Targeting of the rasT24 oncogene to the proximal convoluted tubules in transgenic mice results in hyperplasia and polycystic kidneys.
Targeting of the rasT24 oncogene to the proximal convoluted tubules in transgenic mice results in hyperplasia and polycystic kidneys.
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发表时间：
1993-04
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通讯作者：
David L. Schaffner;Roberto Barrios;Carolyn Massey;Eugene I. Bañez;Ching-nan Ou;Sridharan Rajagopalan;Estuardo Aguilar-Cordova;R. Lebovitz;P. Overbeek;Michael W. Lieberman
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作者：
David L. Schaffner;Roberto Barrios;Carolyn Massey;Eugene I. Bañez;Ching-nan Ou;Sridharan Rajagopalan;Estuardo Aguilar-Cordova;R. Lebovitz;P. Overbeek;Michael W. Lieberman
从注射γ GT-rasT 24的单细胞期胚胎中获得5个转基因小鼠家族，γ GT-rasT 24是一种融合基因，由含有启动子I的γ-谷氨酰转肽酶（γ GT）5'侧翼区与突变的（密码子12）人H-ras癌基因连接组成。通过逆转录-聚合酶链反应、核酸酶保护试验和原位杂交确定，转基因在所有家族的肾脏、眼睛和大脑中选择性表达。在5个家族中的2个家族中，雄性动物出现肾脏病变，包括近端肾小管增生、肾囊肿和微腺瘤;雄性动物还表达较高水平的γ GT/rasT 24 RNA。早期病变包括近端肾小管增生定义的碱性磷酸酶组织化学，γ GT免疫组化和电子显微镜，并可以与存在的rasT 24 RNA内的囊性近端肾小管上皮细胞原位杂交。晚期病变也累及肾单位的其他部分，由扁平的单细胞层变薄上皮内衬的囊肿组成。原位杂交结果显示远端肾单位和集合小管的囊性病变中没有发现rasT 24，它们很可能是由外部压迫引起的。来自两个转基因品系的表现出囊性病变的动物在8月龄开始死于肾衰竭。通过流式细胞术分析，转基因和对照肾脏之间的细胞周期参数或DNA倍性没有差异。未发生肾癌。该模型系统中H-rasT 24癌基因的原发性肾脏效应包括近端肾小管增生和多囊性肾。该模型似乎为研究ras癌基因功能和控制肾细胞增殖提供了一个有用的体内系统。
Five families of transgenic mice were derived from one-cell-stage embryos injected with gamma GT-rasT24, a fusion gene consisting of the gamma-glutamyl transpeptidase (gamma GT) 5' flanking region containing promoter I linked to a mutated (codon 12) human H-ras oncogene. The transgene was expressed selectively in the kidneys, eyes, and brains of all families as determined by reverse transcription-polymerase chain reaction, nuclease protection assays, and in situ hybridization. In two of five families, kidney lesions consisting of proximal tubular hyperplasia, renal cysts, and microadenomas developed in male animals; males also expressed higher levels of gamma GT/rasT24 RNA. Early lesions consisted of proximal tubular hyperplasia as defined by alkaline phosphatase histochemistry, gamma GT immunohistochemistry, and electron microscopy and could be correlated with the presence of rasT24 RNA within the cystic proximal tubular epithelium by in situ hybridization. Advanced lesions also involved other segments of the nephron and consisted of cysts lined by a flattened unicellular layer of attenuated epithelium. No rasT24 could be identified within cystic lesions of the distal nephron and collecting tubules by in situ hybridization, and they most likely arise by external compression. Animals from the two transgenic strains exhibiting cystic lesions die of renal failure beginning at 8 months of age. No difference in cell-cycle parameters or DNA ploidy between transgenic and control kidneys was identified by flow cytometric analysis. No renal carcinomas developed. The primary renal effects of the H-rasT24 oncogene in this model system consist of proximal tubular hyperplasia and polycystic kidneys. This model appears to provide a useful in vivo system for the study of ras oncogene function and control of renal cell proliferation.