Developmental expression of cytochrome P450 aromatase genes (CYP19a and CYP19b) in zebrafish fry (Danio rerio).
Developmental expression of cytochrome P450 aromatase genes (CYP19a and CYP19b) in zebrafish fry (Danio rerio).
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DOI：
10.1002/jez.1090
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发表时间：
2001-09
期刊：
影响因子：
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通讯作者：
J. Trant;S. Gavasso;J. Ackers;B. Chung;A. Place
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文献类型：
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作者：
J. Trant;S. Gavasso;J. Ackers;B. Chung;A. Place
细胞色素P450芳香酶(CYP19)是类固醇合成途径中将雄激素(如睾酮)转化为雌激素(如雌二醇)的末端酶。该基因的调控决定了雄激素和雌激素的比例；因此，该酶的适当表达对生殖至关重要，也是大多数脊椎动物性别分化的关键。据推测，大多数脊椎动物都有一个受多个组织特异性启动子区域调控的CYP19基因。然而，斑马鱼(Danio Rerio)有两个基因(CyP19a和CyP19b)，每个基因编码一种明显不同的蛋白质，并拥有自己的调节机制。本研究的主要目的是确定CYP19基因在发育中的斑马鱼中的表达模式。一种基于荧光的实时定量RT-PCR方法提供了检测受精后0-41天(DPF)单个胚胎/幼体转录丰度的敏感性和特异性，该方法包括性别决定和性腺分化的发育事件。CyP19转录本最早可在3d或4d检测到(分别为CyP19a和CyP19b)，并在第5天检测到丰度高峰。总体而言，CYP19基因在其表达的个体发育过程中存在显著差异。在大多数情况下，性腺形式的细胞色素P19(CYP19a)比脑形式的细胞色素P19b(CyP19b)更丰富；然而，与CYP19a不同的是，细胞色素P19b的表达模式可以明显地分为两个群体，这表明与性别分化有关。药物类固醇(乙炔雌二醇和17α-甲基睾酮)在被检测的所有三天(4、6和10dpf)都增强了CYP19b基因的表达。这些数据表明，适时和适当地表达CYP19在发育过程中很重要，而CYP19b(性腺外形式)的表达可能与性别分化有关，如果不是与性决定有关的话。J.Exp.祖尔。290：475-483,2001。
Cytochrome P450 aromatase (CYP19) is the terminal enzyme in the steroidogenic pathway that converts androgens (e.g., testosterone) into estrogens (e.g., estradiol). Regulation of this gene dictates the ratio of androgens to estrogens; therefore, appropriate expression of this enzyme is critical for reproduction as well as being pivotal in sex differentiation for most vertebrates. It is assumed that most vertebrates have a single CYP19 gene that is regulated by multiple tissue-specific promoter regions. However, the zebrafish (Danio rerio) has two genes (CYP19a and CYP19b), each encoding a significantly different protein and possessing its own regulatory mechanism. The primary purpose of this study was to determine the pattern of expression of each of the CYP19 genes in the developing zebrafish. A fluorescent-based method of real-time, quantitative RT-PCR provided the sensitivity and specificity to determine transcript abundance in single embryos/juveniles harvested at days 0 through 41 days post-fertilization (dpf), which encompasses the developmental events of sex determination and gonadal differentiation. CYP19 transcripts could be detected as early as 3 or 4 dpf, (CYP19a and CYP19b, respectively) and peak abundance was detected on day five. In general, the CYP19 genes differed significantly in the ontogeny of their expression. In most cases, the gonadal form of CYP19 (CYP19a) was more abundant than the brain form (CYP19b); however, unlike CYP19a, the pattern of CYP19b expression could be clearly segregated into two populations, suggesting an association with sex differentiation. Pharmacological steroids (ethinylestradiol and 17 alpha-methyltestosterone) enhanced the expression of the CYP19b gene at all three days examined (4, 6, and 10 dpf). These data suggest that the timely and appropriate expression of CYP19 is important in development and that the expression of CYP19b (the "extra-gonadal" form) may be associated with sexual differentiation if not sexual determination. J. Exp. Zool. 290:475-483, 2001.