Effects of N, N-Dimethylglycine on the Development of In Vitro Produced Bovine Embryos

Abstract

Abstract. This study investigated the effects of N, N-Dimethylglycine (DMG) on the development of in vitro produced (IVP) bovine embryos. IVP embryos were obtained by in vitro fertilization of in vitro matured oocytes for 6 h. In Experiment 1, IVP embryos were cultured in mSOFaa supplemented with bovine serum albumin but without glucose (SOF1) for 4 days, transferred to mSOFaa (with 5% fetal bovine serum and 1.5 mM glucose; SOF2) supplemented with 0 (control), 0.1,1 or 10 μM DMG and cultured for an additional 7 days (11 days in total) to assess their development in vitro. When cultured in the medium with 0.1 μM DMG, a significantly higher number of IVP embryos developed to the blastocyst and hatched blastocyst stages (40.3 and 40.8%, respectively) compared with the other groups (18.7-31.0% and 15.0-28.7%, respectively; P<0.05, analysis of variance). In Experiment 2, IVP embryos were cultured in SOF1 with or without 0.1 μM DMG for 4 days, transferred to SOF2 with or without 0.1 μM DMG and further cultured as in Experiment 1; DMG was added to either SOF1 or SOF2 and to both of them to assess its exposure effects on embryo development. When cultured continuously with DMG for 11 days, significantly higher rates of IVP embryos developed into blastocyst and hatched blastocyst stages (39.0 and 47.7%, respectively) compared with the other groups (31.0-32.2% and 29.5-31.0%, respectively; P<0.05). In Experiment 3, we examined developmental speed of IVP embryos cultured with or without addition of 0.1 μM DMG to IVC medium after 7 days of IVC. When DMG was added to IVC medium, the ratio of embryos developed to advanced developmental stages (No. of embryos developed to the blastocyst and expanded blastocyst stages/No. of embryos developed to the morula stage) was 28.7% (86/3) and 7 times higher than that of those cultured without DMG, 4.0% (52/13). These results suggest that addition of 0.1 μM DMG to mSOFaa during IVC of IVP bovine embryos has a promoting effect on their development.

The metabolomic profiling identifies N, N‐dimethylglycine as a facilitator of dorsal root ganglia neuron axon regeneration after injury

“…To identify the most relevant pathways and elucidate the underlying mechanism of DRG neuron survival and regeneration post injury from the metabolic pathways in a global angle, Metaboanalyst (http://www.metaboanalyst.ca/) was utilized. The analysis results revealed that the primary disturbed pathways in DRG differential metabolites post sciatic nerve injury were ‘Histidine metabolism’, 34,35 ‘Pentose phosphate pathway’, 36,37 ‘Arginine and proline metabolism’, 38 and ‘Glycerolipid metabolism’ (6 h vs. 0 h) 17,39 ; ‘Histidine metabolism’, ‘Glycine, serine and threonine metabolism’, 40–42 ‘Taurine and hypotaurine metabolism’, 43 ‘beta‐Alanine metabolism’, 44–46 ‘Pantothenate and CoA biosynthesis’, 47 and ‘Nicotinate and nicotinamide metabolism’ (1 d vs. 0 h) 48 ; ‘Histidine metabolism’, ‘Glycine, serine and threonine metabolism’, ‘Alanine, aspartate and glutamate metabolism’, 49 and ‘Arginine and proline metabolism’ (4 d vs. 0 h) (Figure 4, Dataset S3). Among these differential metabolic pathways, “Histidine metabolism’ is the only one that was enriched at each time point (vs. 0 h) after injury.…”

N, N-Dimethylglycine decreases oxidative stress and improves <i>in vitro</i> development of bovine embryos

“…Oocytes were collected from follicles (2–5 mm in diameter) of bovine ovaries collected at a meat processing plant by aspirating follicular fluid containing cumulus–oocyte complexes (COCs) with a 5-ml syringe equipped with an 18G needle. The IVM medium was 25 mM HEPES-buffered TCM-199 (TCM-199; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 0.4% (w/v) bovine serum albumin (BSA, Fraction V), 100 μM cysteamine, 50 ng/ml epidermal growth factor, 100 ng/ml transforming growth factor α, 0.05 IU/ml human menopausal gonadotropin (Serono Laboratories, Tokyo, Japan) and 10 μg/ml gentamicin (Gibco) [ 11 ]. The COCs with two or more layers of adhered cumulus cells were washed three times with IVM medium.…”

“…DMG is presumably involved in the betaine pathway, the metabolism of homocysteine and glutathione synthesis [ 9 , 10 ]. In a previous report, addition of 0.1 μM DMG to a culture medium prompted the development of early IVP bovine embryos [ 11 ].…”

Effect of Linoleic Acid Albumin in a Dilution Solution and Long‐term Equilibration for Freezing of Bovine Spermatozoa with Poor Freezability

“…We used samples of control and LT + LAA groups at Examination 1 of good motility after freezing–thawing frozen semen for A and B bulls. We conducted in vitro fertilization (IVF) using methods explained by Takahashi et al. (2009).…”