Levi Liraz, Levavi-Sivan Berta, Lubzens Esther
Department of Marine Biology and Biotechnology, Israel Oceanographic and Limnological Research, Haifa, Israel.
Biol Reprod. 2008 Sep;79(3):570-7. doi: 10.1095/biolreprod.107.066548. Epub 2008 May 7.
In vertebrates producing oviparous eggs, retinoids and their precursor molecules need to be deposited in oocytes during vitellogenesis. While most studies focus on the transport of retinoids and carotenoids formed outside the fish ovary and their deposition within the developing oocyte, recent investigations in mammalian species suggest the ovary is an important site for retinoid and carotenoid metabolism. Therefore, we investigated the expression of six genes (bcmo1, bcdo2, rbp1, lrat, rbp4, and stra6) associated with retinoids and carotenoids in juvenile and adult trout ovaries. Except for bcdo2, these genes were expressed in the ovary. Expression of stra6 was detected in the ovary but not in the liver. Gene expression levels of bcmo1 and stra6 were significantly higher in juvenile ovaries, in contrast to those of rbp1, rbp4, and lrat, which were similar in all tested ovarian stages. The mean values of the relative mRNA levels of the tested genes differed between the ovary and the liver. Gene transcripts of rbp4 and bcmo1 were identified by in situ hybridization in the theca layer, and all five genes were expressed in the granulosa, stromal cells, and only the early vitellogenic oocyte. The occurrence of retinol-binding protein in the theca and granulosa cells and within oocytes at all developmental stages was revealed by immunocytochemistry. These results indicate that ovarian cells express genes putatively associated with cleavage of beta-carotene, storage and mobilization of retinyl-esters, and of retinol-binding protein synthesis, suggesting a novel pathway for providing retinoids and carotenoids to developing fish ovarian follicles.
在产羊膜卵的脊椎动物中,类视黄醇及其前体分子在卵黄发生过程中需要沉积在卵母细胞中。虽然大多数研究集中于鱼类卵巢外形成的类视黄醇和类胡萝卜素的运输及其在发育中卵母细胞内的沉积,但最近对哺乳动物物种的研究表明,卵巢是类视黄醇和类胡萝卜素代谢的重要场所。因此,我们研究了与类视黄醇和类胡萝卜素相关的六个基因(bcmo1、bcdo2、rbp1、lrat、rbp4和stra6)在幼年和成年鳟鱼卵巢中的表达。除bcdo2外,这些基因均在卵巢中表达。在卵巢中检测到stra6的表达,但在肝脏中未检测到。与rbp1、rbp4和lrat在所有测试卵巢阶段的表达相似相比,bcmo1和stra6的基因表达水平在幼年卵巢中显著更高。所测试基因的相对mRNA水平的平均值在卵巢和肝脏之间存在差异。通过原位杂交在卵泡膜层鉴定出rbp4和bcmo1的基因转录本,并且所有五个基因均在颗粒细胞、基质细胞以及仅早期卵黄发生的卵母细胞中表达。免疫细胞化学揭示了在所有发育阶段的卵泡膜细胞、颗粒细胞以及卵母细胞内视黄醇结合蛋白的存在。这些结果表明,卵巢细胞表达可能与β-胡萝卜素裂解、视黄酯的储存和转运以及视黄醇结合蛋白合成相关的基因,提示了一条为发育中的鱼类卵巢卵泡提供类视黄醇和类胡萝卜素的新途径。