新生大鼠神经干细胞分离培养及缺氧损伤模型的建立

目的建立缺氧损伤模型,观察缺氧损伤对神经干细胞分化的影响。方法用MTT,LDH等方法比较95%N2,5%CO2比率的缺氧气体分别培养1,2,4,6,8 h神经干细胞的变化。结果三天内的大鼠海马存在大量神经干细胞,可分化为神经元、胶质细胞等神经细胞类型,95%N2,5%CO2比率的缺氧气体培养6 h后神经干细胞OD值由缺氧前0.369±0.352降至0.270±0.229,LDH 漏出量由缺氧前10.374±0.390 unit/ml.min增加到14.710±1.337 unit/ml.min,差异显著。结论证实了新生大鼠海马内提取的细胞为神经干细胞,神经干细胞缺氧培养6 h可以建立神经干细胞缺氧损伤模型,但短时间缺氧培养对细胞分化类型无明显改变。     

An acetylcholinesterase inhibitor, 3-benzidino-5-methyl-6-phenylpyridazine, blocking outward potassium currents in acutely isolated rat hippocampal pyramidal neurons

3-benzidino-5-methyl-6-phenylpyridazine (BMP) inhibited electric eel acetylcholinesterase (AChE), with IC50 being 0.58 μmol·L-1. As an AChE inhibitor, the effects of BMP on delayed rectifier potassium current (IK(DR)) and transient outward potassium current (IK(A)) in acutely isolated rat hippocampal pyramidal neurons were studied using the whole cell patch-clamp technique. BMP (0.1―50 μmol·L-1) inhibited IK(DR) and IK(A) in a concentration-dependent and voltage-independent manner. The IC50 value for the bl...     

Ethanol impairs Rho GTPase signaling and differentiation of cerebellar granule neurons in a rodent model of fetal alcohol syndrome

Developmental exposure to ethanol impairs fetal brain development and causes fetal alcohol syndrome. Although the cerebellum is one of the most alcohol-sensitive brain areas, signaling mechanisms underlying the deleterious effects of ethanol on developing cerebellar granule neurons (CGNs) are largely unknown. Here we describe the effects of in vivo ethanol exposure on neurite formation in CGNs and on the activation of Rho GTPases (RhoA and Rac1), regulators of neurite formation. Exposure of 7-day-old rat pups to ethanol for 3 h moderately increased blood alcohol concentration (BAC) (∼40 mM) and inhibited neurite formation and Rac1 activation in CGNs. Longer exposure to ethanol for 5 h resulted in higher BAC (∼80 mM), induced apoptosis, inhibited Rac1, and activated RhoA. Studies demonstrated a regulatory role of Rho GTPases in differentiation of cerebellar neurons, and indicated that ethanol-associated impairment of Rho GTPase signaling might contribute to brain defects observed in fetal alcohol syndrome. Received 16 July 2006; received after revision 12 September 2006; accepted 13 October 2006     

慢病毒介导的CBS基因过表达对原代皮质神经元β淀粉样蛋白分泌的影响

目的通过过表达手段上调胱硫醚β-合成酶(Cystathionine-β-synthase,CBS)在原代神经元细胞中的表达,观察其对原代神经元β淀粉样蛋白(Aβ)分泌的影响。方法使用过表达CBS慢病毒载体感染体外培养的原代皮质神经元,设定为慢病毒介导阳性(CBS)组、另设阴性对照慢病毒感染(NC)组、未经慢病毒感染(CON)组。采用实时荧光定量PCR检测CBS基因mRNA的表达,Western blot检测CBS蛋白的表达,采用Elisa检测Aβ40的表达水平。结果使用过表达CBS的慢病毒载体感染体外培养的神经元,实时荧光定量PCR检测其mRNA的表达上调明显升高,与NC组和CON组相比差异有统计学意义(P0.01),Western blot结果显示蛋白表达升高与定量PCR一致,Elisa检测感染72小时后CBS组、NC组、CON组Aβ40的分泌分别为(182.19±24.52)ng/L、(411.90±12.43)ng/L、(390.41±54.53)ng/L,CBS组与NC组和CON组相比差异有统计学意义(P0.05)。结论慢病毒介导的CBS基因过表达可降低原代皮质神经元Aβ40的分泌。     

