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S-nitrosylation of Cdk5 regulates its kinase activity and dendrite growth during neuronal development

S-nitrosylation of Cdk5 regulates its kinase activity and de...

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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： Anthony H.K.Tsang Amy K.Y.Fu Wing-Yu FU Kenny K.Chung Nancy Y.Ip Division of Life Science Molecular Neuroscience Center and State Key Laboratory of Molecular NeuroscienceThe Hong Kong University of Science and TechnologyClear Water Bay

Objective Precise regulation of cyclin-dependent kinase 5(Cdk5),a member of the cyclin-dependent kinase family,is critical for the proper neuronal development and ***5 is activated through its association with the neuron-specific activator p35 or ***,how its kinase activity is regulated in neurons is not well *** the present study,we investigate whether Cdk5 is regulated by the S-nitrosylation,a post-translational modification of protein,and how the S-nitrosylation of Cdk5 modulates its kinase activity and functions during neuronal *** The kinase activity of Cdk5 upon S-nitrosylation was measured using in vitro kinase *** potential physiological role of S-nitrosylation of Cdk5 in neuronal development was studied by overexpressing Cdk5 or its C83A mutant in hippocampal *** change in the neuronal morphology was examined using confocal ***(1)Cdk5 is S-nitrosylated both in vitro and in vivo.(2)S-nitrosylation of Cdk5 inhibits its kinase activity.(3)Inhibition of the S-nitrosylation of Cdk5 enhances dendrite growth in neurons. Conclusion S-nitrosylation of Cdk5 regulates its activity and dendritic development.

关键词： cyclin dependent kinase 5 p35 dendrite nitric oxide S-nitrosylation

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Rolipram,a selective phosphodiesterase-4 inhibitor,decreases ethanol seeking and consumption in alcohol-preferring Fawn Hooded rats

Rolipram,a selective phosphodiesterase-4 inhibitor,decreases...

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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： Andrew J.Lawrence Howard Florey Institute and Centre for Neuroscience ParkvilleVictoria 3010Australia

＜正＞Alcohol dependence is a complex psychiatric disorder demanding development of novel pharmacotherapy. Since the cyclic AMP（cAMP） signaling cascade has been implicated in mediating the behavioral response to alcohol, it would be of interest to determine whether phosphodiesterase-4（PDE4）,an enzyme that selectively hydrolyzes intracellular cAMP,is involved in the regulation of alcohol ***,the selective PDE4 inhibitor rolipram was used to measure its anti-drinking properties in alcohol-preferring Fawn-Hooded（FH/Wjd） *** showed that acute administration of rolipram（0.05,0.1,and 0.2 mg/kg） selectively and dose-dependently reduced 5%ethanol, but not 10%sucrose,operant self-administration;it also decreased voluntary 5%ethanol intake,but not water consumption,for the initial 30 min in rats showing high stable preference for *** course assessment revealed the significant anti-drinking effect of rolipram（0.1,0.2 mg/kg） lasted for 5 h in both continuous- and intermittentaccess to 5%or 10%***,chronic rolipram treatment decreased 5%ethanol consumption and preference during the drug treatment *** effect lasted even after withdrawal of drug *** locomotor activity was inhibited by rolipram at 0.1 and 0.2 mg/kg,the effect was present only in the initial about 20 min,which did not seem to alter long-term ethanol *** data suggest that PDE4 may play a promising role in ethanol drinking and dependent *** may be a potential treatment target for further development of anti-drinking drugs.

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Properties of synaptic responses evoked by tactile stimulus in Purkinje cells in mice cerebellar cortex crus Ⅱ in vivo

Properties of synaptic responses evoked by tactile stimulus ...

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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： Chun-Ping CHU~(1,3),Yan-Hua BING~(1,2),Quan-Ri LIU~(1,2),De-Lai QIU~(1,2,*) 1 Cellular Function Research Center,Yanbian University 2 Department of Physiology and Pathophysiology 3 Function Experiment Center,College of Basic Medicine,Yanbian University,977,GongYuan Road,Yanji 133002,China.

