H-ATPase in the plasma membrane of Arabidopsis pollen cells is involved in extracellular calmodulin-promoted pollen germination

In this paper, the role of the plasma membrane (PM) H⁺-ATPase in extracellular calmodulin (CaM)-promoted pollen germination and in tube growth of Arabidopsis thaliana was investigated. Pollen germination, pollen tube growth rate, free cytosolic Ca²⁺ concentration ([Ca²⁺]_cyt) and Ca²⁺ channel activity in the PM of pollen cells were measured. In response to fusicoccin or CaM treatment, in vitro pollen germination and pollen tube growth rate, [Ca²⁺]_cyt and activity of a hyperpolarization-activated Ca²⁺-permeable channel increased. Sodium vanadate inhibited the promotion of these parameters by extracellular CaM. The results suggest that H⁺-ATPase may be involved in extracellular CaM-regulated pollen germination and in tube growth by modulation of the hyperpolarization-activated Ca²⁺ channel in the PM of A. thaliana pollen cells.     

Effects of nitrogen ion implantation on lily pollen germination and the distribution of the actin cytoskeleton during pollen germination

The effects of low energy nitrogen ion implantation on lily (Lilium davidii Duch.) pollen germination and the distribution of the actin cytoskeleton during pollen germination have been studied. Preliminary results showed that the ratio of pollen germination increased from (16.0± 1.6)% to (27.0±2.1)% when implanted with nitrogen ions by 100 keV and a dose of 10¹³ ions/cm². Further experiments were performed by staining the actin filaments in pollen with rhodamine-phalloidin and detected by using laser confocol microscopy. After hydration for 10 h, the actin filaments in ion implanted pollen grains tended to form thick bundles oriented in parallel or ring shape at the germinal furrow, indicating that the effect of nitrogen ion implantation on the germination of pollen might be mediated by reorganization of the actin cytoskeleton.     

Action mechanisms of a new erythrocyte-derived depressing factor

To investigate the action mechanisms of a new erythrocyte-derived depressing factor (EDDF), the focus is placed on the effect of EDDF on both cytosolic and nuclear free calcium (Ca²⁺) transportation in vascular smooth muscle cell (VSMC), as well as the apoptosis and cell cycle of VSMC of rats. EDDF has been extracted from human erythrocytes. The changes of Ca²⁺ levels in cytoplasm ([Ca²⁺]_i) and nucleus ([Ca²⁺]_n) have been observed using a laser scanning confocal microscope together with fluo-3/AM as a calcium indicator. Flow cytometric technique was used to study the effect of EDDF on cell cycle and apoptosis of VSMC. [Ca²⁺], and [Ca²⁺]_n were significantly decreased through several different pathways: ( i ) it reduced the Ca²⁺ influx by blocking L-type voltage-dependent calcium channel (L-VDC) and R-type voltage-dependent calcium channel (R-VDC); (ii) it inhibited the Ca²⁺ release from inositol 1, 4, 5-trisphosphate (IP₃) sensitive calcium store; and (iii) activated Ca²⁺-ATPase of sarcoplasmic reticulum (SR) and promoted the transportation of Ca²⁺ from cytoplasm to SR. However, EDDF seemed to have little inhibitory effect on the Ca²⁺ release from ryonodine sensitive calcium pool. It was also found that EDDF (10⁻⁴ g/mL) significantly decreased the proportion of S phase of human umbilical vein (HUV) and inhibited the proliferation of VSMC induced by angiotensin II (Angll, 10⁻⁵ mol/L). The apopotosis did not occur when VSMC was cultured under normal condition. While VSMC apoptosis was induced by Angll (10⁻⁵ mol/L) and EDDF (10⁻⁴ g/mL) seemed to have little effect on it. The inhibitory effect of EDDF on the elevation of [Ca²⁺]_i and [Ca²⁺]_n of VSMC might play an essential role in its action mechanisms and the ways it affects the Ca²⁺ handling of VSMC demonstrate that EDDF was different from other endogenous blood pressure regulators and some known antihypertensive drugs. EDDF could inhibit the proliferation of VSMC, which indicated that it might be beneficial to the prevention and treatment of hypertension and arteriosclerosis.     

