Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance (<sup>1</sup>H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS)

Paiva, Carla; Amaral, Alexandra; Rodrı́guez, Miguel A.; Canyellas, Nicolau; Correig, X.; Ballescà, Josep Lluís; Ramalho‐Santos, João; Oliva, Rafael

doi:10.1111/andr.12027

Cited by 56 publications

(63 citation statements)

References 46 publications

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“…The use of both GC/MS and LC/MS for analysis of AGF allowed the identification of 371 metabolites, some of which have been previously described in sperm and seminal plasma of humans (Paiva et al, 2015; Qiao et al, 2017) and bulls (Velho et al, 2018). GC/MS identified fewer compounds than LC/MS, but this was an expected result because LC analysis does not require the separation of thermally stable and volatile molecules, or large metabolites (Issaq et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

Untargeted metabolomic profiling of accessory sex gland fluid from Morada Nova rams

Sousa

Lucini

Ajmone‐Marsan

et al. 2020

Molecular Reproduction Devel

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The present study was conducted to characterize the metabolome of accessory gland fluid (AGF) of locally adapted Morada Nova rams, raised in the Brazilian Northeast. AGF was collected by an artificial vagina from five vasectomized rams. Metabolites were identified by gas chromatography-mass spectrometry (GC/MS) and high-performance liquid chromatography-mass spectrometry (LC/MS), with the support of Human Metabolome Database, PubChem, LIPID Metabolites, Pathways Strategy databases, and MetaboAnalyst platforms. There were 182 and 190 metabolites detected by GC/MS and LC/MS, respectively, with an overlap of one molecule. Lipids and lipid-like molecules were the most abundant class of metabolites in the ram AGF (127 compounds), followed by amino acids, peptides, and analogs(103 metabolites). Considering all GC/MS and LC/MS, fructose, glycerol, citric acid, D-mannitol, D-glucose, and L-(+)-lactic acid were the most abundant single metabolites present in the ram AGF. Meaningful pathways associated with AGF metabolites included glycine, serine and threonine metabolism; pantothenate and CoA biosynthesis; galactose metabolism; glutamate metabolism and phenylalanine metabolism, and so forth. In conclusion, the combined use of LC/MS and GC/MS was essential for getting a holistic view of the compounds embedded in the ram AGF. Chemical analysis of the accessory sex gland secretion is relevant for understanding sperm function and fertilization. K E Y W O R D S accessory sex gland, GC/MS, LC/MS, metabolome, ovine 410 | SOUSA ET AL.

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Section: Discussionmentioning

confidence: 99%

Untargeted metabolomic profiling of accessory sex gland fluid from Morada Nova rams

Sousa

Lucini

Ajmone‐Marsan

et al. 2020

Molecular Reproduction Devel

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“…Studies reported in the literature concerning FA profiling in the various settings of sperm characteristics focused on specific compounds, compared limited categories of subjects, included a limited number of samples, and results varied widely (Gulaya et al ., ; Wathes et al ., ; Chavarro et al ., ). With the advancement of technology, it is possible to explore new markers, and new metabolites have been described recently (Paiva et al ., ).…”

