Automated screening of C. elegans neurodegeneration mutants enabled by microfluidics and image analysis algorithms,Integrative Biology

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Automated screening of C. elegans neurodegeneration mutants enabled by microfluidics and image analysis algorithms
Integrative Biology ( IF 1.5 ) Pub Date : 2018-08-17 , DOI: 10.1039/c8ib00091c
Ivan de Carlos Cáceres _{1,

2,

3,

4} , Daniel A. Porto _{1,

2,

3,

4} , Ivan Gallotta _{5,

6,

7,

8} , Pamela Santonicola _{6,

7,

8,

9} , Josue Rodríguez-Cordero _{4,

10,

11,

12} , Elia Di Schiavi _{5,

6,

7,

8,

9} , Hang Lu _{1,

2,

3,

4,

13}

Affiliation

Spinal muscular atrophy (SMA) is a degenerative disorder that selectively deteriorates motor neurons due to a deficiency of survival motor neuron protein (SMN). The illness is the leading genetic cause of death in infants and is difficult to study in complex biological systems such as humans. A simpler model system, such as the nematode C. elegans, can be used to study potential mechanisms underlying this disease; C. elegans expresses the smn-1 gene, a homologue of SMN; powerful genetic tools in C. elegans research can be used to discover novel genes whose effect on SMN remains unknown or uncharacterized. Currently, conventional screening methods are time-consuming and laborious, as well as being subjective and mostly qualitative. To address these issues, we engineer an automated system capable of performing genetic suppressor screens on C. elegans using microfluidics in combination with custom image analysis software. We demonstrate the utility of this system by isolating 21 alleles that significantly suppress motor neuron degeneration at a screening rate of approximately 300 worms per hour. Many of these mutants also have improved motor function. These isolated alleles can potentially be further studied to understand mechanisms of protection against neurodegeneration. Our system is easily adaptable, providing a means to saturate screens not only implicated in the smn-1 pathway, but also for genes involved in other neurodegenerative phenotypes.

中文翻译：

通过微流控和图像分析算法自动筛选秀丽隐杆线虫神经变性突变体

脊髓性肌萎缩症（SMA）是一种退行性疾病，由于生存运动神经元蛋白（SMN）的缺乏，选择性地使运动神经元退化。该疾病是婴儿死亡的主要遗传原因，很难在人类等复杂的生物系统中进行研究。一个更简单的模型系统，诸如线虫秀丽隐杆线虫，可用于研究这种疾病的潜在基础机制; 秀丽隐杆线虫表达smn-1基因，它是SMN的同系物；秀丽隐杆线虫的强大遗传工具研究可用于发现其对SMN的影响尚未知或未鉴定的新基因。当前，常规的筛查方法既费时又费力，并且是主观的并且大多是定性的。为了解决这些问题，我们设计了一种能够对秀丽隐杆线虫进行基因抑制子筛选的自动化系统将微流体技术与定制图像分析软件结合使用。我们通过分离21个等位基因来证明该系统的实用性，这些等位基因以每小时大约300个蠕虫的筛选速率显着抑制运动神经元变性。这些突变体中的许多也具有改善的运动功能。这些分离的等位基因可以潜在地被进一步研究以了解针对神经变性的保护机制。我们的系统很容易适应，提供了一种不仅使与smn-1途径有关的屏幕饱和，而且使与其他神经变性表型有关的基因饱和的屏幕的方法。

更新日期：2018-08-17

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