Liquid chromatographic-electrospray mass spectrometric determination of 1-methyl-4-phenylpyridine (MPP+) in discrete regions of murine brain.
Liquid chromatographic-electrospray mass spectrometric determination of 1-methyl-4-phenylpyridine (MPP+) in discrete regions of murine brain.
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DOI：
10.3109/15376516.2010.538753
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发表时间：
2011-03
影响因子：
3.2
通讯作者：
Rumbeiha W
中科院分区：
文献类型：
--
作者：
Lehner A;Johnson M;Simkins T;Janis K;Lookingland K;Goudreau J;Rumbeiha W
1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）作为神经毒素被广泛应用于帕金森病小鼠模型中。MPTP被代谢为1-甲基-4-苯基吡啶（MPP+），其是中枢多巴胺（DA）神经元的线粒体毒物。对MPTP的敏感性存在种属、品系和年龄差异。同时测量MPTP活性代谢产物MPP+和多巴胺（DA）在大脑中将有助于这种神经毒素的机制研究。本研究的目的是开发一种液相色谱-质谱（LC/MS）法分析MPTP和MPP+在脑组织中，并将这些在同一样品中与DA的变化，通过HPLC测量加上电化学检测。研究中使用了25只8周龄的C57 BL/6 J7雌性小鼠。给予小鼠单次皮下注射MPTP（20 mg/kg），并在1、2、4或8 h后处死。零时间对照小鼠接受0.9%生理盐水（10 ml/kg）的注射并在1 h后处死。快速收获脑并快速冷冻，将显微切割的脑区域置于含有20%甲醇的0.1 M磷酸盐-柠檬酸缓冲液（pH 2.5）中。成功开发了一种新的LC/MS方法，该方法利用MPP+ m/z 170→127、170→128和170→154碎片的选择反应监测（SRM）进行定量，并利用面积比（m/z 127）/（m/z 128）和（m/z 154）/（128）进行鉴别确认。来自m/z 174的类似SRM策略无法检测到低至0.4 ppb的任何显著水平的MPTP。根据该方法，在中脑核（NA）和纹状体（ST）中检测到MPP+，其中NA中的水平比ST中的水平高3倍。该方法的优点在于，在该过程中使用的组织缓冲液允许同时测量纹状体DA，从而使组织MPP+的积累和脑的离散区域中DA浓度的消耗之间直接相关。
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is widely used as a neurotoxin in several models of Parkinson’s disease in mice. MPTP is metabolized to 1-methyl-4-phenylpyridinium (MPP+), which is a mitochondrial toxicant of central dopamine (DA) neurons. There are species, strain, and age differences in sensitivity to MPTP. Simultaneous measurement of the MPTP active metabolite MPP+ and dopamine (DA) in the brain would be helpful in mechanistic studies of this neurotoxin. The objective of this study was to develop a liquid chromatography–mass spectrometry (LC/MS) method for analysis of MPTP and MPP+ in brain tissue and correlate these in the same sample with changes in DA measured via HPLC coupled with electrochemical detection. Twenty-five C57BL/6J7 8-week old female mice were used in the study. Mice were given a single subcutaneous injection of MPTP (20 mg/kg) and were sacrificed 1, 2, 4, or 8 h later. Zero time control mice received an injection of 0.9% normal saline (10 ml/kg) and were killed 1 h later. Brains were rapidly harvested and quickly frozen, and microdissected brain regions were placed in 0.1 M phosphate-citric acid buffer containing 20% methanol (pH 2.5). A new LC/MS method was successfully developed that utilized selected reaction monitoring (SRM) of MPP+ m/z 170→127, 170→128, and 170→154 fragmentation for quantitation and area ratios (m/z 127)/(m/z 128) and (m/z 154)/(128) for identity confirmation. A similar SRM strategy from m/z 174 was unable to detect any significant levels of MPTP down to 0.4 ppb. According to this method, MPP+ was detected in the nucleus accumbens (NA) and the striatum (ST), with the levels in the NA being 3-times higher than those in the ST. The advantage of this approach is that the tissue buffer used in this procedure allowed concurrent measurement of striatal DA, thus enabling direct correlation between accumulation of tissue MPP+ and depletion of DA concentrations in discrete regions of the brain.
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影响因子：
3.3
作者：
Behrouz, B.;Drolet, R. E.;Goudreau, J. L.
通讯作者：
Goudreau, J. L.
影响因子：
6.1
作者：
FULLER, RW;HEMRICKLUECKE, SK;PERRY, KW
通讯作者：
PERRY, KW
DOI：
10.1016/s1044-0305(03)00216-2
发表时间：
2003-06-01
影响因子：
3.2
作者：
Cox, FJ;Johnston, MV;Dasgupta, A
通讯作者：
Dasgupta, A
影响因子：
3
作者：
Hows, MEP;Ashmeade, TE;Shah, AJ
通讯作者：
Shah, AJ
影响因子：
1.7
作者：
Filipov, Nikolay M.;Norwood, Allison B.;Sistrunk, Shannon C.
通讯作者：
Sistrunk, Shannon C.