Effects of alpha-amanitin on nuclear maturation of porcine oocytes in vitro.
Effects of alpha-amanitin on nuclear maturation of porcine oocytes in vitro.
复制标题
α-鹅膏蕈碱对体外猪卵母细胞核成熟的影响。
DOI：
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发表时间：
1993
期刊：
影响因子：
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通讯作者：
S. Meinecke
中科院分区：
文献类型：
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作者：
B. Meinecke;S. Meinecke
研究了α-鹅膏蕈碱在不同时间（0、0.5、1、2、4、6、8、12和20 h）对卵丘封闭和裸露的猪卵母细胞暴露于这种异质性核RNA（hnRNA）合成抑制剂的影响。另一个目的是确定去除抑制剂后细胞核进展的顺序。在培养开始后0、0.5或1 h，向含有促性腺激素的培养基（TCM 199中10 IU PMSG ml-1）中加入10 μ g α-鹅膏蕈碱ml-1，可阻止卵丘扩张，只有4.4、6.4和3.5%的卵母细胞发生生殖囊泡破裂。通过将药物添加到培养基中延迟2-8 h，这种抑制作用显著降低（2 h：34.9%，4 h：53.5%，6 h：46.9%，8 h：59.2%的生发泡破裂），当卵母细胞复苏后12或20 h加入α-鹅膏蕈碱时，未观察到对核成熟的抑制。当卵丘-卵母细胞复合体在无细胞因子的培养基中培养>或= 2小时，然后转移到补充有α-鹅膏蕈碱的培养基中时，在44小时培养结束时，在所有情况下都观察到完全卵丘扩增。剥脱的卵母细胞培养前在培养基中补充α-鹅膏蕈碱或显微注射的α-鹅膏蕈碱到卵浆中的浓度为1.0和10.0毫克毫升-1仍然没有任何影响核进展。（250字处删节）
The effects of alpha-amanitin on cumulus enclosed and denuded porcine oocytes exposed to this heterogeneous nuclear RNA (hnRNA) synthesis inhibitor at different times (0, 0.5, 1, 2, 4, 6, 8, 12 and 20 h) after the start of culture was investigated. A further objective was to determine the sequence of nuclear progression after removing the inhibitor. The addition of 10 micrograms alpha-amanitin ml-1 to a gonadotrophin containing medium (10 iu PMSG ml-1 in TCM 199) at 0, 0.5 or 1 h after the onset of culture prevented cumulus expansion, and only 4.4, 6.4 and 3.5% of oocytes underwent germinal vesicle breakdown. This inhibitory effect was considerably reduced by delaying the addition of the drug to the culture medium for 2-8 h (2 h: 34.9%, 4 h: 53.5%, 6 h: 46.9%, 8 h: 59.2% germinal vesicle breakdown), and no inhibition of nuclear maturation was observed when alpha-amanitin was added after 12 or 20 h following explantation of the oocytes. When cumulus-oocyte complexes were cultured for > or = 2 h in inhibitor-free medium and then transferred to medium supplemented with alpha-amanitin, full cumulus expansion was observed in all cases, at the end of the 44 h culture. Denudation of the oocytes before culture in either medium supplemented with alpha-amanitin or microinjection of alpha-amanitin into the ooplasm at concentrations of 1.0 and 10.0 mg ml-1 remained without any effect on nuclear progression.(ABSTRACT TRUNCATED AT 250 WORDS)