Elucidate C. elegans Oxygen Sensing with Microfluidics

描述(由申请人提供)： 氧感知在新陈代谢和许多与疾病相关的过程中都很重要，如缺氧和血管生成。本项目的目的是研究线虫氧气感受的机制和神经元的作用机制。线虫是一种简单的多细胞生物体，拥有强大的分子和遗传工具，是一个很好的模式系统。然而，由于技术上的困难，目前对线虫行为研究的方法限制了可以进行的实验类型和对一些结果的解释。微流体有助于解决这些技术挑战，并可以通过对行为的定量评估来推进这些研究。在这个项目中，将开发可靠的微流控氧输送系统，以分析蠕虫对特定氧气浓度或梯度的反应。将使用适当的数学模型来设计这些装置，并将探索使用生物相容性和透氧性聚合物材料的微制造工艺。这些工具允许使用在四个潜在的重要感觉神经元中缺乏特定功能的突变体来定量研究神经回路。这项测试将探索这些神经元是否以及如何定义氧气偏好的特异性。此外，这项研究将阐明氧气偏好是否是一个适应性过程，在此过程中有机体可以根据环境改变其新陈代谢。还将研究一类鸟苷酸环化酶在感知和行为中的作用，以及与经典转录途径的可能联系。这种类型的分析只有在严格控制氧气输送系统的情况下才有可能。这项研究中开发的一般技术将影响线虫的其他感知和行为研究，如气味识别和温度感觉。

DESCRIPTION (provided by applicant): Oxygen sensing is important in metabolism and many disease-related processes, such as hypoxia and angiogenesis. The goal of this project is to study the mechanisms of and neurons involved in oxygen sensing in C. elegans. C. elegans serves as an excellent model system because it is a simple multicellular organism with powerful molecular and genetic tools available. However, current approaches for C. elegans behavior research limit the type of experiments that can be performed and the interpretation of some results due to technical difficulties. Microfluidics lends itself in solving these technical challenges and can advance these studies with quantitative assessment of behaviors. In this project, reliable microfluidic oxygen delivery systems that assay worms' response to specific oxygen concentrations or gradients will be developed. Appropriate mathematical models to design these devices will be used, and microfabrication processes using biocompatible and oxygen permeable polymer materials will be explored. These tools allow the quantitative investigation of the neural circuitry using mutants that lack specific functions in four potentially important sensory neurons. The assay will probe whether and how these neurons define the specificity of oxygen preference. Furthermore, this study will elucidate whether oxygen preference is an adaptable process during which the organism may change its metabolism in accordance with the environment. The roles of a class of guanylyl cyclases in sensing and behaviors will also be studied as well as possible connections to the classical transcriptional pathways. This type of analysis is only possible with a finely controlled oxygen delivery system. The general techniques developed in this study should impact other C. elegans researches of sensing and behaviors such as odor discrimination and thermo-sensation.