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文昌鱼卵巢中性类固醇的体外转化及性类固醇生成酶基因的表达

In vitro conversion of sex steroids and expression of sex steroidogenic enzyme genes in amphioxus ovary.

作者信息

Mizuta Takanobu, Asahina Kiyoshi, Suzuki Miwa, Kubokawa Kaoru

机构信息

Ocean Research Institute, the University of Tokyo, Nakano, Tokyo, Japan.

出版信息

J Exp Zool A Ecol Genet Physiol. 2008 Mar 1;309(2):83-93. doi: 10.1002/jez.438.

Abstract

Sex steroids are essential hormones for reproduction in vertebrates. The existence of a sex steroidogenic pathway in invertebrates is controversial because cytochrome P450 (CYP) genes have not been detected in the genomes of an echinoderm and a urochordate. However, cloning and gene expressions of sex steroid-metabolizing enzymes have been reported in a cephalochordate. In this study, in vitro conversion of sex steroids from pregnenolone (P5) to progesterone (P4), from 17alpha-hydroxyprogesterone (17alpha-P4) to 17alpha, 20beta-dihydroxy-4-pregnen-3-one (17alpha, 20beta-P) and 17alpha, 20alpha-P, and from androstenedione (AD) to estrone (E1), estradiol-17beta (E2), and testosterone (T) were confirmed by an incubation experiment performed using (14)C-labeled precursors and mature ovarian homogenates of the amphioxus Branchiostoma belcheri. In amphioxus, the ability of immature ovaries to synthesize sex steroids was much lower than that of mature ovaries. Post-spawning, the mRNA of CYP11A significantly decreased in females. Transcripts of the CYP19 gene also declined in one-fourth of the females after spawning, although this trend was not supported statistically. The mRNAs of CYP17 and 17beta-HSD did not change before and after spawning. Our results indicate the presence of Delta(4) and another derivative pathway in the amphioxus ovary and suggest that the synthesis of sex steroids, particularly estrogen synthesis, may be low in females after spawning behavior.

摘要

性类固醇是脊椎动物繁殖所必需的激素。由于在棘皮动物和尾索动物的基因组中未检测到细胞色素P450(CYP)基因,因此无脊椎动物中是否存在性类固醇生成途径存在争议。然而,在头索动物中已报道了性类固醇代谢酶的克隆和基因表达。在本研究中,通过使用(14)C标记的前体和文昌鱼文昌鱼成熟卵巢匀浆进行的孵育实验,证实了性类固醇从孕烯醇酮(P5)向孕酮(P4)、从17α-羟孕酮(17α-P4)向17α,20β-二羟基-4-孕烯-3-酮(17α,20β-P)和17α,20α-P以及从雄烯二酮(AD)向雌酮(E1)、雌二醇-17β(E2)和睾酮(T)的体外转化。在文昌鱼中,未成熟卵巢合成性类固醇的能力远低于成熟卵巢。产卵后,雌性中CYP11A的mRNA显著下降。产卵后,四分之一的雌性中CYP19基因的转录本也有所下降,尽管这一趋势在统计学上未得到支持。产卵前后CYP17和17β-HSD的mRNA没有变化。我们的结果表明文昌鱼卵巢中存在Δ4和另一种衍生途径,并表明产卵行为后雌性中性类固醇的合成,特别是雌激素的合成可能较低。

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