大鼠背根神经节神经元的分离方法及电生理特征探讨

在获取大鼠背根神经节神经元后,全细胞膜片钳技术记录动作电位和钠电流,探讨大鼠背根神经节细胞的分离方法和细胞形态以及电生理特征.结果显示,本实验能得到完整圆形或椭圆长条形的大鼠背根神经节体.正常的单个背根神经节神经元呈圆形或椭圆形,大小不等,胞膜清晰,折光性好,隐约可见细胞核.在背根神经节细胞上记录的动作电位呈正立锐角三角形,静息电位小,动作电位时程短.背根神经节神经元的钠通道最大电流密度为(-62.04±4.45)pA/pF,几乎能被河豚毒素(TTX)完全抑制.本实验分离方法简单易行,背根神经节神经元容易获得和辨认,电生理特征明确,适用于神经系统疾病的电生理特征研究和治疗药物观察.     

碘化N-正丁基氟哌啶醇对大鼠缺氧复氧心肌细胞钠钙交换体电流的抑制作用

碘化N-正丁基氟哌啶醇(N-n-butyl haloperidol iodide,F2)为本研究室改造合成的新化合物。前期研究发现F2作为L-型钙通道拮抗剂,能剂量依赖地拮抗缺血再灌注所导致的大鼠心脏损伤。研究F2对缺氧复氧(hypoxia/reoxygenation,H/R)大鼠心肌细胞钠钙交换体电流的作用并探讨其保护机制。采用Langendorff灌流系统灌流SD大鼠心脏,标准酶解法消化分离得到单个心室肌细胞。正常台式液灌流5min,立即灌流充90%N2-10%CO2的缺氧液,建立体外心肌细胞H/R模型,采用全细胞膜片钳技术记录对照、模型以及不同浓度F2(0.1、1、10μmol/L)对心肌细胞钠钙交换体电流,观察H/R状态F2对心肌细胞钠钙交换体电流的影响。结果显示:缺氧抑制钠钙交换体电流主要是抑制外向电流;H/R引起钠钙交换体电流增大,尤其是外向电流的增大。F2呈浓度依赖地抑制钠钙交换体电流,钠钙交换体电流I-V曲线上移。以上表明:F2能抑制钠钙交换体电流,尤其是外向电流,防止H/R时心肌细胞的钙超载,保护心肌细胞。     

基于MEMS技术的微电极阵列细胞传感器

经典的细胞电生理记录方法的缺点是难以实现长时程测量,同时也难以实现对细胞网络的多位点同时测量.近年来,随着胞外电生理研究的不断深入和半导体微机械加工技术的发展,传感器件的微型化和集成化为细胞的微环境测试和电生理研究提供了强有力的手段.基于多年来在细胞芯片上的研究,介绍了用于细胞外电生理测试的微电极阵列(microelectrode array,MEA),分析其原理、设计及加工过程;并对心肌细胞在其表面生长情况和电信号传递过程进行八通道并行动作电位检测;最后对所测得的结果加以讨论.实验证明,该细胞芯片可对多个细胞同时进行长期、无损的胞外电信号测量,制备简单、使用方便、可进一步应用于细胞电生理研究、药物检测等领域.     

小鼠海马神经元急性分离和膜片钳技术的应用

为了获得适用于膜片钳实验技术的单个海马神经元,应用酶消化及机械分离法,急性分离出生10～13d的昆明小鼠得到单个海马神经元通过倒置显微镜直接观察和全细胞膜片钳技术分别对分离得到的海马细胞的形态学和电生理学特征进行研究．通过实验可得,急性分离所得活性较好的海马神经元胞体具有锥形胞体和顸树突特征,立体感强．在全细胞膜片钳记录模式下可记录到全细胞电流及钠、钾通道电流．由实验结果可知,该方法可以得到形态和生理特征良好的单个海马神经元,证实该方法适用于膜片钳技术的研究,对深入探讨药物、物理因子等对海马离子通道及信号转导机制的作用有重要的应用价值．     

