Energy conservation capacity and morphological integrity of mitochondria in hypotonically treated rabbit epididymal spermatozoa.
Energy conservation capacity and morphological integrity of mitochondria in hypotonically treated rabbit epididymal spermatozoa.
复制标题
低渗处理的兔附睾精子线粒体的能量守恒能力和形态完整性。
DOI：
10.1016/0005-2728(73)90075-3
复制
发表时间：
1973
期刊：
影响因子：
--
通讯作者：
B. Storey
中科院分区：
文献类型：
--
作者：
E. Keyhani;B. Storey
在10 mM磷酸盐缓冲液中对兔附睾精子进行低渗处理，可破坏质膜，并从仍然附着的细胞中除去细胞质液滴。然而，对线粒体没有影响，线粒体保持其围绕轴丝复合体的螺旋构型，具有完整的内膜和外膜，并显示与未处理细胞中的线粒体相同的晶体形态。低渗处理的精子在加入苹果酸-丙酮酸、琥珀酸和抗坏血酸+N，N′-四甲基-对苯二胺的情况下呼吸，但在加入果糖或NADH的情况下则不呼吸。寡霉素可抑制呼吸，解偶联剂可刺激呼吸，这表明线粒体仍保持着能量守恒的能力。加入细胞色素不能进一步刺激解偶联呼吸速率。在−196 °C低渗处理的细胞中获得的还原-减去-氧化差谱显示了完整的细胞色素，包括细胞色素c。这一结果表明，哺乳动物精子的细胞色素在贮藏或冷冻过程中丢失[Mann，T.（1951）Biochem.J.48，386-388]是尚未掺入线粒体的细胞质细胞色素。线粒体的细胞色素仍然牢固地保持在完整的外膜内。
Hypotonic treatment of rabbit epididymal spermatozoa in 10 mM phosphate buffer disrupts the plasma membrane and removes the cytoplasmic droplet from those cells to which it is still attached. There is, however, no effect on the mitochondria, which retain their helical configuration around the axial filament complex, have intact inner and outer membranes, and show the same cristal morphology as do the mitochondria in untreated cells. Hypotonically treated spermatozoa respire with added malate-pyruvate, succinate, and ascorbateplusN,N′-tetramethyl-p-phenylenediamine, but not with added fructose or NADH. Respiration is inhibited by oligomycin and stimulated by uncoupler, showing that the mitochondria have retained their capacity for energy conservation. The uncoupled respiration rate is not further stimulated by added cytochromec. Reduced-minus-oxidized difference spectra obtained at −196 °C of the hypotonically treated cells show a full complement of cytochromes, including cytochromec. This result implies that the cytochromeclost from mammalian spermatozoa on storage or chilling [Mann, T. (1951)Biochem. J.48, 386–388] is cytoplasmic cytochromecnot yet incorporated into the mitochondria. The mitochondrial cytochromecremains firmly held inside the intact outer membrane.