快速老化小鼠海马神经元电压门控离子通道特点

目的:观察快速老化小鼠(Senescence-accelerated mouse,SAM)海马神经元的基本离子通道特点,并对抗快速老化亚系(SAM-resistance/1,SAMR1)与快速老化亚系(SAM-prone/8,SAMP8)的基本离子通道特点进行了比较,探讨离子通道变化在衰老中的可能角色.方法:应用全细胞记录方式,观察并比较原代培养SAMR1和SAMP8海马神经元的电压门控离子通道及膜参数.结果:原代培养SAMR1和SAMP8海马神经元电压门控Na+通道电流(INa)和电压门控延迟整流K+通道电流(Ik)的电学特点和幅度基本一致.SAMP8的电压门控Ca2+通道电流(ICa)和瞬时外向K+通道电流(IA)的幅值则大于相同培养天数的SAMR1.经膜电容校正所得的ICa电流密度也表现出增大的变化规律.结论:SAMP8与SAMR1神经元间IA和ICa的差异可能与其神经系统变异而产生的学习记忆功能下降有关.     

快速老化模型小鼠学习记忆行为及有关脑区单胺递质水平的增龄性变化

目的 研究快速老化模型小鼠(senescence accelerated mice，SAM)学习记忆能力及其大脑皮层、海马和下丘脑单胺递质含量的增龄性变化及它们之间的关系。方法 分别采用跳台实验和穿梭箱实验测定SAM的被动和主动回避反应能力，采用高效液相色谱电化学检测法测定脑内单胺递质的含量。结果 2月龄快速老化亚系SAM-prone/8(SAMP8)的被动和主动回避反应能力已较同龄抗快速老化亚系SAM-resistance／1(SAMR1)明显降低，且其主动回避反应能力随增龄进一步降低。同时，SAMP8大脑皮层、海马及下丘脑内单胺递质水平多明显高于同龄SAMR1，且随增龄明显增高。结论 SAMP8学习记忆能力的衰退可能与其相关脑区单胺递质的变化密切相关。     

8-oxo-G在快速老化小鼠SAMP8海马中表达的增龄性变化研究

目的观察8-氧鸟嘌呤核苷(8-oxo-G)在SAMP8品系快速老化小鼠海马不同区域的表达,以探讨其与SAMP8小鼠增龄性变化的关系。方法选用1、4、8、12月龄的快速老化小鼠SAMP8以及同龄抗快速老化小鼠SAMR1(对照组),每组各6只,采用免疫组化方法检测小鼠海马不同区域8-oxo-G的表达。结果8-oxo-G主要在SAM小鼠海马神经元胞浆内表达。对照组SAMR1 1月龄小鼠海马CA1和CA3区8-oxo-G表达显著高于其他月龄组(P<0.05),4、8、12月龄小鼠间其表达差异无统计学意义(P>0.05);SAMP8 12月龄小鼠海马8-oxo-G的表达显著高于1、4、8月龄组(P<0.05);SAMP8和SAMR1小鼠之间比较,1、4月龄间差异无统计学意义(P>0.05),8、12月龄间差异有统计学意义(P<0.05)。结论SAMP8小鼠海马8-oxo-G的表达除1月龄外随月龄增加而增加,提示8-oxo-G的表达增加与SAMP8小鼠的快速老化相关,有可能为增龄,甚至AD的生物学标志。     

8-氧鸟嘌呤脱氧核苷在快速老化小鼠SAMP8海马中表达的增龄性变化研究

目的观察8-氧鸟嘌呤脱氧核苷（8-oxo-7,8-dihydroguanine,8-oxo-dG）在快速老化小鼠SAMP8海马不同区域的表达,探讨其变化与SAMP8增龄的关系。方法选用1、4、8、12月龄快速老化小鼠SAMP8（每组各6只）,对照组为同龄抗快速老化小鼠SAMR1（每组各6只）,用免疫组化法检测海马不同区域内8-oxo—dG的表达水平。结果8-oxo-dG在海马不同区域均有表达,且主要在海马神经细胞胞核内表达。对照组SAMR1小鼠海马各区域8-oxo-dG的吸光度定量结果显示各月龄组间无统计学差异（P〉0．05）;1、4月龄组SAMP8小鼠海马各区域8-oxo-dG的吸光度定量结果与同龄匹配的SAMR1小鼠间无统计学差异（P〉0．05）;8、12月龄组SAMP8小鼠海马各区域8-oxo-dG的吸光度定量结果分别显著高于1月龄和4月龄组（P〈0．05）,也分别显著高于同龄SAMP1对照组（P〈0．05）。结论8-oxo-dG在SAMP8快速老化小鼠海马中的水平随增龄而显著增高。     

