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CFTR在恒河猴气道表面上皮中的表达与定位

Expression and localization of CFTR in the rhesus monkey surface airway epithelium.

作者信息

Dupuit F, Bout A, Hinnrasky J, Fuchey C, Zahm J M, Imler J L, Pavirani A, Valerio D, Puchelle E

机构信息

INSERM Unité 314, Université de Reims, France.

出版信息

Gene Ther. 1995 Mar;2(2):156-63.

PMID:7536618
Abstract

The Rhesus monkey has been used as a model for evaluating the possibility of introducing the CFTR gene into the airway epithelium in vivo. We addressed the question of whether the simian airway surface epithelium exhibits a CFTR distribution and functional activity (ciliary beating frequency) similar to that of human airway surface epithelium. Expression of CFTR mRNA was demonstrated on Rhesus monkey tracheobronchial tissue by reverse transcription polymerase chain reaction (RT-PCR) analysis. By immunofluorescent light microscopy, CFTR was localized on the apical plasma membrane of ciliated cells as we previously described for human tracheobronchial surface epithelium. The ciliary beat frequency (CBF) measured on the explant of Rhesus monkey tracheobronchial tissue appeared to be similar to that of the CBF of human tracheobronchial ciliated cells. To compare the Rhesus monkey CFTR gene with that of the human, we sequenced parts of exon 13 (encoding the R domain) and exon 24 (encoding the C terminal part of the protein) of the Rhesus monkey CFTR gene. The nucleotide sequence identity with the human counterpart was found to be 98% and 94% respectively, although restriction enzyme differences allow discrimination between Rhesus monkey and human CFTR cDNA. Taken together, these results suggest that the airway epithelium of the Rhesus monkey is a suitable model of human respiratory epithelium for analyzing the effect of human CFTR gene transfer.

摘要

恒河猴已被用作评估在体内将CFTR基因导入气道上皮可能性的模型。我们探讨了猿类气道表面上皮是否表现出与人类气道表面上皮相似的CFTR分布和功能活性(纤毛摆动频率)这一问题。通过逆转录聚合酶链反应(RT-PCR)分析,在恒河猴气管支气管组织上证实了CFTR mRNA的表达。通过免疫荧光显微镜检查,CFTR定位于纤毛细胞的顶端质膜上,正如我们之前对人类气管支气管表面上皮所描述的那样。在恒河猴气管支气管组织外植体上测得的纤毛摆动频率(CBF)似乎与人类气管支气管纤毛细胞的CBF相似。为了比较恒河猴CFTR基因与人类的CFTR基因,我们对恒河猴CFTR基因的第13外显子(编码R结构域)和第24外显子(编码蛋白质的C末端部分)的部分进行了测序。发现与人类对应序列的核苷酸序列同一性分别为98%和94%,尽管限制性内切酶差异允许区分恒河猴和人类的CFTR cDNA。综上所述,这些结果表明恒河猴的气道上皮是用于分析人类CFTR基因转移效果的人类呼吸道上皮的合适模型。

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