Halangk W, Bohnensack R, Frank K, Kunz W
Biomed Biochim Acta. 1985;44(3):411-20.
Ejaculated bull spermatozoa separated from seminal plasma by repeated washing were used to study the effects of various substrates on respiration, mitochondrial energization, cellular ATP/ADP ratios, and motility. The respiratory rate in the presence of lactate, pyruvate, acetate or glycerol was about two to three times higher compared to cells oxidizing amino acids, fatty acids, or endogenous substrates. When well oxidizable substrates were present, the uncoupler FCCP produced a marked increase in respiration (two to three times at optimal FCCP concentrations). Mitochondrial energization was monitored in intact spermatozoa by spectrophotometry with the dye safranin. Lactate and pyruvate yielded more pronounced mitochondrial energization than did alanine or endogenous substrates. Cellular ATP/ADP ratios, with lactate, pyruvate, acetate or glycerol, were found to range from 6 to 10, whereas endogenous substrates and amino acids were able to establish ATP/ADP quotients as low as about 1 only. The capacity to oxidize various substrates was clearly reflected by the amount of motile cells. Motility in the presence of lactate, acetate or glycerol was seen in more than 60% of spermatozoa, while spermatozoa in the presence of alanine or butyrate showed a motility not above the endogenous level (approx. 30% motile cells). The experiments demonstrated that cellular performance is highly dependent on the energy state as made possible by the capacity of substrate utilization.
通过反复洗涤从精浆中分离出的射精公牛精子，用于研究各种底物对呼吸、线粒体能量化、细胞ATP/ADP比率和活力的影响。与氧化氨基酸、脂肪酸或内源性底物的细胞相比，在乳酸、丙酮酸、乙酸盐或甘油存在下的呼吸速率高出约两到三倍。当存在易于氧化的底物时，解偶联剂FCCP会使呼吸显著增加（在最佳FCCP浓度下增加两到三倍）。通过使用番红染料的分光光度法监测完整精子中的线粒体能量化。乳酸和丙酮酸产生的线粒体能量化比丙氨酸或内源性底物更明显。发现细胞的ATP/ADP比率在乳酸、丙酮酸、乙酸盐或甘油存在下为6至10，而内源性底物和氨基酸只能建立低至约1的ATP/ADP商。运动细胞的数量清楚地反映了氧化各种底物的能力。在乳酸、乙酸盐或甘油存在下，超过60%的精子具有活力，而在丙氨酸或丁酸盐存在下的精子活力不高于内源性水平（约30%的运动细胞）。实验表明，细胞性能高度依赖于底物利用能力所实现的能量状态。