Mechanisms of peroxynitrite-induced [Ca2+]i increase in single neuronal cell

The mechanism of peroxynitrite (ONOO⁻)-induced [ca²⁺]_i increase in single MN9D cell (Dopaminergic neuroblastoma cell line) was studied by using Fura-2 microfluorometric technique. The results show that ONOO⁻ caused a rapid increase of [Ca²⁺]_i when ONOO₋ was puffed to the cell. Removing Ca²⁺ from the bath or using calcium channel antagonist (CdCl₂, Nifedipine) greatly inhibited the [Ca²⁺]_i increase induced by ONOO⁻¹, suggesting that the opening of L-Ca²⁺ channel makes a great contribution to the [Ca²⁺]_i increase. The effect of sulfhydryl reductive agent (DTT) on ONOO⁻-induced [Ca²⁺]_i increase suggests that ONOO⁻-activating L-Ca²⁺ channel is partly related to its oxidative speciality.     

Role of Ubisch bodies secreted by tapetum in Ca2+ transprot

The distribution of Ca²⁺ in the anthers of wheat was observed using cytochemical method of potassium antimonite. At the later tetrad stage, Ubisch bodies carrying Ca²⁺ were observed on the inner surface of tapetum, in anther locule and on pollen surface. The Ubisch bodies contacted with pollen, and Ca²⁺ began to accumulate on pollen surface. At the uninucleate pollen stage, abundant Ubisch bodies were distributed in anther locule, and the amount of Ca²⁺ on pollen surface increased. At the mature pollen stage a large amount of Ca²⁺ ions were localized on the inner surface of tapetum, the surface of pollen and Ubisch bodies. In the pollen wall, Ca²⁺ precipitates arranged in radial lines. These results demonstrated that Ubisch bodies were involved in Ca²⁺ transport from anther wall to pollen surface at some developmental stages of anther.     

Ca2+ signals induced from calcium stores in pancreatic islet β cells

In single rat pancreatic β cells, using fura-2 microfluorometry to measure [Ca²⁺]i response upon different stimuli, the ways of calcium regulation have been studied. When the extracellular calcium concentration was 2.5 mmol/L, either 60 mmol/L KCl, 20 mmol/L D-glucose or 0.1 mmol/L tolbutamide induced increase in [Ca²⁺]i. Such increase in [Ca²⁺]i was absent when the same stimuli were applied under zero extracellular calcium. These results indicate that the increase of [Ca²⁺]i is induced by the activation of voltage-dependent calcium channels in β cells. The manifold forms of [Ca²⁺]i change induced by glucose imply that the effects of glucose are complex. 5 mmol/L caffeine or 5 mmol/L MCh increase the [Ca²⁺]i, which is independent of the external calcium, suggesting that [Ca²⁺]i can be regulated by Ca²⁺ release from not only the IP₃-sensitive but also the ryanodine sensitive calcium stores in β cells. The latency of Ca responses for IP₃ pathway (5 s) is faster than that for ryanodine pathway (30 s). It is concluded that there are multiple calcium stores in rat pancreatic β cells.     

