Organisciak Daniel, Darrow Ruth, Gu Xiaorong, Barsalou Linda, Crabb John W
Petticrew Research Laboratory, School of Medicine, Wright State University, Dayton, OH, USA.
Photochem Photobiol. 2006 Jul-Aug;82(4):1088-96. doi: 10.1562/2005-06-30-RA-599.
To probe for possible relationships between retinal crystallins and retinal degenerations, protein expression was compared in normal Sprague-Dawley rats, treated or not with intense light, Royal College of Surgeons (RCS) rats and transgenic rats expressing rhodopsin mutations. Rats were reared in dim cyclic light for 21-75 days. Photoreceptor cell DNA levels were determined at various ages to assess the rates of visual cell loss. 1D- and 2D-gel electrophoresis was used to profile retinal protein expression. Crystallins were identified by western analysis and by tandem mass spectrometry. In normal rat retinas, alpha, beta and gamma crystallins were present, although alphaA- and gamma-crystallins exhibited some increase with age. As measured by DNA levels, the rate of genetically induced photoreceptor cell loss was greater in rats with faster degenerating retinas (RCS, S334-ter Line 4, P23H Line 3) than in rats with slower degenerating retinas (S334-ter Line 9, P23H Line 2). In genetic models of retinal degeneration increased levels of immunoreactivity for all crystallins, especially alphaA-insert, correlated with the different rates of photoreceptor loss. In the light induced degeneration model alphaA-insert was unchanged, truncated alphaB-crystallin levels were increased and gamma-crystallins were greatly reduced. In the RCS rat retina 16 different crystallins were identified. Our data suggests that an increase in crystallin expression occurs during various retinal degenerations and that the increases may be related to the severity, type and onset of retinal degeneration.
为探究视网膜晶状体蛋白与视网膜变性之间可能存在的关系,对正常的斯普拉格-道利大鼠(无论是否接受强光照射)、皇家外科学院(RCS)大鼠以及表达视紫红质突变的转基因大鼠的蛋白质表达进行了比较。大鼠在昏暗的循环光照下饲养21 - 75天。在不同年龄段测定光感受器细胞的DNA水平,以评估视觉细胞的损失率。采用一维和二维凝胶电泳分析视网膜蛋白表达情况。通过蛋白质免疫印迹分析和串联质谱法鉴定晶状体蛋白。在正常大鼠视网膜中,存在α、β和γ晶状体蛋白,尽管αA-晶状体蛋白和γ-晶状体蛋白随年龄有所增加。通过DNA水平测量发现,视网膜退化较快的大鼠(RCS大鼠、S334-ter品系4、P23H品系3)中,基因诱导的光感受器细胞损失率高于视网膜退化较慢的大鼠(S334-ter品系9、P23H品系2)。在视网膜变性的遗传模型中,所有晶状体蛋白的免疫反应性水平升高,尤其是αA-插入变体,这与不同的光感受器损失率相关。在光诱导变性模型中,αA-插入变体无变化,截短的αB-晶状体蛋白水平升高,γ-晶状体蛋白水平大幅降低。在RCS大鼠视网膜中鉴定出16种不同的晶状体蛋白。我们的数据表明,在各种视网膜变性过程中晶状体蛋白表达增加,且这种增加可能与视网膜变性的严重程度、类型和发病时间有关。