钙在IAA诱导绿豆下胚轴原生质体膨大过程中的作用

含钙培养液(对照)和仅用IAA处理的原生质体的体积和~(45)Ca~(2 )放射性强度均无变化。IAA处理含钙培养液中的原生质体,5min后~(45)Ca~(2 )积累明显增多,体积开始膨大。处理30min时~(45)Ca~(2 )积累最多,此时原生质体的膨大效应最好;随后~(45)Ca~(2 )积累和膨大效应逐渐下降。K~ 、Zn~(2 )、Ba~(2 )、Mg~(2 )等也可在一定程度上代替Ca~(2 )使原生质体体积膨大。原生质体的吸水在膨大中起着一定作用。EGTA、LaCl_3和verapamil均抑制IAA诱导的原生质体~(45)Ca~(2 )积累和体积膨大。说明Ca~(2 )可能在6-BA诱导原生质体膨大的过程中起着重要作用。     

钙在6-BA诱导黄化绿豆幼苗下胚轴原生质体体积膨大中的作用

对照和仅用6-BA(无Ca~(2 ))处理的原生质体的体积均无变化。含钙培养液中的原生质体经6-BA处理后10min~(45)Ca~(2 )积累明显增多,15min后开始膨大;处理30min时~(45)Ca~(2 )积累最多,此时原生质体的膨大效应最好;随后~(45)Ca~(2 )积累和膨大效应逐渐下降。两者的时间进程十分相似。K~ 、Zn~(2 )、Ba~(2 )、Mg~(2 )等可在不同程度上代替Ca~(2 )的作用。EGTA、verapamil和LaCl_3处理均可使原生质体~(45)Ca~(2 )累积降到对照的水平,膨大效应完全消失。表明Ca~(2 )可能在6-BA诱导原生质体膨大的过程中起着重要作用。     

低钾与哇巴因诱发经低温处理的心室肌节律活动

65个猪心室肌标本经24或48h低温处理后,用玻璃微电极进行细胞内动作电位记录。在经低温处理 48h的心室肌上,电刺激均能诱发节律性电活动,而在经低温处理 24h的标本上,节律性电活动的发生率只有8.3％。经24h低温处理后仍无节律性电活动的标本对低钾与哇巴因的反应和新鲜标本不同。低钾(0.35—0.7mM)台氏液能诱导所有低温处理标本的动作电位出现舒张期自动去极化(SD),从而导致节律性电活动。低温处理标本不仅对哇巴因毒性作用的耐受性大大减弱,而且在哇巴因(2×10~(-7)M)作用下,均出现叠加在缓慢 SD上的振荡电位,这种振荡电位的幅度一般小于10mV。低钾(1.4mM)台氏液灌流能使这种振荡电位增大,并在SD基础上导致自发活动。 实验结果提示,钠钾泵活动的抑制可能与心室肌细胞的节律性电活动有关。     

蜜蜂間接飞翔肌的电生理观察

本文报告我們观察蜜蜂間接飞翔肌肌細胞的电反应和測量膜的一些电性貭的結果。膜电位的数值为35±7毫伏,峯电位的振幅为42±11毫伏。“自发”放电和通过头腹部两点刺激引起的反应伴有延續时間較长(数十至数百毫秒)的負后电位,在它的上面还往往重复出現一些峯电位。直接刺激肌肉引起的反应的負后电位較小,而且沒有重复反应。細胞膜的电阻为1—4×10~3欧姆厘米~2,电容为2—9微法拉/厘米~2。     

L—天冬氨酸和草酰乙酸对豚鼠耳蜗电位的影响

用耳蜗灌流和电生理技术,研究了L—天冬氨酸(L—ASP)和草酰乙酸(Oxa)对豚鼠耳蜗电位的影响。对照灌流液为人工外淋巴液(Elliotts’液),试验液分别为20或25mM的L—ASP或10mM的Oxa,均做鼓阶灌流以进行比较。灌流液由电子微量泵驱动。在灌流过程中记录由短声引起的CM和APN_1,并在微处理机上作常规处理。20mM的L—ASP使APN_1振幅降低49％,P相似文献   

