Polar metabolomics in human muscle biopsies using a liquid-liquid extraction and full-scan LC-MS

Schomakers, Bauke V.; Hermans, Jill; Jaspers, Yorrick R. J.; Salomons, Gajja S.; Vaz, Frédéric M.; Weeghel, Michel van; Houtkooper, Riekelt H.

doi:10.1016/j.xpro.2022.101302

Cited by 22 publications

(16 citation statements)

References 11 publications

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“…Metabolomics and lipidomics were performed as previously described, with minor adjustments (Schomakers et al, 2022). For each sample, approximately 150.000 cumulus cells or exactly 10 oocytes were added to a 2 mL Eppendorf Safe-Lock tube.…”

Section: Methodsmentioning

confidence: 99%

Human ovarian ageing is characterized by oxidative damage and mitochondrial dysfunction

Smits¹,

Schomakers²,

Weeghel³

et al. 2023

Preprint

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Human ovarian ageing encompasses the age-related decline in female fertility. Oxidative stress and mitochondrial dysfunction in oocytes are suggested as causal, but corroborating evidence is limited. Using immunofluorescence imaging on human ovarian tissue, we found oxidative damage by protein and lipid (per)oxidation at the primordial follicle stage. Additionally, using comprehensive metabolomics and lipidomics, a cohort of 150 human germinal vesicles and metaphase I oocytes and 15 corresponding cumulus cell samples displayed a shift in glutathione to oxiglutathione ratio and depletion of phospholipids. Age-related changes in polar metabolites suggested a decrease in mitochondrial function, as demonstrated by NAD+, purine and pyrimidine depletion, while glycolysis substrates and glutamine accumulated with age. Oocytes of advanced maternal age likely used alternative energy sources like glycolysis and the adenosine salvage pathway, and possibly increased ATP production in cumulus cells. These findings indicate that oocytes of advanced maternal age suffer from oxidative damage and mitochondrial dysfunction.

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Section: Methodsmentioning

confidence: 99%

Human ovarian ageing is characterized by oxidative damage and mitochondrial dysfunction

Smits¹,

Schomakers²,

Weeghel³

et al. 2023

Preprint

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“…Metabolomics analysis was performed as described previously. 9,10 In short, a 75 μl aliquot of the following internal standards in water was added to 20 μl of washed erythrocytes: adenosine-15 N 5 -monophosphate (100 μM), adenosine-15 N 5 -triphosphate (1 mM), D 4 -alanine (100 μM), D 7 -arginine (100 μM), D 3 -aspartic acid (100 μM), D 4 -citric acid (100 μM), 13 C 1 -citrulline (100 μM), 13 C 6 -fructose-1, 6-diphosphate (100 μM), guanosine-15 N 5 -monophosphate (100 μM), guanosine-15 N 5 -triphosphate (1 mM), 13 C 6glucose (1 mM), 13 C 6 -glucose-6-phosphate (100 μM), D 3glutamic acid (100 μM), D 5 -glutamine (100 μM), 13 C 6 -isoleucine (100 μM), D 3 -leucine (100 μM), D 4 -lysine (100 μM), D 3 -methionine (100 μM), D 6 -ornithine (100 μM), D 5 -phenylalanine (100 μM), D 7 -proline (100 μM), 13 C 3pyruvate (100 μM), D 3 -serine (100 μM), D 5 -tryptophan (100 μM), D 4 -tyrosine (100 μM), D 8 -valine (100 μM). Subsequently, 405 μl water and 500 μl methanol was added and samples were homogenized with a vortex.…”

Section: Metabolomicsmentioning

confidence: 99%

Multi‐omics in classical galactosemia: Evidence for the involvement of multiple metabolic pathways

