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Quantitative evaluation of extrinsic factors influencing electrical excitability in neuronal networks： Voltage Threshold Measurement Method(VTMM)

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Neural Regeneration Research 2018年第6期13卷 1026-1035页

作者： Shuai An Yong-Fang Zhao Xiao-Ying Lu Zhi-Gong Wang State Key Laboratory of Bioelectronics Southeast University Nanjing Jiangsu Province China Institute of RF- & OE-ICs Southeast University Nanjing Jiangsu Province China Co-innovation Center of Neuroregeneration Nantong University Nantong Jiangsu Province China

The electrical excitability of neural networks is influenced by different environmental factors. Effective and simple methods are required to objectively and quantitatively evaluate the influence of such factors, including variations in temperature and pharmaceutical dosage. The aim of this paper was to introduce ‘the voltage threshold measurement method＇, which is a new method using microelectrode arrays that can quantitatively evaluate the influence of different factors on the electrical excitability of neural networks. We sought to verify the feasibility and efficacy of the method by studying the effects of acetylcholine, ethanol, and temperature on hippocampal neuronal networks and hippocampal brain slices. First, we determined the voltage of the stimulation pulse signal that elicited action potentials in the two types of neural networks under normal conditions. Second, we obtained the voltage thresholds for the two types of neural networks under different concentrations of acetylcholine, ethanol, and different temperatures. Finally, we obtained the relationship between voltage threshold and the three influential factors. Our results indicated that the normal voltage thresholds of the hippocampal neuronal network and hippocampal slice preparation were 56 and 31 m V, respectively. The voltage thresholds of the two types of neural networks were inversely proportional to acetylcholine concentration, and had an exponential dependency on ethanol concentration. The curves of the voltage threshold and the temperature of the medium for the two types of neural networks were U-shaped. The hippocampal neuronal network and hippocampal slice preparations lost their excitability when the temperature of the medium decreased below 34 and 33°C or increased above 42 and 43°C, respectively. These results demonstrate that the voltage threshold measurement method is effective and simple for examining the performance/excitability of neuronal networks.

关键词： nerve regeneration threshold voltage microelectrode array electrical excitability of neural networks acetylcholine alcohol temperature hippocampal neuronal network hippocampal slice electrical stimulation action potentials neural regeneration

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GABA_A Receptor Activity Suppresses the Transition from Interictal to Ictal Epileptiform Discharges in Juvenile Mouse Hippocampus

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Neuroscience Bulletin 2018年第6期34卷 1007-1016页

作者： Yan-Yan Chang Xin-Wei Gong Hai-Qing Gong Pei-Ji Liang Pu-Ming Zhang Qin-Chi Lu Department of Neurology Ren Ji Hospital School of Medicine Shanghai Jiao Tong University School of Biomedical Engineering Shanghai Jiao Tong University

Exploring the transition from inter-ictal to ictal epileptiform discharges(IDs) and how GABAAreceptormediated action affects the onset of IDs will enrich our understanding of epileptogenesis and epilepsy *** used Mg2+-free artificial cerebrospinal fluid(ACSF) to induce epileptiform discharges in juvenile mouse hippocampal slices and used a micro-electrode array to record the discharges. After the slices were exposed to Mg2+-free ACSF for 10 min–20 min, synchronous recurrent seizurelike events were recorded across the slices, and each event evolved from inter-ictal epileptiform discharges(IIDs) to pre-ictal epileptiform discharges(PIDs), and then to *** the transition from IIDs to PIDs, the duration of discharges increased and the inter-discharge interval decreased. After adding 3 lmol/L of the GABAAreceptor agonist muscimol, PIDs and IDs disappeared, and IIDs remained. Further, the application of 10 lmol/L muscimol abolished all the epileptiform discharges. When the GABAAreceptor antagonist bicuculline was applied at 10 lmol/L, IIDs and PIDs disappeared, and IDs remained at decreased intervals. These results indicated that there are dynamic changes in the hippocampal network preceding the onset of IDs, and GABAAreceptor activity suppresses the transition from IIDs to IDs in juvenile mouse hippocampus.

