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:     

Influence of location-dependent protuberance damage on cell viability

The influence of femtosecond laser-induced damages on viability of olfactory ensheathing cells (OECs) is investigated. Several cytokinetic processes including cellular damage, recovery and death are discussed. Using femtosecond laser with the power of 100 μW and cutting speed of 2 μm/s, we cut the cellular protuberance with smaller diameter twice in different locations, and then observe the viability of the damaged cells. Under the same conditions, the root of protuberance with larger diameter is cut six times to observe changes of cellular shape. Whether the damage is located in the end, middle or root of protuberance with smaller diameter, the cell viability can recover within 3 h. When the damage is located in the root of protuberance with larger diameter, the damaged cell will die in the way of oncosis. Cytokinetic phenomena including intracellular high Ca²⁺ concentration, cellular morphologic change, recovery and oncosis are discussed. Meanwhile, high Ca²⁺ concentration is observed after femtosecond laser surgery. Therefore, femtosecond laser surgery is an important tool for establishing cell damage model and studying cytokinetics. Supported by National High Technology Research & Development Program of China (Grant No. 2006AA04Z307), Author of National Excellent Doctoral Dissertation of PR China (Grant No. 2006039), National Natural Science Foundation of China (Grant No. 50775104), Natural Science Foundation of Jiangsu Province (Grant No. BK2006507) and Jiangsu Provincial Research Innovation Program for College Graduates (Grant No. CX07B_086z)     

Mechanism for chronic pain generation

Neuropathic pain and the other abnormalities of sensation induced by axon injury or by peripheral nerve inflammation should result from functional compensations of the injured neurons during their regeneration. Ectopic distribution of proteins related to Na⁺, K⁺ and Ca²⁺ channels as well as of receptors on both membranes of injured axon and its cell body becomes a main pacemaker from which spontaneous ectopic afferent of primary sensatory neurons and crosstalk between neurons occur. Abnormal ectopic afferent activities lead to disorders of the sensation, such as hyperalgesia, allodynia, spontaneous pain and paraesthesia. Administration of some ion channel agents and/or α2-adrenergic blockers has shown efficiency in preventing neuropathic pain development and in relieving neuropathic pain.     

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…     

珍珠贝肉提取液抗衰作用的动物实验

在实验室检测马氏珍珠贝肉提取液对老龄小白鼠分辩学习及记忆保持能力的影响 ,并于行为实验结束后测定海马突触体内游离钙水平变化。小白鼠 1 6～ 1 7月龄 ,体重 (65±4) g,雌雄兼用 ,共 30只 ,分为天然提取液组 1 0只 ,强化提取液组 (天然提取液 +牛磺酸 ) 1 0只 ,对照组 (饮用普通清水 ) 1 0只。平均每只小鼠每天服用量为 5 ml,共服用 40 d。珍珠贝肉天然提取液和牛磺酸强化提取液与对照组相比 ,都明显增强老龄鼠在 Y-迷宫中的分辩学习能力和学习速度 ,减少训练次数 (P <0 .0 5) ;对记忆保持率的提高具有促进作用 ,尤其是牛磺酸强化提取液使记忆保持率大幅度提高 (P <0 .0 1 )。天然提取液与强化提取液都一致降低鼠脑的海马突触体游离钙离子水平 ,提示这种提取液对衰老脑细胞的“钙超载”有拮抗效应     

Ionization and dissociation of CH<Subscript>2</Subscript>I<Subscript>2</Subscript> and CH<Subscript>2</Subscript>ICl in femtosecond intense field

With the development of ultrafast laser technique,the femtosecond laser electric field is comparable to themolecular field experienced by valence electrons. In thiscase, the classical quantum mechanical theory, based onperturbation theory, becomes inapplicable. Theoreticalunderstanding of the interaction of molecules with intenselaser radiation is quite difficult. The Coulomb explosionhas been extensively studied[1—10] and explained by somesemiclassical models, such as the above threshold ion…     

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.     

Effects of lithium chloride on outward potassium currents in acutely isolated hippocampal CA1 pyramidal neurons

Lithium is best known for its therapeutic efficacy in the treatment of manic-depressive illness. Its clinical profile includes the antimanic and antidepressant ac- tions as well as prophylaxis of both mania and depres- sion. Despite its efficacy, the mole…     

Membrane conductance oscillations in astrocytes induced by phorbol ester

Glial cells in the central nervous systems (CNS) have complex functions which are difficult to decipher because of the intimate intertwining of glial cells with neurons. We have therefore developed an essentially neuron-free preparation of CNS astrocytes in the kainic acid lesioned hippocampal slice. With this preparation we have examined the effect of activating protein kinase C in astrocytes with a phorbol ester, TPA (12-O-tetradecanoyl-phorbol-13-acetate). In most cells, TPA induced rhythmic oscillations (0.1-3.0 Hz) of membrane potential which were typically 5-10 mV in amplitude and were associated with increases of up to eightfold in input resistance during the depolarizing phase. These large changes in membrane conductance are the first reported observations of endogenously generated conductance changes in astrocytes of the mammalian CNS and they could influence excitability of surrounding neurons, possibly by altering extracellular ion concentrations.     

Monitor and control of neuronal activities with femtosecond pulse laser

Combined with the fluorescence labeling technique, two-photon microscopy excited with femtosecond pulse laser has become an important tool for neuroscience research. In this research, the calcium signals from neurons in rat cortex slice were monitored by a custom-built two-photon microscopy, and the spontaneous calcium signals and the pharmacological responses as well as the responses to femtosecond pulse laser stimulation were recorded. The results showed that the amplitude of the cal- cium signals increased in direct proportion to the corresponding electrical activities. Glutamate induced a calcium transient, but continuous application resulted in smaller response. Simultaneous monitoring of neuronal populations distinguished the neurons of different microcircuits. The femtosecond pulse laser induced local or global calcium signals in the pyramidal neurons. The approach of interrogation and control of neural activities using femtosecond pulse laser is non-contact, nondestructive, repeatable, and without any additional substrates, which will contribute to the development of neuroscience.     

Separation and analysis of the soluble trimer of Aβ1-40 and its effects on the rise in intracellular calcium

Alzheimer’s disease (AD) is one of the progressive neurodegenerative diseases associated with protein conformational transition, generally known as protein conformational disorders[1]. The general characteriza- tions of AD are the intracellular fibrillar…     

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⁺.     

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...     

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.     

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.     

Cardiac protective role of a novel erythrocyte-derived depressing factor on rats and its Ca^2＋ mechanism

The cardiac protective role of a novel erythro-cyte-derived depressing factor (EDDF) on spontaneous hy-pertensive rats (SHR), calcium overload (CaO) rats and Wistar rats and its mechanism was evaluated. Mean artery pressure (MAP), heart rate (HR) and LVdp/dtmax were measured by physiological recorder. The effect of EDDF on the Ca2+-ATPase activity in myocardial sarcoplasmic reticu-lum (SR) of CaO rats was determined by inorganic phos-phate assay. Calcium transport in myocytes was measured by 45Ca2+ radioactive isotope measurement. The phosphoryla-tion levels of extracellular signal-regulated protein kinases (ERK1/2) in myocardial tissue of SHR and CaO rats were measured by Western blot method. And the ultrastructures of cardiac muscle cells were observed with the transmission electron microscope. The results indicated that EDDF could significantly decrease MAP, HR and LVdp/dtmax in a dose dependent manner (P < 0.05). It seems that the mechanism might relate with activating the Ca2+-APTase, enhancing the uptake and release of Ca2+ from SR (P < 0.05), decreasing the phosphorylation levels of ERK1/2 of myocytes (P < 0.01) and lightening the ultrastructural lesion of cardiac muscle cells. In CaO rats, the Ca2+-ATPase activity decreased clearly com-pared to control (64.99 7.16 vs 94.48 7.68 nmol·min-1 ·mg-1 protein, P < 0.01), while EDDF (100 mg/mL) could significantly increase the activity (87.93 ?9.54 vs 64.99 ?7.16, P < 0.05, n = 7). Both uptake and release rate of Ca2+ (祄ol 45Ca2+/g protein/min) from myocardial SR of CaO rats re-markably decreased compared to control (32.40 ?2.70 and 15.46 ?1.49 vs 61.09 ?10.89 and 25.47 ?4.29, P < 0.05); EDDF (100 mg/mL) could significantly stimulate their activi-ties (50.48 6.76 and 21.76 2.75 vs 32.40 2.70 and 15.46 1.49, P < 0.05). EDDF could evidently down-regulate the phosphorylation of ERK1/2 in myocardial tissue from SHR and CaO rats (P < 0.01), lighten the ultrastructural lesion of cardiac muscle cells of SHR as well. It is concluded that EDDF seems to play protective roles on both structure and function of heart, which closely related with amelioration of Ca2+ transport and inhibition of Ca2+-MAP kinase pathway.     

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.     

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.     

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.