Sensory stimuli evoke responses in cerebellar Purkinje cells（PCs） via mossy fiber-granule cell pathway. However,the properties and characterization of synaptic responses evoked by the tactile stimulus in cerebella PCs are *** the present study,we investigated the synaptic responses of cerebellar PCs in response to the airpuff stimulus on the ipsilateral whisker pad in urethane-anesthetized mice.A total of 33 PCs from 35 adult（6-8 weeks old） ICR mice were studied by somatic or dendritic patch-clamp recording with biolytic staining and pharmacological *** current-clamp conditions（I = 0）,air-puff stimulus evoked strong inhibitory postsynaptic potentials （IPSPs） in PCs *** of SR95531,a specific GABA receptor antagonist,blocked IPSPs,and revealed the stimulus-evoked simple spike *** voltage-clamp conditions,tactile stimulus evoked a sequence of transient inward currents followed by strong inhibitory postsynaptic currents（IPSCs） in both PCs stomata and *** of GABA receptor antagonist,SR95531 blocked IPSCs,revealed the excitatory postsynaptic currents（EPSCs） in somata and a temporal summation of parallel-fiber EPSCs in dendrites of *** also demonstrated that PCs respond to both onset and offset of air-puff *** findings indicated that the tactile stimulus could not evoke the strong EPSCs and fire PCs,but induced the rapid activation of strong GABA rereptormediated IPSCs in both stomata and dendrites of *** propose that the inhibition play a crucial role in controlling the output of sensory information in PCs in cerebellar cortex CrusⅡin mice.

关键词： cerebellar cortex in vivo patch-clamp Purkinje cell tactile stimulation postsynaptic currents GABAA receptor

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Organotypic slice culture of neonatal rat cortex and induced neural stem cell differentiation

Organotypic slice culture of neonatal rat cortex and induced...

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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： Qian JIAO,Hai-Xia ZHANG,Hai-Xia LU~*,Yong LIU~* Institute of Neurobiology,Xi'an Jiaotong University College of Medicine,Xi'an 710061,China

Objective During neural development,neural stem cell（NSC） undergoes a precise proliferation and differentiation *** process is directed by various factors from the NSC *** order to investigate the effect of spatial signal on NSC differentiation,in the present study,the survival and differentiation of CM-DiI labeled NSCs were observed on the organotypic slice cultures from neonatal rat *** SD rats on 3 to 5 postnatal days were sacrificed according to the ethical guidelines of the NIH Regulations for Experimentation on Laboratory Animals and set out by the Xi'an Jiaotong *** slices with 200-μm thickness were cut on a vibrotome microtome and cultured in modified DMEM/F12 serum free medium at 37℃with 95%O and 5%CO. Organotypic slice cultures were observed under inverted phase-contrast microscope every day and the medium was half refreshed every 3 to 4 *** were isolated from the cortex of rat embryos on embryonic day 14.5 and cultured in DMEM/F12 serum free medium supplemented with B27 and *** on passage number 3 were labeled by CM-DiI for 20 min in dark and cultured for 3 to 4 *** labeled cells were transplanted onto the organotypic slices in 2 weeks *** survival of transplanted NSCs was confirmed by visual CM-DiI labeled cells on the *** differentiation of transplanted NSCs was identified by immunofluorescence staining and the DiI/GFAP or DiI/βtubullinⅢdouble labeled cells were considered to be astrocytes or *** The organotypic slice cultures survived well for at least 4 weeks in the modified medium and became clear vision on 10 days later,which was possibly due to the elimination of ***-DiI labeled NSCs were observed on the slice cultures 4 days after *** labeled NSCs differentiated into GFAP glia andβtubullinⅢ *** Our results suggest that the organotypic slice culture might produce some specific signals to induce the transplanted NSCs to different

关键词： organotypic brain slice neural stem cells rat differentiation

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LanCLl functions as a binding partner and regulator of Cystathionine beta-synthase in brain

LanCLl functions as a binding partner and regulator of Cysta...

