ZONE-SPECIFIC ADRENOCORTICAL CELL LINES

高血压是一种多因素疾病，肾脏异常， 心血管和内分泌系统都会产生血压 提升 这些系统之间复杂的相互作用阻碍了 努力查明原发性高血压的原因。 无序 在某些临床上，肾上腺类固醇生成明显诱导高血压。 并有助于更复杂的高血压的发病机制 states. 研究类固醇激素生物合成的正常调节 并确定皮质类固醇的具体作用， 高血压，发展肾上腺皮质细胞是有利 保持分化功能的线条。 老鼠是逻辑 这些细胞系的来源，因为它在整个动物中的卓越作用 类固醇激素的生物合成和遗传模型的研究 高血压 SV 40大T抗原（TAg）的转基因靶向具有以下优点： 被其他人用来生产永生化细胞系， 分化功能。 将TAg靶向表达至 转基因大鼠肾上腺皮质将产生永生化细胞 从离散区域导出的线。 三个基因的启动子区 将使用类固醇生成酶：类固醇11 β-羟化酶， 选择性地在内区表达，即醛固酮合成酶，它是 选择性地表达于肾小球外膜，类固醇21 - 羟化酶，其在所有皮质区中表达。 这些启动子 在体内仅在肾上腺皮质细胞中表达， 细胞保持分化的表型。 两种方法将 用于增加获得分化细胞系的机会。 首先，将在转基因大鼠中诱导肾上腺皮质肿瘤， TAg的靶向表达。 第二，将准备转基因动物 在特定的肾上腺皮质中表达温度敏感性TAg 区界限 单个克隆将来源于肿瘤细胞， （野生型TAg）或肾上腺皮质细胞的原代培养物（温度- 敏感的TAg）和肾上腺皮质功能的各种参数将被 研究了 肾上腺皮质在高血压新遗传模型中的作用， Ren-2转基因大鼠也将被研究。小鼠表达 特定肾上腺皮质区的Ren-2将确定重要部位 治疗高血压 其他实验将确定，如果 在特定的皮质区域中大鼠肾素基因的过度表达也 诱发高血压。 这些研究将扩展以前的结果， 小鼠Ren-2基因，从而增加我们对 高血压的遗传模型中的肾素的影响。

Hypertension is a multifactorial disease, and abnormalities in the renal, cardiovascular, and endocrine systems all produce blood pressure elevation. The complex interactions of these systems have hindered efforts to identify the causes of primary hypertension. Disordered adrenal steroidogenesis clearly induces hypertension in certain clinical settings and contributes to the pathogenesis of more complex hypertensive states. To study the normal regulation of steroid hormone biosynthesis and to define the specific contributions of corticosteroids to hypertension, it would be advantageous to develop adrenocortical cell lines that maintain differentiated function. The rat is the logical source of these cell lines because of its preeminent role in whole-animal studies of steroid hormone biosynthesis and genetic models of hypertension. Transgenic targeting of SV40 large T antigen (TAg) has been used by others to produce immortalized cell lines that retained differentiated function. Targeted expression of TAg to specific zones of the adrenal cortex in transgenic rats will generate immortalized cell lines derived from discrete zones. The promoter regions of three steroidogenic enzymes will be used: steroid 11 beta-hydroxylase, which is selectively expressed in the inner zones, aldosterone synthase, which is selectively expressed in the outer zona glomerulosa, and steroid 21 -hydroxylase, which is expressed in all cortical zones. These promoters are expressed in vivo only in adrenocortical cells, and they may ensure that the cells maintain a differentiated phenotype. Two approaches will be used to increase the chances of obtaining differentiated cell lines. First, adrenocortical tumors will be induced in transgenic rats by targeted expression of TAg. Second, transgenic animals will be prepared that express a temperature-sensitive TAg in specific adrenocortical zones. Individual clones will be derived from either the tumors (wild-type TAg) or primary cultures of adrenocortical cells (temperature- sensitive TAg) and various parameters of adrenocortical function will be studied. The role of the adrenal cortex in a novel genetic model of hypertension, the Ren-2 transgenic rat, will also be investigated. Expression of mouse Ren-2 in specific adrenocortical zones will identify essential site(s) for the hypertension. Additional experiments will establish if over-expression of the rat renin gene in specific cortical zones also induces hypertension. These studies will extend previous results with the mouse Ren-2 gene, and will thus increase our understanding of the effects of renin in this provocative genetic model of hypertension.