Quantifying Tissue-Specific Proteostatic Decline in Caenorhabditis elegans.
Quantifying Tissue-Specific Proteostatic Decline in Caenorhabditis elegans.
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DOI：
10.3791/61100
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发表时间：
2021-09-07
影响因子：
1.2
通讯作者：
Samuelson, Andrew, V
中科院分区：
文献类型：
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作者：
Lazaro-Pena, Maria, I;Cornwell, Adam B.;Samuelson, Andrew, V
在正常衰老过程中，维持蛋白质组正常功能和折叠（蛋白质稳态）的能力下降，促进了越来越多的年龄相关疾病的发生。例如，具有多聚谷氨酰胺扩增的蛋白质易于聚集，如亨廷顿蛋白和伴随的亨廷顿病的发作所例示的。通过使用表达与黄色荧光蛋白（YFP）融合的polyQ重复序列的转基因秀丽隐杆线虫，已广泛研究了与年龄相关的蛋白质组退化。这种polyQ：：YFP转基因动物模型通过对荧光灶的逐渐形成（即，蛋白质聚集体）和随后由于蛋白质组的崩溃而产生的运动缺陷的发生。此外，polyQ：：YFP转基因的表达可以由组织特异性启动子驱动，允许在完整多细胞生物体的背景下评估跨组织的蛋白质稳态。该模型非常适合遗传分析，从而提供了一种方法来量化老化，这是对寿命测定的补充。我们描述了如何准确地测量polyQ：：YFP病灶内的神经元或体壁肌肉在老化过程中形成，以及随后的行为缺陷的发病。接下来，我们将重点介绍这些方法如何适应更高的吞吐量，以及使用其他新兴的C语言策略的潜在未来应用。elegans遗传分析
The ability to maintain proper function and folding of the proteome (protein homeostasis) declines during normal aging, facilitating the onset of a growing number of age-associated diseases. For instance, proteins with polyglutamine expansions are prone to aggregation, as exemplified with the huntingtin protein and concomitant onset of Huntington’s disease. The age-associated deterioration of the proteome has been widely studied through the use of transgenic Caenorhabditis elegans expressing polyQ repeats fused to a yellow fluorescent protein (YFP). This polyQ::YFP transgenic animal model facilitates the direct quantification of the age-associated decline of the proteome through imaging the progressive formation of fluorescent foci (i.e., protein aggregates) and subsequent onset of locomotion defects that develop as a result of the collapse of the proteome. Further, the expression of the polyQ::YFP transgene can be driven by tissue-specific promoters, allowing the assessment of proteostasis across tissues in the context of an intact multicellular organism. This model is highly amenable to genetic analysis, thus providing an approach to quantify aging that is complementary to lifespan assays. We describe how to accurately measure polyQ::YFP foci formation within either neurons or body wall muscle during aging, and the subsequent onset of behavioral defects. Next, we highlight how these approaches can be adapted for higher throughput, and potential future applications using other emerging strategies for C. elegans genetic analysis.
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影响因子：
16
作者：
Labbadia J;Morimoto RI
通讯作者：
Morimoto RI
影响因子：
8.8
作者：
Brehme M;Voisine C;Rolland T;Wachi S;Soper JH;Zhu Y;Orton K;Villella A;Garza D;Vidal M;Ge H;Morimoto RI
通讯作者：
Morimoto RI
影响因子：
1.2
作者：
Cornwel, Adam B.;Llop, Jesse R.;Samuelson, Andrew, V
通讯作者：
Samuelson, Andrew, V
影响因子：
4.5
作者：
Das R;Melo JA;Thondamal M;Morton EA;Cornwell AB;Crick B;Kim JH;Swartz EW;Lamitina T;Douglas PM;Samuelson AV
通讯作者：
Samuelson AV
影响因子：
12.3
作者：
通讯作者：
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