Lindemann C B, Gardner T K, Westbrook E, Kanous K S
Dept. of Biological Sciences, Oakland University, Rochester, MI 48309-4401.
Cell Motil Cytoskeleton. 1991;20(4):316-24. doi: 10.1002/cm.970200407.
Rat sperm, demembranated with 0.1% Triton X-100, were used to explore the reversal in flagellar curvature induced by calcium ion. As reported earlier (Lindemann and Goltz, Cell Motil. Cytoskeleton, 10:420-431, 1988), the radius of curvature of the flagellar midpiece of rat sperm is controlled by the free Ca2+ concentration. A reversal of the direction of curvature (judged by the asymmetric sperm head) takes place at approximately 2.5 x 10(-6) M free Ca2+. In our current study, the time course of the curvature change, after elevating free Ca2+ to 3.5 x 10(-4) M, was utilized to assess the effects of the cAMP-kinase A pathway on the calcium response. In addition, calmodulin's involvement in this response was explored using anti-calmodulin and Cd2+. The activity state of the sperm models (which could be directly influenced through cAMP) was found to control the rate of curvature change in response to increased free Ca2+. In the most extreme case, fully quiescent sperm did not respond to Ca2+ at all, and cAMP-primed sperm models completed the response to Ca2+ in two minutes or less. Anti-calmodulin demonstrated strong inhibitory effects on the curvature reversal. Cadmium ion was also extremely potent at blocking the response to Ca2+, completely eliminating the curvature reversal at 2 x 10(-10) M free Cd2+. Based on these findings, it appears that the Ca(2+)-activated curvature reversal of rat sperm is potentiated by cAMP-dependent kinase and may be mediated through calmodulin.
用0.1% Triton X - 100去除细胞膜的大鼠精子,用于探究钙离子诱导的鞭毛曲率反转。如先前报道(Lindemann和Goltz,《细胞运动与细胞骨架》,10:420 - 431,1988),大鼠精子鞭毛中段的曲率半径由游离Ca2 +浓度控制。曲率方向的反转(通过不对称的精子头部判断)发生在游离Ca2 +浓度约为2.5×10(-6) M时。在我们当前的研究中,将游离Ca2 +升高到3.5×10(-4) M后,利用曲率变化的时间进程来评估cAMP - 蛋白激酶A途径对钙反应的影响。此外,使用抗钙调蛋白和Cd2 +探究钙调蛋白在该反应中的作用。发现精子模型的活性状态(可通过cAMP直接影响)控制着对游离Ca2 +增加的曲率变化速率。在最极端的情况下,完全静止的精子对Ca2 +根本没有反应,而用cAMP预处理的精子模型在两分钟或更短时间内完成对Ca2 +的反应。抗钙调蛋白对曲率反转表现出强烈的抑制作用。镉离子在阻断对Ca2 +的反应方面也极其有效,在游离Cd2 +浓度为2×10(-10) M时完全消除曲率反转。基于这些发现,似乎大鼠精子的Ca(2 +)激活的曲率反转由cAMP依赖性激酶增强,并且可能通过钙调蛋白介导。