海洋来源琼胶酶的分离纯化及酶学性质研究

采用Vibiro sp.ZC-1发酵制备琼胶酶,粗酶液经过中空纤维柱浓缩、硫酸铵沉淀、DEAE-阴离子交换层析,得到一个电泳纯的琼胶酶组分Aga ZC-1,其分子质量约为45k Da,比活力为114.613U/mg。对Aga ZC-1进行酶学性质分析,结果表明,其最适反应p H为7.0,在p H为5.0~9.0时保温1h仍能保持80%以上的酶活力;最适反应温度为50℃,在45℃条件下保温1h酶活力保持在60%以上。在高浓度(5mmol/L)下,Fe~(3+)、Cu~(2+)、Sn~(2+)和Zn~(2+)能完全抑制琼胶酶的活性,在低浓度(1mmol/L)下,Cu~(2+)、Ba~(2+)、Na~+、Zn~(2+)、Ag~+、Sr~(3+)、K+对琼胶酶活性具有明显抑制作用。琼胶酶的动力学参数K_m和V_(max)分别为0.538mg/ml和6.33μmol/(L·min),对琼胶底物具有高度专一性,降解产物主要为新琼四糖和新琼六糖。     

乌头碱对神经肌肉接头电活动和神经干复合电位作用的观察

利用大白鼠离体膈神经膈肌标本和猫在体腓肠神经与腓肠内侧肌神经观察了乌头碱对神经肌肉接头电活动和神经千复合电位的作用,结果如下:1.大白鼠神经肌肉标本在乌头碱(1.7×10~5)作用下,20分钟阻遏间接刺激引起的肌肉收缩,此时肌纤维静息膜电位无显著变化,在神经干上通常仍可记到复合电位。这种阻遏的可逆性很差。标本对乌头碱的最早反应是增大收缩幅度,出现自发活动,这种作用是通过接头的。2.新斯的明对乌头碱阻遏接头传递无解除作用,但高钙(8mM)却能有效地恢复标本对间接刺激的反应。用小鸡颈二腹肌测定乙酰胆碱的敏感性表明,在乌头碱阻遏神经肌肉间传递后,肌肉乙酰胆碱敏感性有所降低,但并不消失。3.乌头碱在使终板电位消失前不显著改变其时程与振幅。小终板电位在乌头碱作用下先是增加发放频率,最后完全消失。终板电位量子含量的变化是在显著下降前可以看到短时稍有上升。4.1×10~(-4)浓度乌头碱可完全取消猫在体腓肠神经与腓肠内侧肌神经复合电位各成份,冲洗以除去药物后只有少部 A 类纤维可重新传导冲动。1×10~(-5)乌头碱的作用是使复合电位各成份的振幅下降,传导时间延长。乌头碱对复合电位各成份的持续时间无影响。本文结果说明,乌头碱对神经肌肉系统的作用首先是阻遏兴奋在神经末梢的传导,高浓     

锂对淡水鲇Parasilurus asotus坛形电感受器活性的阻断作用

在后侧线神经上记录了单单元传入活动并在受此神经支配的感官开口处用测试溶液替换正常溶液,结果是:1.Li~ (>50mM)首先引起反应的激发极性逆转,随后反应被阻断.作为对照,Na~ 仅引起逆转而无阻断作用.2.Li~ 降低自发放电率并使脉冲排列更趋规则,放电率降至最低值所需时间与阻断反应所需时间相同.3.EGTA溶液阻断了反应并抑制自发活动,其结果与Li~ 的作用类似.4.Ca~(2 )可掩盖Li~ 的阻断作用,含Ca~(2 )的Li~ 溶液可引起逆转但无阻断现象出现.已知多种细胞内的三磷酸肌醇可调控细胞内Ca~(2 )水平.考虑到Li对磷酸肌醇酶的抑制作用,本文结果表明磷脂酰肌醇的代谢产物可能参与了鲇鱼电感受器的传感过程.     

多变鱼腥藻锰和放氧的研究

用EPR法测定藻中游离Mn~(2+)量.当降低培养液中锰浓度或用6μM EDTA处理,使藻中游离Mn~(2+)量下降时,放氧活性不变.0.8MTros(pH8.5)使60%的结合锰释放出时,放氧活性完全被抑制.1mM NH_2OH·HCI已抑制放氧.≥20mM NH_2OH·HCI引起结合锰的释放.经Tris和NH_2OH·HCl处理后,藻的荧光发射光谱上F_(686)下降,F_(695)和 F_(730)上升.本文对鱼腥藻的锰库和放氧进行了初步讨论.     

