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在脑片灌注装置中进行局部药物应用和亚秒级伏安法多巴胺释放测量。

Localized drug application and sub-second voltammetric dopamine release measurements in a brain slice perfusion device.

作者信息

Sun Meng, Kaplan Sam V, Gehringer Rachel C, Limbocker Ryan A, Johnson Michael A

机构信息

Department of Chemistry and R. N. Adams Institute for Bioanalytical Chemistry, University of Kansas , Lawrence, Kansas 66045 United States.

出版信息

Anal Chem. 2014 May 6;86(9):4151-6. doi: 10.1021/ac5008927. Epub 2014 Apr 28.

Abstract

The use of fast scan cyclic voltammetry (FSCV) to measure the release and uptake of dopamine (DA) as well as other biogenic molecules in viable brain tissue slices has gained popularity over the last 2 decades. Brain slices have the advantage of maintaining the functional three-dimensional architecture of the neuronal network while also allowing researchers to obtain multiple sets of measurements from a single animal. In this work, we describe a simple, easy-to-fabricate perfusion device designed to focally deliver pharmacological agents to brain slices. The device incorporates a microfluidic channel that runs under the perfusion bath and a microcapillary that supplies fluid from this channel up to the slice. We measured electrically evoked DA release in brain slices before and after the administration of two dopaminergic stimulants, cocaine and GBR-12909. Measurements were collected at two locations, one directly over and the other 500 μm away from the capillary opening. Using this approach, the controlled delivery of drugs to a confined region of the brain slice and the application of this chamber to FSCV measurements, were demonstrated. Moreover, the consumption of drugs was reduced to tens of microliters, which is thousands of times less than traditional perfusion methods. We expect that this simply fabricated device will be useful in providing spatially resolved delivery of drugs with minimum consumption for voltammetric and electrophysiological studies of a variety of biological tissues both in vitro and ex vivo.

摘要

在过去20年里,使用快速扫描循环伏安法(FSCV)来测量活脑组织切片中多巴胺(DA)以及其他生物源分子的释放和摄取变得越来越流行。脑切片具有维持神经网络功能三维结构的优势,同时还能让研究人员从单个动物身上获得多组测量数据。在这项工作中,我们描述了一种简单、易于制造的灌注装置,旨在将药理试剂局部递送至脑切片。该装置包含一个在灌注浴下方运行的微流体通道和一个从该通道向切片供应流体的微毛细管。我们在施用两种多巴胺能兴奋剂可卡因和GBR - 12909之前和之后测量了脑切片中电诱发的DA释放。在两个位置进行测量,一个直接在毛细管开口上方,另一个距离毛细管开口500μm。使用这种方法,证明了将药物可控地递送至脑切片的受限区域以及该腔室在FSCV测量中的应用。此外,药物消耗量减少到几十微升,这比传统灌注方法少数千倍。我们预计,这种简单制造的装置将有助于以最小的消耗量提供空间分辨的药物递送,用于体外和离体的各种生物组织的伏安法和电生理研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c15c/4018083/80d25ab912e7/ac-2014-008927_0001.jpg

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