CONTRACTILE PROTEIN ALTERATIONS IN OBSTRUCTED BLADDER

在男性，膀胱出口梗阻是神经源性的结果， 前列腺病、尿路狭窄症。逼尿肌的初始反应 慢性梗阻是血管平滑肌(SM)的发育 肥大。尽管肌肉质量增加了，但细胞力 代谢率下降，膀胱功能受损。切换到 逼尿肌从分化的收缩表型转变为 去分化的合成表型无疑在 梗阻引起的膀胱功能障碍。尽管肥厚性生长 已在心脏、肾脏、肠道和尿路系统中被描述， 它在分子和细胞水平上的性质仍不清楚。 Caldesmon被认为是Smooth中的一种重要调节蛋白 肌肉收缩和膀胱出口梗阻引起的改变 钙调蛋白表情。我们的假设是，这些变化解释了 最不重要的是，膀胱肥大引起的收缩能力改变。 因此，这项建议的总体目的是研究表达和 钙调素基因在梗阻性膀胱炎中的表达 兔部分性膀胱出口梗阻模型的建立 方法：核径流分析、基因克隆、核酸测序、 Northern分析、Western印迹分析、抗体生成和 免疫组织化学。提出了以下具体目标：(一) 克隆兔h-caldesmon基因，12)，以确定是否存在梗阻- 诱导的逼尿肌肥大改变钙调蛋白基因 表达，(3)建立和鉴定豚鼠多克隆 特异性识别兔h-和L-caldesmon的抗体，以及(4) 确定钙调蛋白表达的细胞定位。

In man, bladder outlet obstruction occurs as result of neurogenic, prostatic, and urethral stricture disease. The detrusor's initial response to chronic obstruction is the development of smooth muscle (SM) hypertrophy. Despite this increase in muscle mass, cellular force generation decreases and bladder function is impaired. The switch of detrusor smooth muscle from a differentiated, contractile phenotype to a dedifferentiated, synthetic phenotype undoubtedly plays a vital role in obstruction-induced bladder dysfunction. Although hypertrophic growth has been described in cardiac, renal, intestinal and urinary tract systems, its nature at the molecular and cellular level still remains unclear. Caldesmon is believed to be an important regulatory protein in smooth muscle contraction and bladder outlet obstruction induces alteration of caldesmon expression. Our hypothesis is that these alterations explain, at least in part, the altered contractility seen with bladder hypertrophy. Therefore, the general aim of this proposal is to study the expression and regulation of the caldesmon gene in the obstructed bladder using the rabbit partial bladder outlet obstruction model utilizing the following methods: nuclear run-off assay, cDNA cloning, nucleic acid sequencing, northern analysis, westernblot analysis, antibody generation and immunohistochemistry. The following specific aims are proposed: (1) to clone rabbit h-caldesmon cDNA, 12) to determine whether obstruction- induced detrusor smooth muscle hypertrophy alters caldesmon gene expression, (3) to develop and characterize guinea pig polyclonal antibodies that specifically recognize rabbit h- and l-caldesmon, and (4) to determine the cellular localization of caldesmon expression.