关键词： Epileptiform discharge Gamma-aminobutyric acid Bicuculline Muscimol Micro-electrode array hippocampal slice

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Duplicate preconditioning with sevoflurane in vitro improves neuroprotection in rat brain via activating the extracellular signal-regulated protein kinase

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Neuroscience Bulletin 2010年第6期26卷 437-444页

作者：王胜代志刚董希玮郭素香刘扬王志萍曾因明新疆石河子大学医学院第一附属医院麻醉科石河子832008 江苏省麻醉学重点实验室徐州医学院徐州221002

Objective Sevoflurane preconditioning has been demonstrated to reduce cerebral ischemia–reperfusion（IR） injury,but the underlying mechanisms have not been fully ***,different protocols would usually lead to different *** objective of this study was to determine whether dual exposure to sevoflurane improves the effect of anesthetic preconditioning against oxygen and glucose deprivation（OGD）injury in *** Rat hippocampal slices under normoxic conditions（95%O2/5%CO2）were pre-exposed to sevoflurane 1,2 and 3 minimum alveolar concentration （MAC）for 30 min,once or twice,with 15-min washout after each *** slices were then subjected to 13-min OGD treatment（95%N2/5%CO2,glucose-free）,followed by 30-min *** population spikes（PSs）were recorded in the CA1 region of rat *** percentage of PS amplitude at the end of 30-min reoxygenation to that before OGD treatment was calculated,since it could indicate the recovery degree of neuronal *** addition,to assess the role of mitogen-activated protein kinases（MAPKs）in preconditioning,U0126,an inhibitor of extracellular signal–regulated protein kinase（MEK-ERK1/2,ERK1/2 MAPK）,and SB203580,an inhibitor of p38 MAPK,were separately added 10 min before sevoflurane *** Preconditioning once with sevoflurane 1,2,and 3 MAC increased the percentage of PS amplitude at the end of 30-min reoxygenation to that before OGD treatment,from（15.13±3.79）%（control）to（31.88±5.36）%, （44.00±5.01）%,and（49.50±6.25）%,respectively,and twice preconditioning with sevoflurane 1,2,and 3 MAC increased the percentage to（38.53±4.36）%,（50.74±7.05）%and（55.86±6.23）%,*** effect of duplicate preconditioning with sevoflurane 3 MAC was blocked by U0126[（16.23±4.62）%].Conclusion Sevoflurane preconditioning can induce neuroprotection against OGD injury in vitro,and preconditioning twice enhances this ***,the activation of extracellular signal–regulated protein kinase（MEK-ER

关键词： electrophysiology hippocampal slice oxygen and glucose deprivation neuronal damage sevoflurane preconditioning mitogen-activated protein kinases

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Dual Isoflurane-induced Preconditioning Improves Neuroprotection in Rat Brain In Vitro and the Role of Extracellular Signal-regulated Protein Kinase

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Chinese Medical Sciences Journal 2011年第1期26卷 36-42页

作者： Sheng Wang Su-xiang Guo Zhi-gang Dai Xi-wei Dong Yang Liu Shan Jiang Zhi-ping Wang Department of Anesthesiology Affiliated Hospital of Shihezi Medical School Shihezi University Shihezi 832008 China Anesthesiology Key Laboratory of Jiangsu Province Xuzhou 221002 China

