School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332-0100, USA.
Lab Chip. 2017 May 16;17(10):1826-1833. doi: 10.1039/c6lc01573e.
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.
动物的长期生存依赖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，然后这些信息可以调节成年行为和发育轨迹。这些变化背后的神经和分子机制还知之甚少。秀丽隐杆线虫是研究这些机制的有力模式生物；然而，传统的基于平板的培养技术在控制和调节动物环境条件方面的能力有限。为了解决这一需求，我们开发了一种基于微流控的实验平台，能够长期培养处于 L1 幼虫阶段到成年的秀丽隐杆线虫群体，同时实现其环境的空间一致性和时间控制。为了防止细菌积累并在超过一百五十小时的运行期间保持最佳的流动特性和营养一致性，对微流控系统和外围设备的几个特征进行了优化。通过在几天内操纵食物和信息素的暴露，我们能够证明生长速度和进入 dauer 的环境依赖性变化， dauer 是一种替代发育状态。我们设想这个系统在研究环境变化对行为和发育的长时间尺度变化的机制方面是有用的。