Rho激酶影响大鼠海马神经元突起生长的实时成像

目的:观察Rho激酶对大鼠海马神经元突起生长的影响.方法:体外培养新生大鼠海马神经元5 d后,连续动态观察溶血性磷脂酸(lysophosphatidic acid,LPA)及Rho激酶抑制剂Y-27632对神经元突起生长的影响.结果:对照组神经元一级突起迅速生长、延伸,不断发分支,形成丰富的二级、三级突起;用LPA处理后,神经元大部分突起进行性塌陷,一级突起逐渐缩短并且变细,二级、三级突起数量减少,且较细小;而预先用Y-27632处理后再加LPA,细胞突起塌陷的现象减少,神经元的突起不断分支、延伸,一级、二级、三级突起数量较LPA组增多且长度也增加.结论:Rho激酶参与LPA诱导海马神经元突起回缩的过程,抑制Rho激酶的活性可抑制LPA诱导的突起回缩.     

Detecting the breakup of spiral waves in small-world networks of neurons due to channel block

The breakup of a spiral wave by blockade of sodium and potassium channels in a small-world network of Hodgkin-Huxley neurons is investigated in detail.The influence of ion channel block in poisoned excitable membrane patches of a certain size is measured,by varying channel noise and channel densities resulting from the change in conductance,For example,tetraethylammonium is known to cause a block(poisoning) of potassium channels,while tetrodotoxin blocks sodium channels.We observed the occurrence of spiral waves,which are ordered waves believed to play an important role in facilitating the propagation of electric signals across quiescent regions of the brain.In this paper,the effect of channel block was measured by the factors xK and xNa,which represent the ratios of unblocked,or active,ion channels,to the overall number of potassium or sodium ion channels,respectively.To quantify these observations,we use a simple but robust synchronization measure,which succinctly captures the transition from spiral waves to other collective states,such as broken segments resulting from the breakup of the spiral wave.The critical thresholds of channel block can be inferred from the abrupt changes occurring in plots of the synchronization measure against different values of xK and xNa.Notably,small synchronization factors can be tightly associated with states where the formation of spiral waves is robust to mild channel block.     

大鼠海马锥体神经元外向电流特征分析

利用全细胞膜片钳技术研究了大鼠海马锥体神经元去极化激活的外向电流的特征.结果表明,短时去极化至-60mV以上电位可引发快速上升的外向电流,之后缓慢衰减至一平台.当保持电位改变时,该峰电流与平台电流的幅度均会发生变化,但前者较后者变化显著.试验中测得峰电流与平台电流的逆转电位分别为-(76.1±5.97)mV和-(83.6±4.13)mV,与用Nerst方程计算出的本试验细胞外液与电极内液的钾离子平衡电位Ek=-88mV接近,说明该外向电流是钾电流,且提示此外向电流包括两种成分.试验结果还表明,此外向电流的衰减过程可用双指数方程很好地拟合,而在500ms预脉冲刺激后再经去极化激活的快成分的失活过程则适合用单指数方程进行拟合,从而进一步证实大鼠海马锥体神经元的外向钾电流包括快慢两种成分(IA和IK).IA的稳态激活和失活过程均可用Boltzmann方程进行拟合,半数激活和半数失活电压分别为(7.40±3.01)mV和-(66.27±3.62)mV,但其稳态失活在测试电压范围内(-100mV至+30mV)不完全.     

Neuro-protective effects of CNTF on hippocampal neurons via an unknown signal transduction pathway

Stress is one of the leading contributing factors for psychosomatic diseases of modern society. Prolonged or strong stress may cause more release of glutamic acid (Glu) transmitter from hippocampal neurons. As most hippocampal neurons are glutaminergic ne…     

读心的模拟说进路

本文从几个方面考察了日常生活中的读心能力实现的主要进路之一模拟说,交待了模拟说作为理论说的对立方出现的背景,复制策略、最小努力原则和离线机制三种模拟形式,几组支持模拟说的有力的实验证据,以及模拟说在遭到质疑时的各种辩护,说明了读心过程和读物过程存在巨大差异,模拟说对于读心研究而言的确存在诸多理论说所不可比拟的解释优势,但读心始终是一个不可将理论因素完全排除在外的过程,它容纳模拟与理论甚至更多进路的结合。