Objective To test the ability of isoflurane-induced preconditioning against oxygen and glucose dep- rivation （OGD） injury in vitro. Methods Rat hippocampal slices were exposed to 1 volume percentage （vol%）, 2vo1% or 3vo1% isoflurane respectively for 20 minutes under normoxic conditions （95% O2/5% CO2） once or twice （12 slices in each group） before OGD, with 15-minute washout after each exposure. During OGD experiments, hippocampus slices were bathed with artificial cerebrospinal fluid （ACSF） lacking glucose and perfused with 95% N2 and 5% CO2 for 14 minutes, followed by a 30-minute reperfusion in normal ACSF. The CA1 population spike （PS） was measured and used to quantify the degree of neuronal function recovery after OGD. To assess the role of mitogen-activated protein kinases （MAPKs） in isoflurane preconditioning, U0126, an inhibitor of extracellular signal-regulated protein kinase （ERK1/2）, and SB203580, an inhibitor of p38 MAPK, were used before two periods of 3vol% isoflurane exposure. Results The degree of neuronal function recovery of hippocampal slices exposed to 1 vol%, 2vol%, or 3vol% isoflurane once was 41.88%±9.23%, 55.05%±11.02%, or 63.18%±10.82% respectively. Moreover, neuronal function recovery of hippocampal slices exposed to 1 vol%, 2vo1%, or 3vo1% isoflurane twice was 53.75%±12.04%, 63.50%±11.06%, or 76.25%±12.25%, respectively. Isoflurane preconditioning increased the neuronal function recovery in a dose-dependent manner. U0126 blocked the preconditioning induced by dual exposure to 3vo1% isoflurane （6.13%±1.56%, P〈0.01） and ERK1/2 activities. Conclusions Isoflurane is capable of inducing preconditioning in hippocampal slices in vitro in a dose-dependent manner, and dual exposure to isoflurane with a lower concentration is more effective in triggering preconditioning than a single exposure. Isoflurane-induced neuroprotection might be involved with ERK 1/2 activities.

关键词： electrophysiology hippocampal slice isoflurane preconditioning oxygen andglucose deprivation mitogen-activated protein kinase

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Effects of Glucocorticoid on Electrical Activities of hippocampal Neurons in vitro

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中国人民解放军军医大学学报(英文版) 1988年第2期 116-119页

作者：华少莹陈宜张

Intracellular recording of the rat hippocampal slices was established and usedto analyse the effects of glucocorticoid on resting membrane potential as well as inputresistance of the cells in CA1 and CA2 of hippocampal *** results indicate thatwithin 1-2 min the steroid hyperpolarized some of the hippocampal neurons with atendency of increasing the cells’ input resistance,suggesting that glucocorticoid caninfluence the function of central neurons in a non-genomic pattern,and the underlyingmechanism involves the change of resting membrane potential.

关键词： glucocorticoid hippocampal slice membrane potential cell input resistance

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Urotensin Ⅱ inhibits electrical activity of hippocampal CA1 neurons by potentiating the GABA_A receptor-mediated Cl^- current

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Neuroscience Bulletin 2006年第2期22卷 110-114页

作者： Yu-Ming WU Ru WANG Rui-Rong HE Department of Physiology Institute of Basic Medicine Hebei Medical University Shijiazhuang 050017China

Objective To examine the effects of urotensin Ⅱ （UⅡ） on the discharges of neurons in CA1 area of hippocampal slices by using extracellular recording technique. Results ①In response to the application of UⅡ （0.3, 3.0, 30.0,300.0 nmol/L, n = 77） into the perfusate for 2 min, the spontaneous discharge rates （SDR） of 63/77 （81.8% ） neurons were significantly decreased in a dose-dependent manner. ②Pretreatment with bicuculline（BIC, 100 μmol/L）, a specific GABAs receptor antagonist, led to a marked increase in the SDR of 6/7 （85.71%） neurons in an epileptiform pattern. The increased discharges were not significantly changed after UⅡ （30.0 nmol/L） was applied into the perfusate for 2 min. ③Pretreatment with picrotoxin （ PIC, 50 μmol/L）, a selective blocker of Cl^- channel, led to an increase in the SDR of all 8/8（ 100% ） neurons. The increased discharges were not influenced by the UⅡ （30.0 nmol/L） applied. ④Application of nitric oxide synthase （NOS） inhibitor N^G nitro-L-arginine methyl ester （L-NAME, 50μmol/L） into the perfusate for 2 min also significantly augmented the SDR of 14/16 （87.5 % ） neurons , then UⅡ （30.0 nmol/L） applied into the perfusate reduced the increased the SDR of all 14/14 （ 100% ） neurons. Conclusion These results suggest that UⅡ may decrease neuronal activity by potentiating GABAA receptor-mediated CI current in hippocampal CA1 neurons, and involved with the mediation of nitric oxide.

关键词： hippocampal slice urotensin Ⅱ bicuculline L-NAME picrotoxin

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