Ijiri Shigeho, Kaneko Hiroyo, Kobayashi Tohru, Wang De-Shou, Sakai Fumie, Paul-Prasanth Bindhu, Nakamura Masaru, Nagahama Yoshitaka
Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan.
Biol Reprod. 2008 Feb;78(2):333-41. doi: 10.1095/biolreprod.107.064246. Epub 2007 Oct 17.
The Nile tilapia, a gonochoristic teleost fish with an XX/XY sex-determining system, provides an excellent model for studying gonadal sex differentiation because genetic all-females and all-males are available. In this study, we used quantitative real-time RT-PCR to determine the precise timing of the gonadal expression of 17 genes thought to be associated with gonadal sex differentiation in vertebrates. Gonads were isolated from all-female and all-male tilapia before (5-15 days after hatching [dah]) and after (25-70 dah) morphological sex differentiation. The transcript of aromatase (cyp19a1a), an enzyme responsible for producing estradiol-17beta, was expressed only in XX gonads at 5 dah, with a marked elevation in expression thereafter. In contrast, mRNA expression of steroid 11beta-hydroxylase (cyp11b2), an enzyme responsible for the synthesis of 11-ketotestosterone (11-KT, a potent androgen in fish), was found in XY gonads from 35 dah only. These results, combined with the presence of transcripts for other steroidogenic enzymes and estrogen receptors in XX gonads at 5-7 dah, are consistent with our earlier suggestion that estradiol-17beta plays a critical role in ovarian differentiation in tilapia, whereas a role for 11-KT in testicular differentiation is questionable. A close relationship between the expression of foxl2, but not nr5a1 (Ad4BP/SF-1), and that of cyp19a1a in XX gonads suggests an important role for Foxl2 in the transcriptional regulation of cyp19a1a. Dmrt1 exhibited a male-specific expression in XY gonads from 6 dah onward, suggesting an important role for Dmrt1 in testicular differentiation. Sox9 and amh (anti-Mullerian hormone) showed a testis-specific expression, being evident only in the later stages of testicular differentiation. It is concluded that the sex-specific expression of foxl2 and cyp19a1a in XX gonads and dmrt1 in XY gonads during early gonadal differentiation (5-6 dah) is critical for undifferentiated gonads to differentiate into either the ovary or testis in the Nile tilapia.
尼罗罗非鱼是一种具有XX/XY性别决定系统的雌雄异体硬骨鱼,因其可获得遗传上的全雌和全雄个体,为研究性腺性别分化提供了一个极佳的模型。在本研究中,我们使用定量实时RT-PCR来确定17个被认为与脊椎动物性腺性别分化相关的基因在性腺中表达的精确时间。在形态学性别分化之前(孵化后5 - 15天[dah])和之后(25 - 70 dah),从全雌和全雄罗非鱼中分离性腺。芳香化酶(cyp19a1a)的转录本,一种负责产生雌二醇-17β的酶,在5 dah时仅在XX性腺中表达,此后表达显著升高。相反,类固醇11β-羟化酶(cyp11b2)的mRNA表达,一种负责合成11-酮睾酮(11-KT,鱼类中的一种强效雄激素)的酶,仅在35 dah后的XY性腺中被发现。这些结果,结合在5 - 7 dah时XX性腺中其他类固醇生成酶和雌激素受体转录本的存在,与我们早期的推测一致,即雌二醇-17β在罗非鱼卵巢分化中起关键作用,而11-KT在睾丸分化中的作用值得怀疑。在XX性腺中,foxl2而非nr5a1(Ad4BP/SF-1)的表达与cyp19a1a的表达之间存在密切关系,表明Foxl2在cyp19a1a的转录调控中起重要作用。Dmrt1从6 dah起在XY性腺中表现出雄性特异性表达,表明Dmrt1在睾丸分化中起重要作用。Sox9和抗苗勒管激素(amh)表现出睾丸特异性表达,仅在睾丸分化的后期才明显。结论是,在性腺早期分化(5 - 6 dah)期间,XX性腺中foxl2和cyp19a1a以及XY性腺中dmrt1的性别特异性表达对于尼罗罗非鱼未分化性腺分化为卵巢或睾丸至关重要。