MOLECULAR REGULATION OF THE PROENKEPHALIN GENE IN VIVO

内源性阿片肽与吗啡相同的受体结合， 其他阿片类化合物，他们被认为是参与缓解疼痛 和鸦片成瘾 存在三大类阿片肽： 脑啡肽内啡肽和强啡肽 每一个都是从一个不同的 基因产生特定的前体肽。 本提案的主题 是脑啡肽原（Penk）基因的调节和 脑啡肽 该策略是确定调节阿片类药物的因素 肽基因表达，并最终开发新的方法， 治疗疼痛和阿片类药物滥用; Penk基因在中枢神经系统和中枢神经系统中都有表达。 肾上腺髓质。 在大鼠肾上腺髓质神经冲动活动中， 抑制和刺激基因表达的基础水平。 这个过程 需要糖皮质激素的存在。 Penk基因 在仓鼠肾上腺中的表达是高的，受神经 活动像大多数哺乳动物和似乎是糖皮质激素的独立。 然而，在发育中的仓鼠中Penk基因表达的增加 肾上腺似乎是糖皮质激素依赖性的。 因此， 神经冲动活动和激素调节Penk基因表达， 随发育、物种和处理而变化。 扩展这些 观察，这项建议将（1）使用生物化学，免疫细胞化学 和原位杂交技术来检测 仓鼠Penk基因表达及评价糖皮质激素的影响 和突触神经支配（2）决定糖皮质激素在脑缺血中的作用。 在仓鼠中诱导Penk基因表达。 [It假定 仓鼠中的基础Penk基因表达不依赖于糖皮质激素， 由于治疗或发育而诱导Penk基因表达是 糖皮质激素依赖。 (3)使用药物操作， 确定多个跨突触和/或受体介导的 调节基因表达的机制。 (4)定义顺式和/或反式 调节Penk基因在成人和发育中表达的元件 仓鼠使用迁移率变动分析、DNA酶I足迹和/或甲基化 干扰测定和（5）开发条件，在体外， 在器官培养中维持仓鼠肾上腺，并在受控条件下检查 调节Penk基因表达的神经营养因子和激素因子 和肽释放。 Penk基因差异的检测 种属（仓鼠和大鼠）之间的表达， 体内治疗的发展和效果将产生新的见解 关于Penk基因调控的具体问题。

The endogenous opioid peptides bind to th e same receptors as morphine and other opiate compounds and they are thought to be involved in pain relief and opiate addiction. Three major groups of opioid peptides exist: enkephalins, endorphins and dynorphins. Each is derived from a distinct gene to produce specific precursor peptides. The subject of this proposal is the regulation of the proenkephalin (Penk) gene and the biosynthesis of enkephalins. The strategy is to determine factors that regulate opioid peptide gene expression and to ultimately develop new approaches for the treatment of pain and opiate drug abuse; The Penk gene is expressed in both the central nervous system and in the adrenal medulla. In the rat adrenal medulla nerve impulse activity both inhibits and stimulates how basal levels of gene expression. This process requires the presence of glucocorticoids. In contrast, Penk gene expression in the hamster adrenals are high, positively regulated by nerve activity like most mammals and appear to be glucocorticoid independent. However, increases in Penk gene expression in the developing hamster adrenal appear to be glucocorticoid dependent. Thus, a combination of nerve impulse activity and hormones regulates Penk gene expression which varies with development, species and treatment. To extend these observations, this proposal will (1) use biochemical, immunocytochemical and in situ hybridization techniques to examine developmental changes in hamster Penk gene expression and evaluate the influence of glucocorticoid and synaptic innervation (2) determine glucocorticoids role int he induction of Penk gene expression in the hamster. [It is hypothesized that basal Penk gene expression in the hamster is glucocorticoid independent and induced Penk gene expression due to treatment or development are glucocorticoids dependent.] (3) use pharmacological manipulation to determine whether multiple transsynaptic and/or receptor mediated mechanisms regulate gene expression. (4) define the cis and/or trans elements that regulate Penk gene expression in the adult and developing hamster using mobility shift analysis, DNase I footprint and/or methylation interference assays and (5) develop conditions, in vitro, that will maintain the hamster adrenal in organ culture and examine under controlled conditions neurotropic and hormonal factors regulating Penk gene expression and peptide release. An examination of the disparity in Penk gene expression between species (hamster and rat), the changes during development and the effects of treatment in vivo will produce new insights on specific issues in Penk gene regulation.