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大鼠卵巢中高密度脂蛋白结合蛋白信使核糖核酸的表达及其受促性腺激素的调节。

Expression of high density lipoprotein-binding protein messenger ribonucleic acid in the rat ovary and its regulation by gonadotropin.

作者信息

Chen Z, Menon K M

机构信息

Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor 48109-0278.

出版信息

Endocrinology. 1994 Jun;134(6):2360-6. doi: 10.1210/endo.134.6.7514997.

Abstract

The presence of high density lipoprotein-binding protein (HBP) mRNA in rat ovarian cells and its possible regulation by gonadotropin were examined in this study. RNA, isolated from pseudopregnant rat ovaries, was amplified by reverse transcriptase-polymerase chain reaction (PCR) with HBP-specific oligonucleotide primers. A distinct and prominent 576-basepair PCR product was generated. Sequence analysis revealed that the sequence homology of the PCR product and the corresponding human HBP cDNA sequence was greater than 90%. Using Northern blot analysis, we identified two species of HBP mRNA transcripts in the ovarian cells (i.e. a major one of 4.5 kilobases and a minor one of 6.0 kilobases). Cellular localization of HBP mRNA in the ovary was determined by in situ hybridization analysis. Prominent specific hybridization of the labeled antisense probe was observed in all cell types of the follicles and corpora lutea, whereas little hybridization was observed in the stroma and the connective tissues separating corpora lutea and follicles. As hCG has been shown to induce high density lipoprotein-binding activity in the rat ovary, we examined the possible regulation of ovarian HBP mRNA by this hormone. Administration of hCG caused a significant time-dependent increase in the steady state levels of HBP mRNA. The induction of HBP mRNA levels by hCG is specific for the ovary, as pretreatment with hCG had no effect on the HBP mRNA levels of the liver, heart, lung, or kidney. The present study, for the first time, shows conclusively the presence of HBP mRNA in the rat ovary and its induction by hCG, implicating a physiological role for HBP in the ovary.

摘要

本研究检测了大鼠卵巢细胞中高密度脂蛋白结合蛋白(HBP)mRNA的存在情况及其可能受促性腺激素的调节。从假孕大鼠卵巢中分离出的RNA,用HBP特异性寡核苷酸引物通过逆转录-聚合酶链反应(PCR)进行扩增。产生了一个清晰且显著的576碱基对的PCR产物。序列分析表明,该PCR产物与相应的人HBP cDNA序列的同源性大于90%。通过Northern印迹分析,我们在卵巢细胞中鉴定出两种HBP mRNA转录本(即一种主要的4.5千碱基转录本和一种次要的6.0千碱基转录本)。通过原位杂交分析确定了HBP mRNA在卵巢中的细胞定位。在卵泡和黄体的所有细胞类型中均观察到标记的反义探针有明显的特异性杂交信号,而在基质以及分隔黄体和卵泡的结缔组织中几乎未观察到杂交信号。由于已证明hCG可诱导大鼠卵巢中的高密度脂蛋白结合活性,我们研究了这种激素对卵巢HBP mRNA的可能调节作用。给予hCG导致HBP mRNA的稳态水平随时间显著增加。hCG对HBP mRNA水平的诱导作用对卵巢具有特异性,因为用hCG预处理对肝脏、心脏、肺或肾脏的HBP mRNA水平没有影响。本研究首次确凿地表明大鼠卵巢中存在HBP mRNA且其可被hCG诱导,这表明HBP在卵巢中具有生理作用。

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