The new sideway of CNTF signal transduction pathway

The action of ciliary neurotrophic factor (CNTF) on intercellular free Ca²⁺ concentrations [Ca²⁺]_i induced by glutamate (Glu) in primary cultured hippocampal neurons were detected with Fura2/AM, a Ca²⁺-sensitive fluorophore, and the morphological influence of G-protein on it was objected. Glu could induce rapid increase of [Ca²⁺]_i in hippocampal neurons. CNTF had no significant action on [Ca²⁺]_i in resting hippocampal neurons. However, after incubation of CNTF for 5 min, the increase of [Ca²⁺]_i in hippocampal neurons rapidly induced by Glu was inhibited. Pretussis toxin (PTX)-sensitive G protein could block the action. These results indicate that a new non-genomic rapid sideway might exist in the upper stream of CNTF signal transduction pathway, which was related to Ca²⁺ signal transduction. Keywords:     

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.     

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.     

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.     

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…     

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.     

Immunohistochemical localization of Ca2+/calmodulin-dependent kinase in tobacco

The existence of Ca²⁺/calmodulin-dependent kinase (CaM kinase, CaMK) in tobacco is verified immuno- logically and its distribution in different tissues of tobacco is studied. It has been demonstrated that CaMK is mainly distributed in early developing anthers, developing ovules and embryos, lateral root primordium, apical meristem and leaf primordium of buds and mesophyll cells and developing vascular bundles of leaves. There is enormous CaM kinase distributed in leaf epidermis fair cells and guard cells of stomas too. Little kinase is found in mature stem or root cells. The distribution properties of CaM kinase in tobacco are consistent with those of CaM, suggesting that there exists the Ca²⁺ signal transduction pathway mediated by CaM kinase in tobacco and it plays an important role in the plant growth and development.     

玉米幼苗热激诱导抗冷性过程中钙的效应

 玉米幼苗在冷胁迫前经过热激处理或CaCl₂浸种后再热激处理,其存活率,抗氧化酶GR的活性、可溶性蛋白质中热稳定蛋白质和游离脯氨酸的含量在的冷胁迫中均发生了变化,发现热激处理幼苗的这些参数高于对照,而最高的是CaCl₂浸种后再热激处理的,表明热激能提高玉米幼苗的抗冷性,而Ca²⁺对上述热激处理有加强作用.     

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.     

Correlation between platelet gelsolin levels and different types of coronary heart disease

Gelsolin is an important cytoskeletal protein of platelets and studies have shown a close relationship between gelsolin and cardiovascular disease.However,the role of gelsolin in the development of coronary heart disease(CHD) is unclear.In this study,we record the distribution of gelsolin in human platelets and plasma and its association with different types of CHD.This study included 114 cases,with 33 stable angina pectoris(SAP) cases,81 acute coronary syndrome(ACS) cases—composed of 39 unstable angina pectoris(UAP) and 42 acute myocardial infarction(AMI) cases,and 31 healthy control participants.Gelsolin concentration in platelet rich plasma(PRP) and platelet poor plasma(PPP),actin filament(F-actin) and Gc-globulin of PPP were determined by enzyme-linked immunoadsorbent assay(ELISA).The fluorescence intensity of CD62p and cytoplasmic calcium([Ca2+] i) in human platelets measured by flow cytometry.We also used turbidimetry to detect the platelet aggregation rate(PAR).We analyzed the correlation between platelet gelsolin concentration and CD62p or plasma F-actin levels among each different patient group.Compared with the control group,the gelsolin level in PRP of UAP and AMI groups increased significantly(P<0.01),while the gelsolin level in PPP of all the three patient groups decreased markedly(P<0.01),and the CD62p,PAR,[Ca2+] i of platelets,F-actin and Gc-globulin of the UAP and AMI groups increased significantly(P<0.01).Compared with the SAP group,the gelsolin level in PRP,the PAR,[Ca2+] i of platelets and CD62p of other two groups increased significantly(P<0.01),F-actin of the AMI group increased markedly(P<0.01).Platelet cytoskeleton protein dynamics vary among the different types of CHD.Platelet gelsolin levels are markedly increased and accompanied by increased platelet activity,F-actin and [Ca2+] i of ACS patients,while gelsolin levels in PPP are markedly lower.Abnormally increased platelet gelsolin levels show high positive correlation with the level of platelet activity.Therefore,platelet gelsolin might be a novel molecular marker and/or a new potential therapeutic target of anti-platelet therapy of ACS.     

