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Effects of NR2A overexpression on prefrontal synaptic plasticity and prefrontal cortex-related memory

Effects of NR2A overexpression on prefrontal synaptic plasti...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Yan-Hong DUAN,Wen SI,Jing MA,Bo WANG,Xiao-Hua CAO~* Shanghai Institute of Brain Functional Genomics,Key Laboratories of MOE and STCSM,East China Normal University,Shanghai 200062,China

Objective N-methyl-D-aspartate receptors(NMDARs) play an important role in learning and memory. NR2A and NR2B subunits are predominant in the prefrontal *** growing evidence implicates that specific subunit-containing NMDARs correlate with subsequent modifications of synaptic plasticity,whether the upregulation of NR2A subunit influences prefrontal synaptic plasticity and prefrontal cortex-related memory is not yet *** In the present study,we combined electrophysiological technique and behavioral method to investigate effects of NR2A overexpression on prefrontal synaptic plasticity and prefrontal cortex-related memory. Results(1) Western blotting results showed that the level of NR2A,but not NR2B protein,was significantly higher in prefrontal cortex of the transgenic mice than wild type mice.(2) Whole cell Patch-clamp recording showed that the ratio of NR2A/NR2B current increased significantly in transgenic mice compared with that in wide type mice;the decay time of evoked NMDAR-EPSC was sharply faster in transgenic mice than in wide type ***,the rise time of evoked NMDAR-EPSC in transgenic mice did not differ significantly from wide type mice.(3) Field potential recording showed that no significant differences were measured in basal synaptic transmission between transgenic mice and wide type mice;however,both prefrontal long-term potentiation(LTP) and long-term depression(LTD) were impaired in transgenic mice.(4) Behavioral tests showed that NR2A transgenic mice exhibited impaired prefrontal cortex-related working memory and reverse learning in the modified water maze and water maze reverse learning task. Conclusion NR2A/NR2B ratio increasing by NR2A overexpression can shorten the decay time of NMDAR-EPSC and impair prefrontal bidirectional synaptic plasticity and prefrontal cortex-related memory.

关键词： NMDA receptor NR2A subunit prefrontal cortex synaptic plasticity learning and memory

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Neuronal damage and blood-retinal barrier disruption in retinal ischemia/reperfusion injury:protective effects of Lycium barbarum polysaccharides

Neuronal damage and blood-retinal barrier disruption in reti...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Amy Cheuk Yin Lo Suk-Yee Li Chung-Man Yeung Raymond Chuen-Chung Chang Kwok-Fai So David Wong Eye Institute Li Ka Shing Faculty of MedicineThe University of Hong Kong Research Center of Heart BrainHormone and Healthy AgingLi Ka Shing Faculty of MedicineThe University of Hong Kong Department of Anatomy Li Ka Shing Faculty of MedicineThe University of Hong Kong

Objective Neuronal cell death,glial cell activation,retinal swelling and oxidative injury are complications in retinal ischemia/reperfusion(I/R) *** barbarum polysaccharides(LBP),extracts from the wolfberries, are good for "eye health" in Chinese *** aim of our present study is to explore the use of LBP in retinal I/R *** Retinal I/R injury was induced by surgical occlusion of the internal carotid *** to induction of ischemia,mice were fed orally with either vehicle(PBS) or LBP(1 mg/kg) once a day for 1 week. Ischemia was maintained for 2 hours and reperfusion was then allowed for 22 ***-embedded retinal sections were *** cells were counted while apoptosis was assessed using TUNEL *** levels of glial fibrillary acidic protein(GFAP),aquaporin-4(AQP4),poly(ADP-ribose)(PAR) and nitrotyrosine (NT) were investigated by *** integrity of blood-retinal barrier(BRB) was examined by IgG extravasation *** Decreased viable cell count and apoptosis were found in the ganglion cell layer (GCL) and the inner nuclear layer(INL) of the vehicle-treated I/R ***,increased retinal thickness, GFAP activation,AQP4 up-regulation,IgG extravasation and PAR expression level were observed in the vehicletreated I/R *** importantly,many of these changes were diminished or abolished in the LBP-treated I/R *** Pre-treatment with LBP for 1 week effectively protected the retina from neuronal death,apoptosis, glial cell activation,aquaporin water channel up-regulation,disruption of BRB and oxidative *** present study suggests that LBP may have a neuroprotective role in ocular diseases in which I/R is a feature.

