BIOLOGY OF THE EPITHELIAL BASEMENT MEMBRANE

该补助金的长期目标（R23转换为FIRST 联合收割机是将形态学、生物化学和分子生物学 生物学来确定功能和结构关系 基底膜、上皮细胞骨架和 上皮代谢 这项工作将在角膜上完成 上皮细胞模型，其可用于研究 基底膜区通过这些技术。 地下室 膜（基底层）在此定义为由中央 富含层粘连蛋白和IV型胶原蛋白的致密层（致密层） 两侧被稀有层包围， 糖蛋白和大部分的蛋白多糖的基础 膜的 在这项资助的第一年，上皮细胞模型被用来 确定基底膜与细胞骨架的关系。 的 细胞骨架被破坏与特定的药物在存在或 缺乏完整的基底膜或额外的细胞外 基质（ECM）分子。 我们确定， 细胞骨架是上皮胶原增加所必需的 基底细胞与基底膜接触产生 分子。 这种竞争性的更新包括具体的目标， 使用胶原蛋白和其他蛋白质的mRNA探针， 确定1）mRNA的位置，2）建立相关性 mRNA水平与蛋白质合成的关系，以及3）蛋白质的类型 合成调节（翻译的转录）发生在 上皮细胞 该提案将直接解决EB的疾病过程， 研究ECM、细胞骨架和 角膜上皮细胞、鸡表皮的蛋白合成， 在未来，人类的表皮。 该提案的结果将 促进我们对角膜上皮代谢的理解， 分化，因此与眼畸形有关， 和其他疾病。 我们希望这些研究能为细胞- 基底膜相互作用，并开辟新的途径， 探讨异常BL对细胞代谢的可能影响 和差异化。

The longterm objective of this grant (R23 conversion to a FIRST award) is to combine morphology, biochemistry and molecular biology to determine the functional and structural relationships between the basement membrane, epithelial cytoskeleton, and epithelial metabolism. This work will be done on a corneal epithelial cell model which can be used to study the biology of the basement membrane zone by these techniques. The basement membrane (basal lamina) is defined here as consisting of a central dense sheet (lamina densa) rich in laminin and type IV collagen surrounded on both sides by laminae rarae containing glycoproteins and most of the proteoglycan of the basement membrane. In the first year of this grant, the epithelia cell model was used to determine basement membrane-cytoskeleton relationships. The cytoskeleton was disrupted with specific drugs in the presence or absence of an intact basement membrane or added extracellular matrix (ECM) molecules. We determined that the actin cytoskeleton was necessary for the increase in epithelial collagen production caused by basal cell contact with basement membrane molecules. This competitive renewal includes specific aims directed toward using probes for the mRNA of collagen and other proteins to determine 1) the location of the mRNA, 2) to make a correlation of mRNA levels with protein synthesis, and 3) the type of protein synthesis regulation (transcription of translation) occurring in epithelial cells. This proposal will address the disease process of EB directly, by studying the relationship between the ECM, cytroskeleton, and protein synthesis of corneal epithelial cells, chick epidermis, and in the future, human epidermis. The results of the proposal will advance our understanding of corneal epithelial metabolism and differentiation and thus and relevant to eye malformations in this and other diseases. We expect these studies to cast light on cell- basement membrane interaction and to open up new pathways for approaching possible effects of abnormal BL on cell metabolism and differentiation.