快速老化痴呆模型小鼠SAMP8学习记忆能力的增龄性变化

目的对快速老化痴呆模型小鼠SAMP8学习记忆能力的增龄性变化进行较系统的研究,为利用该模型进行其他研究提供实验依据。方法此实验选用1、4、8、12月龄的快速老化痴呆模型小鼠SAMP8,与同龄的正常老化小鼠SAMR1作对照,从老化度评分、避暗实验、Morris水迷宫实验和自主活动实验等方面观察了SAMP8小鼠学习记忆能力的增龄性变化。结果与对照组SAMR1相比,SAMP8小鼠随月龄增加老化度评分呈增高趋势,在8、12月龄的老化度评分值显著高于同龄对照组(P<0.05);避暗实验中,8、12月龄的SAMP8小鼠在电击24h后进入暗箱的潜伏期比同龄SAMR1小鼠显著缩短(P<0.05);Morris水迷宫实验中,1、4月龄SAMP8小鼠找到暗台的潜伏时间与同龄SAMR1小鼠相比差异无显著性,而8、12月龄SAMP8小鼠与同龄对照组相比,潜伏时间显著延长(P<0.05);从自主活动实验看,1、4、8月龄SAMP8小鼠单位时间内自主活动次数与同龄SAMR1小鼠相比无显著变化,而12月龄SAMP8小鼠与同龄对照组相比单位时间内自主活动次数显著减少(P<0.05)。结论SAMP8小鼠随月龄增长学习记忆能力逐渐减退;与同龄对照组相比,8、12月龄SAMP8小鼠出现明显衰老特征,表现出学习记忆能力明显低下,故可作为老化痴呆的动物模型用于痴呆有关研究。     

阿尔茨海默病动物模型研究进展

阿尔茨海氏病(Alzheimer`s disease,AD)是老年人常见的痴呆类型,AD动物模型的建立是研究AD的基础,学者们从不同角度探讨了多种制备AD动物模型的方法。以老化为基础的动物模型1.自然衰老认知障碍模型AD是一个年龄相关的疾病,衰老因素在其发病过程中占有重要地位,衰老所特有的病理生理变化及其它病的影响,是用年轻动物制作的动物模型所不能替代的。常见的有灵长类如猴、狒狒[1-2]和鼠的自然衰老模型。2.衰老加速小鼠模型衰老加速小鼠(senescence accelerated mouse,SAM)是一种较理想的衰老模型,SAM分为易快速老化系SAM(senescenceacclerated mouse prone,SAM)和抗快速老化系SAM(senescenceaccelerated mouse resistance,SAM);其中SAMP有9个(P1、P2、P3、P6、P7、P8、P9、P10和P11);其中SAMR有3个(P1、P4、P5);在SAM品系中,SAMP8是以学习记忆功能衰退为特征的加速老化模型,既有自然衰老小鼠的特征,又有类似老年性痴呆的脑部病理及学习记忆功能衰退[3~5]。生后...     

大鼠背根神经节神经元H+-门控离子通道电流分型及其基因型组成

目的根据大鼠背根神经节（DRG）神经元所记录的天然H^＋-门控离子通道电流特征将其进行分型，并探讨各型天然H^＋-门控离子通道与其基因型组成的酸敏感离子通道（ASICs）亚基间的相关性。方法采用全细胞膜片钳技术记录急性分离的大鼠DRG神经元H^＋-门控离子通道电流，结合单细胞免疫组化方法检测其基因型组成的ASICs亚基。结果依据H^＋-门控离子通道电流的激活及失活动力学、电流形状特点、细胞直径大小、pH依赖性等及其它参数可将H^＋-门控离子通道电流分为T-型、S．型、B-型和O-型。测定了四种类型H^＋-门控离子通道电流的激活（10%-90％21升时间）与细胞直径大小的相关性，其中T-型、B-型和O-型三种类型的激活动力学与细胞直径有关（r=0．69，P〈0．01），而S-型电流与细胞直径无关（r=0．12，P〉0．05）。对S-型电流（pH5．0）的浓度一效应关系进行了分析，其阈值在pH6．0左右，最大浓度〉2．0；在pH4．5-2．5之间出现-外向电流，浓度-效应曲线呈钟形。提供了Hi门控离子通道电流表型与其基因型的关系：T-型为ASIC1，ASIC2a，ASIC3；S．型为ASIC2a，ASIC4；B-型为ASIC1，ASIC3；O-型为ASIC1．ASIC3和ASIC4。结论大鼠DRG神经元天然H^＋-门控离子通道随着其亚基组构的改变．其电流特征也发生改变，推测系通道内向移动离子的选择性发生改变所致：其中T-型、B-型和O-型受体的神经元分布与其胞体直径相关。     

