力竭性运动对大鼠骨骼肌Ca2+含量的影响

观察大鼠力竭性运动后即刻组及三小时后组(以下简称3h组)骨骼肌内Ca2+的变化情况,探讨骨骼肌Ca2+在力竭性运动后和恢复中的变化及其机制.力竭性运动后取大鼠骨骼肌,采用原子吸收分光光度计测其Ca2+浓度.运动后即刻组骨骼肌中Ca2+浓度(2068.33±752.02μg/g)比对照组(615.31±546.03μg/g)有极显著增高(P＜0.001),3 h组(734.58±258.70μg/g)与对照组无明显差异,即刻组和3 h组有极显著性差异(P＜0.001)力竭性运动即刻,骨骼肌中Ca2+浓度显著增高,恢复期内Ca2+浓度逐渐恢复.     

大黄酸对IL-2诱发T淋巴细胞增殖和细胞[Ca2+]i变化的作用

研究大黄酸对IL-2引起大鼠T淋巴细胞增殖和细胞内[Ca2+]i变化的影响.采用MTT法检测细胞增殖,用单细胞钙成像系统记录胞内游离Ca2+浓度([Ca2+]i).结果表明,大黄酸对IL-2引起的细胞增殖有显著抑制作用,且抑制效果与剂量有关;IL-2引起[Ca2+]i升高,并持续维持高[Ca2+]i水平,[Ca2+]i升高的机制包括胞内Ca2+释放和胞外Ca2+内流;大黄酸显著抑制IL-2引起的[Ca2+]i升高.结果提示大黄酸可抑制T淋巴细胞增殖,作用机制可能与抑制细胞内[Ca2+]i升高相关.     

间歇性低氧及运动对大鼠红细胞膜Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性的影响

Wistar雄性大鼠分为常氧对照组、常氧运动组、低氧对照组、低氧运动组.分别对常氧运动、低氧运动组进行游泳训练,对低氧对照、低氧运动组进行低氧刺激.用导管法测血压和心率,用比色法测红细胞膜Na+-K+-ATP酶Ca2+-Mg2+-ATP酶活性,探讨低氧和训练对红细胞膜Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性的影响.发现低氧刺激降低大鼠红细胞膜Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶的活性,而适宜负荷运动则可提高以上两种酶的活性.结果表明,间歇性低氧运动对大鼠红细胞膜的Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性具有一定的保护作用.     

滑膜细胞[Ca2＋]i变化的理论模型研究

以滑膜细胞内钙离子浓度变化为研究对象,通过分析整合影响滑膜细胞内钙离子浓度变化的模块来建立滑膜细胞[Ca2+]i变化的理论模型.先考虑参与钙调节的质膜上钙泵、内质网上钙泵和渗漏、IP3受体、Na+/K+泵、延迟整流钾通道、TRPV1通道等各个模块在网络中的作用,确定钙调控过程中各模块的数学模型.在此基础上,引入钙缓冲系统的参量,并重点突出[Ca2+]i和膜电位的相互作用,建立起滑膜细胞上胞浆[Ca2+]i变化的动力学模型.最后,调整模型参数,使得模型处于稳定的初始状态,并保证模型参数的取值在合理的范围,给出滑膜细胞响应刺激所对应的[Ca2+]i随时间变化的模拟结果,并简要分析了某些参数对模型的影响.研究结果表明:IP3受体的调控机制在辣椒素刺激滑膜细胞胞浆[Ca2+]i变化的动力学模型中有明显作用;[Ca2+]i和膜电位的相互作用对滑膜细胞钙网络调控至关重要;质膜上钙泵的密度、活性,IP3受体通道密度,TRPV1通道密度等的变化对[Ca2+]i模拟曲线形状有明显影响.     

不同浓度ET—1引起心肌细胞[Ca^2＋]i升高作用的？…

目的:探讨内皮素-1(ET-1)对心肌细胞内游离钙浓度([Ca2+]i)的作用以及不同浓度ET-1引起[Ca2+]i升高作用的量效关系.方法:采用分离的Sprague-Dawley大鼠心室肌细胞,以Fura-2/AM荧光指示剂负载,检测不同浓度ET-1引起[Ca2+]i变化.结果:ET-1引起[Ca2+]i升高呈双相反应,即起始的短暂快速相和随后的持续相.在1×10-9～5×10-7mol/L范围内,随着ET-1浓度的增加,其升高[Ca2+]i的作用亦增强;并且这种作用可被ETA的特异性受体阻断剂BQ123(2×10-6mol/L)所阻断.结论:ET-1升高[Ca2+]i呈剂量依赖关系,其作用具有特异性,并且是通过ETA受体介导的.     

