Song Wan, Smith Marianne R, Syed Adeela, Lukacsovich Tamas, Barbaro Brett A, Purcell Judith, Bornemann Doug J, Burke John, Marsh J Lawrence
Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, USA.
Methods Mol Biol. 2013;1017:41-57. doi: 10.1007/978-1-62703-438-8_3.
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder. The HD gene encodes the huntingtin protein (HTT) that contains polyglutamine tracts of variable length. Expansions of the CAG repeat near the amino terminus to encode 40 or more glutamines (polyQ) lead to disease. At least eight other expanded polyQ diseases have been described. HD can be faithfully modeled in Drosophila with the key features of the disease such as late onset, slowly progressing degeneration, formation of abnormal protein aggregates and the dependence on polyQ length being evident. Such invertebrate model organisms provide powerful platforms to explore neurodegenerative mechanisms and to productively speed the identification of targets and agents that are likely to be effective at treating diseases in humans. Here we describe an optical pseudopupil method that can be readily quantified to provide a fast and sensitive assay for assessing the degree of HD neurodegeneration in vivo. We discuss detailed crossing schemes as well as factors including different drivers, various constructs, the number of UAS sites, genetic background, and temperature that can influence the result of pseudopupil measurements.
亨廷顿舞蹈症（HD）是一种常染色体显性神经退行性疾病。HD基因编码含有可变长度多聚谷氨酰胺序列的亨廷顿蛋白（HTT）。氨基末端附近CAG重复序列的扩展导致编码40个或更多谷氨酰胺（多聚Q）从而引发疾病。至少还有其他八种多聚Q扩展疾病已被描述。HD可以在果蝇中得到忠实模拟，该疾病的关键特征如发病较晚、进行性缓慢退化、异常蛋白质聚集体的形成以及对多聚Q长度的依赖性都很明显。这种无脊椎动物模型生物为探索神经退行性机制以及高效加速识别可能有效治疗人类疾病的靶点和药物提供了强大的平台。在此，我们描述了一种光学假瞳孔方法，该方法可轻松量化，以提供一种快速且灵敏的检测方法，用于在体内评估HD神经退行性变的程度。我们讨论了详细的杂交方案以及包括不同驱动子、各种构建体、UAS位点数量、遗传背景和温度等可能影响假瞳孔测量结果的因素。