海马脑片锥体神经元钙离子通道的盲法膜片钳研究

目的和方法 :采用大鼠海马脑片盲法膜片钳全细胞记录技术研究CA1区锥体神经元电压门控性Ca2 通道的动力学特征。结果 :大鼠海马脑片CA1区锥体神经元电压门控性Ca2 通道电流具有如下特点 :①激活的阈电位偏低 ,为 (- 4 9.3± 8.6 )mV ,范围为 - 6 5～ - 30mV(n =2 3)。②衰减时间常数τ值较大 ,且变化范围大 (10 0～ 70 0ms) (n =12 ) ,并且衰减具有Ca2 电流幅值的依赖性 ,③稳态失活呈现电压依赖性 ,半失活电压为 (- 5 5 .4± 9.7)mV ,斜率因子为 (5 .3± 0 .9)mV(n =10 )。④当细胞外Ca2 浓度为 2 .5mmol/L时 ,Ca2 通道的反转电位为 (5 5±13)mV(n =10 )。⑤尾电流成分较为单一 ,不表现电压依赖性。另外 ,Ca2 电流对戊脉胺及双氢吡啶类化合物硝苯地平均不敏感。结论 :根据上述Ca2 电流特征 ,海马脑片CA1区锥体神经元上的Ca2 通道主要以N型为主     

Modulating action of barium and other ions on the sensory activity of frog labyrinth

The effect of Ba2+, TEA, 4-AP and CoCl2 on the EPSP and spike discharges recorded from single fibres of the posterior nerve in the isolated frog labyrinth has been investigated. In Ca-free solution Ba2+ preserved, at low concentration (0.3 mM), the resting activity and at higher levels (up to 6 mM) it resulted in a pronounced facilitation of the EPSP and spike discharges. Facilitation increased on increasing Ba2+ concentration up to 4-5 mM and it was more evident in those units exhibiting a low resting spike firing. The effect of Ba2+ (1 mM) was completely antagonized by 10 mM Ca2+ X CoCl2 (3 mM) suppressed the resting rate at the normal external Ca2+ concentration; the Co2+ block was partially relieved by 1.8 mM Ba2+ X TEA (20 mM) evoked a clear-cut increase in the EPSP and spike discharges which, however, was less consistent than that produced by Ba2+. By comparing the effect of TEA on the spike frequency with that obtained at different Ba2+ levels, the Ba2+ capacity to carry the Ca2+ current was dissected. Such an effect is dose-dependent and it is more evident in low-frequency units. Conversely, 4-AP did not affect the resting discharge frequency. These results indicate that either the Ca2+ or the Ba2+ current sustain the transmitter release at the cyto-neural junction. The effect of TEA suggests that the Ca2+-dependent K+ current may play an important role in supporting the neurosecretory process by controlling the membrane potential of the hair cells.     

Purification and characterization of a Ca(2+)-activated thiol protease from Drosophila melanogaster.

A Ca(2+)-activated thiol protease was purified from Drosophila melanogaster. The procedure involves Phenyl-Sepharose, Reactive Red-Agarose and Q-Sepharose fast flow (or MonoQ) chromatographic steps. The enzyme eluting from Q-Sepharose fast flow seems to be homogeneous as judged by silver staining on SDS-PAGE: it consists of a single polypeptide chain of M(r),app = 94K and pI = 5.46. The proteolytic activity of the purified enzyme is absolutely Ca(2+)-dependent, characterized by 0.6 mM free Ca2+ at half-maximal activity. Ca2+ ions cannot be replaced effectively by the divalent cations Mg2+, Mn2+, Zn2+, Ba2+, and Cd2+. The enzyme shows the inhibitor pattern of thiol proteases. Human recombinant calpastatin (domain I) completely inhibits the enzyme at a nearly 1:1 molar ratio. Several of these properties resemble those of vertebrate calpain II. However, various attempts to detect a small subunit of M(r) approximately 30K, common with vertebrate calpains, remained unsuccessful. We suggest that the Drosophila enzyme is a novel calpain II-like protease.     

The effect of divalent cations on bovine spermatozoal adenylate cyclase activity.

