Single-cell metabolomics: changes in the metabolome of freshly isolated and cultured neurons.
Single-cell metabolomics: changes in the metabolome of freshly isolated and cultured neurons.
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DOI：
10.1021/cn300100u
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发表时间：
2012-10-17
影响因子：
5
通讯作者：
Sweedler, Jonathan V.
中科院分区：
文献类型：
--
作者：
Nemes, Peter;Knolhoff, Ann M.;Rubakhin, Stanislav S.;Sweedler, Jonathan V.
关键词：
代谢物参与多种细胞内过程，包括细胞对不断变化的细胞外环境的反应。使用单细胞毛细管电泳与电喷雾电离质谱联用，我们研究了将单个识别的神经元放入培养物中如何影响其代谢特征。首先，使用加州海兔的大脑神经元评估基于甘油的细胞稳定性；与未经甘油稳定化分离的细胞相比，甘油稳定化细胞的测量误差从~24%相对标准偏差降低至~6%。为了确定培养引起的变化，对来自加利福尼亚州加州两种代谢不同的细胞类型（B1 和 B2 颊神经元）的 14 个新鲜分离的神经元和 11 个过夜培养的神经元进行了表征。在检测到的 300 多种独特的细胞相关信号中，根据其已知的生物学作用选择了 35 种化合物，并在每个测量的细胞之间进行比较。无监督的多变量和统计分析揭示了这两种已识别的神经元类型之间存在显着的代谢差异。然后我们比较了过夜培养引起的变化；培养的 B1 细胞中 26 种化合物的代谢浓度不同。相比之下，培养对 B2 神经元代谢谱的影响较小，只有 5 种化合物发生显着变化。由于这些培养诱导的变化，B1 神经元的代谢组成与培养的 B2 细胞难以区分。这一观察结果表明，这两种细胞类型响应环境变化而差异性地调节其体内或体外代谢组。
Metabolites are involved in a diverse range of intracellular processes, including a cell’s response to a changing extracellular environment. Using single-cell capillary electrophoresis coupled to electrospray ionization mass spectrometry, we investigated how placing individual identified neurons in culture affects their metabolic profile. First, glycerol-based cell stabilization was evaluated using metacerebral neurons from Aplysia californica; the measurement error was reduced from ∼24% relative standard deviation to ∼6% for glycerol-stabilized cells compared to those isolated without glycerol stabilization. In order to determine the changes induced by culturing, 14 freshly isolated and 11 overnight-cultured neurons of two metabolically distinct cell types from A. californica, the B1 and B2 buccal neurons, were characterized. Of the more than 300 distinctive cell-related signals detected, 35 compounds were selected for their known biological roles and compared among each measured cell. Unsupervised multivariate and statistical analysis revealed robust metabolic differences between these two identified neuron types. We then compared the changes induced by overnight culturing; metabolite concentrations were distinct for 26 compounds in the cultured B1 cells. In contrast, culturing had less influence on the metabolic profile of the B2 neurons, with only five compounds changing significantly. As a result of these culturing-induced changes, the metabolic composition of the B1 neurons became indistinguishable from the cultured B2 cells. This observation suggests that the two cell types differentially regulate their in vivo or in vitro metabolomes in response to a changing environment.
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影响因子：
7.4
作者：
Mellors, J. Scott;Jorabchi, Kaveh;Smith, Lloyd M.;Ramsey, J. Michael
通讯作者：
Ramsey, J. Michael
影响因子：
19
作者：
Huh, Dongeun;Hamilton, Geraldine A.;Ingber, Donald E.
通讯作者：
Ingber, Donald E.
DOI：
10.1039/c2an16211c
发表时间：
2012-07-07
期刊：
The Analyst
影响因子：
--
作者：
Cecala C;Sweedler JV
通讯作者：
Sweedler JV
影响因子：
16.2
作者：
Fuller, RR;Moroz, LL;Sweedler, JV
通讯作者：
Sweedler, JV
影响因子：
7.4
作者：
Amantonico, Andrea;Urban, Pawel L.;Zenobi, Renato
通讯作者：
Zenobi, Renato