Tissue and Cellular Expression Patterns of Porcine CFTR: Similarities to and Differences From Human CFTR
Tissue and Cellular Expression Patterns of Porcine CFTR: Similarities to and Differences From Human CFTR
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DOI：
10.1369/jhc.2010.955377
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发表时间：
2010-09
影响因子：
3.2
通讯作者：
S. Plog;L. Mundhenk;M. Bothe;N. Klymiuk;A. Gruber
中科院分区：
文献类型：
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作者：
S. Plog;L. Mundhenk;M. Bothe;N. Klymiuk;A. Gruber
新出现的囊性纤维化（CF）猪模型有望比目前的小鼠模型更接近地模拟人类疾病。然而，猪CF跨膜传导调节因子（pCFTR）的组织和细胞表达模式以及与人CFTR（hCFTR）的可能差异知之甚少。在此，在mRNA和蛋白质水平上系统地建立了pCFTR的表达模式。使用特异性抗pCFTR抗体，在石蜡包埋切片和冷冻切片上，在肠隐窝上皮细胞、鼻、气管和支气管上皮细胞以及其他选择的主要是腺上皮细胞的顶端胞质溶胶中，对大多数蛋白质进行免疫化学检测。共聚焦激光扫描显微镜与共定位的高尔基体标记58 K定位的蛋白质之间的高尔基体和顶端细胞膜偶尔点状或弥漫染色的顶端膜的胞质溶胶。在激光捕获显微切割后，通过RT-PCR从全组织裂解物或选择的细胞确认组织和细胞分布模式。因此，pCFTR的表达被发现在很大程度上类似于hCFTR的表达，除了在猪中缺乏表达的肾、脑和皮肤腺。pCFTR和hCFTR之间的物种特异性差异可能与猪模型中CF表型的未来解释相关。
Emerging porcine models of cystic fibrosis (CF) are expected to mimic the human disease more closely than current mouse models do. However, little is known of the tissue and cellular expression patterns of the porcine CF transmembrane conductance regulator (pCFTR) and possible differences from human CFTR (hCFTR). Here, the expression pattern of pCFTR was systematically established on the mRNA and protein levels. Using specific anti-pCFTR antibodies, the majority of the protein was immunohistochemically detected on paraffin-embedded sections and on cryostate sections in the apical cytosol of intestinal crypt epithelial cells, nasal, tracheal, and bronchial epithelial cells, and other select, mostly glandular epithelial cells. Confocal laser scanning microscopy with co-localization of the Golgi marker 58K localized the protein in the cytosol between the Golgi apparatus and the apical cell membrane with occasional punctate or diffuse staining of the apical membrane. The tissue and cellular distribution patterns were confirmed by RT-PCR from whole tissue lysates or select cells after laser capture microdissection. Thus, expression of pCFTR was found to largely resemble that of hCFTR except for the kidney, brain, and cutaneous glands, which lack expression in pigs. Species-specific differences between pCFTR and hCFTR may become relevant for future interpretations of the CF phenotype in pig models.