CONTRACTILE PROTEIN ALTERATIONS IN OBSTRUCTED BLADDER

在人类中，膀胱出口梗阻是神经性的， 前列腺和尿道狭窄疾病。逼尿肌的初始反应 慢性阻塞的主要原因是平滑肌（SM）的发育 肥厚尽管肌肉质量增加，细胞力 产生减少并且膀胱功能受损。的开关 逼尿肌平滑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大性生长 已在心脏、肾脏、肠道和泌尿道系统中描述， 其在分子和细胞水平上的性质仍然不清楚。 Caldesmon被认为是平滑肌细胞中一种重要的调节蛋白 肌肉收缩和膀胱出口梗阻引起的改变， caldesmon表达。我们的假设是，这些变化解释了，在 至少部分是膀胱肥大引起的收缩性改变。 因此，本提案的总体目标是研究表达方式， 使用免疫调节剂调节梗阻膀胱中的钙调蛋白基因， 兔部分膀胱出口梗阻模型，利用以下 方法：核溢流法，cDNA克隆，核酸序列测定， 北方分析、westernblot分析、抗体产生和 免疫组化建议的具体目标如下：（1） 克隆兔h-caldesmon cDNA，12）以确定是否阻塞- 诱导逼尿肌平滑肌肥大改变caldesmon基因 表达，（3）开发和表征豚鼠多克隆 特异性识别兔H-和L-钙调蛋白的抗体，和（4） 以确定钙调素表达的细胞定位。

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.