Effects of La3+ and Gd3+ on Ca2+ influx in rat hepatoma H-35 cells

Effects of La³⁺ and Gd³⁺ on Ca²⁺ influx were investigated in rat hepatoma H-35 cells by measuring the initial rate of⁴⁵Ca²⁺ uptake. It was found that the maximum initial rate of Ca²⁺ uptake was increased six-to ten-fold at low concentrations of La³⁺ and Gd³⁺. Kinetic analyses by measuring the initial rate of Ca²⁺ influx at different external Ca²⁺ concentrations indicated the existence of two intracellular exchangeable components in the basal Ca²⁺ system, with low and high affinities for Ca²⁺, and only one class of Ca²⁺ binding sites was observed in the La³⁺-or Gd³⁺-treated cells. For high affinity, La³⁺ and Gd³⁺ increased both kinetic parametersK _m andV _max of basai Ca²⁺ influx. La³⁺ and Gd³⁺ compete directly with Ca²⁺ for Ca²⁺ binding site for low affinity. The kinetics is competitive.     

Cell cycle and cell signal transduction in marine phytoplankton

tAs unicellular phytoplankton, the growth of a marine phytoplankton population results directly from the completion of a cell cycle, therefore, cell-environment communication is an important way which involves signal transduction pathways to regulate cell cycle progression and contribute to growth, metabolism and primary production and respond to their surrounding environment in marine phytoplankton. Cyclin-CDK and CaM/Ca²⁺ are essentially key regulators in control of cell cycle and signal transduction pathway, which has important values on both basic research and applied biotechnology. This paper reviews progress made in this research field, which involves the identification and characterization of cyclins and cell signal transduction system, cell cycle-control mechanisms in marine phytoplankton cells, cell cycle proteins as a marker of a terminal event to estimate the growth rate of phytoplankton at the species level, cell cycle-dependent toxin production of toxic algae and cell cycle progression regulated by environmental factors.     

叶绿体巯基亚硝基化修饰蛋白质组学研究进展

蛋白质S-亚硝基化是NO与蛋白质的半胱氨酸残基共价连接形成S-亚硝基硫醇的过程.叶绿体内快速变化的氧化还原环境容易导致蛋白质S-亚硝基化修饰发生.综述了植物逆境应答过程中叶绿体蛋白质S-亚硝基化参与调节的光合电子传递、卡尔文循环、抗氧化系统、蛋白质合成、蛋白质加工与周转、Ca~(2+)介导的信号转导、氮同化、硫同化,以及四吡咯化合物合成等途径,为全面理解植物逆境应答过程的NO调控网络机制提供了线索.     

Icariin suppresses bone resorption activity of rabbit osteoclasts in vitro

The effect of icariin on the bone resorption activity of rabbit osteoclasts is assessed in vitro. Osteoclasts were isolated from Japanese white rabbits and cultured on plates with a sterilized bone slice in each well. After treatment with icariin at various concentrations, the bone resorption activity of osteoclasts was evaluated by examining pit areas, superoxide anion (·O2-) generation, size and number of actin rings and intracellular calcium concentration [Ca2 ]i. As revealed by these data, icariin elicited continuous decline of [Ca2 ]i, making actin ring constricted and ·O2- generation decreased. These events resulted in smaller and fewer pits which indicate suppressed bone resorption activity of rabbit osteoclasts by icariin.     

Rearrangements of microtubule cytoskeleton in stomatal closure of <Emphasis Type="Italic">Arabidopsis</Emphasis> induced by nitric oxide

NO （nitric oxide）, known as a key signal molecule in plant, plays important roles in regulation of stomatal movement. In this study, microtubule dynamics and its possible mechanism in the NO signal pathway were investigated. The results were as follows： （i） In vivo stomatsl aperture assays revealed that both vinblastine （microtubule-disrupUng drug） and SNP （exogenous NO donor） prevented stomatsl opening in the light, and vinblastine even could enhance the inhibitory effect of SNP, whereas tsxol （a microtubule-stsbilizing agent） was able to reduce this effect; （ii） microtubules in the opening Arabi-dopsis guard cells expressing GFP：a-tubulin-6 （AtGFP：a-tubulin-6） were organized in parallel, straight and dense bundles, radiating from the ventral side to the dorsal side, and most of them were localized perpendicularly to the ventral wall; （iii） under the same environmental conditions, treated with SNP for 30 min, the radial arrays of microtubules in guard cells began to break down, twisted partially and be- came oblique or exhibited a random pattern; （iv） furthermore, the involvement of cytosolic Ca^2＋ in this event was tested. Stomatal aperture assays revealed that BAPTA-AM （a chelator of Ca^2＋） greatly suppressed the effect of NO on stomatal closure; however, it did not show the same function on stomatal closure induced by vinblastine. When BAPTA-AM was added to the SNP-pretreated solution, the SNP-induced disordered microtubulue cytoskeleton in guard cells underwent rearrangement in a time-dependent manner. After 30 min of treatment with BAPTA-AM, the cortical microtubules resumed the original radial distribution, almost the same as the control. All this indicates that NO may promote rearrangement of microtubule cytoskeleton via elevation of [Ca^2＋]cyt （free Ca^2＋ concentration in the cytoplasm）, finally leading to stomatsl closure.     