ATP刺激巨噬细胞[Ca2+]i升高和氧自由基增加及大黄酸的抑制作用研究

研究了ATP刺激的大鼠腹腔巨噬细胞[Ca2 ]i升高与氧自由基(ROS)产生的关系和大黄酸抑制作用特征.提取大鼠腹腔巨噬细胞,利用Ca2 探针Fura-2检测单细胞胞内自由Ca2 浓度([Ca2 ]i)变化,同时利用NBT还原反应强度检测同一细胞ROS产生能力.结果发现无ATP刺激的巨噬细胞的ROS产生量较低;1mmol/L ATP刺激巨噬细胞单细胞后诱发[Ca2 ]i显著升高由胞内Ca2 释放和胞外Ca2 内流组成,同时ROS产生增强2倍;胞外无Ca2 条件下ROS产生随着[Ca2 ]i下降而减少.10-5和10-4mol/L浓度大黄酸对1mmol/L ATP刺激巨噬细胞单细胞的[Ca2 ]i升高和ROS产生有剂量依赖性的抑制作用;多细胞的统计分析表明10-5和10-4mol/L浓度大黄酸分别抑制了[Ca2 ]i峰值的49%和84%,同时抑制了ROS产生的59%和81%.因此认为ATP刺激大鼠腹腔巨噬细胞诱发的[Ca2 ]i升高介导了ROS的产生,大黄酸剂量依赖性的抑制ATP刺激的细胞[Ca2 ]i升高和ROS产生能力,并提示大黄酸抑制细胞[Ca2 ]i升高是其抑制ROS产生的重要机制.     

龙葵糖蛋白对MCF -7细胞内[Ca2+]i的影响

研究龙葵糖蛋白对乳腺癌MCF -7细胞内[Ca2+]i的影响.通过MTT法测定龙葵糖蛋白对MCF -7细胞的细胞毒作用;Hoechst33258染色荧光显微镜下观察龙葵糖蛋白作用MCF -7细胞的形态学变化;采用Fluo - 3/AM探针标记,激光共聚焦技术观测龙葵糖蛋白对MCF -7细胞内[Ca2+]i的影响.结果表...     

Zn2+、Ca2+及其相互作用对大蒜根尖生长和细胞分裂的影响

用不同浓度的Zn2+、Ca2+和Zn2++Ca2+溶液处理大蒜鳞茎发现;Ca2+能抑制Zn2+的毒害,明显提高细胞分裂比率,促进根尖生长,降低异常细胞比率.基本趋势为根的长度和细胞分裂比率为Ca2+>Zn2++Ca2+>Zn2+,异常细胞比率为Ca2+>Zn2+>Ca2++Zn2+.     

外源Ca对旱柳Cd毒害的缓解作用

为了探究Cd胁迫对旱柳的毒害以及外源Ca的缓解作用,以一年生旱柳枝条为水培材料,设置对照组、Ca处理组(Ca2+浓度为5 mmol/L)、Cd处理组(Cd2+浓度为50μmol/L)和Ca+Cd处理组(Ca2+和Cd2+浓度分别为5μmol/L和50 mmol/L)4个处理组,研究Cd胁迫环境中的旱柳在添加了外源Ca后光合色素含量、光合生理、保护酶系统和丙二醛含量等的变化.结果表明:①Cd胁迫作用下,旱柳叶片的光合色素含量明显减少,在添加外源Ca后光合色素含量显著增加;②Cd处理会降低旱柳光合作用参数进而影响旱柳的光合作用进程,而Ca+Cd处理组中各项光合作用参数有明显恢复;③Cd处理会诱导保护酶活性的升高并增加MDA含量,外源Ca的添加则会降低保护酶活性和MDA的含量.因此认为,外源Ca的添加可以有效缓解Cd胁迫对旱柳的毒害作用.     

新型Ca2+指示剂STDIn双波长比率法测定血清钙

采用自行设计合成的新型Ca2 指示剂STDIn,建立了一种可在生理条件下测定血清中Ca2 的新方法.指示剂STDIn在pH值为7.2的中性缓冲介质中,可与Ca2 络合形成稳定的络合物,试剂自身的吸收峰位于543nm,与Ca2 络合后紫移至423nm,Δλ=120nm,对比明显.在指示剂浓度为10μmol/L时,采用双波长比率法求得测定Ca2 的线性范围分别为2×10-5～2×10-3mol/L(相关系数r=0.9977)和5×10-3～1×10-1mol/L(相关系数r=0.9968),检出限为6×10-6.该法已用于血清中Ca2 的测定.     

