Cellular and subcellular heterogeneity of [Ca2+]i in single heart cells revealed by fura-2

Digital imaging of calcium indicator signals (fura-2 fluorescence) from single cardiac cells has revealed different subcellular patterns of cytoplasmic calcium ion concentration ([Ca2+]i) that are associated with different types of cellular appearance and behavior. In any population of enzymatically isolated rat heart cells, there are mechanically quiescent cells in which [Ca2+]i is spatially uniform, constant over time, and relatively low; spontaneously contracting cells, which have an increased [Ca2+]i, but in which the spatial uniformity of [Ca2+]i is interrupted periodically by spontaneous propagating waves of high [Ca2+]i; and cells that are hypercontracted (rounded up) and that have higher levels of [Ca2+]i than the other two types. The observed cellular and subcellular heterogeneity of [Ca2+]i in isolated cells indicates that experiments performed on suspensions of cells should be interpreted with caution. The spontaneous [Ca2+]i fluctuations previously observed without spatial resolution in multicellular preparations may actually be inhomogeneous at the subcellular level.     

Activation of salivary secretion: coupling of cell volume and [Ca2+]i in single cells

High-resolution differential interference contrast microscopy and digital imaging of the fluorescent calcium indicator dye fura-2 were performed simultaneously in single rat salivary gland acinar cells to examine the effects of muscarinic stimulation on cell volume and cytoplasmic calcium concentration ([Ca2+]i). Agonist stimulation of fluid secretion is initially associated with a rapid tenfold increase in [Ca2+]i as well as a substantial cell shrinkage. Subsequent changes of cell volume in the continued presence of agonist are tightly coupled to dynamic levels of [Ca2+]i, even during [Ca2+]i oscillations. Experiments with Ca2+ chelators and ionophores showed that physiological elevations of [Ca2+]i are necessary and sufficient to cause changes in cell volume. The relation between [Ca2+]i and cell volume suggests that the latter reflects the secretory state of the acinar cell. Agonist-induced changes in [Ca2+]i, by modulating specific ion permeabilities, result in solute movement into or out of the cell. The resultant cell volume changes may be important in modulating salivary secretion.     

Regulation of phagocytosis and [Ca2+]i flux by distinct regions of an Fc receptor.

The binding of multivalent immunoglobulin G complexes to Fc receptors (Fc gamma Rs) on macrophages activates multiple immune functions. A murine macrophage cell line, but not a fibroblast cell line, that was transfected with human Fc gamma RIIA mediated phagocytosis and an intracellular Ca2+ concentration ([Ca2+]i) flux upon cross-linking of human Fc gamma RIIA. Transfected macrophages that expressed a truncated receptor lacking 17 carboxy-terminal amino acids phagocytosed small antibody complexes. However, only wild-type transfectants phagocytosed labeled erythrocytes and fluxed [Ca2+]i. Thus, the cytoplasmic domain of human Fc gamma RIIA contains distinct functional regions.     

Ca2+ and Ca2+-activated K+ currents in mammalian gastric smooth muscle cells

Inward movement of calcium through voltage-dependent channels in muscle is thought to initiate the action potential and trigger contraction. Calcium-activated potassium channels carry large outward potassium currents that may be responsible for membrane repolarization. Calcium and calcium-activated potassium currents were identified in enzymatically isolated mammalian gastric myocytes. These currents were blocked by cadmium and nifedipine but were not substantially affected by diltiazem or D600. No evidence for a tetrodotoxin-sensitive sodium current or an inwardly rectifying potassium current was found.     

Atrotoxin: a specific agonist for calcium currents in heart

A specific label for voltage-dependent calcium channels is essential for the isolation and purification of the membrane protein that constitutes the calcium channel and for a better understanding of its function. A fraction of Crotalus atrox that increases voltage-dependent calcium currents in single, dispersed guinea pig ventricular cells was isolated. In the doses used, neither sodium nor potassium currents were changed. The fraction was active in the absence of detectable phospholipase or protease activity, and the active component, designated atrotoxin, produced its effect rapidly and reversibly. The effect was produced by extracellular but not intracellular application of the agent. The increase in Ca2+ current was blocked by the Ca2+ channel blockers cobalt and nitrendipine. The active fraction completely blocked specific [3H]nitrendipine binding to guinea pig ventricular membrane preparations. The inhibition of nitrendipine binding by atrotoxin was apparently via an allosteric mechanism. Thus atrotoxin was shown to bind to the Ca2+ channel and to act as a specific Ca2+ channel agonist.     

