SYNTHESIS OF GLUTATHIONE IN THE PREIMPLANTATION MOUSE EMBRYO
SYNTHESIS OF GLUTATHIONE IN THE PREIMPLANTATION MOUSE EMBRYO
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DOI：
10.1006/abbi.1995.1200
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发表时间：
1995-04-01
影响因子：
3.9
通讯作者：
REED, DJ
中科院分区：
文献类型：
--
作者：
GARDINER, CS;REED, DJ
本文研究了小鼠二细胞至囊胚期胚胎谷胱甘肽的消耗和补充，提取的胚胎样品经丹磺酰氯衍生后，用荧光HPLC法测定还原型谷胱甘肽（GSH）和氧化型谷胱甘肽（GSSG）的含量。添加马来酸二乙酯（DEM）可使二细胞期和囊胚期胚胎的GSH含量降低，去除DEM后，GSH含量恢复正常，添加BSO或去除培养液中的胱氨酸可阻断GSH的补充。二细胞期胚胎不能恢复GSH水平耗尽后，表现出减少体外发育。向培养基中添加甲硫氨酸不能替代胱氨酸作为谷胱甘肽合成中的半胱氨酸来源，表明胚胎不使用胱硫醚途径。在发育第3天早期收集的胚胎不能在5小时内恢复GSH水平，而在第3天晚期收集的胚胎在5小时内恢复GSH，在发育的第3天中午向培养基中添加放线菌酮降低了囊胚在第3天晚些时候恢复其GSH水平的能力。这些数据表明，GSH周转和合成在两细胞和囊胚阶段之间增加。第3天胚胎合成GSH的能力增加取决于蛋白质合成，卵裂期胚胎合成GSH的能力有限，似乎容易受到毒物或耗尽谷胱甘肽的条件的不利影响。(C)出版社：Academic Press
Depletion and repletion of glutathione in two-cell to blastocyst stage mouse embryos was examined, Reduced (GSH) and oxidized glutathione (GSSG) were measured by fluorimetric HPLC after derivatization of extracted embryo samples with dansyl chloride, Addition of buthionine sulfoximine (BSO) to culture medium for 16 h decreased GSH levels in both two-cell and blastocyst stage embryos; however, GSH decreased more drastically in blastocysts, Addition of diethyl maleate (DEM) to culture medium depleted GSH in both two-cell and blastocyst stage embryos, After removal of DEM, GSH levels returned to normal in blastocysts, and addition of BSO or removal of cystine from medium blocked GSH repletion. Two-cell stage embryos were unable to recover GSH levels after depletion and exhibited decreased in vitro development. Addition of methionine to culture medium was unable to substitute for cystine as a source of cysteine in glutathione synthesis, indicating that the embryos do not use the cystathionine pathway, Embryos collected early on Day 3 of development were unable to recover GSH levels within 5 h, whereas embryos collected late on Day 3 recovered GSH within 5 h, Addition of cycloheximide to culture medium at noon on Day 3 of development decreased the ability of blastocysts to recover their GSH levels late on Day 3, These data indicate that GSH turnover and synthesis increases between the two-cell and blastocyst stages. The increase in the ability of embryos to synthesize GSH on Day 3 is dependent on protein synthesis, Cleavage stage embryos have limited capacity to synthesize GSH and appear susceptible to adverse effects of toxicants or conditions that deplete glutathione. (C) 1995 Academic Press, Inc.