第三讲 电压门控离子通道

在所有可兴奋细胞的细胞膜上都有许多蛋白通道。这些通道打开时 ,允许各种离子通过。其中 ,有些通道的打开由跨膜电压控制 ,称为电压门控离子通道。它们的特点是对某一种离子有特别高的通透性 ,根据离子通透的选择性可鉴别与区分离子通道。神经细胞中第一个被识别的电压门控离子通道是钠通道 (VDSC)和钾通道 (VDKC) ,它们决定着膜动作电位的变化。 Alan Hidgkin和 Andrew Huxley应用电压钳 (Voltageclamping)并结合药理学技术在枪鸟贼轴突上首先研究了钠通道和钾通道 ,证明动作电位由早期流入细胞的钠电流和晚期流出的钾电流组成。钠…     

增龄性脑血管相关改变对阿尔茨海默病模型小鼠SAMP8认知能力的影响

目的 探讨脑血流量、血脑屏障(BBB)通透性、脑葡萄糖跨膜转运蛋白表达情况随年龄增大对阿尔茨海默病(AD)模型小鼠SAMP8认知能力的影响. 方法 选择SAMP8小鼠及正常同源抗快速老化小鼠R1(SAMR1)各10只进行观察.采用Morris水迷宫测定小鼠的学习记忆能力,激光多普勒仪测定脑血流量,荧光分光光度计法测定BBB通透性,Western blotting测定葡萄糖转运蛋白(GLUT)1和3的表达. 结果 与SAMR1相比,SAMP8的认知能力早在4月龄时即已出现明显损伤,表现为思维僵化、学习过程减慢.随着年龄增大,SAMP8脑血流明显下降,BBB渗漏更为严重,皮层和海马的GLUT1和GLUT3表达也有不同程度地改变.脑血流、BBB完整性、GLUT1和GLUT3表达受年龄和品系影响明显,并与认知能力高度相关. 结论 衰老及缺血引起的血管损伤、能量供给不足是造成AD神经元功能异常及导致认知障碍的主要原因.     

硬脑膜炎性刺激对三叉神经节小直径神经元电压门控钾电流的影响

目的通过炎性介质(IM)和降钙素基因相关肽(CGRP)诱发偏头痛反复发作,运用全细胞膜片钳的方法观察大鼠三叉神经节小直径神经元电压门控性钾电流的变化。方法雄性SD大鼠15只,分为空白组(不做任何干预)、生理盐水组和IM+CGRP组(大鼠硬脑膜上埋置PE-10管,连续7 d给予等量生理盐水和IM+CGRP)。用Von Frey毛测定大鼠眶周皮肤机械痛阈。在急性分离的三叉神经节小直径神经元上,通过全细胞膜片钳方法记录延迟外向钾电流(IK)和瞬时外向钾电流(IA)的变化。结果给药7 d后,IM+CGRP组大鼠的眶周机械痛阈明显降低,生理盐水组眶周机械痛阈无明显改变。生理盐水组三叉神经节神经元膜上总钾电流、IK、IA与空白组比较无明显差异;IM+CGRP组三叉神经节神经元膜上总钾电流、IK、IA与空白组和生理盐水组比较明显减小。结论 IM和CGRP诱发偏头痛反复发作模型大鼠的三叉神经节中急性分离神经元的IK和IA明显降低,提示电压门控性钾通道可能参与了外周机械痛阈的降低。     

Whole-cell membrane current and membrane resistance during hypoxic spreading depression.

In rat hippocampal tissue slices we recorded extracellular potential (Vo) and whole-cell patch clamp current of CA1 pyramid cells. During hypoxic spreading depression (SD)-like depolarization, the holding current (Ih) increased sharply. Membrane 'slope' resistance (Rm) decreased to 10-67% (mean 39%) of the resting value. The SD-related membrane current (ISD) reversed near zero mV. With voltage dependent K+ and Na+ currents blocked by Cs+ and QX-314, shifts of Ih and decrease of Rm during SD were not suppressed. We conclude that hypoxic SD of CA1 pyramidal cells is associated with a large non-selective inward current through yet to be identified membrane mechanisms, which cannot fully explain the SD-related Vo shift.     

