Perchec G, Jeulin C, Cosson J, André F, Billard R
Laboratoire d'Ichtyologie, URM no. 3, Muséum National d'Histoire Naturelle, Paris, France.
J Cell Sci. 1995 Feb;108 ( Pt 2):747-53. doi: 10.1242/jcs.108.2.747.
Carp spermatozoa are immotile in seminal plasma or in saline solution of high osmolality (> 400 mosmol kg-1). These 'quiescent' spermatozoa initiate a progressive forward motility when transferred in freshwater or in saline solution with low osmolality (< 160 mosmol kg-1). In this study we investigated 'in vitro' the relationship between sperm ATP content (measured by bioluminescence) and sperm motility (analysed by videomicroscopy). Sperm ATP content remained high in the immobilizing medium (200 mM KCl, Tris 30 mM, pH 8.0) where no flagellar movement occurs. Dilution of these spermatozoa in the activating medium (45 mM NaCl, 5 mM KCl, Tris 30 mM, pH 8.0) triggered forward motility which varied with temperature. At 20 degrees C, sperm ATP content decreased rapidly during the progressive forward motility phase from 12 to 4 nmol/10(8) spermatozoa, concomitantly with decreases in velocity (130 to 10 microns s-1) and the beat frequency (50 to 7 Hz). An inhibitor of mitochondrial respiration (KCN 10 mM) produced a drop in sperm ATP content irrespective of the incubation medium (activating or immobilizing). A second phase of sperm motility in the activating medium was induced following a previous transfer of spermatozoa into a medium of high osmolality for a few minutes prior to the second phase. Within 10 minutes, spermatozoa recover 90% of the initial ATP level as well as forward motility. These results suggest that motility of carp spermatozoa depends on sperm ATP synthesized by mitochondrial respiration mainly stored before activation. In low osmolality conditions, the mitochondrial oxidative phosphorylation is unable to compensate for the ATP hydrolysis required to sustain motility.(ABSTRACT TRUNCATED AT 250 WORDS)
鲤鱼精子在精浆中或高渗盐溶液(>400毫摩尔/千克)中是不活动的。当转移到淡水或低渗盐溶液(<160毫摩尔/千克)中时,这些“静止”的精子会开始进行向前的渐进性运动。在本研究中,我们“体外”研究了精子ATP含量(通过生物发光测量)与精子运动(通过视频显微镜分析)之间的关系。在不发生鞭毛运动的固定培养基(200 mM氯化钾、30 mM Tris、pH 8.0)中,精子ATP含量保持较高。将这些精子在激活培养基(45 mM氯化钠、5 mM氯化钾、30 mM Tris、pH 8.0)中稀释会引发向前运动,该运动随温度变化。在20摄氏度时,精子ATP含量在向前的渐进性运动阶段迅速从12纳摩尔/10⁸个精子减少到4纳摩尔/10⁸个精子,同时速度(从130微米/秒降至10微米/秒)和摆动频率(从50赫兹降至7赫兹)也降低。线粒体呼吸抑制剂(10 mM氰化钾)会导致精子ATP含量下降,而与孵育培养基(激活或固定)无关。在激活培养基中,精子运动的第二阶段是在精子先前转移到高渗培养基中几分钟后诱导产生的。在10分钟内,精子恢复90%的初始ATP水平以及向前运动能力。这些结果表明,鲤鱼精子的运动取决于主要在激活前储存的由线粒体呼吸合成的精子ATP。在低渗条件下,线粒体氧化磷酸化无法补偿维持运动所需的ATP水解。(摘要截断于250字)