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卵泡内氨基酸浓度以及特定成熟培养基中氨基酸对猪卵母细胞成熟、受精和植入前发育的影响。

Intrafollicular amino acid concentration and the effect of amino acids in a defined maturation medium on porcine oocyte maturation, fertilization, and preimplantation development.

作者信息

Hong J, Lee E

机构信息

School of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chunchon, Republic of Korea.

出版信息

Theriogenology. 2007 Sep 15;68(5):728-35. doi: 10.1016/j.theriogenology.2007.06.002. Epub 2007 Jul 20.

Abstract

The objective of this study was to determine the intrafollicular concentrations of free amino acids in pigs and to examine the effect of amino acids in a chemically defined maturation medium on oocyte maturation, in vitro fertilization (IVF), and embryo development in vitro. Pooled follicular fluid aspirated separately from small (<3mm in diameter), medium (3-8mm), and large follicles (>8mm) in three pairs of ovaries was analyzed for amino acid concentration. In addition, oocyte maturation, fertilization, and embryo development were examined after in vitro maturation (IVM) of oocytes in a defined maturation medium supplemented individually with glutamate (GLU), glutamine (GLN), glycine (GLY), aspartate (ASP), asparagine (ASN), arginine (ARG), alanine (ALA), leucine (LEU), lysine (LYS), proline (PRO), and valine (VAL). Irrespective of follicle size, GLY, GLU, ALA, GLN, and PRO were the most abundant amino acids in pig follicular fluid (pFF). Sperm penetration was not altered by amino acid treatment during IVM, but monospermic fertilization was increased (P<0.05) by GLN, ASP, and VAL. All amino acids except ASP and ASN stimulated (P<0.05) male pronuclear formation after IVF. ARG and ALA treatment during IVM improved (P<0.05) blastocyst formation. In conclusion, GLY, GLU, ALA, GLN, and PRO were the most abundant amino acids in pFF and amino acids in a defined medium improved porcine monospermic fertilization, male pronuclear formation, and preimplantation development.

摘要

本研究的目的是测定猪卵泡内游离氨基酸的浓度,并研究化学成分明确的成熟培养液中的氨基酸对卵母细胞成熟、体外受精(IVF)及体外胚胎发育的影响。分别从三对卵巢中的小卵泡(直径<3mm)、中等卵泡(3-8mm)和大卵泡(>8mm)中抽取的混合卵泡液,用于分析氨基酸浓度。此外,将卵母细胞在分别添加谷氨酸(GLU)、谷氨酰胺(GLN)、甘氨酸(GLY)、天冬氨酸(ASP)、天冬酰胺(ASN)、精氨酸(ARG)、丙氨酸(ALA)、亮氨酸(LEU)、赖氨酸(LYS)、脯氨酸(PRO)和缬氨酸(VAL)的化学成分明确的成熟培养液中进行体外成熟(IVM)后,检测卵母细胞成熟、受精及胚胎发育情况。无论卵泡大小,甘氨酸、谷氨酸、丙氨酸、谷氨酰胺和脯氨酸都是猪卵泡液(pFF)中含量最丰富的氨基酸。IVM期间氨基酸处理未改变精子穿透率,但谷氨酰胺、天冬氨酸和缬氨酸使单精子受精率增加(P<0.05)。IVF后,除天冬氨酸和天冬酰胺外的所有氨基酸均刺激(P<0.05)雄原核形成。IVM期间精氨酸和丙氨酸处理使囊胚形成率提高(P<0.05)。总之,甘氨酸、谷氨酸、丙氨酸、谷氨酰胺和脯氨酸是pFF中含量最丰富的氨基酸,化学成分明确的培养液中的氨基酸可改善猪的单精子受精、雄原核形成及植入前发育。

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