Abstract

ObjectiveConsiderable 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. MethodsAmplification 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. ResultsWe 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. ConclusionOur data represent an explicit warning against measuring circulating irisin using available methods. Measuring irisin is akin to chasing shadows.

Highlights

  • The putative myokine irisin has been the subject of debate since its description in 2012 by Boström et al [1]

  • We first amplified three fragments allegedly representative of three fibronectin type III domain containing 5 (FNDC5) transcripts annotated in GenBank (NM_001171941.3, NM_153756.3, and NM_001171940.2; T1-3 in Figure 1) in human skeletal muscle cDNA with primers described by Kim et al [30]

  • We tried to amplify further fragments of transcripts T1-T3 and searched for respective expressed sequence tags (ESTs) in NCBI databases supporting the structure of the identified transcripts

Read more

Summary

Introduction

The putative myokine irisin has been the subject of debate since its description in 2012 by Boström et al [1]. The scientists who discovered irisin addressed some of these points of contention [7]. They reported detection of glycosylated as well as deglycosylated native irisin in human plasma samples using western blotting with a commercial antibody. They used quantitative mass spectroscopy to measure circulating irisin in sedentary and trained individuals. The study involved only a few individuals with a marginal increase in irisin, some considered the data sufficient to settle the debate [8]. Others suggested independently reproducing these results [9]

Methods
Results
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call