关键词： Neuronal damage and blood-retinal barrier disruption in retinal ischemia/reperfusion injury:protective effects of Lycium barbarum polysaccharides

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Variations of ionotropic glutamate receptors mediating epileptiform activity in rat hippocampal CA1 region

Variations of ionotropic glutamate receptors mediating epile...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Bin-Sheng WANG,Zhu-Yan YOU,Min XIE,Zhen LI,Lie-Cheng WANG~* Department of Physiology,School of Basic Medical Science,Anhui Medical University,Hefei 230027,China

Objective The epileptiform activity was induced by perfusing GABAA receptor antagonist bicuculline (Bic,30μmol/L)in normal hippocampal slices and temporal lobe epilepsy(TLE)model was established by kainic acid(KA)injected into the center site of right hippocampus CA3 region with the stereotaxic *** investigate the variations of ionotropic glutamate receptors mediating epileptiform activity in rat hippocampal CA1 region by using extracellular recording in hippocampal slices,and to explore the mechanism of epileptogenesis. Methods Population spike(PS)was recorded in CA1 pyramidal cell layer by stimulating the Shaffer collaterals in stratum radiatum using extracellular field-potential ***(1)Single PS was usually recorded in normal pyramidal cell layer of hippocampal CA1 *** normal hippocampal slices with Bic(30μmol/L)could induce multiple epileptiform discharges,in which the PS number was(5.07±0.30,n=11),the amplitude of the first PS was increased significantly by(150.86±22.56)%(n=11,P<0.01)compared with the *** discharges could recorded in slices of TLE model rats,the PS number was(6.12±0.33,n=9).The PS number was increased significantly in slices of epileptiform activity compared with the control(P<0.01).(2)Variations of ionotropic glutamate receptors mediating PS in hippocampal CA1 *** PS recorded in normal pyramidal cell layer of hippocampal CA1 mainly mediated by non-NMDA *** PS recorded in normal slices after perfusing with Bic was partially mediated by NMDA receptor in addition to non-NMDA *** perfusing with NMDA receptor antagonist,APV(50μmol/L),the amplitude of the first PS was not significantly affected(n=11,P>0.05), however,the amplitude of the fourth and fifth PS was decreased obviously(n=11,P<0.05;n=9,P<0.05).After perfusing APV in slices of TLE model rats,the amplitude of the first PS was not significantly affected(n=9,P>0.05), but the amplitude of the fourth and fifth PS w

关键词： epilepsy hippocampus population spike receptor

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Peripheral inflammation induces promoter demethylation of endogenous hydrogen sulfide synthesis enzyme CBS in rat dorsal root ganglion

Peripheral inflammation induces promoter demethylation of en...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： You-Lang ZHOU~1,Fei-Hu QI~1,Ning-Ang LIU~2,Guang-Yin XU~(1,2,*) 1 Jiangsu Key Laboratory of Neuroregeneration,Nantong University Medical College,Nantong 226001,China 2 Institute ofNeuroscience,Key Laboratory of Pain Research & Therapy,Soochow University,Suzhou 215123,China

Objective Chronic pain associated with inflammation is a common clinical problem,and the underlying mechanisms have only begun to be *** sulfide（HS）,an endogenous gaseotransmitter/modulator, is becoming appreciated that it may be involved in a wide variety of processes including inflammation and nociception. However,the molecular and epigenetic mechanisms for expression of HS synthesizing enzyme cystathioninebeta -synthetase（CBS） in peripheral nociceptive processing remain *** present study is to explore the role of CBS gene promoter methylation following peripheral *** Complete Freund's adjuvant（CFA） was injected into plantar surface（100 ml each） of hindpaw of adult male Sprague Dawley rats（6-8 week old）.Realtime polymerase chain reaction（qPCR） and Western blot analysis were employed to measure expression of mRNA and protein in lumbar dorsal root ganglion（DRGs,L4-L6） from control and CFA-treated rats,*** PCR（MSP） and bisulfite sequencing（BSP） were carried out for determining methylation status of the cbs gene *** Peripheral inflammation significantly reduced hind paw withdrawal threshold and remarkably upregulated expression of CBS at both protein and mRNA levels in DRGs when compared with control *** ratio of methylated and unmethylated CpG island for MSP primers（+325～+425;+657～+767） was markedly reduced in inflammatory rats when compared with control *** sequencing of the bisulfite-treated cbs promoter dem- onstrated that the fragment（F210bp,+496 bp～+705 bp） of cbs promoter region,which contains 11 CpG sites was significantly demethylated in DRGs 3 days after CFA ***,expression of MBD4 and Gadd45a was highly enhanced in CFA-treated rats while expression of DNMT3a and 3b was not significantly *** Our data suggest that peripheral inflammation leads to an active DNA demethylation in cbs gene promoter region,thus contributing to the enhanced C

