Irisin: Still chasing shadows
Irisin: Still chasing shadows
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DOI：
10.1016/j.molmet.2020.01.016
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发表时间：
2020-04-01
影响因子：
8.1
通讯作者：
Maak, Steffen
中科院分区：
文献类型：
--
作者：
Albrecht, Elke;Schering, Lisa;Maak, Steffen
目的：在其最初描述后的七年多时间里，关于测量肌因子虹膜素的准确性仍然存在相当大的不确定性。未解决的问题包括跨物种的鸢尾素前体纤连蛋白III型结构域5（FNDC5）基因的转录的性质，商业酶联免疫吸附测定（ELISA）测量的鸢尾素水平的可靠性，以及最近发表的定量质谱法测量的人血清参考值的整体有效性。我们利用多个物种和措施，以评估常用的试剂和方法报告irisin.Methods的稳健性：扩增cDNA被用来评估在人类和小鼠的FNDC5转录模式。免疫印迹法检测不同鸢尾素抗体的特异性和敏感性。循环天然鸢尾素的定量进行质谱使用的绝对定量肽为irisin.Results：我们表明，有一个更大的人类FNDC5的转录比目前注释的多样性，但没有迹象表明的表达的转录导致的截断形式的鸢尾素。可用的鸢尾素抗体仍然结合非特异性血清蛋白的模式，这损害了用ELISA法对鸢尾素的可靠测量。绝对定量的鸢尾素与标记肽质谱是一种先进的方法，但需要多步骤的样品制备引入不可控的变化在measurement.Conclusion：我们的数据代表了一个明确的警告，对测量循环鸢尾素使用现有的方法。测量Irisin类似于追踪阴影。(C)2020作者（S）由爱思唯尔有限公司出版。
Objective: Considerable uncertainty remains regarding the veracity of measuring myokine irisin more than seven years after its original description. Unresolved issues include the nature of transcription of the irisin precursor fibronectin type III domain containing 5 (FNDC5) gene across species, the reliability of irisin levels measured with commercial enzyme-linked immunosorbent assays (ELISAs), and the overall validity of the recently published reference values for human serum measured with quantitative mass spectrometry. We utilized multiple species and measures to evaluate the robustness of commonly used reagents and methods for reporting irisin.Methods: Amplification of cDNA was used to assess the FNDC5 transcript patterns in humans and mice. The specificity and sensitivity of different irisin antibodies were examined via western blotting. Quantification of circulating native irisin was conducted with mass spectrometry using an absolute quantification peptide for irisin.Results: We show that there is a greater transcript diversity of human FNDC5 than currently annotated, but no indication of the expression of transcripts leading to a truncated form of irisin. Available irisin antibodies still bind to patterns of unspecific serum proteins, which compromise reliable measurements of irisin with ELISAs. Absolute quantification of irisin with labeled peptides by mass spectrometry is an advanced method but requires a multi-step sample preparation introducing uncontrollable variations in the measurement.Conclusion: Our data represent an explicit warning against measuring circulating irisin using available methods. Measuring irisin is akin to chasing shadows. (C) 2020 The Author(s). Published by Elsevier GmbH.