Reconstitution of Ca2+-regulated membrane fusion by synaptotagmin and SNAREs

We investigated the effect of synaptotagmin I on membrane fusion mediated by neuronal SNARE proteins, SNAP-25, syntaxin, and synaptobrevin, which were reconstituted into vesicles. In the presence of Ca2+, the cytoplasmic domain of synaptotagmin I (syt) strongly stimulated membrane fusion when synaptobrevin densities were similar to those found in native synaptic vesicles. The Ca2+ dependence of syt-stimulated fusion was modulated by changes in lipid composition of the vesicles and by a truncation that mimics cleavage of SNAP-25 by botulinum neurotoxin A. Stimulation of fusion was abolished by disrupting the Ca2+-binding activity, or by severing the tandem C2 domains, of syt. Thus, syt and SNAREs are likely to represent the minimal protein complement for Ca2+-triggered exocytosis.     

Carcinogen and microsomal membrane interactions: changes in membrane density and ability to bind nucleic acids

DNA and synthetic copolymer polyribocytidylic-polyriboguanylic acid bind to microsomal membrane. The nucleic acid-membrane complex may be detected by centrifugation in cesium chloride density gradients. The density of the nucleic acid-membrane complex and, in certain cases, the amount of nucleic acid associated with the membrane was changed in the presence of various carcinogenic chemicals.     

Muscle postjunctional membrane: changes in chemosensitivity produced by calcium

Increases in the extracellular concentration of calcium ions above 1.8 millimoles per liter caused a reversible decrease in the sensitivity of tmuscle postiunctional membrane to carbamylcholine. A quantitative study of the inhibitory effect of calcium ions on membrane depolarization produced by carbamylcholine indicates that calcium ions compete with carbamylcholine for some common binding sites on the postjunctional membrane. Calcium ions (20 millimoles per liter) caused a neuromuscular block wherein prolonged end plate potentials were produced after nerve stimulation. Calcium ions applied ionophoretically to the postjunctional membrane decreased the amplitude and prolonged the time course of the transient depolarization produced by ionophoretically applied carbamylcholine.     

NaCl胁迫下钙对小麦幼苗保护酶及膜质过氧化的影响

:以抗盐和盐敏感2种不同基因型的冬小麦为试验材料,研究Ca2+对NaCl胁迫下不同生长阶段的小麦幼苗保护酶系统和膜脂过氧化的影响.结果表明,1% NaCl胁迫对小麦幼苗造成了一定程度的氧化伤害.对抗盐品种的氧化伤害晚于敏感品种,且程度轻.适当浓度外源Ca2+处理能够提高NaCl胁迫下小麦幼苗体内超氧化物歧化酶(SOD)、过氧化物酶(POD)的活性,抑制过氧化作用产物丙二醛(MDA)的积累,表明适当浓度的Ca2+处理对提高小麦抗盐能力是有益的,缓解盐害的作用与基因型、钙浓度和胁迫时间等因素有关.     