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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： Weixia ZHONG~#,Yubin WANG~#,Zhenxue GE,Lin PENG,Jianhong LUO~* Institute of Neuroscience,School of Medicine,Zhejiang University,Hangzhou 310058,China

Eukaryotic lanthionine synthetase C-like protein 1（LanCL1）,a glutathione binding protein,is homologous to prokaryotic lanthionine cyclases（LanC）.LanCL1 is involved in amyotrophic lateral sclerosis,wherein its biochemical functions remains *** beta-synthase（CBS）,the only source of hydrogen sulfide in brain,catalyzes the condensation of serine with homocysteine（Hcy） to form *** Hcy was detected in neurodegenerative disorders such us AD and ***,we identified both LanCL1 and CBS were highly expressed in brain,and we observed that LanCL1 was a CBS-binding protein in brain tissue and heterogeneous expression system by *** further confirmed that LanCL1 N-terminal 131-197 and CBS C-terminal 409-551 were necessary for this physical *** limited digestion with trypsin,Co-IP analysis showed that LanCL1 binded less full-length *** we investigated the regulation of LanCL1 on CBS activity using transgenic mouse and cell ***,ELISA assays revealed that hcy level in LanCL1 mice was downregulated due to increased CBS ***,CBS of LanCL1 mice brain lysates was more sensitive to trypsin ***,after incubation with brain lysates,purified recombinant protein his-LanCL1 directly binded endogenous CBS,downregulated CBS activity during enzymatic reaction in vitro,partly preventing CBS from trypsin *** last,HEK293 cell injury induced by oxygen-glucose deprivation/rehabilitation facilitated LanCL1-CBS interaction and impaired CBS *** summary,we identified that LanCL1 regulated CBS activity through direct binding,suggesting this functional association may be involved in some neurodegenerative diseases.

关键词： CBS LanCLl functions as a binding partner and regulator of Cystathionine beta-synthase in brain

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ATP-dependent retrograde axonal transport of P2X3 receptor in primary sensory neurons

ATP-dependent retrograde axonal transport of P2X3 receptor i...

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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 WANG~1,Xu-Qiao CHEN~1,Cheng-Biao WU~2,Jin PAN~1,Bo YUAN~3,Yuan-Yuan SU~1,Xing-Yu JIANG~3, Xu ZHANG~4,Lan BAO~(1,*) 1 Laboratory of Molecular Cell Biology,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai 200031,China 2 Department of Neurosciences,School of Medicine,University of California,San Diego,La Jolla,California 92093- 0662,USA 3 National Center for Nanoscience and Technology,Beijing 100080,China 4 Institute of Neuroscience and State Key Laboratory of Neuroscience,Institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai 200031,China

Objective P2X receptors are ATP-gated cation channels and play variable roles in pathophysiological *** P2X receptors,P2X receptor is abundantly expressed in dorsal root ganglion（DRG）*** is well established that P2X receptor modulates synaptic transmission and is involved in pain ***, the mechanism of P2X receptor-mediated purinergic signaling in long distance is *** Transport of P2X receptor is measured by sciatic nerve ligation and live imaging *** injection and axonal agonist application in microfluidic chamber were used to detect retrograde transport of P2X ***（1） P2X receptors are retrogradely transported in sciatic nerve and DRG neuron axon;（2）Live cell imaging in cultured DRG neurons showed that small GTPase Rab7-endosomes mediated the long-distance retrograde transport of P2X receptors;（3）The retrograde transport of P2X receptor is *** These results demonstrate that retrograde transport of P2X receptor is regulated by ATP in primary sensory neurons.

关键词： primary sensory neuron P2X3 receptor axonal retrograde transport

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Calcineurin mediated CREB phosphorylation regulate hippocampal neuron death in excitotoxic rats

Calcineurin mediated CREB phosphorylation regulate hippocamp...