Ca2+-dependent and Ca2+-independent phagocytosis in human neutrophils

The phagocytic function of neutrophils is a crucial element in host defence against invading microorganisms. Two main specific receptor-mediated mechanisms operate in the phagocyte plasma membrane, one recognizing the C3b/bi fragment of complement and the other the Fc domain of immunoglobulin G (ref. 1). There is evidence that phagocytosis mediated by these receptors differs in the number and nature of the intracellular signals generated. However, the mechanisms by which receptor binding is transduced into a signal that generates the formation of the phagocyte pseudopod is not known, although extensive biochemical evidence has allowed the postulate that calcium ion gradients in the peripheral cytoplasm, by interacting with calcium-sensitive contractile proteins, initiate the process of engulfment. Using the high-affinity fluorescent calcium indicator quin2 both to measure and to buffer intracellular calcium ([Ca2+]i), we show here that in human neutrophils two mechanisms of phagocytosis coexist: a [Ca2+]i-dependent and modulated phagocytosis, triggered by activation of the Fc receptor, and a [Ca2+]i-independent mechanism triggered by the activation of the C3b/bl receptors.     

Open structure of the Ca2+ gating ring in the high-conductance Ca2+-activated K+ channel

High-conductance voltage- and Ca(2+)-activated K(+) channels function in many physiological processes that link cell membrane voltage and intracellular Ca(2+) concentration, including neuronal electrical activity, skeletal and smooth muscle contraction, and hair cell tuning. Like other voltage-dependent K(+) channels, Ca(2+)-activated K(+) channels open when the cell membrane depolarizes, but in contrast to other voltage-dependent K(+) channels, they also open when intracellular Ca(2+) concentrations rise. Channel opening by Ca(2+) is made possible by a structure called the gating ring, which is located in the cytoplasm. Recent structural studies have defined the Ca(2+)-free, closed, conformation of the gating ring, but the Ca(2+)-bound, open, conformation is not yet known. Here we present the Ca(2+)-bound conformation of the gating ring. This structure shows how one layer of the gating ring, in response to the binding of Ca(2+), opens like the petals of a flower. The degree to which it opens explains how Ca(2+) binding can open the transmembrane pore. These findings present a molecular basis for Ca(2+) activation of K(+) channels and suggest new possibilities for targeting the gating ring to treat conditions such as asthma and hypertension.     

Calmodulin bifurcates the local Ca2+ signal that modulates P/Q-type Ca2+ channels

Acute modulation of P/Q-type (alpha1A) calcium channels by neuronal activity-dependent changes in intracellular Ca2+ concentration may contribute to short-term synaptic plasticity, potentially enriching the neurocomputational capabilities of the brain. An unconventional mechanism for such channel modulation has been proposed in which calmodulin (CaM) may exert two opposing effects on individual channels, initially promoting ('facilitation') and then inhibiting ('inactivation') channel opening. Here we report that such dual regulation arises from surprising Ca2+-transduction capabilities of CaM. First, although facilitation and inactivation are two competing processes, both require Ca2+-CaM binding to a single 'IQ-like' domain on the carboxy tail of alpha1A; a previously identified 'CBD' CaM-binding site has no detectable role. Second, expression of a CaM mutant with impairment of all four of its Ca2+-binding sites (CaM1234) eliminates both forms of modulation. This result confirms that CaM is the Ca2+ sensor for channel regulation, and indicates that CaM may associate with the channel even before local Ca2+ concentration rises. Finally, the bifunctional capability of CaM arises from bifurcation of Ca2+ signalling by the lobes of CaM: Ca2+ binding to the amino-terminal lobe selectively initiates channel inactivation, whereas Ca2+ sensing by the carboxy-terminal lobe induces facilitation. Such lobe-specific detection provides a compact means to decode local Ca2+ signals in two ways, and to separately initiate distinct actions on a single molecular complex.     

