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 立面。这些系统的复杂相互作用阻碍了 努力查明原发性高血压的病因。无序的 肾上腺类固醇激素合成在某些临床上明显诱发高血压 更复杂的高血压的发病机制及其作用机制 各州。类固醇激素生物合成正常调控的研究 并确定皮质类固醇对 高血压时，有利于肾上腺皮质细胞的发育 保持差异化功能的线路。老鼠是合乎逻辑的 这些细胞系的来源是因为它在整个动物中的卓越作用 类固醇激素生物合成及遗传模型的研究进展 高血压。SV40大T抗原(Tag)的转基因靶向 被其他人用来生产永生细胞系，保留了 差异化功能。标签在特定区域的靶向表达 转基因大鼠的肾上腺皮质将产生永生化细胞 从离散区域派生的线。三种基因的启动子区域 将使用类固醇生成酶：类固醇11β-羟基酶，这是 在内区选择性表达的是醛固酮合成酶，它是 选择性表达于外肾小球带和类固醇21 -羟基酶，在所有皮质区都有表达。这些推动者 仅在体内表达于肾上腺皮质细胞，它们可能确保 细胞保持分化的表型。有两种方法将 用于增加获得分化细胞系的机会。 首先，转基因大鼠将通过以下方式诱发肾上腺皮质肿瘤 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标签 区域。单独的克隆将来自肿瘤 (野生型TAG)或肾上腺皮质细胞原代培养(温度- 敏感标签)和肾上腺皮质功能的各种参数 学习。 肾上腺皮质在一种新的高血压遗传模型中的作用 Ren-2转基因大鼠也将接受调查。小鼠的表情 特定肾上腺皮质区的Ren-2将确定必要的部位(S) 治疗高血压。更多的实验将确定是否 大鼠肾素基因在特定皮质区的过度表达 会导致高血压。这些研究将对先前的结果进行扩展 小鼠的REN-2基因，从而将增加我们对 肾素在这一刺激性高血压遗传模型中的作用。

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.