Tracing the nitrogen metabolites of glycine using <sup>15</sup>
N-glycine and mass spectrometry

Liu, Linna; Fu, Tingting; Xu, Xun-fu; Fu, Chuan; Fang, Meijuan; Liu, Yan; Xu, Pengxiang; Zhao, Yufen

doi:10.1002/rcm.7132

Cited by 5 publications

(6 citation statements)

References 33 publications

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“…Amino acids are present in oviductal and uterine fluids [ 1 – 3 ] and serve a variety of physiological functions in the preimplantation embryo. Aside from being substrates for protein synthesis, amino acids are important for ATP production [ 4 , 5 ], purine and pyrimidine synthesis [ 5 ], methylation [ 6 ], ammonium detoxification [ 7 , 8 ], maintaining the REDOX balance of the cell [ 9 ], and as signaling molecules [ 10 , 11 ]. It is perhaps not surprising then that their inclusion in embryo culture media has profound, beneficial effects on embryonic development and viability [ 12 – 16 ].…”

Section: Introductionmentioning

confidence: 99%

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Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos

et al. 2016

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Concentrations of glycine (Gly) in embryo culture media are often lower (~0.1 mM) than those in oviductal or uterine fluids (≥1.2 mM). The objective of this study was to determine direct and osmolarity-dependent effects of physiological concentrations of Gly on blastocyst formation and hatching, cell allocation to the trophectoderm (TE) and inner cell mass (ICM), and metabolic activity of bovine embryos. In experiment 1, zygotes were cultured with 100 or 120 mM NaCl and 0 or 1 mM Gly for the first 72 h of culture. Blastocyst formation and hatching were improved (P<0.05) when embryos were cultured with 100 compared to 120 mM NaCl. Inclusion of 1 mM Gly improved (P<0.05) blastocyst formation compared to 0 mM Gly, but this effect was only significant (P<0.05) for embryos cultured with 120 mM NaCl, suggesting bovine embryos can utilize Gly as an osmolyte. In experiment 2, embryos were cultured with 0.1, 1.1, 2.1, or 4.1 mM Gly (100 mM NaCl) for the final 96 h of culture. Blastocyst development was not affected (P>0.05) by Gly, but hatching (0.1 mM Gly, 18.2%) was improved (P<0.05) when embryos were cultured with 1.1 (31.4%) or 2.1 (29.4%) mM Gly. Blastocyst, TE, and ICM cell numbers were not affected (P>0.05) by Gly in either experiment. Blastocysts produced alanine, glutamine, pyruvate, and urea and consumed aspartate, but this metabolic profile was not affected (P>0.05) by Gly. In conclusion, Gly (1.0 mM) improves the development of both early and late stage embryos, but beneficial effects are more pronounced for early embryos exposed to elevated osmolarity.

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Section: Introductionmentioning

confidence: 99%

“…However, this is not the only role of Gly. Glycine is necessary for the synthesis of purines, S-adenosylmethionine, and glutathione, [ 5 ]. Glycine is also involved in one carbon metabolism, which maintains intracellular pools of methyl donors and influences epigenetic alterations during early development [ 6 , 30 ].…”

Section: Introductionmentioning

confidence: 99%

Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos

et al. 2016

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“…Furthermore, more blastocysts formed when bovine zygotes were cultured in a 247 mOsm medium (34.6%) than in a 286 mOsm medium (17.0%) in the presence of 1 mM glycine [28]. The beneficial effect of organic osmolytes can be explained by previous findings that amino acids, including osmolytes, are present in oviductal and uterine fluid [29][30][31] and carry out various physiological functions in the preimplantation embryo, including ATP production [32], ammonium detoxification [33,34] and maintaining the redox balance [35].…”

Section: Discussionmentioning

confidence: 94%

Effect of evaporation-induced osmotic changes in culture media in a dry-type incubator on clinical outcomes in in vitro fertilization-embryo transfer cycles