关键词： cystathionine-beta-synthetase dorsal root ganglion inflammatory pain methylation-specific polymerase chain reaction and bisulfite sequencing

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Understanding structure and function of acid-sensing ion channels

Understanding structure and function of acid-sensing ion cha...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Tian-Le XU~(1,2) 1 Institute of Neuroscience and State Key Laboratory of Neuroscience,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences,Shanghai 200031,China 2 Neuroscience Division,Department of Biochemistry and Molecular Cell Biology,Institute of Medical Sciences, Shanghai Jiaotong University School of Medicine,Shanghai 200025,China

The acid-sensing ion channel（ASIC） has a prominent role in tissue pH-sensing and neuronal signalling. Subunits of ASICs are members of the degenerin/epithelial Na channel protein *** date,seven ASIC subunits have been identified:ASIC1a,ASIC1b,ASIC1b2,ASIC2a,ASIC2b,ASIC3,and *** subunit contains cytosolic N and C termini,two alpha-helical transmembrane domains（M1-M2）,and a disulfide-rich multidomain extracellular *** form a variety of homo- and heterotrimeric ion channels that are activated by extracellular acidosis associated with neurotransmission,injury,inflammation and ischemia,triggering membrane depolarization via Na influx into target *** the operating mechanisms of these ion channels could therefore provide the catalyst for further studying ASIC physiology and pharmacology,as well as promoting the development of experimental techniques for other families of ion channels and/or *** this talk,I will present our current work on ASIC structure and function,in particular those of ASIC1a and ASIC3 *** in this area has been stimulated in recent years by the publication of high-resolution structures of ASIC1,which have permitted the structure-directed approaches and the creation of ASIC homology models.I will first discuss the crucial roles of ASICs in mediating neuronal injury and pain *** a series of computational,chemical and functional approaches,and nonproton ligands（GMQ,agmatine and amiloride） as probes,I will then delineate the nonproton ligand sensing domain in ASIC3 channels at the atomic level and describe how chemical biology,homology modeling and ligand docking can provide useful information about ligand interactions.I will also consider some of the ongoing projects in my laboratory that are investigating the pathophysiological roles of ASICs using ASIC subunit gene knockout *** report ends with a discussion on the generality of ASICs as a model system for the study of struc

关键词： Understanding structure and function of acid-sensing ion channels

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Study of puerarin in the signalling of myocardial ischemia mediated by P2X7 receptor of superior cervical ganglion

Study of puerarin in the signalling of myocardial ischemia m...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Shuang-Mei LIU,Chang-Shui XU,Shang-Dong LIANG~* Department of Physiology,Basic Medical College of Nanchang University,Nanchang 330006,China

Objective Myocardial ischemia activates cardiac sympathetic afferent nerve endings and elicits chest pain,which is often associated with a sympathoexcitatory reflex characterized by an increase in blood pressure and sympathetic nerve activity.P2X channel is responsible for the release of cardioprotectants induced by ischemic preand ***（a major active ingredient extracted from the traditional Chinese medicine Ge-gen）is widely used in the treatment of cardiovascular disease in *** present study was aimed to observe the effect of puerarin on sympathoexcitatory reflex induced by myocardial ischemic nociceptive signaling via P2X receptor in rat superior cervical ganglion（SCG）.Methods Myocardial ischemia model was ***-Dawley rat were randomly divided into groupⅠ（Sham group）,groupⅡ（Puerarin,intraperitoneal injection）,groupⅢ（Myocardial ischemia injury）,groupⅣ（Myocardial ischemia injury + Puerarin）and group V（Blank control）.Electrocardiogram （ECG）of the rats were *** blood pressure,heart rate and respiration were measured by non-invasive blood pressure determinator with different *** expression of P2X mRNA and protein was analyzed by PCR,Western blotting and *** The results showed that the systolic blood pressure,heart rate and respiration in groupⅢwere higher than those in groupⅠ,Ⅱ,Ⅴ（P<0.01）.There was no difference in group I,ⅡandⅤ（P>0.05）.After the treatment with puerarin in the myocardial ischemic rats,systolic blood pressure,heart rate and respiration in groupⅣwere lower than those in groupⅢ（P<0.05）.The expression of P2X protein and mRNA in SCG of myocardial ischemic injury rats were increased compared with those in control *** decreased the P2X protein and mRNA expression in SCG of myocardial ischemic injury *** According to these results,we conclude that puerarin can reduce the transmission of myocardial ischemic signalling mediated by P2X receptor in ra

