IMPACT OF ETHANOL ON FEMALE RODENT REPRODUCTION

当过量使用时，乙醇（ETOH）是滥用的主要药物 对男性和女性的下丘脑-垂体- 性腺轴 这些分子和细胞机制的细节， 我们的实验室已经并正在深入研究这些影响， 另一些在雄性大鼠中。 然而，相对而言， 关于乙醇对下丘脑-垂体-肾上腺皮质的影响的深入研究很少， 成年、青春期后雌性大鼠的性腺轴。 这些研究 有证据表明ETOH也有很强的作用。 待检验的特定假设为：急性和慢性乙醇 给药通过降低稳态干扰发情周期 下丘脑前促黄体生成素释放激素水平 （LHRH）mRNA和通过损害LHRH原转化为生物活性LHRH。 这会导致垂体促性腺激素功能衰竭， 失败 证明或反驳这一假设的数据将在研究过程中获得。 实现两个具体目标。 A. 为了确定在发情周期的每一天， IP注射乙醇（与IP盐水相比）对前体proLHRH基因的影响 表达（通过逆转录/聚合酶链反应），proLHRH 对成熟的生物活性LHRH十肽进行处理（通过RIA， 前体或产物特异性抗体），垂体 促性腺激素基因表达（通过北方分析）和含量，血清 促性腺激素水平和卵巢重量、组织学和激素功能 (as通过血清雌二醇和孕酮以及子宫重量进行评估， 组织学）（实验1）。 B。 为了确定两个月的液体ETOH喂养的影响， 含饲料（与配对饲喂的液体对照饲料相比）， 具体目标A（实验2）中的参数。 在第一个实验中，单次IP注射ETOH（或生理盐水对照） 将在发情周期的每一天下午1点给予。 此后， 动物将在发情前期于下午3点、4点、5点、6点、7点和9点处死， 以捕获LH和FSH的发情前期激增。 在其他日子里， 发情周期，下午1：00注射乙醇后，动物将 在实验2中，牺牲时间将是类似的 在实验1中。

When used in excess quantities, ethanol (ETOH) is a major drug of abuse with deleterious effects on the male and female hypothalamic-pituitary- gonadal axis. The molecular and cellular mechanistic details of such effects have been and are being studied intensively by our laboratory and others in the male rat. In contrast, however, there has been relatively little in depth study of ethanol's effects on the hypothalamic-pituitary- gonadal axis of the adult, post-pubertal female rat. The studies that do exist suggest that there is a potent effect of ETOH as well. The specific hypothesis to be tested is: ACUTE AND CHRONIC ETHANOL ADMINISTRATION DISRUPTS THE ESTROUS CYCLE BY DIMINISHING STEADY STATE LEVELS OF HYPOTHALAMUS PRE-PRO LUTEINIZING HORMONE RELEASING HORMONE (LHRH) mRNA AND BY IMPAIRING CONVERSION OF PRO LHRH TO BIOACTIVE LHRH. THIS LEADS TO PITUITARY GONADOTROPIN FAILURE AND SECONDARY GONADAL FAILURE. Data to prove or refute this hypothesis will be obtained in the course of accomplishing TWO specific aims. A. To determine on each day of the estrous cycle the effect of a single IP injection of ethanol (compared to IP saline) on pre-proLHRH gene expression (by reverse transcription/polymerase chain reaction), proLHRH processing to the mature bioactive LHRH decapeptide (by RIA with antibodies specific for either precursor or product), pituitary gonadotropin gene expression (by Northern analysis), and content, serum gonadotropin levels, and ovarian weight, histology and hormonal function (as assessed by serum estradiol and progesterone and uterine weight and histology) (Experiment 1). B. To determine the effects of two months of feeding of a liquid ETOH containing diet (compared to pair fed liquid control diet) on the same parameters as in Specific Aim A (Experiment 2). In the first experiment, a single IP injection of ETOH (or saline control) will be given at 1:00 p.m. on each day of the estrous cycle. Thereafter, animals will be sacrificed on proestrous at 3, 4, 5, 6, 7 and 9 p.m. in order to capture the proestrous surge of LH and FSH. On other days of the estrous cycle, after ethanol injection at 1:00 p.m., animals will be killed only at 4:00 p.m. In Experiment 2, sacrifice times will be similar to those in Experiment 1.