Shi Y L, Ma X H
Shanghai Institute of Physiology, Chinese Academy of Sciences, P.R. China.
Mol Reprod Dev. 1998 Jul;50(3):354-60. doi: 10.1002/(SICI)1098-2795(199807)50:3<354::AID-MRD12>3.0.CO;2-4.
Ion environment and ionic fluxes through membrane are thought to be important in the spermatozoa's maturation, capacitation, and the initiating process of gamete interaction. In this work, the membrane proteins isolated from human sperm plasma membrane were reconstituted into planar lipid bilayers via fusion, and the ion channels activities were observed under voltage clamp mode. In cis 200//trans 100 mM KCl solution, a TEA-sensitive cation-selective channel with a unit conductance of 40 pS was recorded. In a gradient of 200//100 mM NaCl solutions, a Na(+)-selective channel with a unit conductance of 26 pS was recorded. In both cases, reversal potential was about-18 mV, which is close to the predicated value of a perfect Nernst K+ or Na+ electrode. In 50//10 mM CaCl2 solution, a cation channel activity with a unit conductance of 40 pS and reversal potential of about -20 mV was usually observed. In 200//100 mM NMDG(N-methyl-D-glucamine)-Cl solution, where the cation ions were substituted with NMDG, a 30-pS anion-selective channel activity was also detected. The variety in the types of ion channels observed in human spermatozoa plasma membrane suggests that ion channels may play a range of different roles in sperm physiology and gamete interaction.
离子环境以及离子通过细胞膜的通量被认为在精子的成熟、获能以及配子相互作用的起始过程中起着重要作用。在这项研究中,从人精子质膜分离出的膜蛋白通过融合被重组到平面脂质双分子层中,并在电压钳模式下观察离子通道活性。在顺式200//反式100 mM KCl溶液中,记录到一个单位电导为40 pS的TEA敏感阳离子选择性通道。在200//100 mM NaCl溶液梯度中,记录到一个单位电导为26 pS的Na(+)选择性通道。在这两种情况下,反转电位约为 -18 mV,这接近于完美能斯特K+或Na+电极的预测值。在50//10 mM CaCl2溶液中,通常观察到一个单位电导为40 pS且反转电位约为 -20 mV的阳离子通道活性。在200//100 mM NMDG(N-甲基-D-葡萄糖胺)-Cl溶液中,阳离子被NMDG取代,还检测到一个30-pS的阴离子选择性通道活性。在人精子质膜中观察到的离子通道类型的多样性表明,离子通道可能在精子生理和配子相互作用中发挥一系列不同的作用。