Effect of Aβ1-40 on membrane permeability and intracellular free Ca2+ of nerve cells

The effects of soluble and fibrillar Aβ1-40 on membrane permeability and intracellular free Ca^2 of nerve cells were investigated by the laser confocal microscopy. Results indicate that: i) Effects of soluble and fibrillar Aβ1-40 on cell membrane permeability are both concentration-dependent. Soluble Aβ1-40 increases membrane permeability only at concentration of 3μmol/L, while the toxic effect of fibrillar Aβ1-40 is much stronger, its evident effect begins from 1μmol/L. When its concentration rose to 3μmol/L, not only the membrane permeability increased, but also the nuclear membrane broke seriously, ii) Both soluble and fibrillar Aβ1-40 at high concentrations increased the intracellular free Ca^2 , and the increased amplitudes are concentration-dependent. However, the fibrillar one induces the increase of intracellular Ca^2 much quicker and synchronously.These results indicate that some correlation exists between the neurotoxicity of high concentration soluble and fibrillar Aβ1-40 and the change of physico-chemical properties and intracellular Ca ion imbalance.     

Effects of ytterbium on outward K<Superscript>+</Superscript> currents in NIH3T3 cell

Using the whole cell patch-clamp technique, we observed the outward K+ currents and studied for the first time the effects of Yb3+ on the currents and kinetics of activation and inactivation in non-excitable NIH3T3 cell. Our results show that the outward K+ currents were promoted with increasing concentration of Ca2+ in pipette solution and saturated at the concentration of 100 μmol/L Ca2+. Yb3+ in bath solution inhibited the currents in a concentration-dependent manner. At the concentration of 1 μmol/L, Yb...     

A dynamic model of calcium signaling in mast cells and LTC4 release induced by mechanical stimuli

Mast cells （MCs） play an important role in theimmune system. It is known that mechanical stimuli caninduce intracellular Ca2＋ signal and release a variety ofmediators, including leukotriene C4 （LTC4）, leading toother cellular and physiological changes. In this paper, wepresent a mathematical model to explore signalling path-ways in MCs, by including cellular mechanisms for intra-cellular Ca2＋ increase and LTC4 release in response tomechanical stimuli, thapsigargin （TG, SERCA pumpinhibitor）, and LTC4 stimuli. We show that （i） mechanicalstimuli activate mechano-sensitive ion channels and induceinward ion fluxes and Ca2＋ entry which increases intra-cellular Ca2＋ concentration and releases LTC4; （ii） TGinhibits SERCA pumps, empties the internal Ca2＋ stores,which activates Ca2＋ release-activated Ca2＋ channels andresults in sustained intracellular Ca2＋ increase; and （iii）LTC4 activates receptors on MCs surface and increasesintracellular Ca2＋ concentration. Our results are consistentwith experimental observations, and furthermore, they alsoreveal that mechanical stimuli can increase intracellularCa2＋ even when LTC4 release is blocked, which suggests afeed forward loop involved in LTC4 production. This studymay facilitate our understanding of the mechanotransduc-tion process in MCs and provide a useful modeling tool forquantitatively analyzing immune mechanisms involvingMCs.     

Mitochondrial response and calcium ion change in apoptotic insect cells induced by SfaMNPV

Mitochondrial responses and changes of calcium ions in apoptotic insect SL-1 cells induced by Syngrapha falcifera multiple nuclear polyhedrosis virus (SfaMNPV) are reported in this paper. By using Rhodamine 123 as a fluorescent labeling probe, flow cytometry analysis and confocal laser scanning microscope observation we observed that the mitochondrial transmembrane potential (△ψm) began to decrease in SL-1 cells at 4 h post infection and △ψm reduced continuously with the extension of virus infection. Western blotting indicated that the Bcl-2 level in the mitochondria gradually declined and was down- regulated. Cells undergoing apoptosis were found to have an elevation of cytochrome c in the cytosol and a corresponding decrease in the mitochondria, which indicated that cytochrome c was released from mitochondria into cytosol. These results suggest that mitochondrion-mediated apoptotic signal transduction pathway exists in apoptotic insect cell induced by SfaMNPV. Cytosolic free calcium ([Ca^2 ]i) concentration rapidly increased after SfaMNPV infection and the elevated calcium was tested to come partly from extracelllular calcium ion influx. Flow cytometry analysis indicated that the apoptosis in SL-1 cells was not influenced by established cytosolic calcium clamped conditions and the EGTA inhibiting calcium influx. Therefore, neither the elevation of cytosolic calcium ion nor extracellular calcium entry was the inducing factor of apoptosis, which hinted that the depletion of ER Ca^2 store contributed to SL-1 cell apoptosis induced by SfaMNPV.     

