Kato Ryo, Maeda Tomoji, Akaike Toshihiro, Tamai Ikumi
Faculty of Pharmaceutical Sciences, Dept. of Molecular Biopharmaceutics, Tokyo Univ. of Science, 2641 Yamasaki, Noda, Chiba, 278-8510, Japan.
Am J Physiol Endocrinol Metab. 2006 May;290(5):E968-75. doi: 10.1152/ajpendo.00160.2005. Epub 2005 Dec 20.
In the testis, nucleosides and nucleobases are important substrates of the salvage pathway for nucleotide biosynthesis, and one of the roles of Sertoli cells is to provide nutrients and metabolic precursors to spermatogenic cells located within the blood-testis barrier (BTB). We have already shown that concentrative and equilibrative nucleoside transporters are expressed and are functional in primary-cultured rat Sertoli cells as a BTB model, but little is known about nucleobase transport at the BTB or about the genes encoding specific nucleobase transporters in mammalian cells. In the present study, we examined the uptake of purine ([3H]guanine) and pyrimidine ([3H]uracil) nucleobases by primary-cultured rat Sertoli cells. The uptake of both nucleobases was time and concentration dependent. Kinetic analysis showed the involvement of three different transport systems in guanine uptake. In contrast, uracil uptake was mediated by a single Na+-dependent high-affinity transport system. Guanine uptake was inhibited by other purine nucleobases but not by pyrimidine nucleobases, whereas uracil uptake was inhibited only by pyrimidine nucleobases. In conclusion, it was suggested that there might be purine- or pyrimidine-selective nucleobase transporters in rat Sertoli cells.
在睾丸中,核苷和核碱基是核苷酸生物合成补救途径的重要底物,而支持细胞的作用之一是为位于血睾屏障(BTB)内的生精细胞提供营养和代谢前体。我们已经表明,在作为BTB模型的原代培养大鼠支持细胞中,浓缩型和平衡型核苷转运体均有表达且具有功能,但对于BTB处的核碱基转运或哺乳动物细胞中编码特定核碱基转运体的基因知之甚少。在本研究中,我们检测了原代培养的大鼠支持细胞对嘌呤([3H]鸟嘌呤)和嘧啶([3H]尿嘧啶)核碱基的摄取。两种核碱基的摄取均呈时间和浓度依赖性。动力学分析表明,鸟嘌呤摄取涉及三种不同的转运系统。相比之下,尿嘧啶摄取由单一的Na+依赖性高亲和力转运系统介导。鸟嘌呤摄取受到其他嘌呤核碱基的抑制,但不受嘧啶核碱基的抑制,而尿嘧啶摄取仅受到嘧啶核碱基的抑制。总之,提示大鼠支持细胞中可能存在嘌呤或嘧啶选择性核碱基转运体。