Label-free CARS microscopy reveals similar triacylglycerol acyl chain length and saturation in myocellular lipid droplets of athletes and individuals with type 2 diabetes

Sabine Daemen, Anne Gemmink, Alexandra Paul, Nils Billecke, Katrina Rieger, Sapun H. Parekh*, Matthijs K. C. Hesselink*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Web of Science)


Aims/hypothesis Intramyocellular lipid (IMCL) content associates with development of insulin resistance, albeit not in insulin-sensitive endurance-trained athletes (trained). Qualitative and spatial differences in muscle lipid composition may underlie this so-called athlete's paradox. Here we studied triacylglycerol (TAG) composition of individual myocellular lipid droplets (LDs) in trained individuals and individuals with type 2 diabetes mellitus.

Methods Trained ((V) over dot(2max) 71.0 +/- 1.6 ml O-2[kg lean body mass (LBM)](-1) min(-1)), normoglycaemic (fasting glucose 5.1 +/- 0.1 mmol/l) individuals and untrained ((V) over dotO(2max) 36.8 +/- 1.5 ml O-2[kg LBM](-1) min(-1)) individuals with type 2 diabetes (fasting glucose 7.4 +/- 0.5 mmol/l), with similar IMCL content (3.5 +/- 0.7% vs 2.5 +/- 0.3%,p = 0.241), but at opposite ends of the insulin sensitivity spectrum (glucose infusion rate 93.8 +/- 6.6 vs 25.7 +/- 5.3 mu mol [kg LBM](-1) min(-1) for trained individuals and those with type 2 diabetes, respectively) were included from our database in the present study. We applied in situ label-free broadband coherent anti-Stokes Raman scattering (CARS) microscopy to sections from skeletal muscle biopsies to measure TAG acyl chain length and saturation of myocellular LDs. This approach uniquely permits examination of individual LDs in their native environment, in a fibre-type-specific manner, taking into account LD size and subcellular location.

Results Despite a significant difference in insulin sensitivity, we observed remarkably similar acyl chain length and saturation in trained and type 2 diabetic individuals (chain length: 18.12 +/- 0.61 vs 18.36 +/- 0.43 number of carbons; saturation: 0.37 +/- 0.05 vs 0.38 +/- 0.06 number of C=C bonds). Longer acyl chains or higher saturation (lower C=C number) could be detected in subpopulations of LDs, i.e. large LDs (chain length: 18.11 +/- 0.48 vs 18.63 +/- 0.57 carbon number) and subsarcolemmal LDs (saturation: 0.34 +/- 0.02 vs 0.36 +/- 0.04 C=C number), which are more abundant in individuals with type 2 diabetes.

Conclusions/interpretation In contrast to reports of profound differences in the lipid composition of lipids extracted from skeletal muscle from trained and type 2 diabetic individuals, our in situ, LD-specific approach detected only modest differences in TAG composition in LD subpopulations, which were dependent on LD size and subcellular location. If, and to what extent, these modest differences can impact insulin sensitivity remains to be elucidated.

Original languageEnglish
Pages (from-to)2654-2664
Number of pages11
Issue number12
Early online date3 Sep 2020
Publication statusPublished - Dec 2020


  • Athlete's paradox
  • CARS microscopy
  • Intramyocellular lipid storage
  • Lipid composition
  • Lipid droplet chemical composition
  • Lipid droplets
  • Type 2 diabetes

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