Adam10基因条件剔除小鼠的培育及ADAM10在胸腺细胞发育中调控Notch信号通路的研究

发布时间：2018-07-10 13:21

本文选题：ADAM10 + 基因条件剔除　；参考：《复旦大学》2007年博士论文

【摘要】： Notch信号通路在哺乳动物胸腺细胞发育过程的多个阶段发挥关键作用。该通路的失调将导致T细胞淋巴瘤等疾病的发生。跨膜蛋白Notch受体与配体结合后经过受严格调控的多步蛋白裂解,释放出胞内转录活性片段(IntracellularNotch,ICN),并进入细胞核,引发Notch下游靶基因的转录。在果蝇中的研究表明,kuzbanian(kuz)基因编码一个能裂解激活Notch受体的蛋白酶。因此推测在哺乳动物中KUZ的同源蛋白ADAM10对胸腺细胞发育过程中Notch通路的激活也起重要作用。为验证这一推测,我们利用基因打靶技术,成功培育了Adam10基因条件剔除小鼠,并在小鼠胸腺细胞中特异性地剔除Adam10基因。我们发现Adam10基因失活导致小鼠胸腺明显变小、胸腺细胞总数和双阳性(DP)细胞数量明显减少,这些都与以前在相同条件下剔除Notch1基因的小鼠表型相似。进一步研究显示,缺乏Adam10的胸腺细胞中Notch1及其下游靶基因的活化受到抑制。我们的研究表明,ADAM10在胸腺细胞发育中能通过调节Notch的裂解活化过程来调控胸腺细胞的分化与成熟。作为一种定位于细胞膜上的蛋白水解酶,ADAM10也是一个潜在的治疗Notch信号失调相关疾病的药物靶点。
[Abstract]:Notch signaling pathway plays a key role in multiple stages of mammalian thymocyte development. The imbalance of this pathway will lead to T cell lymphoma and other diseases. After binding to the ligand, the transmembrane protein Notch receptor cleavage through the tightly regulated multistep protein, releasing the intracellular transcriptional active fragment (ICN) and entering the nucleus to initiate the transcription of the Notch downstream target gene. Studies in Drosophila have shown that the Kuzbanian (kuz) gene encodes a protease that cleavage and activates Notch receptors. It is suggested that the KUZ homologous protein ADAM10 may also play an important role in the activation of Notch pathway during the development of thymocytes in mammals. To verify this hypothesis, we successfully culled Adam10 gene conditioned culling mice by gene targeting technique, and specifically culled Adam10 gene in mouse thymocytes. We found that inactivation of the Adam10 gene resulted in a significant decrease in the number of thymocytes and the number of double positive (DP) cells in the thymus of mice, which were similar to the phenotypes of mice that had previously eliminated the Notch1 gene under the same conditions. Further studies showed that the activation of Notch1 and its downstream target genes in thymocytes lacking Adam10 was inhibited. Our results suggest that ADAM10 can regulate the differentiation and maturation of thymocytes by regulating Notch cleavage and activation during the development of thymocytes. ADAM10, a proteolytic enzyme located on the cell membrane, is also a potential target for the treatment of Notch signaling disorders.
【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2007
【分类号】：R392

【相似文献】