Spatial memory performance and hippocampal neuron number in osteoporotic SAMP6 mice

The senescence-accelerated mouse strain P6 (SAMP6) is an inbred mouse that represents a clinically relevant model of senile osteoporosis. However, whether osteoporotic SAMP6 mice have cognitive deficits remains largely unexplored. Here, we used Morris water maze to assess reference memory and working memory performance in SAMP6 mice and SAMR1 controls, at 4 and 8 months of age. In addition, unbiased stereological techniques were used to estimate total neuron number in hippocampal CA1 subfield of the mice used in the behavioral study. Morris water maze test revealed impairments in working memory but not in reference memory of the 4- and 8-month-old SAMP6 mice compared with the SAMR1 mice at the same age. However, there were no significant differences in the total numbers of neurons in hippocampal CA1 subfield when comparing 4-month-old SAMR1 and 4-month-old SAMP6 and 8-month-old SAMR1 and 8-month-old SAMP6, which indicate that, in SAMP6 mice, the structural correlates of working memory deficits are to be found in parameters other than the number of neurons in hippocampal CA1 subfield. These findings suggest that SAMP6 mice exhibit selective cognitive deficits and highlight the importance of this mouse model for studying the brain alterations associated with osteoporosis.     

Variation in the pattern of [Ca2+]i change induced by acetylcholine in cultured hippocampal neurons

Acetylcholine (ACh) caused various patterns of change in the intracellular Ca2+ concentration ([Ca2+]i) in cultured rat hippocampal neurons. We studied the underlying mechanisms of the [Ca2+]i changes with simultaneous recording of [Ca2+]i and membrane potential/current. In most cases, [Ca2+]i rise was accompanied by a membrane depolarization. The [Ca2+]i change was significantly reduced when the membrane was voltage clamped, which implies that most of the [Ca2+]i rise results from the Ca2+ influx through the voltage-gated Ca2+ channel activated by the membrane depolarization. The membrane depolarizations were classified into two types, one associated with membrane conductance decrease and the other associated with membrane conductance increase. The former results from potassium conductance ((gK+) decrease, and the latter may result from the activation of a Na(+)-permeable channel. However, [Ca2+]i elevation was also observed in some neurons showing membrane hyperpolarization in response to ACh. This seems to show that ACh liberates Ca2+ from the intracellular Ca2+ store, resulting in the activation of a calcium-dependent K+ channel (KCa). The variations of ACh response in the hippocampal neurons seem to result from a variety of muscarinic acetylcholine receptors and various species of ion channels governed by those receptors.     

Ion currents and spiking properties of identified subtypes of locust octopaminergic dorsal unpaired median neurons

Efferent dorsal unpaired median (DUM) neurons are key elements of an insect neuromodulatory system. In locusts, subpopulations of DUM neurons mediate octopaminergic modulation at specific targets depending on their activity during different behaviours. This study investigates whether in addition to synaptic inputs, activity in DUM neurons depends on intrinsic membrane properties. Intracellular in situ recordings and whole-cell patch-clamp recordings from freshly isolated somata characterize somatic voltage signals and the underlying ion currents of individual subtypes of DUM neurons identified beforehand by a vital retrograde tracing technique. Na(+), Ca(2+), K(+) currents and a hyperpolarization-activated (I(h)) current are described in detail for their (in-)activation properties and subtype-specific current densities. In addition, a Ca(2+)-dependent K(+) current is demonstrated by its sensitivity to cadmium and charybdotoxin. This complex current composition determines somatic excitability similar in all subtypes of DUM neurons. Both Na(+) and Ca(2+) currents generate overshooting somatic action potentials. Repolarizing K(+) currents, in particular transient, subthreshold-activating A-currents, regulate the firing frequency and cause delayed excitation by shunting depolarizing input. An opposing hyperpolarization-activated (I(h)) current contributes to the resting membrane potential and induces rebound activity after prolonged inhibition phases. A quantitative analysis reveals subtype-specific differences in current densities with more inhibitory I(K) but less depolarizing I(Na) and I(h) - at least in DUM3 neurons promoting a reliable suppression of their activity as observed during behaviour. In contrast, DUM neurons that are easily activated during behaviour (DUM3,4,5 and DUMETi) express less I(K) and a pronounced depolarizing I(h) promoting excitability.     