The effect of divalent cations on bovine sperm adenylate cyclase activity was studied. Mn2+, Co2+, Cd2+, Zn2+, Mg2+ and Ca2+ were found to satisfy the divalent cation requirement for catalysis of the bovine sperm adenylate cyclase. These divalent cations in excess of the amount necessary for the formation of the metal-ATP substrate complex were found to stimulate the enzyme activity to various degrees. The magnitude of stimulation at saturating concentrations of the divalent cations was strikingly greater with M2+ than with either Ca2+, Mg2+, Zn2+, Cd2+ or Co2+. The apparent Km was lowest for Zm2+ (0.1 - 0.2 mM) than for any of the other divalent cations tested (1.2 - 2.3 mM). The enzyme stimulation by Mn2+ was decreased by the simultaneous addition of Co2+, Cd2+, Ni2+ and particularly Zn2+ and Cu2+. The antagonism between Mn2+ and Cu2+ or Zn2+ appeared to have both competitive and non-competitive features. The inhibitory effect of Cu2+ on Mn2+-stimulated adenylate cyclase activity was prevented by 2,3-dimercaptopropanol, but not by dithiothreitol, L-ergothioneine, EDTA, EGTA or D-penicillamine. Ca2+ at concentrations of 1-5 mM was found to act synergistically with Mg2+, Zn2+, Co2+ and Mn2+ in stimulating sperm adenylate cyclase activity. The Ca2+ augmentation of the stimulatory effect of Zn2+, Co2+, Mg2+ and Mn2+ appeared to be specific.     

The effect of calcium channel blockers on blood platelet function, especially calcium uptake

The effects of organic and inorganic calcium antagonists on washed platelets from rat and human have been studied. Platelet aggregation was assessed by turbidimetry. Endogenous serotonin release was measured on the same sample by means of electrochemically treated carbon fiber electrodes. The organic calcium antagonist, nitrendipine, and the inorganic calcium channel blockers (Co2+, Mn2+, Cd2+, La3+) drastically inhibited rat and human platelet aggregation induced by thrombin, ADP or adrenaline in the presence of 0.32 mM Ca2+. In our conditions, the thrombin-induced release of endogenous serotonin was found to be external Ca2+-dependent and completely inhibited by 20 microM nitrendipine or 1 mM Cd2+. In addition, Ba2+ or Sr2+ ions can be substituted for Ca2+ to bring about platelet aggregation as well as endogenous serotonin secretion. In Ba2+ or Sr2+-containing media, rat platelet aggregation and/or serotonin secretion can be inhibited by either nitrendipine or Cd2+. Finally, we have also studied the thrombin- and external Ca2+-dependence of radiolabeled calcium uptake by rat platelets. We found that the thrombin-induced 45Ca uptake was inhibited by either 18 microM nitrendipine or 1 mM Cd2+. These results provide strong evidence for the existence of an influx of divalent cations (Ca2+, Sr2+, Ba2+) triggering platelet function. They also suggest, although they do not prove, that the translocation of these cations occurs through an agonist-operated channel as proposed by Hallam and Rink (FEBS Lett. 186 (1986) 175-179).     

Interconversion of androgen receptor forms by divalent cations and 8 S androgen receptor-promoting factor. Effects of Zn2+, Cd2+, Ca2+, and Mg2+

The effects of divalent cations (Zn2+, Cd2+, Ca2+, Mg2+) on the cytosol androgen receptor were determined by sedimentation into sucrose gradients. At low ionic strength (25 mM KCl, 50 mM Tris, pH 7.4), Zn2+ (200 microM total, which calculates to 130 nM free Zn2+ in 10 mM mercaptoethanol) causes a shift in the sedimentation coefficient of the rat Dunning prostate tumor (R3327H) cytosol receptor and rat ventral prostate cytosol receptor from 7.5 +/- 0.3 S to 8.6 +/- 0.3 S. Zn2+ stabilizes the 8.6 S receptor form in salt concentrations up to 0.15 M KCl in 50 mM Tris, pH 7.2. In low ionic strength gradients containing Ca2+ (greater than or equal to 200 microM) or Mg2+ (greater than or equal to 1 mM), the receptor sediments as 4.7 +/- 0.3 S. The dissociating effects of Ca2+ and Mg2+ can be fully reversed by sedimentation into gradients containing Zn2+ (200 microM total) or Cd2+ (10 microM total). In the presence of Zn2+ (200 microM total), Ca2+ (10 microM to 3 mM) converts the receptor to an intermediate form with sedimentation coefficient 6.2 +/- 0.2 S, Stokes radius 73 A, and apparent Mr approximately 203,000. The potentiating effect of Zn2+ on formation of the 8.6 S receptor (in the absence of Ca2+) and the 6.2 S receptor (in the presence of Ca2+) requires both the 4.5 S receptor and the 8 S androgen receptor-promoting factor. Sodium molybdate stabilizes the untransformed cytosol receptor but, unlike Zn2+, does not promote reconstitution of the 8.6 S receptor from its partially purified components. These results indicate that divalent cations alter the molecular size of the androgen receptor in vitro and thus may have a role in altering the state of transformation of the receptor.     

