Ohno Tomoko, Hiroi Hisahiko, Momoeda Mikio, Hosokawa Yumi, Tsutsumi Ryo, Koizumi Minako, Nakazawa Fumiko, Yano Tetsu, Tsutsumi Osamu, Taketani Yuji
Department of Obstetrics and Gynecology, University of Tokyo, Japan.
Endocr J. 2008 Mar;55(1):83-90. doi: 10.1507/endocrj.k07-082. Epub 2008 Jan 24.
The endometrium is one of the target tissues of the ovarian steroid hormones, estrogen and progesterone. In order to elucidate the mechanism of gene regulation in the endometrium, suppressive subtraction hybridization was performed to isolate the candidate genes controlled by progesterone in rat uterus. Alcohol dehydrogenase (ADH) class I gene was one of the candidate genes. Here we investigated the expression and regulation of ADH class I gene in rat uterus. The mRNA of ADH class I was detected in uterus by RT-PCR using specific primers. Using specific probe for ADH class I, in situ hybridization was performed to investigate localization in rat uterus. Positive signals were detected in the endometrial stromal cells of rat uterus by in situ hybridization and were not detected in endometrial epithelial cells and myometrium in rat uterus. Ovariectomized rats were treated with 17-beta estradiol and progesterone and the uteri of these rats were used for Northern blot analysis and assay of the ADH activity. Northern blot analysis revealed that the expression of ADH class I mRNA in rat uteri was up-regulated approximately two-fold after progesterone treatment, but not estrogen. Likewise, ADH activity was approximately two-fold higher in progesterone-treated rat uteri compared with controls. This study demonstrated that ADH class I gene is progesterone-responsive in the uterus. This implies that progesterone might be involved with retinoic acid synthesis in the uterus, since ADH is the key enzyme for retinoic acid synthesis.
子宫内膜是卵巢甾体激素雌激素和孕激素的靶组织之一。为阐明子宫内膜中的基因调控机制,采用抑制性消减杂交技术分离大鼠子宫中受孕激素调控的候选基因。Ⅰ类乙醇脱氢酶(ADH)基因是候选基因之一。在此,我们研究了大鼠子宫中Ⅰ类ADH基因的表达及调控。使用特异性引物通过逆转录聚合酶链反应(RT-PCR)在子宫中检测到了Ⅰ类ADH的信使核糖核酸(mRNA)。使用Ⅰ类ADH的特异性探针,进行原位杂交以研究其在大鼠子宫中的定位。原位杂交在大鼠子宫的子宫内膜基质细胞中检测到阳性信号,而在大鼠子宫的子宫内膜上皮细胞和子宫肌层中未检测到。对去卵巢大鼠给予17-β雌二醇和孕激素处理,取这些大鼠的子宫用于Northern印迹分析及ADH活性检测。Northern印迹分析显示,孕激素处理后大鼠子宫中Ⅰ类ADH mRNA的表达上调约两倍,而雌激素处理后未上调。同样,与对照组相比,孕激素处理的大鼠子宫中ADH活性约高两倍。本研究表明Ⅰ类ADH基因在子宫中对孕激素有反应。这意味着孕激素可能参与子宫中视黄酸的合成,因为ADH是视黄酸合成的关键酶。