Genetic Disruption of 21-Hydroxylase in Zebrafish Causes Interrenal Hyperplasia.
Genetic Disruption of 21-Hydroxylase in Zebrafish Causes Interrenal Hyperplasia.
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DOI：
10.1210/en.2017-00549
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发表时间：
2017-12-01
期刊：
影响因子：
4.8
通讯作者：
Krone N
中科院分区：
文献类型：
--
作者：
Eachus H;Zaucker A;Oakes JA;Griffin A;Weger M;Güran T;Taylor A;Harris A;Greenfield A;Quanson JL;Storbeck KH;Cunliffe VT;Müller F;Krone N
先天性肾上腺增生症是一组常见的遗传性疾病，导致糖皮质激素缺乏。大多数病例由21-羟化酶缺乏症（21OHD）引起。严重的21OHD导致的类固醇激素生物合成不平衡的系统性后果仍然知之甚少。因此，我们建立了一个21OHD斑马鱼模型，重点研究糖皮质激素生物合成的损伤。基于序列同源性，在斑马鱼基因组中注释了单个21-羟化酶基因（cyp21a2）。我们对21-羟化酶（Cyp21a2）蛋白序列的计算机分析表明，斑马鱼Cyp21a2在体内用于模拟人类21OHD的各个方面具有足够的相似性。我们通过全安装原位杂交和逆转录聚合酶链反应确定了cyp21a2在早期发育中的时空表达模式。早期cyp21a2的表达仅限于肾间腺（斑马鱼肾上腺对应体）和大脑。为了进一步探索21OHD在体内的影响，我们使用转录激活子样效应核酸酶基因组工程策略创建了几个cyp21a2零等位基因斑马鱼系。纯合子突变斑马鱼幼虫表现出下丘脑-垂体-肾间轴（HPI）上调和肾间增生。此外，cyp21a2缺陷的幼虫具有典型的类固醇特征，皮质醇浓度降低，17-羟基孕酮和21-脱氧皮质醇浓度升高。受累幼虫表现为HPI轴上调和肾间增生。糖皮质激素应答基因pck1和fkbp5的下调表明全身性糖皮质激素缺乏。我们的工作证明了Cyp21a2在斑马鱼幼鱼糖皮质激素生物合成中的关键作用，并建立了一个体内模型，允许研究改变类固醇激素合成的全身后果。21-羟化酶紊乱的斑马鱼幼鱼有肾间增生和全身糖皮质激素缺乏。
Congenital adrenal hyperplasia is a group of common inherited disorders leading to glucocorticoid deficiency. Most cases are caused by 21-hydroxylase deficiency (21OHD). The systemic consequences of imbalanced steroid hormone biosynthesis due to severe 21OHD remains poorly understood. Therefore, we developed a zebrafish model for 21OHD, which focuses on the impairment of glucocorticoid biosynthesis. A single 21-hydroxylase gene (cyp21a2) is annotated in the zebrafish genome based on sequence homology. Our in silico analysis of the 21-hydroxylase (Cyp21a2) protein sequence suggests a sufficient degree of similarity for the usage of zebrafish cyp21a2 to model aspects of human 21OHD in vivo. We determined the spatiotemporal expression patterns of cyp21a2 by whole-mount in situ hybridization and reverse transcription polymerase chain reaction throughout early development. Early cyp21a2 expression is restricted to the interrenal gland (zebrafish adrenal counterpart) and the brain. To further explore the in vivo consequences of 21OHD we created several cyp21a2 null-allele zebrafish lines by using a transcription activator–like effector nuclease genomic engineering strategy. Homozygous mutant zebrafish larvae showed an upregulation of the hypothalamic–pituitary–interrenal (HPI) axis and interrenal hyperplasia. Furthermore, Cyp21a2-deficient larvae had a typical steroid profile, with reduced concentrations of cortisol and increased concentrations of 17-hydroxyprogesterone and 21-deoxycortisol. Affected larvae showed an upregulation of the HPI axis and interrenal hyperplasia. Downregulation of the glucocorticoid-responsive genes pck1 and fkbp5 indicated systemic glucocorticoid deficiency. Our work demonstrates the crucial role of Cyp21a2 in glucocorticoid biosynthesis in zebrafish larvae and establishes an in vivo model allowing studies of systemic consequences of altered steroid hormone synthesis. Zebrafish larvae with disrupted 21-hydroxylase have interrenal hyperplasia and systemic glucocorticoid deficiency.
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DOI：
10.1242/dev.138305
发表时间：
2016-07-15
期刊：
Development (Cambridge, England)
影响因子：
--
作者：
Muthu V;Eachus H;Ellis P;Brown S;Placzek M
通讯作者：
Placzek M
影响因子：
4.8
作者：
Griffin A;Parajes S;Weger M;Zaucker A;Taylor AE;O'Neil DM;Müller F;Krone N
通讯作者：
Krone N
影响因子：
11
作者：
Stewart, A. M.;Ullmann, J. F. P.;Norton, W. H. J.;Parker, M. O.;Brennan, C. H.;Gerlai, R.;Kalueff, A. V.
通讯作者：
Kalueff, A. V.
影响因子：
14.9
作者：
Doyle EL;Booher NJ;Standage DS;Voytas DF;Brendel VP;Vandyk JK;Bogdanove AJ
通讯作者：
Bogdanove AJ
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF