Young G P, Young J D, Deshpande A K, Goldstein M, Koide S S, Cohn Z A
Proc Natl Acad Sci U S A. 1984 Aug;81(16):5155-9. doi: 10.1073/pnas.81.16.5155.
Plasma membrane fractions from Xenopus laevis oocytes were incorporated into planar lipid bilayers. We show the existence of numerous Ca2+-activated nonspecific channels that are more permeable to anions. These channels are activated by Ca2+ at micromolar concentration but not by Mg2+, Zn2+, or Mn2+, even at millimolar concentrations. Decreasing Ca2+ concentration to less than 1 microM decreases the time of channel opening until channels close completely in the absence of Ca2+ and in the presence of EGTA. I- and Br- are more permeable through this channel than Cl-. The time during which the channels remain open is also voltage-dependent, with the channels switching off at higher voltages in both polarities. Single-channel activity shows a conductance of 380 pS in 1 M NaCl and 1 mM CaCl2, with an average open lifetime of 1.5 s at 40 mV. Similar channels are found in different stages of oocyte maturation. These observations support the hypothesis that an increase in oocyte-free Ca2+ activates directly these channels, and the resultant Cl- efflux forms the ionic basis for the fertilization potential in X. laevis.
非洲爪蟾卵母细胞的质膜组分被整合到平面脂质双分子层中。我们发现存在大量Ca2+激活的非特异性通道,这些通道对阴离子的通透性更高。这些通道在微摩尔浓度的Ca2+作用下被激活,但即使在毫摩尔浓度下,Mg2+、Zn2+或Mn2+也不能激活它们。将Ca2+浓度降低到小于1微摩尔会减少通道开放时间,直到在没有Ca2+和存在乙二醇双(2-氨基乙基醚)四乙酸(EGTA)的情况下通道完全关闭。碘离子(I-)和溴离子(Br-)通过该通道的通透性比氯离子(Cl-)更高。通道保持开放的时间也依赖于电压,在两种极性下,通道在较高电压时关闭。单通道活性在1 M氯化钠(NaCl)和1 mM氯化钙(CaCl2)中显示出380皮安(pS)的电导,在40毫伏(mV)时平均开放寿命为1.5秒。在卵母细胞成熟的不同阶段发现了类似的通道。这些观察结果支持这样的假说,即卵母细胞中游离Ca2+的增加直接激活这些通道,并且由此产生的Cl-外流形成了非洲爪蟾受精电位的离子基础。