Licht Peter, von Wolff Michael, Berkholz Alexander, Wildt Ludwig
Division of Gynecological Endocrinology and Reproductive Medicine, Department of Obstetrics and Gynecology, University of Erlangen-Nürnberg, Erlangen, Germany.
Fertil Steril. 2003 Mar;79 Suppl 1:718-23. doi: 10.1016/s0015-0282(02)04822-7.
To investigate the expression of full-length and truncated hCG/LH-receptor mRNA in human endometrium and decidua.
In vitro experiment.
Tertiary university center.
PATIENT(S): Premenopausal women undergoing hysterectomy because of benign diseases or induced abortions.
INTERVENTION(S): Isolation of RNA from endometrial samples, reverse transcription, selective preamplification of full-length hCG/LH receptor mRNA and several shorter fragments of the receptor gene (exons 1-11, 1-10, and 1-5), nested polymerase chain reaction with internal primers.
MAIN OUTCOME MEASURE(S): Appropriately sized cDNA product confirmed by sequencing.
RESULT(S): All samples derived from the proliferative as well as from the early and mid-luteal phases were positive for all four amplification products, suggesting the expression of a full-length hCG/LH receptor mRNA. Only 5 of 8 samples derived from the late secretory phase and 2 of 12 samples derived from early decidua amplified the entire receptor sequence. In contrast, the shortest fragment (exons 1-5), coding for part of the extracellular receptor domain, was amplified in all samples.
CONCLUSION(S): The data suggest cycle-dependent regulation of hCG/LH-receptor mRNA by changes in the alternative splicing pattern and down-regulation of full-length hCG/LH receptor mRNA in early decidua. The major splicing site appears to be located between introns 5 and 9. Alternative splicing may be a mechanism regulating hCG/LH-receptor down-regulation.
研究全长和截短的人绒毛膜促性腺激素/促黄体生成素受体(hCG/LH-receptor)mRNA在人子宫内膜和蜕膜中的表达。
体外实验。
三级大学中心。
因良性疾病接受子宫切除术的绝经前妇女或人工流产者。
从子宫内膜样本中分离RNA,逆转录,对全长hCG/LH受体mRNA和该受体基因的几个较短片段(外显子1 - 11、1 - 10和1 - 5)进行选择性预扩增,用内部引物进行巢式聚合酶链反应。
通过测序确认大小合适的cDNA产物。
来自增殖期以及黄体早期和中期的所有样本对所有四种扩增产物均呈阳性,提示全长hCG/LH受体mRNA的表达。仅8份来自分泌晚期的样本中有5份以及12份来自早期蜕膜的样本中有2份扩增出整个受体序列。相比之下,编码部分细胞外受体结构域的最短片段(外显子1 - 5)在所有样本中均被扩增出来。
数据提示hCG/LH受体mRNA通过可变剪接模式的变化呈现周期性依赖调节,且在早期蜕膜中全长hCG/LH受体mRNA下调。主要剪接位点似乎位于内含子5和9之间。可变剪接可能是调节hCG/LH受体下调的一种机制。