Chi¹,

Park²,

Yoo³

et al. 2020

Clin Exp Reprod Med

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Objective: This study investigated whether adding outer-well medium to inhibit osmotic changes in culture media in a dry-type incubator improved the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET) cycles. Methods: In culture dishes, the osmotic changes in media (20 µL)-covered oil with or without outer-well medium (humid or dry culture conditions, respectively) were compared after 3 days of incubation in a dry-type incubator. One-step (Origio) and G1/G2 (Vitrolife) media were used. Results: The osmotic changes in the dry culture condition (308 mOsm) were higher than in the humid culture conditions (285–290 mOsm) after 3 days of incubation. In day 3 IVF-ET cycles, although the pregnancy rate did not significantly differ between the dry (46.2%) and humid culture (52.2%) groups, the rates of abortion and ongoing pregnancy were significantly better in the humid culture group (2.3% and 50.2%, respectively) than in the dry culture group (8.3% and 37.8%, respectively, p<0.05). In day 5 IVF-ET cycles, the abortion rate was significantly lower in the humid culture group (2.2%) than in the dry culture group (25.0%, p<0.01), but no statistically significant difference was observed in the rates of clinical and ongoing pregnancy between the dry (50% and 25.0%, respectively) and humid culture groups (59.5% and 57.3%, respectively) because of the small number of cycles. Conclusion: Hyperosmotic changes in media occurred in a dry-type incubator by evaporation, although the medium was covered with oil. These osmotic changes were efficiently inhibited by supplementation of outer-well medium, which resulted in improved pregnancy outcomes.

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“…Glycine is a small, versatile, and conditionally essential amino acid 1. It is known that extracellular glycine can also be utilised directly for purine and glutathione (GSH) synthesis therefore contributing to DNA replication and antioxidant reactions [ 4 , 11 , 12 , 13 , 14 , 15 ]. In vivo and in vitro experiments with radiolabelled glycine showed that extracellular glycine was incorporated directly into purine [ 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

“…It is known that extracellular glycine can also be utilised directly for purine and glutathione (GSH) synthesis therefore contributing to DNA replication and antioxidant reactions [ 4 , 11 , 12 , 13 , 14 , 15 ]. In vivo and in vitro experiments with radiolabelled glycine showed that extracellular glycine was incorporated directly into purine [ 14 , 15 ]. Rats with fibrocarcinoma and rats with mammary tumour infused with 13C-glycine presented a high concentration of radiolabelled GSH in tumour and also indicated that glycine was utilised in serine and cysteine synthesis [ 11 , 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

Reduction of Rapid Proliferating Tumour Cell Lines by Inhibition of the Specific Glycine Transporter GLYT1

Bierhals¹,

Howard²,

Hirst³

2021

Biomedicines

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Studies have highlighted the relevance of extracellular glycine and serine in supporting high growth rates of rapidly proliferating tumours. The present study analysed the role of the specific glycine transporter GLYT1 in supplying glycine to cancer cells and maintaining cell proliferation. GLYT1 knockdown in the rapidly proliferating tumour cell lines A549 and HT29 reduced the number of viable cells by approximately 30% and the replication rate presented a decrease of about 50% when compared to cells transfected with control siRNA. In contrast, when compared to control, GLYT1 siRNA had only a minimal effect on cell number of the slowly proliferating tumour cell line A498, reducing the number of viable cells by 7% and no significant difference was observed when analysing the replication rate between GLYT1 knockdown and control group. When utilising a specific GLYT1 inhibitor, ALX-5407, the doubling time of rapidly proliferating cells increased by about 8 h presenting a significant reduction in the number of viable cells after 96 h treatment when compared to untreated cells. Therefore, these results suggest that GLYT1 is required to maintain high proliferation rates in rapidly proliferating cancer cells and encourage further investigation of GLYT1 as a possible target in a novel therapeutic approach.

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Tracing the nitrogen metabolites of glycine using ¹⁵ N-glycine and mass spectrometry

Abstract: The method developed here could be applied to investigations of metabolism of other amino acids, and for drug discovery studies targeting the enzymes related to amino acid metabolism.

Cited by 5 publications

References 33 publications

Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos

Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos

Effect of evaporation-induced osmotic changes in culture media in a dry-type incubator on clinical outcomes in in vitro fertilization-embryo transfer cycles

Reduction of Rapid Proliferating Tumour Cell Lines by Inhibition of the Specific Glycine Transporter GLYT1

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Tracing the nitrogen metabolites of glycine using 15 N-glycine and mass spectrometry

Abstract: The method developed here could be applied to investigations of metabolism of other amino acids, and for drug discovery studies targeting the enzymes related to amino acid metabolism.

Cited by 5 publications

References 33 publications

Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos

Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos

Effect of evaporation-induced osmotic changes in culture media in a dry-type incubator on clinical outcomes in in vitro fertilization-embryo transfer cycles

Reduction of Rapid Proliferating Tumour Cell Lines by Inhibition of the Specific Glycine Transporter GLYT1

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Tracing the nitrogen metabolites of glycine using ¹⁵ N-glycine and mass spectrometry