Spontaneous Neurotransmitter Release Depends on Intracellular Rather than ER Calcium Stores in Cultured Xenopus NMJ

Introduction During development, many cell types exhibit sponta-neous neurotransmitter release, with synaptic transmis-sions crucial for normal nervous system activity. Syn-aptic transmissions are initiated when an action poten-tial triggers the neurotran…     

P物质对大鼠星状神经节细胞的影响

应用细胞内生物电记录方法，观察神经肽Ｐ物质（ＳＰ）对大鼠星状神经节能细胞的影响，用灌流或用加压向细胞周围施加ＳＰ可使部分细胞发生膜除极反应，用低钙或用含河豚毒素克氏液灌流神经节，并不影响ＳＰ引起的除极反应的幅度和时程。ＳＰ引起除极反应的同时常伴有膜电阻增大，当膜电位增大时，除极化反应幅度变小，反转电位为－８０ｍＶ至－１００ｍＶ，研究表明，ＳＰ对部分星状神经节细胞具有直接兴奋作用，并且ＳＰ对细胞膜的     

Temporal and spatial changes in Ca2+ distribution during the programmed cell death of tracheary elements

The changes in Ca²⁺ distribution in the tracheary elements (TEs) of the pepper leaves were studied using the cytochemical method of potassium antimonate. At the early stage of TEs formation, the vacuole and the nucleus held large volume, and antimonate Ca²⁺ deposits were observed mainly in the intercellular space and the cell wall. As the thickening of secondary wall occurred, the vacuole, nucleus and other organelles began to rupture, concomitant with the increase of calcium deposits in the cytosol, showing the influx of Ca²⁺ into the cell. With the further rupture of cytoplasm and other organelles, the number of calcium deposits at the non-thickening cell wall increased, but declined at the thickening bands of the secondary wall. When the cytoplasmic contents disappeared completely, the level of Ca²⁺ decreased at the non-thickening wall, but by contrast, increased at the thickening bands of the secondary wall. These observations indicated that the dynamic changes in Ca²⁺ distribution spatially and temporarily might have a close correlation with its distinct roles played during the formation of the secondary walls.     

Ca_9Fe(PO_4)_6(OH)_2的电子结构与形成能的第一性原理计算

基于第一性原理和热力学统计原理计算了单个Fe~(2+)替代羟基磷灰石晶胞中Ca~(2+)后晶体(Ca_9Fe(PO_4)_6(OH)_2)的电子结构和能够说明替代后结构稳定性的替位缺陷形成能.电子结构计算得出(Ca_9Fe(PO4)_6(OH)_2)的带隙值是1.85eV,而没有掺杂的是4.93e V,两者态密度特征相似,只是前者峰值变小了.理论形成能和溶液环境中的形成能的计算结果都表明Ca(B)更易被取代,但是两种形成能的值都大于0eV,具体是理论缺陷形成能的值都在7e V到8e V之间,溶液环境中的缺陷形成能主要和替代的位置、溶液的pH值以及Fe~(2+)在溶液中的浓度有关,且其值大于0eV,说明它们需要吸热才能发生替代,所以在羟基磷灰石中Fe~(2+)替代Ca~(2+)不容易发生.     