关键词： superior cervical ganglia P2X7 receptor puerarin

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Electrostatic interaction in the NH2-terminus accelerates inactivation of the Kvl.4 channel

Electrostatic interaction in the NH2-terminus accelerates in...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Zhuo FAN~(1,2,3),Li-Jun BI~(1,*),Gang JIN~(1,*),Zhi QI~(1,2,*) 1 National Laboratory of Biomacromolecules and State Key Laboratory of Brain and Cognitive Science,Institute of Biophysics,CAS,15 Datun Rd,Beijing 100101,China 2 Department of Physiology,Medical College of Xiamen University,168 Daxue Rd,Xiamen 361005,China 3 Graduate University of Chinese Academy of Sciences,19A Yuquan Rd,Beijing 100039,China

Objective The rapid inactivation of Kv1.4 channel was assumed to be mediated through the "ball and chain" model which was perfectly suitable for ShB ***,some investigations have suggested that inactivation properties of Kv1.4 channel are different from ShB channel in that it has not an apparent chain structure. There are two highly charged regions(carrying charges opposite to each other) in the "chain" region of Kv1.4 channel, one is rich of positive charges(residues 83-98:segment A) and the other rich of negative charges(residues 123-137: segment B).The objective of this study is to examine the electrostatic interactions between these two regions and explain the difference in inactivation property between Kv1.4 and ShB *** We construct a series of mutants by standard molecular method,including neutralizing or deleting the positively charged segment A;replacing the positively charged segment A with negatively charged segment B(and vice versa);exchanging positively charged segment A with negatively charged segment *** inactivation properties of these mutants were examined by voltageclamp ***(1) Neutralizing or deleting positively charged segment A slowed the inactivation rate of Kv1.4 channel;(2) The mutants,which carried both positive or negative charges at these two regions,inactivated much more slowly compared with Kv1.4WT;(3) Exchanging these two segments with each other did not affect the inactivation *** All these results indicate that there are electrostatic attractions between positively charged segment A and negatively charged segment B and the electrostatic interaction between them accelerates the inactivation rate of Kv1.4 channel.

关键词： Kvl.4 channel inactivation electrostatic interaction "ball and chain" model

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Roles of protein kinase C and RhoA in the opening of blood-tumor barrier induced by low-dose endothelial monocyte-activating polypeptide-Ⅱ

Roles of protein kinase C and RhoA in the opening of blood-t...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Li-Bo LIU~1,Li ZHEN~2,Yun-Hui LIU~2,Yi-Xue XUE~(1,*) 1 Department of Neurobiology,College of Basic Medicine,China Medical University,Shenyang 110001,China 2 Department of Neurosurgery,Shengjing Hospital,China Medical University,Shenyang 110004,China

Objective This study was performed with an in vitro blood-tumor barrier(BTB) model to investigate whether protein kinase C(PKC) and RhoA are involved in the low-dose endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)-induced BTB *** The permeability of BTB model was detected by transendothelial electric resistance(TEER) and Horseradish peroxidase(HRP) *** total PKC activity was measured by using PKC Assay *** blot assay was used to test the protein expression of total PKC,PKC isoforms(a,p,y,e, 8,and Q,Rho A and tight junction(TJ)-associated proteins(occludin,cluadin-5 and ZO-1).The distribution and expression changes of TJ associated proteins and cytoskeleton protein(F-actin) were tested by immunofluorescence *** In BMECs treated with EMAP-II(0.05 nmol/L) for 0.5 h and 1 h,the significant increases in total PKC activity and the protein expression of total PKC were ***,the protein expression of 3 PKC isoforms(PKC-a,P and Q and RhoA significantly increased at 0.5 h and 1 h after EMAP-Ⅱadministration and then recovered at 4 *** and HRP flux assays revealed that pretreatment with PKC inhibitor H7,RhoA inhibitor C3 exoenzyme or a combination of the two inhibitors significantly blocked the EMAP-Ⅱ-induced BTB permeability ***,our data showed that the EMAP-Ⅱ-induced downregulation of occludin,claudin-5,ZO-1 as well as F-actin was also significantly inhibited by pretreatment with H7,C3 exoenzyme or a combination of H7 and C3 *** In summary,our present study suggests that PKC and RhoA play crucial roles in the process of EMAP-Ⅱinducing progressive changes to BMECs of BTB,including downregulation of tight junction proteins, reorganization of actin cytoskeleton,and increasing of BTB permeability.

