Wilson M E, Marshall M T, Bollnow M R, McGivern R F, Handa R J
Program in Molecular Biology, Loyola University-Chicago, Stritch School of Medicine, Maywood, Illinois 60153, USA.
Alcohol Clin Exp Res. 1995 Oct;19(5):1211-8. doi: 10.1111/j.1530-0277.1995.tb01603.x.
Previous studies have demonstrated that exposure of female rats to ethanol in utero results in long-term deficits in reproductive function, including a delayed onset of puberty and an early onset of acyclicity. In the present studies, we determined if changes in reproduction are correlated with changes in gonadotropin-releasing hormone (GnRH) mRNA expression in the brain or gonadotropin subunit mRNA expression in the anterior pituitary gland. We used in situ hybridization histochemical techniques to examine the density of GnRH mRNA and the distribution of GnRH mRNA-containing cells in the basal forebrain, and reverse transcription-polymerase chain reaction (RT-PCR) to quantitate the beta-subunit mRNA of luteinizing hormone (LH beta) and follicle-stimulating hormone (FSH beta) in the anterior pituitary gland of adult (3 months of age) fetal alcohol-exposed (FAE) female rats. For GnRH mRNA measurements, animals were gonadectomized 4 days before use. Three groups of animals were examined. FAE females were derived from pregnant dams fed a liquid diet containing 35% ethanol-derived calories from gestational day 14 until parturition. Dams of control animals were either pair-fed (PF) an isocaloric diet with sucrose substituted for ethanol or maintained on normal laboratory rat chow [chow-fed (CF)]. Serial blood samples taken by indwelling right atrial cannulae demonstrated significantly smaller pulses of LH (p < 0.05) and FSH (p < 0.05) in ovariectomized FAE females at 3 months of age, compared with PF and CF controls. Distribution of GnRH mRNA-containing cells was mapped throughout the forebrain, and the number of autoradiographic silver grains/cell was determined.(ABSTRACT TRUNCATED AT 250 WORDS)
先前的研究表明,雌性大鼠在子宫内接触乙醇会导致生殖功能出现长期缺陷,包括青春期延迟开始和无发情周期提前出现。在本研究中,我们确定生殖方面的变化是否与大脑中促性腺激素释放激素(GnRH)mRNA表达的变化或垂体前叶中促性腺激素亚基mRNA表达的变化相关。我们使用原位杂交组织化学技术来检测GnRH mRNA的密度以及含GnRH mRNA的细胞在基底前脑的分布,并使用逆转录聚合酶链反应(RT-PCR)来定量成年(3月龄)胎儿酒精暴露(FAE)雌性大鼠垂体前叶中促黄体生成素(LHβ)和促卵泡生成素(FSHβ)的β亚基mRNA。对于GnRH mRNA测量,动物在使用前4天进行性腺切除。检查了三组动物。FAE雌性大鼠来自于怀孕母鼠,这些母鼠从妊娠第14天到分娩期间喂食含35%乙醇热量的液体饮食。对照动物的母鼠要么成对喂食(PF)用蔗糖替代乙醇的等热量饮食,要么维持正常实验室大鼠饲料喂养[正常饲料喂养(CF)]。通过留置右心房插管采集的系列血样显示,与PF和CF对照组相比,3月龄去卵巢FAE雌性大鼠的LH(p<0.05)和FSH(p<0.05)脉冲明显较小。绘制了含GnRH mRNA细胞在整个前脑的分布图,并确定了每个细胞的放射自显影银粒数量。(摘要截断于250字)