补充小鼠的全身和肺血流动力学测量值无差异。然而，缺氧性肺血管收缩（HPV）在左主干支气管闭塞受损，并与减少全身氧合在null小鼠，但不是在null小鼠轴承人转基因。给野生型小鼠施用环氧合酶抑制剂也会损害HPV。这些发现表明Cyp 2 j基因产物调节肺血管对缺氧的反应。在小鼠和人类中，CYP 2 J类细胞色素P450环氧合酶将花生四烯酸（AA）代谢为环氧二十碳三烯酸（Epoxyeicosatrienoic Acids，EAE），EAE是对肺和全身血管系统均有影响的短寿命介质。迄今为止，CYP 2 J功能的遗传解剖因啮齿动物基因组中的等位基因扩增而变得复杂。在这项研究中，小鼠染色体基因座共线的人CYP 2 J2，含有8个假定的基因和两个假基因，删除通过同源和位点特异性重组步骤，连接两个前体细菌人工染色体（BAC）创建的重组模板的生成。随后通过转基因递送编码人CYP 2 J2的BAC序列补充Cyp 2 j缺失小鼠。出乎意料地发现，在空小鼠中，局部肺泡缺氧期间的肺血管收缩功能减退（HPV）和全身氧合受损，但在携带转基因人等位基因的空小鼠中未受损，这表明Cyp 2 j产物有助于肺血管对缺氧的反应。
Epoxyeicosatrienoic acids (EETs) confer vasoactive and cardioprotective functions. Genetic analysis of the contributions of these short-lived mediators to pathophysiology has been confounded to date by the allelic expansion in rodents of the portion of the genome syntenic to human CYP2J2, a gene encoding one of the principle cytochrome P450 epoxygenases responsible for the formation of EETs in humans. Mice have eight potentially functional genes that could direct the synthesis of epoxygenases with properties similar to those of CYP2J2. As an initial step towards understanding the role of the murine Cyp2j locus, we have created mice bearing a 626-kb deletion spanning the entire region syntenic to CYP2J2, using a combination of homologous and site-directed recombination strategies. A mouse strain in which the locus deletion was complemented by transgenic delivery of BAC sequences encoding human CYP2J2 was also created. Systemic and pulmonary hemodynamic measurements did not differ in wild-type, null, and complemented mice at baseline. However, hypoxic pulmonary vasoconstriction (HPV) during left mainstem bronchus occlusion was impaired and associated with reduced systemic oxygenation in null mice, but not in null mice bearing the human transgene. Administration of an epoxygenase inhibitor to wild-type mice also impaired HPV. These findings demonstrate that Cyp2j gene products regulate the pulmonary vascular response to hypoxia. In mice and humans, the CYP2J class of cytochrome P450 epoxygenases metabolizes arachidonic acid (AA) to epoxyeicosatrienoic acids (EETs), short-lived mediators with effects on both the pulmonary and systemic vasculature. Genetic dissection of CYP2J function to date has been complicated by allelic expansion in the rodent genome. In this study, the mouse chromosomal locus syntenic to human CYP2J2, containing eight presumed genes and two pseudogenes, was deleted via generation of a recombinant template created by homologous and site-specific recombination steps that joined two precursor bacterial artificial chromosomes (BACs). The Cyp2j null mice were subsequently complemented by transgenic delivery of BAC sequences encoding human CYP2J2. Hypoxic pulmonary vasoconstriction (HPV) and systemic oxygenation during regional alveolar hypoxia were unexpectedly found to be impaired in null mice, but not in null mice bearing the transgenic human allele, suggesting that Cyp2j products contribute to the pulmonary vascular response to hypoxia.
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DOI：
10.1164/rccm.200501-034oc
发表时间：
2005-08-01
影响因子：
24.7
作者：
Caironi, P;Ichinose, F;Zapol, WM
通讯作者：
Zapol, WM
影响因子：
19.6
作者：
SCHWARTZMAN, ML;MARTASEK, P;ABRAHAM, NG
通讯作者：
ABRAHAM, NG
影响因子：
2.8
作者：
Fox T;Gotlinger KH;Dunn MW;Lee OL;Milman T;Zaidman G;Schwartzman ML;Bellner L
通讯作者：
Bellner L
影响因子：
4.8
作者：
Keserue, Benjamin;Barbosa-Sicard, Eduardo;Fleming, Ingrid
通讯作者：
Fleming, Ingrid
DOI：
10.1152/ajplung.1997.272.5.l823
发表时间：
1997-05-01
影响因子：
4.9
作者：
Birks, EK;Bousamra, M;Jacobs, ER
通讯作者：
Jacobs, ER