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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： Shi-Wei WANG,Zi-Li YOU,Yu-Bo CHEN~* Key Laboratory for Neurolnformation of Ministry of Education,University of Electronic Science and Technology of China,Chengdu 610054,China

Objective Excessive release of excitatory amino acid and over increased intracellular Ca could lead to hippocampus neurons over-excited,which contribute to excitotoxic cell death and finally developed severe cognitive impairment and motor deficits in epilepsy,stroke and other neurodegenerative *** the mechanisms of neurons damage through proteins phosphorylation have not yet be *** this study,we used kainic acid（KA）-induced neuronal excitotoxic model,investigate the effects of calcineurin（CaN） on regulation of the transcription factor CREB phosphorylation,which involves hippocampus structural and functional *** KA-induced excitotoxic rats were given CaN inhibitors FK506 treatment and compare with *** water maze（MWM） was used to detect the functional changes of hippocampus in learning and memory,and toluidine blue staining to observe the structural changes in *** phosphorylation was detected by Western *** The learning and spatial memory of over-excited rats were significantly improved after CaN inhibition compared with the control *** were considerable cell losses within hippocampal CA3 area of kainic acid over-excited *** neuronal cell losses were rescued significantly after treatment with *** blot analysis showed that rats received CaN inhibition have a significant increasing of p-CREB compare with control *** Treatment with FK506 can reduce hippocampus neuronal damages of excitotoxic rats,thus improve learning and memory of rats. FK506 treatment also led to a significant increased level of CaN downstream targets p-CREB,which paralleled with the decrease of CaN *** that the activation of CaN may be involved in the regulation of cell survival, and play an important role in neuronal excitotoxic injury.

关键词： excitotoxic cell death calcineurin phosphorylation CREB FK506

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Overexpression of α-synuclein caused mitochondria vacuolization and mitochondrial membrene potential to decrease by interacting with voltage-dependent anion channel

Overexpression of α-synuclein caused mitochondria vacuoliza...

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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： Lingling LU,Yuangang ZHU,Chunyan ZHANG,Xiaohong SUN,Shasha ZHAO,Pusheng GONG,Hui YANG~* Beijing Institute for Neuroscience,Capital Medical University(CMU),Beijing Center of Neural Regeneration and Repairing,Key Laboratory for Neurodegenerative Diseases of Ministry of Education,Beijing 100069,China

We previously found that alpha-Synuclein(Syn) localized not only in neuclei but also in axons and cytoplasm,especially enriched in *** of accumulating evidence showing that overexpression of Syn could partly mimic the symptom of Parkison's disease(PD),the exact mechanism remains *** find out the subcelluar localization of Syn in substantia nigra(SN) neurons especially on mitochondria and its effect on mitochondria in Syn induced PD rats model may help explain the pathogenesis of Syn in *** the present study,the synuclein gene was introduced into the SN of rats using recombinant adeno-associtaed viral(rAAV) *** immunogold electron microscopic technique,we found that the density of Syn-positive gold particles was greater in both substantia nigra and striatum in PD rat model than in the control *** compare the distribution of Syn in different cell compartment with the control group,the density of Syn-positive gold particles was greater in neuclei, axons,cytoplasms,especially mitochondria in SN of PD rat model,while the density of gold particles decreased to zero in pre-synaptic *** with the SN regeon,the density of gold particles increased mainly in pre-synaptic terminals,cytoplasm and neucleus in PD rat model,with no difference in axons and mitochondria. The changed distribution of Syn in different cell compartments in PD rat model suggested that high content Syn may be involved in axon transport,plasticity of synapse and gene ***,we also observed that mitochondria were damaged when its membrene potential *** an interaction between Syn and voltagedependent anion channel(VDAC) was also *** together,the current data suggested that Syn may be involved in the maintainance of mitochondria membrene potential and energy metabolism by interacting with permeability transition pore(PTP) protein VDAC,which may play a role in PD pathogenesis.

