Partridge R J, Leese H J
Department of Biology, University of York, UK.
Reprod Fertil Dev. 1996;8(6):945-50. doi: 10.1071/rd9960945.
Bovine embryos produced in vitro from the putative zygote stage to the blastocyst stage, and blastocysts freshly flushed from the uterus, were cultured in a physiological mixture of amino acids. Depletion of amino acids from the medium and, in a few cases, their appearance, was measured by high performance liquid chromatography. Amino acids were depleted at widely differing rates. The depletion of amino acids was higher when embryos at later developmental stages were cultured, implying an increase in amino acid requirement with development. Threonine was the only amino acid to be depleted at all stages of development; depletion increased from 0.18 +/- 0.07 pmol embryo-1 h-1 at the putative zygote stage to 1.96 +/- 0.49 pmol embryo-1 h-1 at the blastocyst stage. Glutamine was depleted at the putative zygote stage and the 4-cell stage (0.76 +/- 0.05 and 0.94 +/- 0.10 pmol embryo-1 h-1 respectively), but was not significantly depleted at the later stages. Alanine was the only amino acid that appeared consistently in the medium and its production increased progressively throughout development. Aspartate, glutamate, threonine and lysine were depleted significantly by blastocysts derived both in vitro and in vivo; the embryos in vivo also depleted arginine, phenylalanine, isoleucine and tyrosine. These results indicate that individual amino acids are depleted at different rates by bovine preimplantation embryos and suggest that amino acid requirements change during development.
将假定合子期至囊胚期体外生产的牛胚胎以及从子宫中新鲜冲洗出的囊胚,在氨基酸生理混合物中培养。通过高效液相色谱法测定培养基中氨基酸的消耗情况,在少数情况下也测定其出现情况。氨基酸的消耗速率差异很大。培养后期发育阶段的胚胎时,氨基酸的消耗更高,这意味着随着发育氨基酸需求增加。苏氨酸是在所有发育阶段都会被消耗的唯一氨基酸;其消耗从假定合子期的0.18±0.07 pmol胚胎-1 h-1增加到囊胚期的1.96±0.49 pmol胚胎-1 h-1。谷氨酰胺在假定合子期和4细胞期被消耗(分别为0.76±0.05和0.94±0.10 pmol胚胎-1 h-1),但在后期阶段消耗不显著。丙氨酸是唯一在培养基中持续出现的氨基酸,其产生在整个发育过程中逐渐增加。天冬氨酸、谷氨酸、苏氨酸和赖氨酸在体外和体内来源的囊胚中都被显著消耗;体内的胚胎还消耗精氨酸、苯丙氨酸、异亮氨酸和酪氨酸。这些结果表明,牛植入前胚胎对单个氨基酸的消耗速率不同,提示氨基酸需求在发育过程中会发生变化。