Wang Deshou, Kobayashi Tohru, Zhou Linyan, Nagahama Yoshitaka
Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki, Japan.
Biochem Biophys Res Commun. 2004 Jul 16;320(1):83-9. doi: 10.1016/j.bbrc.2004.05.133.
A Foxl2 cDNA was cloned from the Nile tilapia ovary by RT-PCR and subsequent RACE. Alignment of known Foxl2 sequences from vertebrates confirmed the conservation of the Foxl2 open reading frame and protein sequences, especially the forkhead domain and C-terminal region, while some homopolymeric runs of amino acids are found only in mammals but not in non-mammalian vertebrates. RT-PCR revealed that Foxl2 is expressed in the tilapia brain (B), pituitary (P), gill, and gonads (G), with the highest level of expression in the ovary, reflecting the involvement of Foxl2 in B-P-G axis. Northern blotting and in situ hybridization also revealed an evident sexual dimorphic expression pattern in the gonads. Foxl2 mRNA was mainly detected in the granulosa cells surrounding the oocytes. The ovarian expression of Foxl2 in tilapia begins early during the differentiation of the gonads and persists until adulthood, implying the involvement of Foxl2 in fish gonad differentiation and the maintenance of ovarian function.
通过逆转录聚合酶链反应(RT-PCR)及随后的快速扩增cDNA末端(RACE)技术,从尼罗罗非鱼卵巢中克隆出了Foxl2 cDNA。对已知脊椎动物的Foxl2序列进行比对,证实了Foxl2开放阅读框和蛋白质序列的保守性,尤其是叉头结构域和C末端区域,不过一些氨基酸同聚物仅在哺乳动物中发现,而非哺乳类脊椎动物中未发现。RT-PCR显示,Foxl2在罗非鱼的脑(B)、垂体(P)、鳃和性腺(G)中表达,其中在卵巢中的表达水平最高,这反映了Foxl2参与脑-垂体-性腺(B-P-G)轴。Northern印迹法和原位杂交也揭示了性腺中明显的性别二态性表达模式。Foxl2 mRNA主要在卵母细胞周围的颗粒细胞中检测到。罗非鱼卵巢中Foxl2的表达在性腺分化早期开始,并持续到成年期,这意味着Foxl2参与鱼类性腺分化及卵巢功能的维持。