Provost P R, Tremblay Y, el-Amine M, Bélanger A
Medical Research Council Group in Molecular Endocrinology, CHUL Research Center Quebec, Canada.
Mol Cell Endocrinol. 1995 Apr 1;109(2):225-36. doi: 10.1016/0303-7207(95)03506-3.
We have isolated and characterized two molecular types of guinea pig (GP) apolipoprotein D (apoD) cDNA. The sequences of cDNA clones GP APO D-20 and -38 are 100 % homologous in their putative exons 2-5, as determined by analogy within human apoD gene, but they differ totally in their putative exon 1. RNase protection assays showed the presence of both apoD RNA types 20 and 38 in cauda epididymis. Northern blot analysis revealed four polyadenylated apoD bands at 3.2, 2.7, 1.7, and 1.0 kb. Types 20 and 38 specific probes hybridized with the major 1-kb mRNA and two of the three other minor RNA transcripts, respectively. Southern blot analysis revealed that the guinea pig genome probably contains one apoD gene. Our data also demonstrated that the cauda epididymis and fallopian tubes had an apoD mRNA concentration 100-fold higher than the liver, suggesting that the apoD gene expression could be associated with the presence of steroids. The levels of the 1-kb mRNA increased in the fallopian tubes and ovaries during gestation and were lower in fetal reproductive tissues and liver than in mature animals. No positive correlation was found between apoD and 3 beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase (3 beta-HSD) mRNA levels in these tissues, thus suggesting that high amounts of apoD mRNA are not necessarily associated with in situ progesterone synthesis. Taken together, our results indicate that both the guinea pig epididymis and fallopian tubes are excellent models to study the local role of apoD in steroid target tissues.
我们已经分离并鉴定了两种分子类型的豚鼠(GP)载脂蛋白D(apoD)cDNA。根据人类apoD基因的类推,cDNA克隆GP APO D - 20和 - 38在其推定的外显子2 - 5中具有100%的同源性,但在其推定的外显子1中完全不同。核糖核酸酶保护试验表明,在附睾尾部同时存在apoD RNA类型20和38。Northern印迹分析显示在3.2、2.7、1.7和1.0 kb处有四条多聚腺苷酸化的apoD条带。类型20和38特异性探针分别与主要的1-kb mRNA以及其他三个次要RNA转录本中的两个杂交。Southern印迹分析表明豚鼠基因组可能包含一个apoD基因。我们的数据还表明,附睾尾部和输卵管中的apoD mRNA浓度比肝脏高100倍,这表明apoD基因表达可能与类固醇的存在有关。在妊娠期间,输卵管和卵巢中1-kb mRNA的水平升高,而在胎儿生殖组织和肝脏中比成熟动物低。在这些组织中,未发现apoD与3β-羟基类固醇脱氢酶/δ5-δ4异构酶(3β-HSD)mRNA水平之间存在正相关,因此表明大量的apoD mRNA不一定与原位孕酮合成相关。综上所述,我们的结果表明,豚鼠附睾和输卵管都是研究apoD在类固醇靶组织中局部作用的优秀模型。