Ca2+ signalling between single L-type Ca2+ channels and ryanodine receptors in heart cells

Ca2+-induced Ca2+ release is a general mechanism that most cells use to amplify Ca2+ signals. In heart cells, this mechanism is operated between voltage-gated L-type Ca2+ channels (LCCs) in the plasma membrane and Ca2+ release channels, commonly known as ryanodine receptors, in the sarcoplasmic reticulum. The Ca2+ influx through LCCs traverses a cleft of roughly 12 nm formed by the cell surface and the sarcoplasmic reticulum membrane, and activates adjacent ryanodine receptors to release Ca2+ in the form of Ca2+ sparks. Here we determine the kinetics, fidelity and stoichiometry of coupling between LCCs and ryanodine receptors. We show that the local Ca2+ signal produced by a single opening of an LCC, named a 'Ca2+ sparklet', can trigger about 4-6 ryanodine receptors to generate a Ca2+ spark. The coupling between LCCs and ryanodine receptors is stochastic, as judged by the exponential distribution of the coupling latency. The fraction of sparklets that successfully triggers a spark is less than unity and declines in a use-dependent manner. This optical analysis of single-channel communication affords a powerful means for elucidating Ca2+-signalling mechanisms at the molecular level.     

Ca2+-activated ATPase and ATP-dependent calmodulin-stimulated Ca2+ transport in islet cell plasma membrane

Calcium is known to play an essential part in the regulation of insulin secretion in the pancreatic beta cell. Calcium influx/efflux studies indicate that glucose promotes an accumulation of calcium by the beta cell. However, interpretation of such data is particularly difficult due to the complex compartmentalization of calcium within the cell. Although indirect evidence using chlorotetracycline suggests that control of calcium homeostasis at the plasma membrane may be central to insulin secretion, the mechanism by which secretagogues influence the handling of calcium remains unknown. Despite its continuous diffusive entry, intracellular calcium is maintained in the submicromolar range by energy-dependent mechanisms. One such process which has been well characterized in erythrocytes is a plasma membrane calcium extrusion pump whose enzymatic basis is a high affinity (Ca+2 + Mg+2)ATPase. A similar mechanism regulated by insulin has recently been identified in adipocyte plasma membranes. We report here the presence of a high affinity (Ca+2 + Mg+2)ATPase and ATP-dependent calmodulin-stimulated calcium transport system in rat pancreatic islet cell plasma membranes.     

钙离子对学习记忆的影响

Ca^2 作为神经信号传递的重要信使，参与学习记忆的神经机制。突触前和突触后细胞内钙离子在长时程突触的可塑性中发挥重要的信息传递作用。研究表明衰老性记忆障碍与中枢神经系统的Ca^2 稳态调节失衡有关。神经细胞内游离钙水平[Ca^2 ]i受多种机制的调节，主要包括Ca^2 的跨膜转运、细胞内钙池摄取与释放Ca^2 等过程；最近的研究表明神经胶质细胞也参与其调节。     

龙柚果实总钙、可溶性Ca2+含量及Ca2+-ATPase活性的变化

研究以柚类品种龙柚为材料,对发育过程中果皮、果肉总钙和可溶性Ca~(2 )含量及Ca~(2 )-ATPase活性进行了测定.结果表明:龙柚花前至花期子房总钙含量相对较低,花后有明显上升;龙柚果皮总钙和可溶性Ca~(2 )均在果实增大期保持持续增长的趋势,此时的果肉总钙呈明显下降趋势,而对应可溶性Ca~(2 )的变化幅度较大;龙柚Ca~(2 )-ATPase活性在花前较低,花期开始上升;而果皮和果肉的可溶性Ca~(2 )含量和Ca~(2 )-ATPase活性在增大期内均有一明显增长的过程或相对较高的水平.     

Intracellular Ca indicator Quin-2 inhibits Ca2+ inflow via Na/Ca exchange in squid axon

Until recently, intracellular free calcium has been amenable to measurement and investigation only in cells large enough to permit either microinjection of a suitable Ca sensor such as a aequorin or arsenazo III or insertion of a Ca-sensitive microelectrode. This constraint on cell size was removed by the development of the fluorescent Ca2+ -sensitive dye Quin-2 and its acetoxymethyl ester, which can be introduced into a wide range of cell types. A major requirement of any intracellular Ca2+ indicator is that it should not disturb intracellular Ca2+ homeostasis and Quin-2 is generally considered to be satisfactory in this respect. We now report that injection of Quin-2 into squid (Loligo forbesi) axons can almost completely abolish one component of Ca2+ entry--intracellular Na+ (Nai)-dependent Ca2+ inflow, which occurs via Na/Ca exchange. Mixtures of Ca and Quin-2 that buffer an ionized Ca2+ at close to physiological concentrations also block Nai-dependent Ca2+ influx but these same mixtures fail to block the extracellular Na+ (Na0)-dependent extrusion of Ca2+, showing that Quin-2 acts specifically on Ca2+ inflow.