CYCLIC ADENOSINE-MONOPHOSPHATE AND OVARIAN FOLLICULAR-FLUID ACT SYNERGISTICALLY TO INHIBIT MOUSE OOCYTE MATURATION
CYCLIC ADENOSINE-MONOPHOSPHATE AND OVARIAN FOLLICULAR-FLUID ACT SYNERGISTICALLY TO INHIBIT MOUSE OOCYTE MATURATION
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DOI：
10.1210/endo-114-2-418
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发表时间：
1984-01-01
期刊：
影响因子：
4.8
通讯作者：
EPPIG, JJ
中科院分区：
文献类型：
--
作者：
DOWNS, SM;EPPIG, JJ
研究了猪卵泡液（PFF）两种部分纯化组分抑制小鼠卵母细胞成熟能力与cAMP的关系。使用的级分为：来自PFF（PM10）的Amicon PM10过滤的低MW滤液（<10，000），和来自PM10滤液（Bio-Gel）的Bio-Gel P2层析的级分，其在猪卵母细胞中具有减数分裂抑制活性。PM10单独产生一个短暂的抑制卵丘细胞封闭和裸露的小鼠卵母细胞的成熟，表现为生殖囊泡破裂。添加FSH或（Bu）2cAMP的培养基中含有的PFF馏分导致显着的协同作用的PFF单独在卵丘细胞封闭的卵母细胞的抑制作用。毛喉素或（Bu）2cAMP促进了类似的协同反应与任何PFF馏分在裸露的卵母细胞。抑制的程度始终是更大的卵丘细胞封闭比裸露的卵母细胞。推定的抑制剂可通过标称MW截止值为1，000的管路透析。PM10的蛋白水解、酸水解或乙醚提取均未降低其与（Bu）2cAMP的协同抑制作用。活性炭提取完全消除抑制。类固醇激素没有模仿PM10诱导的协同作用时，添加到（Bu）2cAMP的培养基中的浓度至少14倍以上，目前在PM10补充培养基。证明了PFF中存在与cAMP依赖性过程协同作用以抑制卵母细胞体外成熟的因子。此外，虽然PFF组分抑制裸露和卵丘细胞封闭的卵母细胞的成熟，卵丘细胞似乎介导PFF的抑制活性。显然，PFF抑制剂是一种小的（molMW <1，000）疏水分子，而不是肽或非极性脂质。
The relationship between cAMP and the capacity of two partially-purified fractions of porcine follicular fluid (PFF) to suppress mouse oocyte maturation was investigated. The fractions used were: a low MW filtrate (< 10,000) derived from Amicon PM10 filtration of PFF (PM10), and a fraction from Bio-Gel P2 chromatography of the PM10 filtrate (Bio-Gel) that has meiotic inhibitory activity in porcine oocytes. PM10 alon produced a transient inhibition of the maturation of both cumulus cell-enclosed and denuded mouse oocytes, as manifested by germinal vesicle breakdown. The addition of FSH or (Bu)2cAMP to medium containing either of the PFF fractions resulted in dramatic synergism of the inhibitory effect of PFF alone in cumulus cell-enclosed oocytes. Forskolin or (Bu)2cAMP promoted a similar synergistic response with either PFF fraction in denuded oocytes. The degree of inhibition was consistently greater in cumulus cell-enclosed than in denuded oocytes. The putative inhibitor was dialyzable through tubing having a nominal MW cutoff of 1,000. Proteolysis, acid hydrolysis or ether extraction of PM10 did not reduce its inhibitory synergism with (Bu)2cAMP. Inhibition was completely abolished by charcoal extraction. Steroid hormones did not mimic the PM10-induced synergism when added to (Bu)2cAMP-containing medium in a concentration at least 14-fold greater than that present in PM10-supplemented medium. The presence of a factor(s) in PFF that acts synergistically with a cAMP-dependent process to inhibit oocyte maturation in vitro was demonstrated. Furthermore, although PFF fractions suppressed the maturation of both denuded and cumulus cell-enclosed oocytes, cumulus cells appear to mediate the inhibitory activity of PFF. Apparently the PFF inhibitor is a small (molMW < 1,000) hydrophobic molecule but not a peptide or nonpolar lipid.