关键词： endothelial monocyte-activating polypeptide-Ⅱ blood-tumor barrier permeability protein kinase C RhoA

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GABAB receptors are involved in ventrolateral orbital cortex-evoked antinociception in rats with neuropathic and inflammatory pain

GABAB receptors are involved in ventrolateral orbital cortex...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Yan ZHAO~2,Wen-Jin XU~1,Feng-Yi XU~1,Fu-Quan HUO~(1,*),Chun-Xia YAN~2,Jing-Shi TANG~1 1 Department of Physiology,2 Department of Forensic Medicine,Xi'an Jiaotong University School of Medicine,Xi'an, Shaanxi 710061,China

Objective Previous studies have shown that the ventrolateral orbital cortex（VLO）is part of the endogenous analgesic system,which consists of an ascending pathway from the spinal cord to the VLO through the thalamic nucleus submedius and a descending pathway to the spinal cord via the periaqueductal gray（PAG）.And the GABAergic interneurons tonically depress this antinociceptive pathway via GABA *** present study examined the role of GABA receptor,an autoreceptor,in the VLO-evoked antinociception in rats with neuropathic pain and inflammatory *** The paw withdrawal threshold（PWT）was measured for 60-min in response to mechanical stimulation（von Frey hairs）using the up-down method in a 10-min interval in spared nerve injury（SNI） rats,and the number of flinches and the durations of licking/biting of the affected paw during each 5-min period were record for 60-min as well as the expression of Fos in the spinal dorsal horn was counted by two experimenters blind to the different groups in formalin test ***（1）Microinjection of baclofen（0.5,1,2μg/0.5μL）,a GABA receptor agonist,into the VLO attenuated the SNI-induced mechanical allodynia in a dose-dependent *** effect was completely blocked by the GABA receptor antagonist CGP35348（2.5μg/0.5μL）.（2）Microinjection of baclofen（2μg/0.5μL）into the VLO also decreased the formalin-induced nociceptive behavior and the expression of Fos in the spinal dorsal *** effects were also completely reversed by the CGP35348（2.5μg/0.5μL）.（3） When CGP35348 was injected alone into the VLO,there was no effect on the SNI-induced mechanical allodynia and formalin-induced nociceptive behavior as well as the expression of Fos in the spinal dorsal horn,suggesting that the blocking effects of CGP35348 on the baclofen-induced inhibition are not a result of facilitation of the nociceptive responses *** GABA receptors are involved in the VLO-evoked antinociception in rats with neuropathic and inflammatory

关键词： GABAB receptor neuropathic pain inflammatory pain ventrolateral orbital cortex

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Overexpression of the transcription factor Sp8 induces ectopic motor neurons in the p3 domain of chick spinal cord

Overexpression of the transcription factor Sp8 induces ectop...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Xiaosu LI~1,Chao LIN~1,Sijing SONG~2,Zhengang YANG~(1,*) 1 Institutes of Brain Science and State Key Laboratory of Medical Neurobiology,Fudan University,Shanghai 200032, China 2 College of Life Science and Technology,Hua Zhong Agricultural University,Wuhan 436700,China

During the period of neurogenesis in the spinal cord,particular types of neurons are generated from specific progenitor cells located in different dorsoventral(DV) regions of the ventricular *** graded morphogens along the DV axis of the neural tube confer positional information to the progenitor *** achieve through induction or inhibition of a set of transcription factors expression in a concentration dependent *** combinatorial expression of distinct transcription factors,the DV axis of the neural tube can be divided into multiple domains and cross-repressive interactions between complementary pairs of transcription factors further refine and sharpen the boundaries between adjacent *** action of these transcription factors directs different progenitor populations to generate particular subtypes of ***8 is a zinc finger transcription factors belonging to specificity protein(Sp) family and expression of Sp8 in the basal plate of spinal cord has been *** there is no study addressing the function role of Sp8 during spinal cord *** the present study,we first confirmed Sp8 expression in the spinal cord using *** neurogenesis in the spinal cord,Sp8 expression shares the same ventral limit with O1ig2 and is complementary to Nkx2.2.O1ig2+ domain and Nkx2.2+ domain are neighboring with each other,and give rise to Isll+ motor neurons and Siml+ V3 interneurons respectively. When mouse Sp8 was overexpressed in the chick spinal cord using either retrovirus or in ovo electroporation,we found that ectopic motor neurons were induced in the Nkx2.2+ p3 ***,Nkx2.2 expression was cellautonomously repressed by Sp8 *** suggests that Sp8 participates in patterning the ventral spinal cord.

关键词： Sp8 motor neuron Nkx2.2 dorsoventral patterning spinal cord

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