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首页> 外文期刊>Metabolomics >Comparison of tissue harvest protocols for the quantitation of acylcarnitines in mouse heart and liver by mass spectrometry
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Comparison of tissue harvest protocols for the quantitation of acylcarnitines in mouse heart and liver by mass spectrometry

机译：质谱法比较小鼠心脏和肝脏中酰基肉碱的组织收获方案

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Profiling of acylcarnitines (ACs) in tissues and biological fluids by mass spectrometry is a powerful approach to examine the impact of genetic, pharmacological, and environmental factors on intermediary metabolism. The AC pool exhibits rapid changes in composition and abundance in response to altered cellular fuel catabolism. However, the mercurial nature of the AC pool makes it prone to spurious influences arising from experimental variables related to tissue harvesting and processing. We evaluated the impact of various strategies to anesthetize, sedate, or euthanize (A/S/E) mice which included Nembutal, Beuthanasia, isoflurane, ketamine/xylazine (Ket/Xy) and CO2/cervical dislocation (CD) on the tissue AC profiles. ACs extracted from heart and liver were derivatized to methyl esters and quantitated by mass spectrometry. Marked differences were seen in the tissue AC profiles, especially in heart, depending upon the choice of the A/S/E strategy. Most importantly, a uniform A/S/E protocol must be employed. While tissue AC profiles in situ cannot be unambiguously defined, use of Nembutal appears to be superior to other A/S/E strategies especially when assessing the AC levels in the heart. We also showed that it is preferable to expeditiously harvest and flash-freeze tissues to avoid procedure-related perturbation of the AC profile. A more protracted tissue harvest, when recovering numerous tissues from the same animal, can alter the AC pool. In conclusion, this investigation provides key guidance for harvesting heart and liver from mice in order to minimize the procedure-associated change of the AC pool which can mask the influence of the intended experimental variables.
机译：通过质谱分析组织和生物体液中的酰基肉碱（AC）是检查遗传，药理和环境因素对中间代谢的影响的有效方法。 AC池响应于改变的细胞燃料分解代谢而表现出组成和丰度的快速变化。然而，AC池的汞性质使其易于受到与组织收获和加工有关的实验变量引起的虚假影响。我们评估了麻醉，镇静或安乐死（A / S / E）小鼠的各种策略的影响，其中包括Nembutal，Beuthanasia，异氟烷，氯胺酮/甲苯噻嗪（Ket / Xy）和CO 2 /宫颈组织AC轮廓上的位错（CD）。从心脏和肝脏中提取的AC衍生化为甲酯，并通过质谱定量。根据A / S / E策略的选择，在组织AC曲线中，尤其是在心脏中观察到明显差异。最重要的是，必须采用统一的A / S / E协议。虽然不能明确定义原位组织AC曲线，但使用Nembutal似乎优于其他A / S / E策略，尤其是在评估心脏的AC水平时。我们还表明，最好尽快收获和速冻组织，以避免与操作相关的AC轮廓扰动。当从同一只动物中恢复大量组织时，更持久的组织收获会改变交流池。总之，这项研究为从小鼠体内采集心脏和肝脏提供了关键指导，以最大程度地减少与过程相关的AC池变化，从而可以掩盖预期实验变量的影响。

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