CONTRACTILE PROTEIN ALTERATIONS IN OBSTRUCTED BLADDER

膀胱梗阻中收缩蛋白的变化

基本信息

批准号：
2518347
负责人：
VICTOR K LIN
金额：
$ 14.35万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-09-30 至 1998-08-31
项目状态：
已结题

项目摘要

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.

在人中，由于神经源而发生膀胱出口阻塞，前列腺和尿道狭窄疾病。逼人的初步回应慢性阻塞是平滑肌的发展（SM）肥大。尽管肌肉质量增加了，但细胞力量生成降低，膀胱功能受损。开关从分化的收缩表型到A 推断，合成表型无疑在阻塞引起的膀胱功能障碍。尽管肥厚的生长具有在心脏，肾脏，肠道和尿路系统中描述，它在分子和细胞水平上的性质仍然不清楚。 Caldesmon被认为是平滑的重要调节蛋白肌肉收缩和膀胱出口阻塞会导致改变 Caldesmon表达。我们的假设是这些改变解释了至少部分是膀胱肥大看到的收缩性的改变。因此，该提议的一般目的是研究表达和使用caldesmon基因在受阻膀胱中调节Caldesmon基因利用以下内容的兔子部分膀胱出口阻塞模型方法：核径流测定，cDNA克隆，核酸测序，北方分析，西部印迹分析，抗体产生和免疫组织化学。提出了以下特定目标：（1）克隆兔H-Caldesmon cDNA，12），以确定是否阻塞诱导的逼尿肌平滑肌肥大改变了Caldesmon基因表达，（3）发展和表征豚鼠多克隆专门识别兔子H和L-Caldesmon的抗体，以及（4）确定Caldesmon表达的细胞定位。