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甘氨酸对牛植入前胚胎的直接及渗透压依赖性作用

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

作者信息

Herrick Jason R, Lyons Sarah M, Greene Alison F, Broeckling Corey D, Schoolcraft William B, Krisher Rebecca L

机构信息

National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO, 80124, United States of America.

Proteomics and Metabolomics Facility, Colorado State University, C-121 Microbiology Building, 2021 Campus Delivery, Fort Collins, CO, 80523, United States of America.

出版信息

PLoS One. 2016 Jul 26;11(7):e0159581. doi: 10.1371/journal.pone.0159581. eCollection 2016.

Abstract

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.

摘要

胚胎培养基中甘氨酸(Gly)的浓度通常低于输卵管或子宫液中的浓度(~0.1 mM比≥1.2 mM)。本研究的目的是确定生理浓度的甘氨酸对牛胚胎囊胚形成、孵化、滋养外胚层(TE)和内细胞团(ICM)的细胞分配以及代谢活性的直接和渗透压依赖性影响。在实验1中,受精卵在培养的前72小时用100或120 mM NaCl和0或1 mM甘氨酸培养。与120 mM NaCl相比,用100 mM NaCl培养胚胎时,囊胚形成和孵化得到改善(P<0.05)。与0 mM甘氨酸相比,添加1 mM甘氨酸可改善(P<0.05)囊胚形成,但这种效应仅在120 mM NaCl培养的胚胎中显著(P<0.05),表明牛胚胎可将甘氨酸用作渗透剂。在实验2中,胚胎在培养的最后96小时用0.1、1.1、2.1或4.1 mM甘氨酸(100 mM NaCl)培养。甘氨酸对囊胚发育没有影响(P>0.05),但当胚胎用1.1(31.4%)或2.1(29.4%)mM甘氨酸培养时,孵化率(0.1 mM甘氨酸时为18.2%)得到改善(P<0.05)。在两个实验中,甘氨酸对囊胚、TE和ICM细胞数量均无影响(P>0.05)。囊胚产生丙氨酸、谷氨酰胺、丙酮酸和尿素,并消耗天冬氨酸,但这种代谢谱不受甘氨酸影响(P>0.05)。总之,甘氨酸(1.0 mM)可改善早期和晚期胚胎的发育,但对暴露于高渗透压的早期胚胎有益效果更明显。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c407/4961386/5102182010dc/pone.0159581.g001.jpg

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