PILOT--GENERATION OF CUTANEOUS MAST CELL DEFICIENT MICE

正常皮肤是肥大细胞的丰富来源。因为肥大细胞释放一种 各种生物活性分子(包括细胞因子， 白三烯、前列腺素和蛋白水解酶) 高亲和力的IgE受体，它们被认为在 某些皮肤炎性反应，如荨麻疹。一位少校 在解决肥大细胞在其他方面的作用方面存在障碍 皮肤病理生理学是缺乏一种动物模型 只有肥大细胞。 最近，我们发现了一些肥大细胞特异的基因， 编码颗粒状蛋白酶。我们已经证明了小鼠肥大细胞蛋白酶-5 (mMCP-5)和小鼠肥大细胞羧基肽酶-A(-A)选择性地 表达于皮肤内的肥大细胞。使用来自这些基因的启动子区域 编码这些酶的两个基因与白喉毒素基因WE连锁 建议培育选择性缺乏皮肤肥大的转基因小鼠 细胞。 在本研究的第一阶段，mMCP-5的5“侧翼区域 MMCCP-A基因将分别与β-半乳糖苷酶相连 记者基恩。将产生转基因小鼠来确定哪一个5‘ 侧翼区域在报告实验中表现最好。在第二阶段 在研究中，mMCP-5基因的5‘侧翼区或mMCCP-A 将基因与野生型或突变的白喉毒素基因连锁，以 培育出皮肤肥大细胞缺陷的转基因小鼠。由此产生的 然后将对动物进行角色化。这些都将提供一种强大的试剂 对于进一步研究肥大细胞在正常和非肥大细胞中的作用 病态皮肤。

Normal skin is a rich source of mast cells. Because mast cells release a diverse array of biologically active molecules (including cytokines, leukotrienes, prostaglandins, and proteases) when activated through their high affinity IgE receptors, they are believed to play a central role in certain cutaneous inflammatory responses such as urticaria. A major obstacle in addressing the role of the mast cell in other aspects of cutaneous pathophysiology is the absence of an animal model deficient in just mast cells. Recently we have identified a number of mast cell-specific genes that encode granule proteases. We have shown that mouse mast cell protease-5 (mMCP-5) and mouse mast cell carboxypeptidase-A (-A) are selectively expressed incutaneous mast cells. Using the promoter regions from these two genes encoding these enzymes linked to the diphtheria toxin gene we propose to generate transgenic mice which selectively lack cutaneous mast cells. In the first phase of this research, the 5" flanking regions of the mMCP-5 and mMCCP-A genes will be separately linked to the beta-galactosidase reporter gene. Transgenic mice will be generated to determine which 5' flanking region performs best in the reporter assay. In the second phase of the research, the 5' flanking region of the mMCP-5 gene or the mMCCP-A gene will be linked to the wild-type or mutated diphtheria toxin gene to generate transgenic mice deficient in cutaneous mast cells. The resulting animals will then be characterized. These will provide a powerful reagent for future studies that address the role of mast cells in normal and pathologic skin.