INDUCTION BY HEPARIN OF A NEW VESSEL WALL COLLAGEN

类肝素糖胺聚糖是血管的正常成分 WALL可能在平滑肌细胞的调节中起重要作用 增长、迁移和差异化。我们最近已经证明， 肝素及其相关分子对大鼠分泌表型的调节作用 体外培养的主动脉平滑肌细胞(SMC)。在这些细胞中，肝素诱导 合成了一种先前未描述的低分子量(MR= 60,000)胶原蛋白。此申请中描述的研究是 针对60Kd的特定分子探针的开发 胶原蛋白和利用这些探针来研究生产， 这种分子的分布、结构和调节。具体来说，我们 打算开发针对45Kd胃蛋白酶抗性形式的单抗 从肝素处理的SMC细胞中分离60Kd胶原蛋白 层次感。这些抗体将被用来研究这种病毒的分布 正常大鼠组织光谱中体内的胶原蛋白。此外， 将分析60Kd胶原在不同时间段的主动脉中的分布。 球囊插管损伤后的时间点。这一方法将 允许我们采样处于不同功能状态的SMC(静态， 迁移性、增殖性、后增殖性，并参与连接 组织合成)。 使用差异杂交选择技术，我们将克隆 与60Kd胶原蛋白基因序列互补的质粒载体。 这些DNA将被测序，以产生60Kd上的结构数据 胶原蛋白。最后，我们将研究其调控的分子基础。 肝素作用60Kd的胶原蛋白，用cDNAs作为特异性探针 胶原蛋白基因表达水平。 希望对这种新的血管壁进行详细的调查 胶原蛋白及其肝素的调节将有助于我们理解 在正常和动脉粥样硬化的情况下，SMC的功能。

Heparin-like glycosaminoglycans are normal components of the blood vessel wall and may play an important role in the regulation of smooth muscle cell growth, migration, and differentiation. We have recently shown that heparin and related molecules can regulate the secretory phenotype of rat aortic smooth muscle cells (SMC) in vitro. In these cells, heparin induces the synthesis of a previously undescribed, low molecular weight (Mr = 60,000) collagenous protein. The research described in this application is directed toward the development of specific molecular probes to the 60Kd collagen and the utilization of these probes to study the production, distribution, structure, and regulation of this molecule. Specifically, we intend to develop monoclonal antibodies to the 45Kd pepsin-resistant form of the 60Kd collagen following its isolation from heparin-treated SMC cell layers. These antibodies will be used to study the distribution of this collagen in vivo in a spectrum of normal rat tissues. In addition, the distribution of the 60Kd collagen will be analyzed in the aorta at various time points after injury by balloon catheterization. This approach will allow us to sample SMC in different functional states (quiescent, migratory, proliferative, post-proliferative, and involved in connective tissue synthesis). Using a differential hybridization selection technique, we will clone in plasmid vectors gene sequences complementary to the 60Kd collagen mRNA. These cDNAs will be sequenced to generate structural data on the 60Kd collagen. Finally, we will study the molecular basis of regulation of the 60Kd collagen by heparin, using a cDNA as a specific probe for 60Kd collagen mRNA levels. It is hoped that a detailed investigation of this new vascular wall collagen and its regulation by heparin will contribute to our understanding of SMC function in both normal and atherosclerotic conditions.