REQUIREMENT FOR GLUCOSE IN LIGAND-STIMULATED MEIOTIC MATURATION OF CUMULUS CELL-ENCLOSED MOUSE OOCYTES
REQUIREMENT FOR GLUCOSE IN LIGAND-STIMULATED MEIOTIC MATURATION OF CUMULUS CELL-ENCLOSED MOUSE OOCYTES
复制标题
DOI：
10.1530/jrf.0.0960681
复制
发表时间：
1992-11-01
期刊：
影响因子：
--
通讯作者：
DOWNS, SM
中科院分区：
文献类型：
--
作者：
FAGBOHUN, CF;DOWNS, SM
本文研究了Eagle最低必需培养基中不同能量对小鼠卵母细胞减数分裂成熟的影响。葡萄糖（5.5 mmol L-1），丙酮酸（0-23 mmol L-1）和谷氨酰胺（2 mmol L-1）在不同的组合的影响进行了测试，在存在或不存在300 μ mol的二丁酰cAMP L-1的培养过程中17-18小时的裸露卵母细胞的成熟。在不存在环核苷酸的情况下，只有来自含有丙酮酸盐的那些组的卵母细胞以高频率（99-100%的萌发囊泡破裂）恢复成熟;所有其他组合导致小于或等于54%的萌发囊泡破裂。当引入二丁酰cAMP时，所有含精氨酸的组表现出约50%的成熟频率，而所有其他组中的成熟可忽略不计（小于或等于10%GVB）。丙酮酸盐对于维持裸露卵母细胞的活力也很重要（在含丙酮酸盐的培养基中活力大于或等于86%;在无丙酮酸盐的组中活力小于或等于35%）。当卵丘细胞封闭的卵母细胞培养在培养基中没有抑制剂，所有的能量底物的组合支持高频率的成熟（大于或等于89%的胚泡破裂）和活力（大于或等于91%）。添加二丁酰cAMP导致在除单独的谷氨酸盐组（97%的胚芽囊泡破裂）之外的所有培养物中减数分裂成熟的抑制（5-33%的胚芽囊泡破裂）。卵丘细胞封闭的卵母细胞的活力是最大的两个或两个以上的能源基板存在于培养基中。卵泡刺激素（FSH）产生的减数分裂成熟的刺激减数分裂阻滞卵丘细胞封闭的卵母细胞的所有文化，但最大的诱导germinal囊泡破裂依赖于D-葡萄糖。伴刀豆球蛋白A（ConA）诱导的减数分裂成熟也依赖于D-葡萄糖。卵丘细胞对D-葡萄糖的摄取和代谢在介导这些配体对卵母细胞成熟的刺激作用中是重要的，因为（1）FSH和ConA都刺激D-葡萄糖和2-脱氧葡萄糖的摄取，但不刺激3-O-甲基葡萄糖的摄取：（2）根皮素在抑制配体诱导的D-葡萄糖摄取的浓度下阻止FSH和ConA对胚泡破裂的刺激作用;（3）2-脱氧葡萄糖，一种抑制糖酵解的己糖，阻止FSH和ConA诱导减数分裂成熟;（4）D-甘露糖，一种糖酵解糖，在支持配体效应方面与D-葡萄糖一样有效。因此，虽然丙酮酸似乎是最有效的支持裸露的卵母细胞的自发成熟，D-葡萄糖的存在是重要的卵丘细胞封闭的卵母细胞的减数分裂阻滞和配体诱导的减数分裂成熟的维护。
In this study, the effect of different energy sources used in Eagle's minimum essential medium on the meiotic maturation of mouse oocytes in culture was examined. The effects of glucose (5.5 mmol l-1), pyruvate (0-23 mmol l-1) and glutamine (2 mmol l-1) in different combinations were tested on the maturation of denuded oocytes in the presence or absence of 300 mumol dibutyryl cAMP l-1 during 17-18 h of culture. In the absence of cyclic nucleotide, only oocytes from those groups containing pyruvate resumed maturation at a high frequency (99-100% germinal vesicle breakdown); all other combinations resulted in less-than-or-equal-to 54% germinal vesicle breakdown. When dibutyryl cAMP was introduced, all pyruvate-containing groups exhibited maturation frequencies of about 50%, whereas maturation in all other groups was negligible (less-than-or-equal-to 10% GVB). Pyruvate was also important for the maintenance of viability in denuded oocytes (greater-than-or-equal-to 86% viability in pyruvate-containing medium; less-than-or-equal-to 35% viability in pyruvate-free groups). When cumulus cell-enclosed oocytes were cultured in medium without inhibitor, all combinations of energy substrates supported high frequencies of maturation (greater-than-or-equal-to 89% germinal vesicle breakdown) and viability (greater-than-or-equal-to 91%). The addition of dibutyryl cAMP resulted in inhibition of meiotic maturation (5-33% germinal vesicle breakdown) in all cultures except the pyruvate-alone group (97% germinal vesicle breakdown). Viability in cumulus cell-enclosed oocytes was greatest when two or more energy substrates were present in the medium. Follicle-stimulating hormone (FSH) produced a stimulation of meiotic maturation in all cultures of meiotically arrested cumulus cell-enclosed oocytes, but maximal induction of germinal vesicle breakdown was dependent upon D-glucose. Concanavalin A (ConA)-induced meiotic maturation was also dependent upon D-glucose. Uptake and metabolism of D-glucose by the cumulus cells is important in mediating the stimulatory effects of these ligands on oocyte maturation because (1) both FSH and ConA stimulated uptake of D-glucose and 2-deoxyglucose but not 3-O-methylglucose; (2) phloretin prevented the stimulatory action of FSH and ConA on germinal vesicle breakdown at a concentration that suppressed ligand-induced uptake of D-glucose; (3) 2-deoxyglucose, a hexose that suppresses glycolysis, prevented the induction of meiotic maturation by FSH and ConA and (4) D-mannose, a glycolysable sugar, was as effective as D-glucose in supporting the ligand effects. Thus, while pyruvate appeared to be most effective in supporting the spontaneous maturation of denuded oocytes, the presence of D-glucose is important in cumulus cell-enclosed oocytes for both the maintenance of meiotic arrest and ligand-induced meiotic maturation.