Visualization of fibrillar amyloid deposits in living, transgenic Caenorhabditis elegans animals using the sensitive amyloid dye, X-34
Visualization of fibrillar amyloid deposits in living, transgenic Caenorhabditis elegans animals using the sensitive amyloid dye, X-34
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DOI：
10.1016/s0197-4580(00)00237-2
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发表时间：
2001-03-01
影响因子：
4.2
通讯作者：
Klunk, WE
中科院分区：
文献类型：
--
作者：
Link, CD;Johnson, CJ;Klunk, WE
转基因秀丽隐杆线虫动物可以被改造成表达高水平的人类β淀粉样肽（Abeta）。这些动物固定组织的组织化学显示沉积物与淀粉样蛋白特异性染料刚果红和硫黄素S反应（Fay et al.， J. Neurochem 71:1616, 1998）。我们通过免疫电镜显示，这些动物含有典型淀粉样纤维超微结构的细胞内免疫反应性沉积物。这些沉积物可以使用新开发的，强荧光，淀粉样蛋白特异性染料X-34在活体动物中可视化。这种体内染色可以监测单个动物的淀粉样蛋白沉积。这种染色的特异性是通过检测表达高水平非纤原性β肽变体（β单链二聚体）的转基因动物来证明的。这些动物具有抗β抗体免疫反应的沉积物，但没有X-34沉积物或纤原性超微结构沉积物。X-34也可以在体内用于观察由人甲状腺转蛋白（另一种淀粉样蛋白）积累引起的假定的淀粉样沉积。体内用X-34进行淀粉样蛋白染色可能是实时监测抗淀粉样蛋白治疗或筛选影响淀粉样蛋白形成的遗传改变的有用工具。(C) 2001爱思唯尔科学公司版权所有。
Transgenic Caenorhabditis elegans animals can be engineered to express high levels of the human beta amyloid peptide (Abeta). Histochemistry of fixed tissue from these animals reveals deposits reactive with the amyloid-specific dyes Congo Red and thioflavin S (Fay et al., J. Neurochem 71:1616, 1998). Here we show by immune-electron microscopy that these animals contain intracellular immunoreactive deposits with classic amyloid fibrillar ultrastructure. These deposits can be visualized in living animals using the newly developed, intensively fluorescent, amyloid-specific dye X-34. This in vivo staining allows monitoring of amyloid deposition in individual animals over time. The specificity of this staining is demonstrated by examining transgenic animals expressing high levels of a non-fibrillar beta peptide variant, the beta single-chain dimer, These animals have deposits immunoreactive with anti-beta antibodies, but do not have X-34 deposits or deposits with a fibrillar ultrastructure. X-34 can also be used in vivo to visualize putative amyloid deposits resulting from accumulation of human transthyretin, another amyloidic protein. In vivo amyloid staining with X-34 may be a useful tool for monitoring anti-amyloidic treatments in real time or screening for genetic alterations that affect amyloid formation. (C) 2001 Elsevier Science Inc. All rights reserved.