Respiratory activity of oocytes isolated from ovarian follicles of the rat.
Respiratory activity of oocytes isolated from ovarian follicles of the rat.
复制标题
从大鼠卵巢卵泡中分离的卵母细胞的呼吸活性。
DOI：
10.1530/acta.0.0780751
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
K. Ahrén
中科院分区：
文献类型：
--
作者：
T. Hillensjö;L. Hamberger;K. Ahrén
为了研究底物依赖性呼吸作用，从青春期前大鼠卵巢的腔卵泡中分离出卵母细胞。使用的卵母细胞要么被1至3层颗粒细胞（卵母细胞+辐射冠）包围，要么被机械地从周围细胞剥离（剥离的卵母细胞）。利用笛卡儿微潜水员技术，可以测定单个卵母细胞+辐射冠细胞复合物或少数脱落卵母细胞样品的呼吸速率。卵母细胞在等渗的Tris-HCL缓冲液或低渗的磷酸盐缓冲液中孵育。在不添加外源底物和添加不同浓度的不同底物的情况下测定呼吸速率。在没有底物和底物存在的情况下，细胞呼吸2 ~ 4 h呈线性。两种类型的卵母细胞制剂都不能利用葡萄糖作为底物。乳酸对脱毛卵母细胞的呼吸有轻微的刺激作用，而丙酮酸和草酸盐对脱毛卵母细胞的呼吸活性均有显著的促进作用。琥珀酸盐仅在低渗培养基中对卵母细胞+辐射冠有影响，而在两种培养基中都容易被脱毛的卵母细胞利用。通过一系列初步的体外实验，研究了牛促黄体生成素（LH）对细胞呼吸的影响。无论有无辐射电晕，摄氧量都显著下降，在添加激素后30分钟内持续出现。由于这一现象与卵子成熟的关系尚未研究，其可能的生物学意义仍有待阐明。
In order to study substrate dependent respiration oocytes were isolated from antrum follicles obtained from ovaries of prepubertal rats. The oocytes used were either surrounded by 1 to 3 layers of granulosa cells (oocyte + corona radiata), or mechanically denuded from surrounding cells (denuded oocytes). By use of Cartesian micro-diver technique the respiratory rate could be determined in single oocyte + corona radiata cell complexes or in samples of a few denuded oocytes. The oocytes were incubated either in an isotonic Tris-HCL buffer or in a hypotonic phosphate buffer. The rate of respiration was determined without addition of exogenous substrate and with addition of different substrates in varying concentrations. Cellular respiration was linear 2-4 h both in absence and presence of substrate. Both types of oocyte preparations were unable to utilize glucose as substrate. Lactate stimulated slightly the respiration of the denuded oocyte while pyruvate and oxalcetate increased markedly the respiratory activity of both types of oocyte preparations. Succinate had an influence on the oocyte + corona radiata only when the hypotonic medium was used while it was easily utilized in both media by the denuded oocyte. In a preliminary series of experiments the in vitro influence of bovine lutienizing hormone (LH) on the cellular respiration was studied. Both in presence and absence of corona radiata a significant decrease in oxygen uptake was registered, consistently appearing within 30 min after the addition of the hormone. As the relationship of the phenomenon to ovum maturation was not studied, the possible biological meaning remains to be elucidated.