Murdoch R N, Armstrong V L, Clulow J, Jones R C
The Department of Biological Sciences, The University of Newcastle, Callaghan, NSW, Australia.
Reprod Fertil Dev. 1999;11(2):87-94. doi: 10.1071/rd99039.
The oxygen consumption of rat sperm was low (2.7 microL O2 10(8) sperm(-1) h(-1)) in caudal epididymal semen (CES) when stimulation of motility was avoided. The addition of 1 microL of Krebs Ringer phosphate buffer (KRP) to 40 microL of CES (CES:KRP = 40:1) did not activate motility, but stimulated oxygen consumption 2-fold. Inclusion of 1-5 mM glucose, acetate, pyruvate or lactate in the KRP further stimulated respiration rate (up to 4.3-fold) without activating motility, but respiration was reduced when 2-deoxyglucose replaced energy substrates. Inclusion of dibutyryl cAMP (1 mM) activated sperm motility in all samples and stimulated oxygen consumption 2.9-fold. Dilution of CES at the ratio of CES:KRP = 40:1000 also activated sperm motility and stimulated respiration rate 2.9-fold. The combined effect of dibutyryl cAMP and glucose in stimulating respiration was greater than their individual effects. However, the response to cAMP or substrates was not altered by incubation in KRP containing either 0 or 0.5 mM Ca2+. It was concluded that the motility and metabolism of rat epididymal sperm are suppressed in vivo. Respiration can be stimulated by a small (1.025-fold) dilution and further stimulated by the inclusion of energy substrate, without activating motility. However, a larger dilution or inclusion of cAMP activated motility and simultaneously stimulated metabolism, with exogenous substrate being required to stimulate respiration to the maximum rate. This suggests that prior to activation, the rate of oxygen consumption and sperm motility are not coupled.
在避免刺激精子运动的情况下,大鼠附睾尾精液(CES)中精子的耗氧量较低(2.7微升O₂ 10⁸精子⁻¹ 小时⁻¹)。向40微升CES中添加1微升磷酸 Krebs - Ringer缓冲液(KRP)(CES:KRP = 40:1)并未激活精子运动,但刺激耗氧量增加了2倍。在KRP中加入1 - 5毫摩尔葡萄糖、乙酸盐、丙酮酸盐或乳酸盐进一步刺激呼吸速率(高达4.3倍),且未激活精子运动,但当2 - 脱氧葡萄糖替代能量底物时,呼吸作用减弱。加入二丁酰cAMP(1毫摩尔)可激活所有样本中的精子运动,并刺激耗氧量增加2.9倍。以CES:KRP = 40:1000的比例稀释CES也可激活精子运动,并刺激呼吸速率增加2.9倍。二丁酰cAMP和葡萄糖联合刺激呼吸的效果大于它们各自单独作用的效果。然而,在含有0或0.5毫摩尔Ca²⁺的KRP中孵育,对cAMP或底物的反应并未改变。得出的结论是,大鼠附睾精子的运动和代谢在体内受到抑制。小幅度稀释(1.025倍)可刺激呼吸作用,加入能量底物可进一步刺激呼吸作用,而不激活精子运动。然而,更大幅度的稀释或加入cAMP可激活精子运动并同时刺激代谢,需要外源底物才能将呼吸作用刺激到最大速率。这表明在激活之前,耗氧率和精子运动并不耦合。