The neuropeptide egg-laying hormone modulates multiple ionic currents in single target neurons of the abdominal ganglion of Aplysia

The bag cell neurons of the abdominal ganglion of Aplysia are a useful system for the study of peptidergic neurotransmission. A 20 min burst of impulse activity in the bag cells induces or augments repetitive firing in LB and LC neurons in the abdominal ganglion for up to several hours. Previous experiments have indicated that this effect is mediated by the putative bag cell transmitter egg-laying hormone (ELH). Using voltage-clamp analysis we found that bag cell bursts (BCBs) evoke long-lasting changes in membrane current in these neurons that are mimicked by the application of ELH. The combined ELH-evoked current is inward at all membrane potentials between -110 and -10 mV and consists of 3 separable currents persisting for 30-120 min. They include (1) a depolarizing current that is activated at membrane potentials above -40 mV. This current, termed ISI, is blocked by prolonged exposure to 10 mM Ni2+/0 mM Ca2+ and is not abolished by 0 mM Na+ or 100 mM TEA+/0 mM Na+ in the bathing medium. It is therefore a Ca2+-sensitive current and does not involve Na+ as a charge carrier. (2) There is a hyperpolarizing current that is activated at membrane potentials below approximately -70 mV. This current, termed IR, is blocked by external Rb+ (5 mM) and Cs+ (10 mM) and has a chord-conductance that shifts with the external [K+] according to the Nernst potential for potassium. It is therefore an inwardly rectifying K+ current. (3) There is a small, steady depolarizing current, termed Ix. This current is the only one that remains after prolonged exposure to 10 mM Ni2+/0 mM Ca2+-containing bathing medium. It is Na+ dependent and is associated with a small increase in membrane conductance that is largely independent of membrane voltage. All 3 currents are slow to inactivate; they appear to sum algebraically to produce the net BCB- or ELH-evoked current.     

Analysis of the expression of endogenous murine leukemia viruses in the brains of senescence-accelerated mice (SAMP8) and the relationship between expression and brain histopathology

Many studies have explored the premature aging of accelerated senescence-prone (SAMP8) mice. However, the cause of premature aging in this strain remains unknown. We analyzed the expression of ecotropic, xenotropic, and polytropic murine leukemia viruses (MuLVs) in the brains of accelerated senescence-resistant (SAMR1) and SAMP8 mice. No ecotropic mRNA was detected in SAMR1 mice, and only Akv-type ecotropic MuLV mRNA was detected in SAMP8 mice. Restriction mapping of the full-length infectious E-MuLV genome from SAMP8 confirmed its identity as Akv. mRNAs corresponding to a prototypical polytropic MuLV and to an unusual xenotropic MuLV were detected at equal levels in SAMP8 and SAMR1 mice, but no infectious virus of either host range type was detected. In order to determine the cellular localization of Akv expression in SAMP8 mice, we used immunohistochemistry and electron microscopy to detect expression of the E-MuLV capsid gag (CAgag) gene in striatum, brainstem, hippocampus, and cerebellum of 12-month-old SAMR1 and SAMP8 mice. The CAgag antigen was seen in the neurons, oligodendroglia, and vascular endothelium of these brain regions of SAMP8 mice, but not in SAMR1 mice. To evaluate the correlation between activation of astrocytes and expression of Akv, we performed double-immunohistochemical staining for both glial fibrillary acidic protein (GFAP) and CAgag in SAMR1 and SAMP8 mice. Strong astrocytic activation and extensive vacuolation were observed around CAgag-positive neurons in SAMP8 mice, whereas in SAMR1 mice neither astrocytosis nor vacuolation were present. CAgag antigen was also localized in astrocytes of the hippocampus region of SAMP8 mice. Electron micrography showed that a number of vacuoles were found in the cytoplasm of MuLV-positive neurons and the extracellular space surrounding these neurons showed lytic changes. These results suggest that endogenous Akv provirus is expressed in neurons, astrocytes, vascular endothelium, and oligodendroglia in the brains of SAMP8 and that this virus could play an important role in the brain aging processes in this mouse strain.     

Expression of ion channels and mutational effects in giant Drosophila neurons differentiated from cell division-arrested embryonic neuroblasts.

