Roberts K P, Griswold M D
Program in Biochemistry and Biophysics, Washington State University, Pullman 99164.
Mol Endocrinol. 1990 Apr;4(4):531-42. doi: 10.1210/mend-4-4-531.
A 3.4 kilobase cDNA complementary to rat transferrin receptor mRNA has been isolated from an adult rat testis cDNA library. The rat transferrin receptor nucleotide sequence was shown to be 82% similar to the human transferrin receptor sequence over the amino acid coding region and over 90% similar in the sequences known to be responsible for iron regulation in the human mRNA. The mRNA was shown by Northern blot analysis to be regulated by iron levels in Sertoli cells in culture. Iron depletion resulted in at least a 5-fold increase in receptor message in Sertoli cells, as well as in an actively growing testicular cell line (S10-7). The level of transferrin receptor mRNA in cultured Sertoli cells was not influenced by hormones; however, chronic administration of testosterone or FSH to hypophysectomized rats resulted in increased transferrin receptor mRNA levels in the testis. Northern blot analysis of mRNAs from testes of rats synchronized at various stages of the cycle of the seminiferous epithelium showed that transferrin receptor mRNA was differentially regulated throughout the cycle. Northern blots of mRNA from germinal cell populations derived from synchronized tests showed that the message was regulated in the nongerminal cell components of the tubule, most likely the Sertoli cell. The comparison of transferrin receptor mRNA levels in normal testes and testes from hypophysectomized rats, as well as in isolated germinal cells and cultured Sertoli cells, suggested that transferrin receptor mRNA levels were considerably higher in Sertoli cells than in other cell types of the seminiferous tubules.
从成年大鼠睾丸cDNA文库中分离出了一段与大鼠转铁蛋白受体mRNA互补的3.4千碱基cDNA。在氨基酸编码区域,大鼠转铁蛋白受体核苷酸序列与人类转铁蛋白受体序列的相似度为82%,在已知负责人类mRNA中铁调节的序列中,相似度超过90%。通过Northern印迹分析表明,该mRNA受培养的支持细胞中铁水平的调节。缺铁导致支持细胞以及一种活跃生长的睾丸细胞系(S10 - 7)中受体信息至少增加5倍。培养的支持细胞中转铁蛋白受体mRNA的水平不受激素影响;然而,对垂体切除的大鼠长期给予睾酮或促卵泡激素会导致睾丸中转铁蛋白受体mRNA水平升高。对处于生精上皮周期不同阶段同步化的大鼠睾丸mRNA进行Northern印迹分析表明,转铁蛋白受体mRNA在整个周期中受到不同调节。对来自同步化睾丸的生殖细胞群体的mRNA进行Northern印迹分析表明,该信息在小管的非生殖细胞成分中受到调节,最有可能是支持细胞。正常睾丸与垂体切除大鼠睾丸中转铁蛋白受体mRNA水平的比较,以及分离的生殖细胞和培养的支持细胞中转铁蛋白受体mRNA水平的比较表明,支持细胞中转铁蛋白受体mRNA水平比生精小管的其他细胞类型高得多。