Laboratory of Nematology, Wageningen University & Reseach, Droevendaalsesteeg 1, 6708PB Wageningen, NL, The Netherlands.
Biomolecular Research Group, Canterbury Christ Church University, North Holmes Road, Canterbury CT1 1QU, UK.
Genes (Basel). 2020 Jul 11;11(7):778. doi: 10.3390/genes11070778.
Different genetic backgrounds can modify the effect of mutated genes. Human α-synuclein () gene encodes α-synuclein, and its oligomeric complexes accumulate with age and mediate the disruption of cellular homeostasis, resulting in the neuronal death that is characteristic of Parkinson's Disease. Polymorphic variants modulate this complex pathologic mechanism. Previously, we constructed five transgenic introgression lines of a model of α-synuclein using genetic backgrounds that are genetically diverse from the canonical wild-type Bristol N2. A gene expression analysis revealed that the α-synuclein transgene differentially affects genome-wide transcription due to background modifiers. To further investigate how complex traits are affected in these transgenic lines, we measured the α-synuclein transgene expression, the overall accumulation of the fusion protein of α-synuclein and yellow fluorescent protein (YFP), the lysosome-related organelles, and the body size. By using quantitative PCR (qPCR), we demonstrated stable and similar expression levels of the α-synuclein transgene in different genetic backgrounds. Strikingly, we observed that the levels of the a-synuclein:YFP fusion protein vary in different genetic backgrounds by using the COPAS™ biosorter. The quantification of the Nile Red staining assay demonstrates that α-synuclein also affects lysosome-related organelles and body size. Our results show that the same α-synuclein introgression in different backgrounds can produces differing effects on complex traits due to background modifiers.
不同的遗传背景可以修饰突变基因的作用。人类α-突触核蛋白（）基因编码α-突触核蛋白，其寡聚体复合物随年龄积累并介导细胞内稳态的破坏，导致帕金森病的特征性神经元死亡。多态变体调节这种复杂的病理机制。先前，我们使用遗传背景构建了五个α-突触核蛋白的转基因导入系，这些背景与经典的野生型 Bristol N2 遗传上不同。基因表达分析表明，由于背景修饰物，α-突触核蛋白转基因会对全基因组转录产生不同的影响。为了进一步研究这些转基因系中复杂性状是如何受到影响的，我们测量了α-突触核蛋白转基因的表达、α-突触核蛋白和黄色荧光蛋白（YFP）融合蛋白的总积累、溶酶体相关细胞器和体型。通过使用定量 PCR（qPCR），我们证明了α-突触核蛋白转基因在不同遗传背景下具有稳定且相似的表达水平。引人注目的是，我们观察到在不同的遗传背景下，通过 COPAS™生物分拣器，α-突触核蛋白：YFP 融合蛋白的水平存在差异。尼罗红染色测定法的定量表明，α-突触核蛋白还影响溶酶体相关细胞器和体型。我们的结果表明，由于背景修饰物，相同的α-突触核蛋白导入不同背景会对复杂性状产生不同的影响。
