Microfabricated Liquid Chromatography System for Analysis of Neurotransmitters

 描述（由申请人提供）：拟议的研究旨在帮助了解脑深部电刺激（DBS）期间发生的潜在神经传递，DBS是一种用于缓解帕金森病患者震颤症状的常见治疗技术。虽然DBS的好处是众所周知的，但它如何发生的机制仍在争论中。预计用于分析神经递质的改进技术将能够帮助揭示DBS期间发生的脑化学变化，然后可以用于支持这种有助于减少帕金森病衰弱性质的治疗方法的进步。这将通过开发小型化液相色谱分析系统来实现。目前用于测量神经递质摄取和释放的微透析技术需要限制这种采样的空间和时间分辨率的探针。微型样品探针现在可以提高空间分辨率，因此必须对当前的分析技术（基于利用LC的分离）进行改进，以便也提高时间分辨率。通过普通的微加工技术来制造LC系统，可以开发出整个集成的微流体装置，以帮助提高该方法的速度和灵敏度。通过利用最近可用的表面多孔颗粒的分离柱，分段流动技术的样品液滴操作，和集成的电喷雾质谱检测，更快，更有效的LC系统将可用于生物分析。一旦开发出来，微型LC将用于监测帕金森病大鼠模型中丘脑底核DBS刺激期间几种神经递质（包括GABA和谷氨酸）的浓度，以测试哪些神经通路与治疗益处最相关。

 DESCRIPTION (provided by applicant): The proposed research is designed to help understand the underlying neurotransmission that occurs during deep brain stimulation (DBS), a common therapeutic technique used to relieve tremor symptoms in patients suffering from Parkinson's Disease. While the benefits of DBS are well known, the mechanisms of how it occurs are still in debate. It is anticipated that improved techniques for the analysis of neurotransmitters will be able to help uncover changes in brain chemistry that occur during DBS that could then be used to support advances to this therapeutic method that helps reduce the debilitating nature of Parkinson's Disease. This will be achieved through the development of a miniaturized liquid chromatography (LC) analysis system. Current microdialysis techniques used for measuring neurotransmitter uptake and release require probes that limit both the spatial and temporal resolution for such sampling. Miniaturized sample probes are now available that improve spatial resolution, so advances must be made to current analytical techniques (based on separations utilizing LC) in order to also improve temporal resolution. By miniaturizing a LC system through common microfabrication techniques, an entire integrated microfluidic device can be developed to help enhance both the speed and sensitivity of the method. By utilizing recently available superficially porous particles in the separation column, segmented flow techniques for sample droplet manipulation, and integrated electrospray for mass spectrometric detection, a faster and more efficient LC system will be available for biological analyses. Once developed, the miniaturized LC will be used to monitor concentrations of several neurotransmitters (including GABA and glutamate) during DBS stimulation of the subthalamic nucleus in parkinsonian rat models in order to test which neural pathways are most relevant to the therapeutic benefits.