淡水鱼肌球蛋白Ca2+-ATPase热稳定性的季节变化

对来自春季（4月份）、夏季（8月份）、秋季（11月份）和冬季（1月份）的草鱼（Ctenopharyngodon idellus）和鲢（Hypophthalmichthys molitrix）骨骼肌肌球蛋白进行提取和纯化,通过测定肌球蛋白在不同热变性温度下Ca^2＋-ATPase的变性速率常数（KD）,考察了两种淡水鱼肌球蛋白热稳定性的季节变化.研究结果表明,两种鱼肌球蛋白Ca^2＋-ATPase的活性均显示夏季鱼＞春季鱼＞秋季鱼＞冬季鱼.在35 ℃、40 ℃、45 ℃和50 ℃的热变性温度下,测定了两种鱼的肌球蛋白Ca^2＋-ATPase变性速率常数（KD）,证明了肌球蛋白的热变性符合一级反应速度方程.经35 ℃热变性的肌球蛋白的KD值显示了冬季鱼约为夏季鱼的4倍,而秋季鱼和春季鱼介于冬季鱼和夏季鱼之间,且秋季鱼的KD值略低于冬季鱼,而春季鱼的KD值略高于夏季鱼.在40 ℃、45 ℃和50 ℃的热变性温度下,对各种肌球蛋白测得的KD值的变化趋势与上述结果相似.本研究证明了淡水鱼肌球蛋白的热稳定性夏季鱼明显优于冬季鱼,而春季鱼和秋季鱼分别与夏季鱼和冬季鱼的相似.     

淡水鱼肌球蛋白Ca2+-ATPase热稳定性的季节变化

对来自春季（4月份）、夏季（8月份）、秋季（11月份）和冬季（1月份）的草鱼（Ctenopharyngodon idellus）和鲢（Hypophthalmichthys molitrix）骨骼肌肌球蛋白进行提取和纯化,通过测定肌球蛋白在不同热变性温度下Ca^2＋-ATPase的变性速率常数（KD）,考察了两种淡水鱼肌球蛋白热稳定性的季节变化.研究结果表明,两种鱼肌球蛋白Ca^2＋-ATPase的活性均显示夏季鱼＞春季鱼＞秋季鱼＞冬季鱼.在35 ℃、40 ℃、45 ℃和50 ℃的热变性温度下,测定了两种鱼的肌球蛋白Ca^2＋-ATPase变性速率常数（KD）,证明了肌球蛋白的热变性符合一级反应速度方程.经35 ℃热变性的肌球蛋白的KD值显示了冬季鱼约为夏季鱼的4倍,而秋季鱼和春季鱼介于冬季鱼和夏季鱼之间,且秋季鱼的KD值略低于冬季鱼,而春季鱼的KD值略高于夏季鱼.在40 ℃、45 ℃和50 ℃的热变性温度下,对各种肌球蛋白测得的KD值的变化趋势与上述结果相似.本研究证明了淡水鱼肌球蛋白的热稳定性夏季鱼明显优于冬季鱼,而春季鱼和秋季鱼分别与夏季鱼和冬季鱼的相似.     

Plasma membrane: substructural changes correlated with electrical resistance and pinocytosis

Inducers of pinocytosis in amoeba cause as much as a 50-fold decrease in the electrical resistance of the plasma membrane prior to the formation of the typical tunnels and vacuoles. In this state the thickness of the electron-transparent core or lamella of the unit membrane is at least twice as thick as that of the control. The changes in structure and resistance as well as the induction of pinocytosis are dependent on the initial external concentration of calcium. These changes are rapidly reversed when the concentration of calcium in the external medium is increased.     

Permeability of a nuclear membrane: changes during normal development and changes induced by growth hormone

The ion permeability of the nuclear membrane envelope of salivary gland cells (of the midge Chironomus thummi) undergoes changes during development. Following the early fourth instar stage, permeability falls to about one-fifth of its base value over a period of 3 to 5 days of development of the animal, and then rises again over the next 2 to 4 days. The falling phase of the change can be reproduced within 1 hour by injections of the growth hormone ecdyson.     

Cytoprotective role of Ca2+- activated K+ channels in the cardiac inner mitochondrial membrane

Ion channels on the mitochondrial inner membrane influence cell function in specific ways that can be detrimental or beneficial to cell survival. At least one type of potassium (K+) channel, the mitochondrial adenosine triphosphate-sensitive K+ channel (mitoKATP), is an important effector of protection against necrotic and apoptotic cell injury after ischemia. Here another channel with properties similar to the surface membrane calcium-activated K+ channel was found on the mitochondrial inner membrane (mitoKCa) of guinea pig ventricular cells. MitoKCa significantly contributed to mitochondrial K+ uptake of the myocyte, and an opener of mitoKCa protected hearts against infarction.     

