MECHANISMS REGULATING MORPHOGENESIS OF OCULAR EPITHELIA

描述：表面外胚层直接覆盖在发育的视神经上。 水泡形成眼睛的两层上皮组织，即 角膜上皮和晶状体。脊椎早期形态发生 晶状体囊泡与眼的进一步发育和分化 上皮细胞涉及严密调控的细胞决定机制和 差异化。调节分子的性质和特定作用 控制这些基因表达程序在很大程度上是未知的。近期 文献中的证据表明，一个转录因子家族 被称为AP-2(激活蛋白-2)因子，是 颅面形态发生。这个实验室的初步发现 提示转录因子AP-2(激活蛋白2)在 在眼睛发育中发挥重要作用，特别是调节参与 晶状体和角膜上皮的形态发生。拟议的研究将 验证AP-2转录因子调控基因表达的假设 确定：a)晶状体囊泡的早期图案形成和 B)角膜上皮发育和分化的后期 还有人工晶状体。三种AP-2基因表达模式的确定 眼上皮细胞分化和再生中的蛋白质(A、B、Y) 使用免疫定位和原位杂交技术将使我们 与候选下游基因的位置进行比较。去调查 AP-2a基因在形态发生过程中的特殊功能要求 对晶状体囊泡进行详细的眼部形态检查 将进行AP-2a/-基因敲除小鼠的缺陷实验。此外，新推出的 衍生透镜互补系统(LCS)将用于直接确定 晶状体的形成是否需要AP-2A。要确定 AP-2因子在晶状体纤维细胞和角膜上皮细胞中的特异性作用 差异化。AP-2活性将在转基因小鼠和 体外培养模型。利用野生型P-2微型基因和显性负性 旨在干扰AP-2活性的迷你基因，AP-2活性将 在体外活细胞和转基因小鼠中进行改变，以便 确定AP-2如何调控上皮细胞生物学的特定方面， 包括细胞黏附。

DESCRIPTION: The surface ectoderm directly overlying the developing optic vesicle gives rise to two stratified epithelial tissues of the eye, the corneal epithelium and the crystalline lens. Early morphogenesis of the lens vesicle and further development and differentiation of the ocular epithelia involve tightly regulated mechanics of cell determination and differentiation. the nature and specific role of regulatory molecules controlling these gene expression programs are largely unknown. Recent evidence in the literature suggests that a family of transcription factors known as AP-2 (Activating Protein - 2) factors, are necessary for craniofacial morphogenesis. preliminary findings from this laboratory suggest that transcription factor AP-2 (Activating Protein 2) plays an important role in eye development, specifically regulating genes involved in morphogenesis of the lens and corneal epithelium. The proposed study will test the hypothesis that AP-2 transcription factors regulate gene expression programs that determine: a) early pattern formation of the lens vesicle and b) later stages of development and differentiation of the corneal epithelium and ocular lens. Determination of the expression pattern of three AP-2 proteins (a, B, y) in the differentiating and regenerating ocular epithelia using immunolocalization and in situ hybridization techniques will enable us to compare with the location of candidate downstream genes. To investigate the specific functional requirement of the AP-2a gene during morphogenesis of the lens vesicle, a detailed morphological examination of the ocular defects of AP-2a-/- knockout mice will be performed. In addition, the newly derived Lens complementation system (LCS) will be used to directly determine whether AP-2A is required for formation of the lens. To determine the specific role of AP-2 factors during lens fiber cell and corneal epithelial differentiation. AP-2 activity will be targeted in transgenic mice and in vitro culture models. Using a wild-type P-2 minigene and dominant negative minigenes designed to interfere with AP-2 activity, AP-2 activity will be altered in living cells in vitro and in transgenic mice in order to determine how AP-2 regulates specific aspects of epithelial cell biology, including cell adhesion.