ELASTIC FIBER STRUCTURE, FIBROGENESIS DISEASE

弹性纤维在结构完整性中起着至关重要的作用 和大血管的功能。 它们的退化 与临床上显著的 心血管疾病 至少有两种主要的蛋白质 弹性纤维的组分称为弹性蛋白， 微纤维 本提案的总体目标是界定 参与调节弹性纤维的机制， 发展和修复状态。 使用鸡的主动脉组织， 细胞，原弹性蛋白和微纤维的合成将是 在转录、翻译和翻译后 程度. 弹性蛋白原分子的细胞外沉积， 将通过定量锁链素来测量形式不溶性弹性蛋白 和羟醛缩合交联。 特异性抗体和cDNA 对于不同的弹性蛋白原亚型和微纤维 来定义差异合成和细胞外沉积。 卵内和体外（器官培养和平滑肌细胞） 培养物）的研究。 为了确定荷尔蒙的调节和对任何伤害的反应- 诱导情况下，细胞培养基质将接受 用弹性蛋白酶进行有限消化。 弹性纤维的响应 将研究这种损伤的合成， 缺乏激素，分离的弹性蛋白肽和合成的 原弹性蛋白肽。 可能的自分泌反应包括 胰岛素样生长因子I（IGF-I）及其协同作用 糖皮质激素和生长因子之间的关系。 为了进行必要的定量和细胞外 免疫电镜定位，特异性寡核苷酸 肽和抗体将根据获得的数据产生 来自原弹性蛋白和微纤维mRNA的核苷酸测序。 自动化DNA和肽合成仪的使用将促进这些 问题研究

Elastic fibers play an essential role in the structural integrity and function of large blood vessels. Their degradation has been implicated in the pathogenesis of clinically significant cardiovascular diseases. There are at least two major protein components of elastic fibers referred to as elastin and microfibril. The overall aims of this proposal are to define the mechanisms involved to the regulation of elastic fibers in development and repair states. Using chick aortic tissue and cells, the syntheses of tropoelastin and microfibril will be examined at transcriptional, translational and post-translational levels. The extracellular deposition of tropoelastin molecules to form insoluble elastin will be measured by quantitating desmosine and aldol condensation crosslinks. Antibodies and cDNAs specific for different tropoelastin isoforms and microfibril will be used to define differential synthesis and extracellular deposition. Both in ovo and in vitro (organ culture and smooth muscle cell cultures) studies are proposed using different aged embryos. In order to define hormonal regulation and response to any injury- induced situation, cell culture matrices will be subjected to limited digestion with elastase. The response of elastic fiber synthesis to this injury will be studied in the presence and the absence of hormones, isolated elastin peptides and synthetic tropoelastin peptides. Possible autocrine responses involving insulin-like growth factor I (IGF-I) and synergistic actions between glucocorticords and growth factors will be explored. In order to perform the necessary quantitations and extracellular immunoelectron microscopic localizations, specific oligonucleotide peptides and antibodies will be generated based on data obtained from nucleotide sequencing of tropoelastin and microfibril mRNAs. Use of automated DNA and peptide synthesizers will facilitate these studies.