A transgenic mouse model for human autosomal dominant cataract
A transgenic mouse model for human autosomal dominant cataract
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DOI：
10.1167/iovs.05-0524
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发表时间：
2006-05-01
影响因子：
4.4
通讯作者：
Petrash, J. Mark
中科院分区：
文献类型：
--
作者：
Hsu, Cheng-Da;Kymes, Steven;Petrash, J. Mark
目的。表达α - a -晶体蛋白突变型和野生型亚基的转基因小鼠的晶状体特征。建立了一系列在晶状体纤维细胞中表达突变体（R116C）和野生型人α - A-晶体蛋白的转基因小鼠品系。对解剖晶状体的存在和混浊程度、纤维细胞形态和后缝合形态进行表型评分。通过逆转录- pcr检测整合转基因的基因转录本。用免疫组化染色法测定晶状体组织切片表达的转基因蛋白的分布。定量Western blot分析内源性和转基因晶状体蛋白的丰度。R116C突变α - a -晶体蛋白亚基的表达导致后皮质白内障和与后缝合相关的异常。晶状体异常的严重程度在9周至30周之间没有增加。在不透明和形态异常方面，表达野生型人α - a -晶体蛋白亚基的转基因小鼠的晶状体与年龄匹配的非转基因对照小鼠没有区别。在R116C转基因小鼠的不同独立品系中观察到相似的表型，突变的转基因蛋白的表达水平至少存在两个数量级的差异。结果表明，在野生型内源性小鼠α -晶体蛋白背景下表达突变体R116C α - a-晶体蛋白亚基时，晶状体混浊和后缝线缺损发生。低水平的R116C α - a -晶体蛋白亚基足以诱导晶状体混浊和缝合线缺陷。
PURPOSE. To characterize lenses from transgenic mice designed to express mutant and wild-type alpha A-crystallin subunits.METHODS. A series of transgenic mouse strains was created to express mutant (R116C) and wild-type human alpha A-crystallin in fiber cells of the lens. Dissected lenses were phenotypically scored for the presence and extent of opacities, fiber cell morphology, and posterior suture morphology. Gene transcripts derived from integrated transgenes were detected by reverse transcriptase-PCR. Distribution of expressed transgenic protein was determined by immunohistochemical staining of lens tissue sections. The abundance of endogenous and transgenic lens proteins was estimated by quantitative Western blot analysis.RESULTS. Expression of R116C mutant alpha A-crystallin subunits resulted in posterior cortical cataracts and abnormalities associated with the posterior suture. The severity of lens abnormalities did not increase between the ages of 9 and 30 weeks. With respect to opacities and morphologic abnormalities, lenses from transgenic mice that express wild-type human alpha A-crystallin subunits were indistinguishable from age-matched nontransgenic control mice. Similar phenotypes were observed in different independent lines of R116C transgenic mice that differed by at least two orders of magnitude in the expression level of the mutant transgenic protein.CONCLUSIONS. The results show that lens opacities and posterior sutural defects occur when mutant R116C alpha A-crystallin subunits are expressed on the background of wild-type endogenous mouse alpha-crystallins. Low levels of R116C alpha A-crystallin subunits are sufficient to induce lens opacities and sutural defects.