Mechanisms of the Ba2+-induced contraction in smooth muscle cells of the rabbit mesenteric artery

The mechanism of the Ba2+-induced contraction was investigated using intact and saponin-treated skinned smooth muscle (skinned muscle) strips of the rabbit mesenteric artery. After depletion of Ca2+ stored in the caffeine-sensitive site, greater than 0.65 mM Ba2+ evoked contraction in muscle strips depolarized with 128 mM K+ in Ca2+-free solution in a dose-dependent fashion, and the ED50 values for Ca2+ and Ba2+ were 0.5 mM and 1.2 mM in intact muscle strips, respectively. Nisoldipine (10 nM) blocked the contraction evoked by high K+ or 10 microM norepinephrine (NE) in the presence of 2.6 mM Ba2+, but did not block the contraction evoked in the presence of 2.6 mM Ca2+. These results may indicate that Ba2+ permeates the voltage-dependent Ca2+ channel. In skinned muscle strips, the ED50 values for Ca2+ and Ba2+ were 0.34 and 90 microM, respectively, as estimated from the pCa- and pBa-tension relationships. Calmodulin enhanced and trifluoperazine inhibited the Ba2+- and Ca2+-induced contractions. After the application of Ba2+ or Ca2+ with ATP gamma S in rigor solution, myosin light chain (MLC) was irreversibly thiophosphorylated, as estimated from the Ba2+- or Ca2+-independent contraction. Furthermore, both divalent cations phosphorylated MLC, as measured using two-dimensional gel electrophoresis, to the extent expected from the amplitudes of the contraction evoked by these cations. Thus, Ba2+ is capable of activating the contractile proteins as Ca2+ does. The amount of Ca2+ or Ba2+ stored in cells was estimated from the caffeine response evoked in Ca2+-free solution in intact and skinned muscle strips. After the application of 0.3 microM Ca2+ or 0.1 mM Ba2+ for 60 s to skinned muscle strips after the depletion of Ca2+ stored in cells, caffeine produced a contraction only upon pretreatment with Ca2+ but not with Ba2+. When Ba2+ was applied successively just after the application of Ca2+, the subsequently evoked caffeine-induced contraction was much smaller than that evoked by pretreatment with Ca2+ alone. The above results indicate that Ba2+ permeates the voltage-dependent Ca2+ channel but may not permeate the receptor-operated Ca2+ channel, it releases Ca2+ from store sites but is not accumulated into the store site, and it directly activates the contractile proteins via formation of a Ba2+-calmodulin complex.     

Differential effects of Ba2+, Sr2+, and Ca2+ on stimulation-induced changes in transmitter release at the frog neuromuscular junction

Endplate potentials (EPP) were recorded from the frog sartorius neuromuscular junction under conditions of low quantal content to study the effect of Ba2+, Sr2+, and Ca2+ on the changes in evoked transmitter release that occur during and after repetitive stimulation. The addition of 0.1-1 mM Ba2+ or Sr2+ to the Ca2+-containing bathing solution, or the replacement of Ca2+ with 0.8-1.4 mM Sr2+, led to a greater increase in EPP amplitudes during and immediately after repetitive stimulation. These changes in release were analyzed in terms of the four apparent components of increased transmitter release that have previously been distinguished on the basis of their kinetic properties. The Ba2+-induced increase in EPP amplitudes was associated with an increase in the magnitude but not the time constant of decay of augmentation. Ba2+ had little effect on potentiation or the first and second components of facilitation. The Sr2+-induced increase in EPP amplitudes was associated with an increase in the magnitude and the time constant of decay of the second component of facilitation. Sr2+ had little effect on potentiation, augmentation, or the first component of facilitation. The selective effects of Ba2+ on augmentation and of Sr2+ on the second component of facilitation were reversible and could be obtained in the presence of the other ion. The addition of 0.1-0.3 mM Ca2+ to the bathing solution had little effect on potentiation, augmentation, or the two components of facilitation. These results provide pharmacological support for the proposal that there are four different components of increased transmitter release associated with repetitive stimulation and suggest that the underlying factors in the nerve terminal that give rise to these components can act somewhat independently of one another.     

