EFFECT OF TRITON X-100 ON ULTRASTRUCTURE, REACTIVATION, AND MOTILITY CHARACTERISTICS OF RAM SPERMATOZOA
EFFECT OF TRITON X-100 ON ULTRASTRUCTURE, REACTIVATION, AND MOTILITY CHARACTERISTICS OF RAM SPERMATOZOA
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DOI：
10.1002/mrd.1120150408
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发表时间：
1986-01-01
期刊：
影响因子：
--
通讯作者：
WHITE, IG
中科院分区：
文献类型：
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作者：
VISHWANATH, R;SWAN, MA;WHITE, IG
为了获得最佳的公羊精子提取和再激活，需要在pH为7.9的培养基中添加谷氨酸、二硫苏糖醇、镁和环AMP。0.01% Triton X-100萃取后，公羊精子仅部分脱膜，质膜大部分保持完整，尤其是中间部分。0.1% Triton X-100处理去除所有质膜，提取线粒体膜和基质。在没有ATP的情况下，16.6% +-。部分去膜的精子中有0.4个精子有活动能力，而0.1% Triton X-100提取的精子完全不活动。加入ATP后，部分脱膜精子的活化率提高了81.6%。在0.1% Triton X-100中（39.5% .+-），精子完全脱膜。4.6)。细胞内LDH的释放量随去污剂浓度的增加呈线性上升（从0.01 ~ 0.05%），并趋于平稳。经ATP处理后，部分脱膜精子的跳动频率和前进速度显著增加。部分去膜的精子有完整的线粒体，可能仍然能够产生ATP，尽管精子的运动受到外源ATP的刺激。
For optimal extraction and reactivation of ram sperm, glutamate, dithiothreitol, magnesium, and cyclic AMP were required in a medium of pH 7.9. On extraction with 0.01% Triton X-100, ram sperm were only partially demembranated, and extensive areas of plasma membrane remained intact especially in the midpiece region. Treatment with 0.1% Triton X-100 removed all plasma membranes and extracted the mitochondrial membrane and matrix. In the absence of ATP, 16.6% .+-. 0.4 of the partially demembranated sperm were motile, but sperm extracted with 0.1% Triton X-100 were completely immotile. On adding ATP partially demembranated sperm reactivated better (81.6% .+-. 2.8) than sperm completely demembranated in 0.1% Triton X-100 (39.5% .+-. 4.6). The release of intracellular LDH rose linearly with increasing concentrations of the detergent from 0.01 to 0.05%, at which it plateaued. There was a significant increase in beat frequency and forward velocity of partially demembranated sperm when treated with ATP. Partially demembranated sperm had intact mitochondria that presumably were still able to produce ATP, although the spermatozoan movement was stimulated by exogenous ATP.