Evidence for the prion hypothesis: induction of the yeast [PSI+] factor by in vitro- converted Sup35 protein

Starting with purified, bacterially produced protein, we have created a [PSI(+)]-inducing agent based on an altered (prion) conformation of the yeast Sup35 protein. After converting Sup35p to its prion conformation in vitro, we introduced it into the cytoplasm of living yeast using a liposome transformation protocol. Introduction of substoichiometric quantities of converted Sup35p greatly increased the rate of appearance of the well-characterized epigenetic factor [PSI+], which results from self-propagating aggregates of cellular Sup35p. Thus, as predicted by the prion hypothesis, proteins can act as infectious agents by causing self-propagating conformational changes.     

Ca(2+)-induced Ca2+ release in sea urchin egg homogenates: modulation by cyclic ADP-ribose

Calcium-induced calcium release (CICR) may function widely in calcium-mediated cell signaling, but has been most thoroughly characterized in muscle cells. In a homogenate of sea urchin eggs, which display transients in the intracellular free calcium concentration ([Ca2+]i) during fertilization and anaphase, addition of Ca2+ triggered CICR. Ca2+ release was also induced by the CICR modulators ryanodine and caffeine. Responses to both Ca2+ and CICR modulators (but not Ca2+ release mediated by inositol 1,4,5-trisphosphate) were inhibited by procaine and ruthenium red, inhibitors of CICR. Intact eggs also displayed transients of [Ca2+]i when microinjected with ryanodine. Cyclic ADP-ribose, a metabolite with potent Ca(2+)-releasing properties, appears to act by way of the CICR mechanism and may thus be an endogenous modulator of CICR. A CICR mechanism is present in these nonmuscle cells as is assumed in various models of intracellular Ca2+ wave propagation.     

Nodal quasiparticles and antinodal charge ordering in Ca2-xNaxCuO2Cl2

Understanding the role of competing states in the cuprates is essential for developing a theory for high-temperature superconductivity. We report angle-resolved photoemission spectroscopy experiments which probe the 4a0 x 4a0 charge-ordered state discovered by scanning tunneling microscopy in the lightly doped cuprate superconductor Ca2-xNaxCuO2Cl2. Our measurements reveal a marked dichotomy between the real- and momentum-space probes, for which charge ordering is emphasized in the tunneling measurements and photoemission is most sensitive to excitations near the node of the d-wave superconducting gap. These results emphasize the importance of momentum anisotropy in determining the complex electronic properties of the cuprates and places strong constraints on theoretical models of the charge-ordered state.     

肉鸡腹水综合征患鸡肺脏组织钙沉积和Ca2＋-ATPase活性变化

肺动脉收缩和重塑在肉鸡AS发生过程中起着重要作用.本实验经采用右心导管法研究发现AS患鸡PASP 、PADP和mPAP极显著高于对照组(P <0.01).同时,采用焦锑酸钾沉淀法、电镜酶细胞化学法研究AS患鸡肺脏组织Ca2+和Ca2+-ATPase活性变化,发现AS患鸡肺脏组织中钙沉积量显著增多,且Ca2+-ATPase的电子密度颗粒显著减少或缺失.表明AS患鸡具有明显的肺动脉高压,其可能与肺脏组织,特别是肺动脉平滑肌细胞钙处理能力异常导致的钙离子浓度升高和肌浆网Ca2+-ATPase酶活性降低有关.     

不同木薯品种气体交换特性及光合酶活性的变化

研究了不同木薯品种生长发育过程中,田间气体交换特性、光合酶活性的变化规律及它们之间的相互关系。结果表明,木薯在衰老过程中,净光合速率急剧下降,光合作用减弱与其内部光合机构及其光合酶变化密切相关,在整个生长发育期,气孔导度、胞间CO2浓度、净光合速率及蒸腾速率之间均呈极显著的正相关关系;Ca2+-ATP酶活性、光合磷酸化活力与净光合速率呈显著的正相关关系,乙醇酸氧化酶活性与净光合速率呈极显著的正相关关系。