A culture system of "giant" Drosophila neurons derived from cytokinesis-arrested embryonic neuroblasts was developed to overcome the technical difficulties usually encountered in studying small Drosophila neurons. Cytochalasin B-treated neuroblasts differentiated into giant multinucleated cells that displayed neuronal morphology and neuron-specific markers (Wu et al., 1990). Here, we report that these giant neurons express different excitability patterns and membrane channels similar to those reported in excitable tissues of Drosophila. Individual neurons exhibited distinct all-or-none or graded voltage responses upon current injection. Both current- and voltage-clamp recordings could be performed on the same neuron because of the large cell size, thus making it possible to elucidate the functional role of the individual types of channels. By using pharmacological agents and ion substitution, the following currents were identified in these giant neurons: inward Na+ and Ca2+ currents and outward voltage-activated (the A-type and delayed rectifier) and Ca(2+)-activated K+ currents. In addition, we found a tetrodotoxin (TTX)-sensitive, Na(+)-dependent outward K+ current and a persistent component of an inward Na+ current, which have not been reported in Drosophila previously. This culture system can be used to analyze the mutational perturbations in ion channels and the resultant alterations in membrane excitability. Neurons from the mutant slowpoke (slo), which is known to lack a component of the Ca(2+)-activated K+ currents in muscles, exhibited prolonged action potentials associated with defects in the Ca(2+)-activated K+ current. This abnormality appeared to be more severe in the neurites than in the soma.     

Sodium conductance in cultured myenteric AH-type neurons from guinea-pig small intestine

Whole-cell patch clamp methods were used to investigate sodium conductance in after-hyperpolarization-type (AH) enteric neurons in culture after dissociation from the myenteric plexus of guinea-pig small intestine. Inward current carried by Na+ (I(Na)) was identified and its current-voltage characteristics were compared with those for inward Ca2+ current (I(Ca)). The I(Na) current was a rapidly inactivating current relative to I(Ca). Application of tetrodotoxin (TTX) blocked I(Na) with an EC50 of 10.7 nM. Activation curves for I(Na) showed a rapid decrease in time to peak for test potentials from holding potentials of -80 mV to between -40 and -10 mV. Voltage-dependence of steady-state inactivation curves for I(Na) was fit to the Boltzmann equation with potential for half-inactivation (V(1/2)) = -55.6 mV and slope factor (k) = 6.4 mV. Steady-state inactivation for I(Ca) fit the Boltzmann equation with a V(1/2) = -38.9 mV and k= 14.4 mV. Kinetics for inactivation of I(Na) were voltage dependent at potentials between -70 and -30 mV and accelerated and became less voltage-dependent at more positive potentials. The time constant (tau) for inactivation at -70 mV was tau = 161 +/- 23 ms and decreased to tau = 2.3 +/- 0.2 ms at -30 mV. Rapid acceleration of inactivation occurred between -50 and -40 mV. This was also the range where activation began. Recovery from inactivation with the membrane potential clamped at -100 or -80 mV was rapid and fit by a single exponential with tau = 7.3 +/- 1.1 ms for -100 mV and 21.5 +/- 5.1 ms for -80 mV. The results suggest that AH-type enteric neurons have only one type of Na+ channel that behaves like the "classical" voltage-gated tetrodotoxin-sensitive fast channel. The findings support the hypothesis that I(Na) current is an important factor in determination of excitability and firing behavior in AH neurons. I(Na) and I(Ca) together determine the properties of the rising phase of the spike and thereby contribute to global determinants of excitability as the neurons are exposed to multiple depolarizing and hyperpolarizing stimuli from synaptic inputs and mediators released from enteroparacrine cells.     

Potassium-induced enhancement of persistent inward current in hippocampal neurons in isolation and in tissue slices

Previous work suggested a role for the voltage-dependent persistent sodium current, I(Na,P), in the generation of seizures and spreading depression (SD). Ordinarily, I(Na,P) is small in hippocampal neurons. We investigated the effect of raising external K(+) concentration, [K(+)](o), on whole-cell persistent inward current in freshly isolated hippocampal CA1 pyramidal neurons. I(Na,P) was identified by TTX-sensitivity and dependence on external Na(+) concentration. When none of the ion channels were blocked, I(Na,P) was not usually detectable, probably because competing K(+) current masked it, but after raising [K(+)](o) I(Na,P) appeared, while K(+) currents diminished. With K(+) channels blocked, I(Na,P) could usually be evoked in control solution and raising [K(+)](o) caused its reversible increase in most cells. The increase did not depend on external calcium [Ca(2+)](o). In CA1 pyramidal neurons in hippocampal slices a TTX-sensitive persistent inward current was always recorded and when [K(+)](o) was raised, it was reversibly enhanced. Strong depolarization evoked irregular current fluctuations, which were also augmented in high [K(+)](o). The findings support a role of potassium-mediated positive feedback in the generation of seizures and spreading depression.