30mT恒定磁场可延长神经细胞的存活时间

为了研究中等强度恒定磁场对神经细胞存活能力的影响,采用急性分离的小鼠额叶皮层锥体神经细胞暴露于30.mT恒定磁场中,应用全细胞膜片钳技术研究神经元钠离子通道电流的记录时间(细胞形成全细胞模式后的存活时间,反映细胞存活能力).实验发现,30.mT恒定磁场作用中的细胞钠通道电流的平均记录时间较未被磁场作用的细胞钠电流的平均记录时间显著增加,分别为33.77.min和15.13.min.在磁场作用中的部分神经细胞即使在全细胞模式形成60.min后仍然有20%的神经细胞可以记录到很好的钠通道电流曲线,而对照组神经细胞在形成全细胞模式后超过40.min细胞就已经失去活性.结果表明,30.mT恒定磁场可一定程度上延长皮层神经细胞钠离子通道电流的存活时间,增强了神经细胞在恶劣环境下的存活能力.     

鹌鹑心肌细胞动作电位参数的测量分析

本文采用浮置微电极技术,记录了鹌鹑在体心肌细胞的动作电位,并用相平面轨迹法对其有关参数进行了分析,摄制了心肌细胞电生理信号的相平面轨迹图.     

兔窦房结细胞的分离方法及其起搏电流特征性探讨

 探讨了家兔窦房结(Sino-Atrial Node,SAN)细胞的分离方法及其形态和起搏电流(Funny Current,I_f)的特征.通过打结定位法确定了带有"中心结区"的家兔SAN组织条,双酶法分离SAN细胞,利用全细胞穿孔膜片钳技术记录SAN细胞的I_f.结果显示,SAN组织为上宽下窄的枣核型,区域体积约为6.25mm×2.23mm×0.26mm,或6mm×2mm×0.1mm—6mm×2mm×3.3mm.SAN细胞成狭长梭形,无清晰横纹,细胞长(65.3±7.8)μm,宽为(7.2±3.1)μm.在正常Tyrode液中,可恢复规则自律性搏动,频率为(187.3±5.6)次/min,典型SAN细胞数量占整个分离细胞的2%—5%.根据电压钳制方案,I_f电流被引出,呈内向性斜坡状,随着钳制电压降低,起搏电流逐渐增大,并随钳制时间延长,倾斜度也增大.I_f带有明显尾电流,也随着超级化激活电位降低,逐渐增大.根据二者电流大小拟合的电流-电压曲线,呈典型横向Y形.本法获得SAN细胞形态典型,状态和活性良好,能够记录具有特征性的I_f,可供心血管领域有关针对SAN细胞I_f研究的技术借鉴.     

膜片钳全细胞记录法在研究谷氨酰胺转运蛋白分子机制中的应用

为了研究中性氨基酸转运蛋白-谷氨酰胺转运蛋白的结构与功能,采用膜片钳全细胞记录法测定谷氨酰胺转运蛋白2介导的电流.结果表明:谷氨酰胺转运蛋白诱导了底物氨基酸和Na+浓度依赖性电流,可用于测定底物氨基酸和Na+对转运蛋白的表观亲和常数Km,SNAT2野生型对丙氨酸的表现亲和常数KmAla为(200±5)×10-6mol/L,对Na+的表观亲和常数KmNa为(100±7)×10-3mol/L.谷氨酸转运蛋白转运底物氨基酸过程是生电的过程,在膜电势负值时氨基酸转运电流增大,可用于转运机制的研究.因此膜片钳全细胞记录法是研究谷氨酰胺转运蛋白分子机制的有效方法.     

快速复极化电流对心肌细胞电生理特征的影响

针对增大快速复极化电流对心肌细胞动作电位、钾离子浓度及电生理特征的影响问题,沿用毕勒-路特心肌细胞数理模型,以一阶龙格-库达法进行计算,研究心肌细胞外加刺激使动作电位、钾离子浓度呈现周期性,结果更具有实际意义.研究结果表明,增大复极化电流使动作电位短期内出现震荡,钾离子电流密度初期减小,心肌细胞兴奋性、传导性、自律性降低,进而诱发心律失常.     

多磷酸肌醇对细胞膜钾通道协同作用的比较研究

本工作用膜片钳全细胞电流记录技术,研究了各种多磷酸肌醇在小白鼠泪腺单细胞的内外灌流实验中对膜钙依赖性钾通道的激活作用,证明了(1,4,5)三磷酸肌醇和(1,3,4,5)四磷酸肌醇具有协同激活作用,并比较了多种磷酸肌醇联用时的特异性。     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with     

Journal of Donghua University ( English Edition) Contents of Volume 31

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