Gevedon Olivia, Bolus Harris, Lye Shu Hui, Schmitz Keaton, Fuentes-González Jesualdo, Hatchell Kathryn, Bley Lyndsey, Pienaar Jason, Loewen Carin, Chtarbanova Stanislava
Department of Biological Sciences, University of Alabama.
Department of Genetics, University of Wisconsin-Madison.
J Vis Exp. 2019 Jul 11(149). doi: 10.3791/59720.
There is much to understand about the onset and progression of neurodegenerative diseases, including the underlying genes responsible. Forward genetic screening using chemical mutagens is a useful strategy for mapping mutant phenotypes to genes among Drosophila and other model organisms that share conserved cellular pathways with humans. If the mutated gene of interest is not lethal in early developmental stages of flies, a climbing assay can be conducted to screen for phenotypic indicators of decreased brain functioning, such as low climbing rates. Subsequently, secondary histological analysis of brain tissue can be performed in order to verify the neuroprotective function of the gene by scoring neurodegeneration phenotypes. Gene mapping strategies include meiotic and deficiency mapping that rely on these same assays can be followed by DNA sequencing to identify possible nucleotide changes in the gene of interest.
关于神经退行性疾病的发病和进展，包括相关的潜在基因，仍有许多需要了解的地方。利用化学诱变剂进行正向遗传筛选是一种有用的策略，可将果蝇和其他与人类具有保守细胞通路的模式生物中的突变表型映射到基因上。如果感兴趣的突变基因在果蝇的早期发育阶段不是致死性的，则可以进行攀爬试验，以筛选脑功能下降的表型指标，如低攀爬率。随后，可以对脑组织进行二次组织学分析，以便通过对神经退行性变表型进行评分来验证该基因的神经保护功能。基因定位策略包括减数分裂和缺失定位，这些策略依赖于相同的试验，随后可进行DNA测序，以识别感兴趣基因中可能的核苷酸变化。
