Microfluidic platform with spatiotemporally controlled micro-environment for studying long-term C. elegans developmental arrests.
Microfluidic platform with spatiotemporally controlled micro-environment for studying long-term C. elegans developmental arrests.
复制标题
具有时空控制微环境的微流体平台，用于研究长期秀丽隐杆线虫发育停滞。
DOI：
10.1039/c6lc01573e
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发表时间：
2017
期刊：
影响因子：
6.1
通讯作者：
McGrath,PatrickT
中科院分区：
文献类型：
--
作者：
Zhuo,Weipeng;Lu,Hang;McGrath,PatrickT
动物的长期生存取决于它们在多个时间尺度上感知、过滤和响应环境的能力。例如，在发育过程中，动物整合环境信息，然后可以调节成年人的行为和发育轨迹。这些变化背后的神经和分子机制知之甚少。C.秀丽线虫是研究这种机制的一种强有力的模式生物;然而，常规的基于平板的培养技术在其持续控制和调节动物的环境条件的能力方面受到限制。为了满足这一需求，我们开发了一种基于微流体的实验平台，能够长期培养发育中的C。线虫覆盖L1幼虫阶段到成年期，同时实现空间一致性和时间控制的环境。为了防止细菌积累并在超过150小时的操作期间保持最佳的流动特性和营养物一致性，优化了微流体系统和外围设备的几个特征。通过操纵食物和信息素暴露在几天内，我们能够证明环境依赖的变化，生长速度和进入dauer，一个替代的发展状态。我们设想这个系统是有用的，在研究长期的行为和发展的变化，以应对环境变化的机制。
Animals' long-term survival is dependent on their ability to sense, filter and respond to their environment at multiple timescales. For example, during development, animals integrate environmental information, which can then modulate adult behavior and developmental trajectory. The neural and molecular mechanisms that underlie these changes are poorly understood. C. elegans is a powerful model organism to study such mechanisms; however, conventional plate-based culturing techniques are limited in their ability to consistently control and modulate an animal's environmental conditions. To address this need, we developed a microfluidics-based experimental platform capable of long-term culture of populations of developing C. elegans covering the L1 larval stage to adulthood, while achieving spatial consistency and temporal control of their environment. To prevent bacterial accumulation and maintain optimal flow characteristics and nutrient consistency over the operational period of over one hundred and fifty hours, several features of the microfluidic system and the peripheral equipment were optimized. By manipulating food and pheromone exposure over several days, we were able to demonstrate environmental-dependent changes to growth rate and entry to dauer, an alternative developmental state. We envision this system to be useful in studying the mechanisms underlying long timescale changes to behavior and development in response to environmental changes.
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