Section: Introductionmentioning

confidence: 97%

Fatty acids profiling reveals potential candidate markers of semen quality

et al. 2016

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Previous reports showed altered fatty acid content in subjects with altered sperm parameters compared to normozoospermic individuals. However, these studies focused on a limited number of fatty acids, included a short number of subjects and results varied widely. We conducted a case-control study involving 155 patients allocated into four groups, including normozoospermia (n = 33), oligoasthenoteratozoospermia (n = 32), asthenozoospermia (n = 25), and varicocoele (n = 44). Fatty acid profiling, including 30 species, was analyzed by a validated gas chromatography (GC) method on the whole seminal fluid sample. Multinomial logistic regression modeling was used to identify the associations between fatty acids and the four groups. Specimens from 15 normozoospermic subjects were also analyzed for fatty acids content in the seminal plasma and spermatozoa to study the distribution in the two compartments. Fatty acids lipidome varied markedly between the four groups. Multinomial logistic regression modeling revealed that high levels of palmitic acid, behenic acid, oleic acid, and docosahexaenoic acid (DHA) confer a low risk to stay out of the normozoospermic group. In the whole population, seminal fluid stearic acid was negatively correlated (r = -0.53), and DHA was positively correlated (r = 0.65) with sperm motility. Some fatty acids were preferentially accumulated in spermatozoa and the highest difference was observed for DHA, which was 6.2 times higher in spermatozoa than in seminal plasma. The results of this study highlight complete fatty acids profile in patients with different semen parameters. Given the easy-to-follow and rapid method of analysis, fatty acid profiling by GC method can be used for therapeutic purposes and to measure compliance in infertility trials using fatty acids supplements.

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“…The association between glycolytic substrates and energy production, which is essential for motility, was determined using metabolomics approach in mouse spermatozoa (Goodson et al, 2012). More recently, using both NMR and gas chromatographymass spectrometry (GC-MS), total of 96 metabolites and more than 10 biological pathways were identified in human sperm (Paiva et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Amino Acids of Seminal Plasma Associated With Freezability of Bull Sperm

Ugur

Hitit

Kaya

et al. 2020

Front. Cell Dev. Biol.

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Sperm cryopreservation is an important technique for fertility management, but postthaw viability of sperm differs among breeding bulls. With metabolites being the end products of various metabolic pathways, the contributions of seminal plasma metabolites to sperm cryopreservation are still unknown. These gaps in the knowledge base are concerning because they prevent advances in the fundamental science of cryobiology and improvement of bull fertility. The objective of this study was to test the hypothesis that seminal plasma amino acids are associated with freezability of bull sperm. To accomplish this objective, amino acid concentrations in seminal plasma from seven bulls of good freezability (GF) and six bulls of poor freezability (PF) were quantified using gas chromatography-mass spectrometry (GC-MS). Multivariate and univariate analyses were performed to identify potential freezability biomarkers. Pathways and networks analyses of identified amino acids were performed using bioinformatic tools. By analyzing and interpreting the results we demonstrated that glutamic acid was the most abundant amino acid in bull seminal plasma with average concentration of 3,366 ± 547.3 nM, which accounts for about 53% of total amino acids. The other most predominant amino acids were alanine, glycine, and aspartic acid with the mean concentrations of 1,053 ± 187.9, 429.8 ± 57.94, and 427 ± 101.3 nM. Pearson's correlation analysis suggested that phenylalanine concentration was significantly associated with post-thaw viability (r = 0.57, P-value = 0.043). Significant correlations were also found among other amino acids. In addition, partial least squares-discriminant analysis (PLS-DA) bi-plot indicated a distinct separation between GF and PF groups. Phenylalanine had the highest VIP score and was more abundant in the GF groups than in the PF groups. Moreover, pathway and network analysis indicated that phenylalanine contributes to oxidoreductase and antioxidant reactions. Although univariate analysis did not yield significant differences in amino acid concentration between the two groups, these findings are significant that they indicate the potentially important roles of amino acids in seminal plasma, thereby building a foundation for the fundamental science of cryobiology and reproductive biotechnology.

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Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance (¹H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS)

Cited by 56 publications

References 46 publications

Untargeted metabolomic profiling of accessory sex gland fluid from Morada Nova rams

Untargeted metabolomic profiling of accessory sex gland fluid from Morada Nova rams

Fatty acids profiling reveals potential candidate markers of semen quality

Amino Acids of Seminal Plasma Associated With Freezability of Bull Sperm

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Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance (1H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS)

Cited by 56 publications

References 46 publications

Untargeted metabolomic profiling of accessory sex gland fluid from Morada Nova rams

Untargeted metabolomic profiling of accessory sex gland fluid from Morada Nova rams

Fatty acids profiling reveals potential candidate markers of semen quality

Amino Acids of Seminal Plasma Associated With Freezability of Bull Sperm

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Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance (¹H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS)