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     

Depolarization Induced by Substance P on Rat Stellate Ganglion Neurons

应用细胞内生物电记录技术，观察神经肽Ｐ物质（ＳＰ）对大鼠星状神经节细胞的影响。ＳＰ在１μｍｏｌ～１０μｍｏｌ或更高的浓度范围内，供试３５个细胞，有２８个细胞发生膜除极反应。用低钙（０．２５ｍｍ）或用含河豚毒素（ＴＴＸ，１μｍｏｌ）克氏液灌流神经节，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兴奋作用，使通过这些细胞的信息传递增强；ＳＰ对细胞膜的除极作用是由于其引起细胞膜钾导降低所致。     

孤啡肽对大鼠背根神经节神经元膜的作用

应用细胞内微电极记录技术,观察了孤啡肽(orphanin OF,OFQ)对2~3周龄大鼠背根神经节(dorsal root gan-glion,DRG)神经元的作用及OFQ受体的特异性阻断剂Nphe对OFQ引起的DRG神经元膜反应的影响。受检的271个细胞,静息膜电位-45.00±7.63mV,当加3×10-7~3×10-10mol/L OFQ时可观察到如下三种膜电位变化:1)超极化;2)去极化;3)双相反应。实验中还可观察到膜电位无反应的现象,其未列入受检细胞个数中。用Nphe(3×10-7~3×10-9mol/L)灌流DRG标本,能分别阻断OFQ引起的膜的3种反应,表明了DRG神经元膜上有OFQ受体的存在,并且OFQ引起DRG神经元膜电位多样性变化。     

乙酰胆碱对大鼠离体输精管平滑肌细胞膜电位的作用

采用细胞内微电极技术对大鼠离体输精管平滑肌细胞进行记录,观察其静息电位、膜阻抗等被动膜电性质及ACh对平滑肌细胞膜电位的作用,在此基础上运用微电泳技术对所记录细胞进行标记,从而分析不同走行方向的平滑肌细胞的电生理学特性。实验测得输精管平滑肌细胞静息电位为-48.56±0.46mV(n=1141),膜输入阻抗为1638.70±205.20MΩ(n=45)。实验用ACh(0.3～100μmol L)灌流平滑肌细胞,可引起膜发生浓度依赖性超极化。10μmol LACh引起超极化幅值为13.67±0.98mV(n=116),膜电导平均增加2.31%(n=10)。标本经1μmol L硫酸阿托品(n=10)和100nmol L2' deoxyadenosine5' monophosphate(n=11)预处理后,ACh引起的超极化幅值明显减小。经荧光染料(0.1%propidiumiodide)标记鉴别后,统计分析外纵走行细胞的静息电位为-53.56±3.88mV(n=37),膜输入阻抗为2245.60±372.50MΩ(n=13);内环走行细胞的静息电位为-51.62±4.27mV(n=17),膜输入阻抗为2101.50±513.50MΩ(n=10);并且2种走形方向的平滑肌细胞对ACh的反应形式亦无显著性差异。实验表明:输精管平滑肌细胞的电生理特性以及对ACh的反应与其走形方向无关。     

Baclofen对大鼠新鲜分离DRG神经元GABA和NMDA激活电流的抑制作用

用全细胞膜片箝技术在大鼠新鲜分离的背根神经节 (DRG)细胞上观察到baclofen对GABA和NMDA激活电流有调制作用。单独给予DRG细胞baclofen (1～ 10 0 μmol/L)未记录到可测的电流改变 ,但预加baclofen对GABA和NMDA激活电流具有明显的抑制作用 ,且呈剂量依赖性。 10 0 μmol/Lbaclofen对GABA和NMDA激活电流分别抑制(5 1.2± 10 .9) % (X±SE ,n=8,P <0 .0 1)和 (6 4 .1± 2 1.1) % (X±SE ,n=6 ,P <0 .0 1) ,此抑制作用是可逆的 ,可被GABAB 受体拮抗剂saclofen(10 0 μmol/L ) 所取消 (n =4 )。     

副交感神经节细胞5─羟色胺去极化反应的离子基础

使用离体细胞内记录，研究了猫副交感性胰腺神经节细胞的５－羟色胺（５－ＨＴ）去极化反应及其离子基础。５－ＨＴ使大部份细胞（４７／５１）产生去极化反应。反应只有两种类型：快去极化（６／４７）及慢去极化（３５／４７），另有６个细胞出现先快、后慢的双相去极化反应；５－ＨＴ导致的快去极化伴有膜电阻减小，向细胞内通以超极化直流电形成条件性膜超极时其幅度增大，提示Ｎａ＋导增大是其离子基础；５－ＨＴ导致的不同细胞的慢去极化分别伴随膜电阻增大、减小或不变，条件性膜超极时其幅度分别增大或不变，提示不是单一离子、而是多种离子参与其形成。分别用低Ｎａ＋、高Ｋ＋溶液灌注神经节，５－ＨＴ慢去极化均明显减小，而低Ｃｌ－溶液则无明显效应，表明Ｎａ＋导增大和／或Ｋ＋导减少是５－ＨＴ慢去极化的离子基础。     

副交感神经节细胞5─羟色胺去极化反应的离子基础

使用离体细胞内记录，研究了猫副交感性胰腺神经节细胞的５－羟色胺（５－ＨＴ）去极化反应及其离子基础。５－ＨＴ使大部份细胞（４７／５１）产生去极化反应。反应只有两种类型：快去极化（６／４７）及慢去极化（３５／４７），另有６个细胞出现先快、后慢的双相去极化反应；５－ＨＴ导致的快去极化伴有膜电阻减小，向细胞内通以超极化直流电形成条件性膜超极时其幅度增大，提示Ｎａ＋导增大是其离子基础；５－ＨＴ导致的不同细胞的慢去极化分别伴随膜电阻增大、减小或不变，条件性膜超极时其幅度分别增大或不变，提示不是单一离子、而是多种离子参与其形成。分别用低Ｎａ＋、高Ｋ＋溶液灌注神经节，５－ＨＴ慢去极化均明显减小，而低Ｃｌ－溶液则无明显效应，表明Ｎａ＋导增大和／或Ｋ＋导减少是５－ＨＴ慢去极化的离子基础。     

P物质对GABAA受体介导电流的作用

应用全细胞膜片箝技术,在大鼠新鲜分离的背根神经节细胞上观察到:SP对GABA激活电流有调制作用。实验结果表明:多数受检的大鼠背根神经节神经元对SP(28/41,68.5％)和GABA(36/41,88.2％)敏感,100μmol/LSP和100μmol/L GABA激活内向电流的幅值分别为243.8±82.7pA(x±s,n=9)和1.76±0.48nA(x±s,n=13)。GABA与SP激活的内向电流明显不同,前者具有明显的去敏感现象。实验预加SP(0.001～1μmol/L)30s后再加GABA,则GABA激活电流的幅值明显减小,而且GABA激活电流减小的程度与预加SP的浓度呈量效依赖关系。SP主要抑制GABA激活电流的峰电流,对其稳态电流的抑制不明显。预加0.1μmol/L SP之后约4min左右,SP对GA-BA激活电流的抑制效率最大,为49.8±7.2％(x±s,n=7,P≤0.01),而预加SP 12min之后,SP才失去对GABA激活电流的抑制作用。     

几种抗氧化剂对DNA的损伤和保护作用

采用单细胞凝胶电泳法研究了几种抗氧化剂对人外周血单核细胞DNA的损伤及对由H2O2引起的DNA损伤的保护作用.结果表明,槲皮素可引起明显的细胞DNA损伤,并呈浓度依赖性;芦丁在较高浓度时,引起损伤.7,8-二羟基-4-甲基香豆素,7-羟基-4-甲基香豆素即使浓度达100μmol/L也不引起细胞DNA损伤.Vc、亚硒酸钠和甘露醇,在较高浓度时均可引起微弱的损伤,只有Vc引起的损伤较严重.当预先用抗氧化剂保护细胞后再用50 μmol/L H2O2处理,发现7,8-二羟基-4-甲基香豆素的保护作用最强,槲皮素次之,保护作用具浓度依赖性.而芦丁、7-羟基-4-甲基香豆素、Vc和亚硒酸钠在浓度≤50 μmol/L时,无任何保护作用.甘露醇在低浓度可起明显的保护作用,在较高浓度25 μmol/L和50 μmol/L时则保护作用丧失.     

铅污染对藻细胞膜电位和膜电阻的影响研究

运用细胞外表面技术和电化学方法,构建了一种藻细胞膜电位传感系统,并研究了Pb2+污染对淡水藻细胞膜电位和膜电阻的影响.结果表明,膜电位和膜电阻两指标互为补充,快速而灵敏反应Pb2+对藻细胞的毒性影响,响应时间均控制在30 min内,明显响应的最低浓度为0.10 mmol·L-1.本实验浓度范围内,低浓度Pb2+(0.10 mmol·L-1)导致藻细胞超极化、膜电阻增大;而高浓度Pb2+(0.20～1.00 mmol·L-1)则引起细胞去极化和膜电阻减小.此外,高浓度Pb2+对藻细胞膜电位、膜电阻的影响不可逆,而低浓度Pb2+对藻细胞的影响具有可逆性;而对比各高浓度组影响差异性,毒性剂量效应不显著,表明藻细胞对高浓度Pb2+具有一定抗性.     

The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels

The enzyme NADPH oxidase in phagocytes is important in the body's defence against microbes: it produces superoxide anions (O2-, precursors to bactericidal reactive oxygen species). Electrons move from intracellular NADPH, across a chain comprising FAD (flavin adenine dinucleotide) and two haems, to reduce extracellular O2 to O2-. NADPH oxidase is electrogenic, generating electron current (I(e)) that is measurable under voltage-clamp conditions. Here we report the complete current-voltage relationship of NADPH oxidase, the first such measurement of a plasma membrane electron transporter. We find that I(e) is voltage-independent from -100 mV to >0 mV, but is steeply inhibited by further depolarization, and is abolished at about +190 mV. It was proposed that H+ efflux mediated by voltage-gated proton channels compensates I(e), because Zn2+ and Cd2+ inhibit both H+ currents and O2- production. Here we show that COS-7 cells transfected with four NADPH oxidase components, but lacking H+ channels, produce O2- in the presence of Zn2+ concentrations that inhibit O2- production in neutrophils and eosinophils. Zn2+ does not inhibit NADPH oxidase directly, but through effects on H+ channels. H+ channels optimize NADPH oxidase function by preventing membrane depolarization to inhibitory voltages.     

Voltage-induced membrane movement

Thermodynamics predicts that transmembrane voltage modulates membrane tension and that this will cause movement. The magnitude and polarity of movement is governed by cell stiffness and surface potentials. Here we confirm these predictions using the atomic force microscope to dynamically follow the movement of voltage-clamped HEK293 cells in different ionic-strength solutions. In normal saline, depolarization caused an outward movement, and at low ionic strength an inward movement. The amplitude was proportional to voltage (about 1 nm per 100 mV) and increased with indentation depth. A simple physical model of the membrane and tip provided an estimate of the external and internal surface charge densities (-5 x 10(-3) C x m(-2) and -18 x 10(-3) C x m(-2), respectively). Salicylate (a negative amphiphile) inhibited electromotility by increasing the external charge density by -15 x 10(-3) C x m(-2). As salicylate blocks electromotility in cochlear outer hair cells at the same concentration, the role of prestin as a motor protein may need to be reassessed.     

Subthreshold Na+-dependent theta-like rhythmicity in stellate cells of entorhinal cortex layer II

The oscillation of membrane potential in mammalian central neurons is of interest because it relates to the role of oscillations in brain function. It has been proposed that the entorhinal cortex (EC), particularly the stellate cells of layer II (ECIIscs), plays an important part in the genesis of the theta rhythm. These neurons occupy a key position in the neocortex-hippocampus-neocortex circuit, a crucial crossroad in memory functions. Neuronal oscillations typically rely on the activation of voltage-dependent Ca2+ conductances and the Ca2+ -dependent K+ conductance that usually follows, as seen in other limbic subcortical structures generating theta rhythmicity. Here we report, however, that similar oscillations are generated in ECIIscs by a Na+ conductance. The finding of a subthreshold, voltage-gated, Na+ -dependent rhythmic membrane oscillation in mammalian neurons indicates that rhythmicity in heterogeneous neuronal networks may be supported by different sets of intrinsic ionic mechanisms in each of the neuronal elements involved.     

Requirement of TRPC channels in netrin-1-induced chemotropic turning of nerve growth cones

Ion channels formed by the TRP (transient receptor potential) superfamily of proteins act as sensors for temperature, osmolarity, mechanical stress and taste. The growth cones of developing axons are responsible for sensing extracellular guidance factors, many of which trigger Ca2+ influx at the growth cone; however, the identity of the ion channels involved remains to be clarified. Here, we report that TRP-like channel activity exists in the growth cones of cultured Xenopus neurons and can be modulated by exposure to netrin-1 and brain-derived neurotrophic factor, two chemoattractants for axon guidance. Whole-cell recording from growth cones showed that netrin-1 induced a membrane depolarization, part of which remained after all major voltage-dependent channels were blocked. Furthermore, the membrane depolarization was sensitive to blockers of TRP channels. Pharmacological blockade of putative TRP currents or downregulation of Xenopus TRP-1 (xTRPC1) expression with a specific morpholino oligonucleotide abolished the growth-cone turning and Ca2+ elevation induced by a netrin-1 gradient. Thus, TRPC currents reflect early events in the growth cone's detection of some extracellular guidance signals, resulting in membrane depolarization and cytoplasmic Ca2+ elevation that mediates the turning of growth cones.     

Effects of nitrogen ion implantation on Ca2+ concentration and membrane potential of pollen cell

The effects of low energy nitrogen ion implantation on Ca²⁺ concentration and membrane potential of lily (lilium davidii Duch) pollen cell have been studied. The results showed that the Ca²⁺ concentration was increased when pollen grain was implanted by nitrogen ion with energy 100 keV and dose 10¹³ ions/cm². However, the increase of Ca²⁺ concentration was partly inhibited by the addition of Ca²⁺ channel inhibitor depending on dose. And nitrogen ion implantation caused depolarization of pollen cell membrane potential. In other words, membrane potential was increased, but the effect decreased by adding Ca²⁺ channel inhibitor. However, it was still significantly higher than the membrane potential of control cells. It was indicated that the depolarization of cell membrane potential opened the calcium channel on the membrane that caused the increasing of intracellular calcium concentration. This might be an earlier step of the effect of low energy nitrogen ion implantation on pollen germination.     

一种高性能的BiCMOS带隙基准电压源

提出了一种新颖的利用负反馈环路以及RC滤波器提高电源抑制比的高精密CMOS带隙基准电压源．采用上海贝岭的1．2μm BiCMOS工艺进行设计和仿真,spectre模拟表明该电路具有较高的精度和稳定性,带隙基准的输出电压为1．254V,在2．7V-5．5V电源电压范围内基准随输人电压的最大偏移为0．012mV,基准的最大静态电流约为11．27μA;当温度-40℃-120℃范围内,基准温度系数为1mV;在电源电压为3．6V时,基准的总电流约为10．6μA,功耗约为38．16μW;并且基准在低频时具有100dB以上的电源电压抑制比（PSRR）,基准的输出启动时间约为39μs．     

Support for a synaptic chain model of neuronal sequence generation

In songbirds, the remarkable temporal precision of song is generated by a sparse sequence of bursts in the premotor nucleus HVC. To distinguish between two possible classes of models of neural sequence generation, we carried out intracellular recordings of HVC neurons in singing zebra finches (Taeniopygia guttata). We found that the subthreshold membrane potential is characterized by a large, rapid depolarization 5-10 ms before burst onset, consistent with a synaptically connected chain of neurons in HVC. We found no evidence for the slow membrane potential modulation predicted by models in which burst timing is controlled by subthreshold dynamics. Furthermore, bursts ride on an underlying depolarization of ～10-ms duration, probably the result of a regenerative calcium spike within HVC neurons that could facilitate the propagation of activity through a chain network with high temporal precision. Our results provide insight into the fundamental mechanisms by which neural circuits can generate complex sequential behaviours.