Cation and sequence effects on stability of intermolecular pyrimidine-purine-purine triplex.

A differential effect is found of various bivalent cations (Ba2+, Ca2+, Mg2+, Cd2+, Co2+, Mn2+, Ni2+, Zn2+ and Hg2+) on stability of intermolecular Py-Pu-Pu triplex with different sequence of base triads. Ca2+, Mg2+, Cd2+, Co2+, Mn2+, Ni2+ and Zn2+ do stabilize the d(C)n d(G)n d(G)n triplex whereas Ba2+ and Hg2+ do not. Ba2+, Ca2+, Mg2+ and Hg2+ destabilize the d(TC)n d(GA)n d(AG)n triplex whereas Cd2+, Co2+, Mn2+, Ni2+ and Zn2+ stabilize it. The complexes we observe are rather stable because they do not dissociate during time of gel electrophoresis in the co-migration experiments. Chemical probing experiments with dimethyl sulfate as a probe indicate that an arbitrary homopurine-homopyrimidine sequence forms triplex with corresponding purine oligonucleotide in the presence of Mn2+ or Zn2+, but not Mg2+. In the complex the purine oligonucleotide has antiparallel orientation with respect to the purine strand of the duplex. Specifically, we have shown the formation of the Py-Pu-Pu triplex in a fragment of human papilloma virus HPV-16 in the presence of Mn2+.     

Divalent cations and nifedipine action on Ca(2+) transport into myometrial plasmalemma vesicles

While using 45Ca2+ on the model of "outside-out configuration" vesicules of the myometrium cells sarcolemma an investigation of Cd2+, Zn2+, Co2+ and niphedipin on Ca2+ transport into the vesicules in the conditions of protons gradient transmembrane dissipation has been conducted. The above listed substances blocking effect corresponds to their physicochemical properties. Cadmium and zinc ions are considerably more effective in suppressing Ca2+ transport into the vesicules under the dissipation of delta pH on the membrane if compare with the case of delta pH = 0. In the case of niphedipin inhibiting action an opposite result is observed. The hypothesis has been made, that dissipation of delta pH on the sarcolemma is capable to strengthen the transmembrane Ca2+ transport by means of changing the channel structures conformation.     

Blockade of current through single calcium channels by Cd2+, Mg2+, and Ca2+. Voltage and concentration dependence of calcium entry into the pore

We studied the blocking actions of external Ca2+, Mg2+, Ca2+, and other multivalent ions on single Ca channel currents in cell-attached patch recordings from guinea pig ventricular cells. External Cd or Mg ions chopped long-lasting unitary Ba currents promoted by the Ca agonist Bay K 8644 into bursts of brief openings. The bursts appear to arise from discrete blocking and unblocking transitions. A simple reaction between a blocking ion and an open channel was suggested by the kinetics of the bursts: open and closed times within a burst were exponentially distributed, the blocking rate varied linearly with the concentration of blocking ion, and the unblocking rate was more or less independent of the blocker concentration. Other kinetic features suggested that both Cd2+ and Mg2+ lodge within the pore. The unblocking rate was speeded by membrane hyperpolarization or by raising the Ba concentration, as if blocking ions were swept into the myoplasm by the applied electric field or by repulsive interaction with Ba2+. Ca ions reduced the amplitude of unitary Ba currents (50% inhibition at approximately 10 mM [Ca]o with 50 mM [Ba]o) without detectable flicker, presumably because Ca ions exit the pore very rapidly following Ba entry. However, Ca2+ entry and exit rates could be resolved when micromolar Ca blocked unitary Li+ fluxes through the Ca channel. The blocking rate was essentially voltage independent, but varied linearly with Ca concentration (rate coefficient, 4.5 X 10(8) M-1s-1); evidently, the initial Ca2+-pore interaction is outside the membrane field and much faster than the overall process of Ca ion transfer. The unblocking rate did not vary with [Ca]o, but increased steeply with membrane hyperpolarization, as if blocking Ca ions were driven into the cell. We suggest that Ca is both an effective permeator and a potent blocker because it dehydrates rapidly (unlike Mg2+) and binds to the pore with appropriate affinity (unlike Cd2+). There appears to be no sharp dichotomy between "permeators" and "blockers," only quantitative differences in how quickly ions enter and leave the pore.