CONTROL OF URNIARY BLADDER FUNCTION--MOLECULAR APPROACH

膀胱对特定的病理和手术反应 膀胱质量、容量、顺应性 和对药物的反应。 例如，局部出口 梗阻引起膀胱质量的快速和显著增加， 膀胱容量和顺应性显著降低， 膀胱功能（收缩和/或排空的能力）。 单侧缺血 引起与出口阻塞所观察到的变化相似的变化。 急性 膀胱过度膨胀引起许多病理变化 收缩和功能与出口阻塞发生，虽然这些 变化在过度膨胀后两周内迅速消退。 通过 相比之下，膀胱转流术导致膀胱重量迅速减少， （萎缩），能力和顺应性下降， 对场刺激和药物的收缩反应。 消除应力（消除出口阻塞或重新植入 输尿管改道后）引起快速和几乎完全的 上述变化的逆转。 到目前为止，我们的研究已经描述了尿动力学， 生理、药理、形态和代谢反应 几种实验诱导的病理，包括部分出口 梗阻、单侧缺血、急性过度扩张和泌尿系统 转移注意力 最近，我们在亚细胞水平上开展了一系列研究， 这些级别旨在识别早期信号， 引发对压力的各种反应。 我们建议将时间 基因激活的初始事件的过程和幅度 蛋白质化学、细胞生长和细胞分裂的变化。 这些研究的长期目标是确定细胞机制 膀胱的结构和功能被特定的 病理学 我们对基因表达的初步研究表明， 部分出口阻塞刺激了多种表达 控制细胞生长/分裂和蛋白质合成的特定基因。 这些基因包括N-Ras、C-Myc、β-肌动蛋白、热休克蛋白-70（HSP-70）、 70），和碱性成纤维细胞生长因子与减少 转化生长因子β 本提案的短期目标包括：1）确定 部分出口刺激的基因表达变化序列 梗阻：诱导后1、2、4、8、16、24小时、3、5、7和14天 的病理学。 2)向上的组织/细胞区域的证明- 或下调特定细胞成分中的基因表达， 膀胱底部和身体（例如，粘膜、粘膜下层、肌层和浆膜） 使用原位杂交和免疫细胞化学。3)定量 改变的转录与翻译事件的相关性 组织/细胞的物质，如碱性成纤维细胞生长因子（bFGF）， 血小板衍生生长因子（PDGF）、热休克蛋白70（HSP-70）和 90（HSP-90）和特异性细胞骨架蛋白波形蛋白、结蛋白和 肌动蛋白。4)特定基因产物对细胞生长和分裂的影响 （使用定量形态测定法和胸苷掺入DNA）。

The urinary bladder responds to specific pathological and surgical situations with major alterations in bladder mass, capacity, compliance, and response to pharmacological agents. For example, partial outlet obstruction induces a rapid and marked increase in bladder mass, significant decreases in bladder capacity and compliance, and a decrease in bladder function (ability to contract and/or empty). Unilateral ischemia induces changes similar to those observed for outlet obstruction. Acute overdistention initiates many of the pathological changes in bladder contraction and function that occur with outlet obstruction, although these changes quickly resolve within two weeks after overdistension. By contrast, bladder diversion induces a rapid decrease in bladder mass (atrophy), decreases in capacity and compliance, and a decreased contractile response to field stimulation and pharmacological agents. Removal of the stress (removal of the outlet obstruction or re-implantation of the ureters following diversion) induces a rapid and nearly complete reversal of the changes described above. Our studies up to this time have characterized the urodynamic, physiological, pharmacological, morphological, and metabolic responses to several experimentally induced pathologies including partial outlet obstruction, unilateral ischemia, acute overdistension, and urinary diversion. Recently, we have initiated a series of studies at sub-cellular levels which are directed at the identification of the early signals which initiate the various responses to stress. We propose to correlate the time course and magnitude of the initial events in gene activation with cellular changes in protein chemistry, cell growth, and cell division. The long term goal of these studies is to determine the cellular mechanisms by which bladder structure and function are altered by specific patholologies. Our initial studies on gene expression demonstrated that partial outlet obstruction stimulated the expression of a variety of specific genes which control cell growth/division, and protein synthesis. These genes included N-Ras, C-Myc, beta-Actin, heat shock protein-70 (HSP- 70), and basic fibroblast growth factor in association with a decrease in transforming growth factor beta. The short term goals of the current proposal include: 1) Determination of the sequence of changes in gene expression stimulated by partial outlet obstruction: 1, 2, 4, 8, 16, 24 hours, 3, 5, 7, and 14 days after induction of the pathology. 2) Demonstration of the tissue/cellular locale(s) of up- or downregulated gene expression in specific cellular elements in the bladder base and body (eg. in mucosa, submucosa, muscularis, and serosa) using in situ hybridization, and immunocytochemistry. 3) Quantitative correlations of transcriptional with translational events in altered tissue/cells for substances such as basic fibroblast growth factor (bFGF), platelet derived growth factor (PDGF), heat shock proteins 70 (HSP-70) and 90 (HSP-90), and the specific cytoskeletal proteins vimentin, desmin, and actin. 4) Effects of specific gene products with cell growth and division (using quantitative morphometry and thymidine incorporation into DNA).