关键词： Parkinson's disease a-Synuclein electronmicroscope subcellular localizations mitochondria dysfunction

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The time course of attentional disengagement from emotional stimuli

The time course of attentional disengagement from emotional ...

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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： Shan CHEN,Jun-Feng SUN,Shan-Bao TONG~* Neural Engineering Laboratory,School of Biomedical Engineering,Shanghai Jiao Tong University,Shanghai 200240, China

Objective Negative information has the priority to capture and maintain humans' *** results have shown that people with emotional disorders(e.g.,depression,anxiety) would not automatically pay attention to negative information compared to healthy counterparts,but have difficulties in disengaging attention from it once the negative information is detected,sμggesting that the delayed disengagement hypothesis may underlie the mechanism of negative attentional ***,how the disengagement component of attention would be influenced by different emotions is not clear *** this consideration,the present study is to explore the time course of attentional disengagement from emotional *** An object-based cue-target paradigm was adopted in this study:an emotional(happy/sad) or a neutral facial cue was firstly presented at the fixation,followed by a digit target(3 or 5) requiring participants to *** interval between cue and target(CTI) was manipulated at five values,including 17,350,1000,600 and 1500 ms(the last two CTIs were conducted in a following complementary experiment).The reaction times(RTs) to the subsequent targets were measured to reflect the disengagement from the previous emotional cues *** right-handed students(aged 21.9±2.0 years,26 males) from the Shanghai Jiao Tong University took part in the ***(1) RTs decreased significantly as CTIs increased until up to 1000 ms;(2) Responses after positive cues were significantly faster than those after neutral ones in the 17 ms CTI conditions,but no difference in longer CTI conditions;(3) Responses after negative cues were significantly slower than those after positive and neutral ones during short and long CTIs,even at 1500 ms after cues *** RTs to the subsequent targets have been significantly influenced by CTIs and the emotion type of *** takes a relatively short time ranging from 350 to 1000 ms to disengage attention from pre

关键词： attentional disengagement negative attentional bias emotional cues CTI

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Direction-selective circuitry in rat retina develops independently of GABAergic,cholinergic and action potential activity

Direction-selective circuitry in rat retina develops indepen...

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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： Le SUN~(1,**),Xu HAN~(1,**),Shi-Gang HE~(1,*) State Key Laboratory of Brain and Cognitive Sciences,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China

Objective Motion detection is an important function of visual *** the retina,three types （ON-,OFF-,ONOFF-）of direction-selective（DS）ganglion cells（DSGCs）have been *** the ON-and ONOFFDSGCs, accumulated results show that they receive DS synaptic input from a kind of interneuron,starburst amacrine cells（SACs）.The asymmetrical inhibition between SACs and DSGCs is established around *** results indicate that visual experience,propagating cholinergic retinal wave and signal mediated by GABA receptor after P7 are not crucial for the establishment of DS *** how the specific connection between DSGCs and SACs formed and what kind of mechanism took a part in the formation of this circuitry,are still *** present work is to study the role of correlated spontaneous activity between DSGCs and SACs during the development of DS circuitry. Methods（1）Intravitreal injections of bicuculline methiodide（BMI）,TTX and/or epibatidine were used to block GABA receptor,retina spontaneous spikes and/or cholinergic waves during the development of rat retina from PO. （2）CTB-488 or CTB-594 was injected into the eyes to visualize the projection from retina to LGN.（3）Patch clamp recording was used to indentify DSGCs and collect data for ***（1）Daily injection is sufficient to block GABA receptor,retina spontaneous spikes and/or cholinergic wave.（2）Projection area from treated eye has altered significantly in both side of LGN.（3）Both physiological and morphological properties of DSGCs have no significant changes in the treated *** Direction selectivity is established independently of correlated spontaneous activity in the retina.

关键词： retina direction selectivity(DS) circuitry development correlated spontaneous activity direction-selective ganglion cell(DSGCs) starburst amacrine cell(SAC) lateral geniculate nucleus(LGN)

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