NEUROENDOCRINE REGULATION--EXPRESSION OF CRF

这个项目的主要目标是研究基因的调控 在哺乳动物下丘脑中表达。 初步研究将侧重于 促肾上腺皮质激素释放因子（CRF），一种下丘脑释放激素 产生于室旁核（PVH）的神经分泌细胞。 CRF的生产是合成 糖皮质激素是生命所必需的， 哺乳动物碳水化合物代谢的关键。 糖皮质激素也是 对生理应激如出血的反应的主要部分， 创伤和感染 此外，糖皮质激素治疗广泛应用于 用于治疗自身免疫性、炎症性和风湿性疾病， 通常导致对内源性糖皮质激素的长期影响 表达和调节。 通用报告格式的产生受下列因素的影响： 糖皮质激素，这一复杂的多器官途径的最终产物， 以及表现出基因的昼夜节律和组织特异性模式 表情 关于CRF表达的两个主要假设将使用 现有的异源细胞系和转基因小鼠 CRF转基因。 压力诱导的转录因子 与Fos相关的CRF表达对糖皮质激素耐药 反馈调节将使用异源的转染来测试。 细胞系 灵长类特有的顺式元件决定 胎盘中的hCRF表达将通过检查 用于表达hCRF转基因的转基因小鼠。 目前，许多技术用于研究的分子细节， 细胞调节，依赖于组织培养系统的使用。 然而，在这方面， 某些类型的分化组织根本不存在于细胞中， 文化，因此不能用于研究。 在这些细胞中， 不存在的类型是神经内分泌器官的许多细胞。 这个项目的第三个目标是开发一种永生的、受调控的细胞 表示CRF的行。 这将通过靶向癌基因来实现， 转基因小鼠下丘脑的CRF产生细胞，或使用 体细胞融合 将对所有开发的细胞系进行检测， CRF、神经元特异性标志物基因的表达和 调节表达，特别是负调节， 糖皮质激素一种细胞系的发展， 代表产生CRF的神经内分泌细胞将提供强大的 为今后的研究提供了一个工具。 本项目中检验监管假设所获得的数据应 有助于更好地了解下丘脑调节， CRF的表达，包括糖皮质激素调节的基因表达， 组织特异性基因表达，以及整合的其他成分 神经内分泌调节

The major goal of this project is to investigate regulation of genes expressed in the mammalian hypothalamus. Initial studies will focus on Corticotropin Releasing Factor, (CRF), a hypothalamic releasing hormone produced in neurosecretory cells of the paraventricular nucleus (PVH). The production of CRF is a critical first step in the synthesis of glucocorticoids, which are essential for life and an integral component of mammalian carbohydrate metabolism. Glucocorticoids also are a principal part of the response to physiologic stress such as hemorrhage, trauma, and infection. Furthermore, glucocorticoid therapy is widely used for treatment of auto-immune, inflammatory, and rheumatic diseases, often resulting in long-lasting effects on endogenous glucocorticoid expression and regulation. Production of CRF is modulated by levels of glucocorticoids, the end product of this complex multi-organ pathway, as well as exhibiting circadian and tissue specific patterns of gene expression. Two major hypotheses regarding CRF expression will be tested using existing heterologous cell lines and transgenic mice which express human CRF transgenes. The hypothesis that stress-induced transcription factors related to Fos dictate CRF expression that is resistant to glucocorticoid feedback regulation will be tested using transfection of heterologous cell lines. The hypothesis that primate-specific cis elements dictate hCRF expression in placenta will be tested by examining the placentas of transgenic mice for expression of the hCRF transgenes. At present, many techniques used to study the molecular details of cellular regulation, rely on the use of tissue culture systems. However, certain types of differentiated tissues simply do not exist in cell culture, and therefore are not available for study. Among these cell types which do not exist are many cells of neuroendocrine organs. A third goal of this project is to develop an immortal, regulated cell line expressing CRF. This will be done by targeting an oncogene to the CRF producing cells of the hypothalamus in transgenic mice, or by using somatic cell fusion. All cell lines developed will be tested for expression of CRF, neuron specific marker genes, and retention of regulated expression, particularly negative regulation by glucocorticoids. The development of a cell line that faithfully represents CRF-producing neuroendocrine cells would provide a powerful tool for a host of future studies. Data obtained testing the regulatory hypotheses in this project should contribute to a greater understanding of hypothalamic regulation and expression of CRF, including glucocorticoid regulated gene expression, tissue-specific gene expression, and other components of integrated neuroendocrine regulation.