Phosphatidylglycerol effect on oxygen-evolving activity in Ca2+-depleted photosystem II

The effect of anionic phosphatidylglycerol (PG) on oxygen evolution in a photosystem II (PS II ) particle depleted of Ca²⁺ (designated d_CaPSII ) has been investigated. The major finding is the observation of a new role of PG in the PSII function. That is, PG restores nearly the lost oxygen evolution in d_caPS II particle owing to Ca²⁺ depletion to the levels in intact PS II. Furthermore, there is a stimulation of oxygen-evolving activity in the d_CaPSII complexed with PG in the presence of exogenous CaCl₂, which PG enhances increasingly oxygen evolution with increasing CaCl₂ concentration. It is suggested that PG-induced oxygen evolution recovery of d_Ca PS II particle results from resumption of normal structure in protein by PG effect, whereas the enhancement of oxygen evolution in complex subject to CaCl₂ is ascribed to the optimization of such a structure due to coordination complex formation of Ca²⁺ ions with PG.     

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⁺.     

Nitric oxide suppresses stomatal opening by inhibiting inward-rectifying K<Stack><Subscript>in</Subscript><Superscript>+</Superscript></Stack> channels in <Emphasis Type="Italic">Arabidopsis</Emphasis> guard cells

We explore nitric oxide （NO） effect on K^＋in, channels in Arabidopsis guard cells. We observed NO inhibited K^＋in, currents when Ca^2＋ chelator EGTA （Ethylene glycol-bis（2-aminoethylether）-N,N,N′,N;tetraacetic acid） was not added in the pipette solution; K^＋in currents were not sensitive to NO when cytosolic Ca^2＋ was chelated by EGTA. NO inhibited the Arabidopsis stomatal opening, but when EGTA was added in the bath solution, inhibition effect of NO on stomatal opening vanished. Thus, it implies that NO elevates cytosolic Ca^2＋ by activating plasma membrane Ca^2＋ channels firstly, then inactivates K^＋in, chartnels, resulting in stomatal opening suppressed subsequently.     

Activities and properties of calcineurin catalytic domain

Calcineurin (CN) is the only protein phosphatase known to be under the control of calcium (Ca²⁺) and calmodulin (CaM). The enzyme consists of two subunits, the catalytic A subunit of 61 ku (CNA) and a regulatory B subunit of 19 ku (CNB). In this study, we used PCR amplication to construct a truncation consisting of only the CNA catalytic domain. The truncation was induced by IPTG and expressed inE. coli. PNPP was used as a substrate to study the phosphatase activity of the CNA catalytic domain. The findings show that its activity is 20 times greater than CNA in the presence of CNB and CaM. The optimum reaction temperature for the CNA catalytic domain protein is 40°C, and the optimum reaction pH value is 8.0. Mn²⁺ is still an effective activator for the CNA catalytic domain, but its activity is not controlled by Ca²⁺. In the presence of 6 mmol/L Mg²⁺, adding either Ca²⁺ or EGTA did not change the activity of the CNA catalytic domain.     

Involvement of Ca<Superscript>2+</Superscript>/CaM in the signal transduction of acetylcholine regulating stomatal movement

It has been known that the neurotransmitter acetylcholine (ACh) also exists in plants and is able to regulate the movement of stomata.In another aspect,Ca^2 /CaM as the second messengers have a critical role of signal transduction in stomatal guard-cell,Here we showed that Ca^2 /CaM were also involved in the ACh regulated stomatal movement,In the medium containing Ca^2 ,the Ca^2 channel blockers (NIF and Ver)and CaM inhibitors (TFP and W7 ) could neutralize the ACh induced stomatal opening,however,they are ineffective in the medium containing K^ ,Those results indicated that Ca^2 /CaM were involved in the signal transduction pathway of ACh regulating stomatal movement.