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。它们的退化一直是 牵连发病机制的临床意义 心血管疾病。至少有两种主要的蛋白质 称为弹性蛋白和弹性蛋白的弹性纤维成分 微细纤维。这项提案的总体目标是定义 弹力纤维调节机制的研究进展 开发和修复状态。使用鸡主动脉组织和 细胞，原弹性蛋白和微原纤维的合成将是 从转录、翻译和翻译后的角度进行考察 级别。原弹性蛋白分子在细胞外的沉积 不溶于水的弹性蛋白将通过定量桥联蛋白来测定 羟醛缩合交联剂。特异性抗体和cDNA 对于不同的原弹性蛋白亚型和微原纤维将被使用 定义差异合成和胞外沉积。 在卵细胞和体外(器官培养和平滑肌细胞 培养)，建议使用不同年龄的胚胎进行研究。 为了确定荷尔蒙调节和对任何伤害的反应- 诱导情况下，细胞培养基质会受到 用弹性酶进行有限消化。弹性纤维的响应 对这种损伤的合成将在现场和 缺乏激素、分离的弹性蛋白多肽和人工合成的 原弹性蛋白多肽。可能的自分泌反应包括 胰岛素样生长因子-I及其协同作用 将探索糖皮质激素和生长因子之间的关系。 为了进行必要的量化和细胞外 免疫电子显微镜定位、特异性寡核苷酸 将根据获得的数据生成多肽和抗体 来自原弹性蛋白和微原纤维mRNAs的核苷酸序列。 使用自动DNA和多肽合成器将促进这些 学习。

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.