BIOLOGY OF THE EPITHELIAL BASEMENT MEMBRANE

这笔赠款的长期目标(R23转换为第一 奖)是将形态、生物化学和分子 确定功能和结构关系的生物学 在基底膜、上皮细胞骨架和 上皮新陈代谢。这项工作将在角膜上完成 一种可用于研究牙周炎生物学的上皮细胞模型 基底膜带采用这些技术。地下室 膜(基膜)在这里被定义为由中央 富含层粘连蛋白和IV型胶原的致密层(致密层) 两侧被稀有板层包围，包含 糖蛋白和大多数基底膜的蛋白多糖 薄膜。 在这笔赠款的第一年，上皮细胞模型被用于 确定基底膜-细胞骨架的关系。这个 细胞骨架在特定药物存在时被破坏 没有完整的基底膜或增加了细胞外 基质(ECM)分子。我们确定肌动蛋白 细胞骨架是上皮胶原蛋白增加所必需的 基底细胞与基底膜接触产生的产物 分子。 这种竞争更新包括针对以下目标的具体目标 使用胶原和其他蛋白质的mRNA探针来 确定1)mRNA的位置，2)进行关联 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(翻译转录)发生在 上皮细胞。 这项提案将通过以下方式直接解决EB的疾病过程 研究细胞外基质、细胞骨架和细胞外基质之间的关系 角膜上皮细胞、鸡上皮和鸡的蛋白质合成 在未来，人类的表皮。该提案的结果将是 促进我们对角膜上皮代谢的了解 分化，从而与眼部畸形有关 以及其他疾病。我们希望这些研究能为细胞-- 基底膜相互作用和开辟新的途径 BL值异常对细胞代谢可能影响的探讨 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.