Hermans

Weeghel

Vaz

et al. 2022

J of Inher Metab Disea

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Classical galactosemia (CG) is one of the more frequent inborn errors of metabolism affecting approximately 1:40.000 people. Despite a life-saving galactose-restricted diet, patients develop highly variable long-term complications including intellectual disability and movement disorders. The pathophysiology of these complications is still poorly understood and development of new therapies is hampered by a lack of valid prognostic biomarkers. Multiomics approaches may discover new biomarkers and improve prediction of patient outcome. In the current study, (semi-)targeted mass-spectrometry based metabolomics and lipidomics were performed in erythrocytes of 40 patients with both classical and variant phenotypes and 39 controls. Lipidomics did not show any significant changes or deficiencies. The metabolomics analysis revealed that CG does not only compromise the Leloir pathway, but

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“…Metabolomics was performed as previously described, with minor adjustments 93 . In a 2 mL tube, the following amounts of internal standard dissolved in water were added to each sample of freeze-dried gastrocnemius muscle: adenosine- 15 N 5 -monophosphate (5 nmol), adenosine- 15 N 5 -triphosphate (5 nmol), D 4 -alanine (0.5 nmol), D 7 -arginine (0.5 nmol), D 3 -aspartic acid (0.5 nmol), D 3 -carnitine (0.5 nmol), D 4 -citric acid (0.5 nmol), 13 C 1 -citrulline (0.5 nmol), 13 C 6 -fructose-1,6-diphosphate (1 nmol), 13 C 2 -glycine (5 nmol), guanosine- 15 N 5 -monophosphate (5 nmol), guanosine- 15 N 5 -triphosphate (5 nmol), 13 C 6 -glucose (10 nmol), 13 C 6 -glucose-6-phosphate (1 nmol), D 3 -glutamic acid (0.5 nmol), D 5 -glutamine (0.5 nmol), D 5 -glutathione (1 nmol), 13 C 6 -isoleucine (0.5 nmol), D 3 -lactic acid (1 nmol), D 3 -leucine (0.5 nmol), D 4 -lysine (0.5 nmol), D 3 -methionine (0.5 nmol), D 6 -ornithine (0.5 nmol), D 5 -phenylalanine (0.5 nmol), D 7 -proline (0.5 nmol), 13 C 3 -pyruvate (0.5 nmol), D 3 -serine (0.5 nmol), D 6 -succinic acid (0.5 nmol), D 4 -thymine (1 nmol), D 5 -tryptophan (0.5 nmol), D 4 -tyrosine (0.5 nmol), D 8 -valine (0.5 nmol).…”

Section: Methodsmentioning

confidence: 99%

Alterations in Peroxisomal-Mitochondrial Interplay in Skeletal Muscle Accelerates Muscle Dysfunction

Scalabrin,

Turco,

Nogara

et al. 2024

Preprint

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Whole-body energy expenditure, as well as glucose and lipid metabolism, are regulated by skeletal muscles, which account for 40-50% of human body mass. Peroxisomes are dynamic organelles that play a crucial role in lipid metabolism and clearance of reactive oxygen species, however their role in muscles remains poorly understood. To clarify this issue, we generated a muscle-specific transgenic mouse line with peroxisome import deficiency resulting from deletion of peroxisomal biogenesis factor 5 (Pex5). Pex5 inhibition disrupted the tethering between peroxisomes and mitochondria, impaired lipid metabolism and reduced muscle force and exercise performance. Moreover, mitochondrial content and function were also altered, accelerating age-related structural defects, neuromuscular junction degeneration, and muscle atrophy. Altogether, our findings show the importance of preserving peroxisomal function and their contact sites with mitochondria to maintain muscle health during aging.

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Polar metabolomics in human muscle biopsies using a liquid-liquid extraction and full-scan LC-MS

Cited by 22 publications

References 11 publications

Human ovarian ageing is characterized by oxidative damage and mitochondrial dysfunction

Human ovarian ageing is characterized by oxidative damage and mitochondrial dysfunction

Multi‐omics in classical galactosemia: Evidence for the involvement of multiple metabolic pathways

Alterations in Peroxisomal-Mitochondrial Interplay in Skeletal Muscle Accelerates Muscle Dysfunction

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