α-LTX and α-LTXN4C induce [Ca2+]i elevation through different mechanisms in pancreatic β cells

α-latrotoxin (α-LTX) is the only neurotoxin from black-widow spider which has secretagogue effects in the vertebrates. It causes massive neurotransmitter and hormone release via two instinct mechanisms after binding with its high-affinity membrane recep…     

Heat and hyposmotic stimulation increase in [Ca^2＋]i by Ca^2＋ influx in rat synoviocytes

Rheumatoid arthritis （RA）, which is marked by inflammatory synovitis, is a common, chronic autoimmune-disease, whose pathogenesis is complex and still unclear. In order to explore the effects of heat and hyposmotic stimuli on synoviocytes in rheumatoid arthritis, the changes of [Ca^2＋]i induced by heat, hyposmotic and 4α-PDD stimuli were observed in synoviocytes. [Ca^2＋]i elevation induced by heat 28℃, hyposmotic and 4α-PDD stimuli is found to be positively relative to increasing temperature, decreasing osmolality and rising concentration of 4α-PDD. Results show that there is reciprocity among these stimuli and desensitization, and that [Ca^2＋]i elevation depends on Ca^2＋ influx, but not necessarily links to Ca^2＋ release from intracellular stores and voltage-dependent Ca^2＋ channel in synoviocytes. The above characteristics of Ca^2＋ influx are similar to those of TRPV4. A probable mechanism has been suggested that heat and hyposmotic stimulation might increase the level of [Ca^2＋]i by activating the TRPV4-like channel and Ca^2＋ influx in the synoviocytes.     

Actin and myosin during pollen germination

Actin and myosin from pollen tubes of Lilium davidii were studied by using immunoblotting, Dot_Blot and myosin Ca 2+_ATPase analysis. On immunoblotting of the total soluble pollen tube proteins, anti_α_actin antibody labelled a polypeptide approximately 43 ku, which is considered to be the actin of lily. The mRNA encoding actin in ungerminated pollen and germinated pollen were both undetectable in our experiments. A myosin exhibited Ca 2+_ATPase activity, with a native molecular weight of 460 ku has been identified by using immunoblotting. A polypeptide of about 205 ku and a polypeptide of about 20 ku were the heavy chain and a set of light chain of the myosin, which can crossreact with anti_skeletal muscle myosin heavy chain monoclonal antibody and anti_skeletal muscle myosin light chain (20 ku) monoclonal antibody, respectively. The Ca 2+_ATPase activities of myosin in crude extracts of germinated pollen were positively related to the growth rates of pollen tubes.     

Neodymium cations Nd3+ were transported to the interior of Euglena gracilis 277

Euglena gracilis 277, a unicellular green alga, demonstrated remarkable ability to transport Nd³⁺ to the cell compartments. For a given amount of Nd³⁺ and cells, the results of ICP-AES indicated that the cellular uptake of Nd³⁺ was independent of Nd³⁺ concentration in the bulk solution. The average uptake of Nd³⁺ per cell (m _Nd) is proportional to a parameter ξ — the ratio of neodymium content to the cell counts of the system. A novel approach for probing cellular neodymium by tetraiodotetra chlorofluorescein (I4TCF) has been devised. Data derived from the cryosections of I4TCF-Nd³⁺ stained cells and EDAX of the fast freezing ultrathin cryosections indicate that Nd³⁺ is distributed over the cell compartments. Chloroplasts are the major compartments as the residence of Nd³⁺ in the alga. The transport should be against a concentration gradient of Nd³⁺ on the order of five, even higher. It is proposed that a calcium ion channel would play an important role in the Nd³⁺ transportation.     

Effect of Ca on CO2 corrosion properties of X65 pipeline steel

The effect of Ca²⁺ on CO₂ corrosion to X65 pipeline steel was investigated in the simulated stratum water of an oil field containing different concentrations of Ca²⁺. It is found that Ca²⁺ can enhance the corrosion rate, especially in the Ca²⁺ concentration from 256 to 512 mg/L, which can be attributed to the growing grain size and loosing structure of corrosion scales with increasing Ca²⁺ concentration. X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) investigations reveal that a complex carbonate (Fe, Ca)CO₃ forms at high Ca²⁺ concentration due to the gradual replacement of Fe²⁺ in FeCO₃ by Ca²⁺.