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Cofilin rod formation and its regulation:a pilot study by using live cell imaging system

Cofilin rod formation and its regulation:a pilot study by us...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Yun WANG institutes of brain science and state key laboratory for medical neurobiology,Fudan University,Shanghai 200032,China

＜正＞Neurite transport defects have been linked to very early stages of many neurodegenerative ***,a protein regulates dynamics of the actin cytoskeleton,has been reported to form rod structure altering neurite transport in pathological condition such as Alzheimer’s *** aim of this study is by using live cell station to investigate the dynamic change of the cofilin rod formation and it’s regulating cellular *** hippocampal neurons were either transfected with cofilin-GFP alone or co-transfected with cofilin-GFP and LIM JCinases/RFP and then were under long term continuous observation（＞10 h） by using Ultraview VoX live cell scanning unit（PerkinElmer Life sciences）.Timelapse confocal image showed neurons transfected with cofilin developed to form rod structure,in distal dendrites but not axon,in a dynamic way:rods elongating,splitting,merging,and moving in a local area,in first 24 h after *** more mature neurons and 2-3 days after transfection,cofilin rods were much more *** cofilin rods were stably formed,neurons co-transfected with LIMK/RFP showed time-dependent dissolving of formed rods in cultured *** conclusion,our results demonstrate that cofilin rod is formed in a dynamic way in distal dendrites and regulated by LIM *** relation between cofilin rod and some neurological disorders are currently under investigation.

关键词： cofilin cofilin rod LIM Kinases live cell station

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Neural stem/progenitor cells in the postnatal subventricular zone give rise not only to granular and periglomerular interneurons,but also to interneurons in the external plexiform layer of the rat olfactory bulb

Neural stem/progenitor cells in the postnatal subventricular...

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

作者： Zhengang YANG institutes of brain science and state key laboratory of medical neurobiology,Fudan University,Shanghai 200032,China

＜正＞Interneurons in the granule cell layer（GCL） and glomerular layer（GL） of the olfactory bulb（OB） are generated from neural stem/progenitor cells in the subventricular zone（SVZ） of the lateral ***,little is known about the origin of interneurons in the external plexiform layer（EPL） of the *** the basis of the concept of corticogenesis,we hypothesized that interneurons in the EPL of the rodent OB also originate in the *** the present study,replicationincompetent retroviruses encoding the marker gene,human placental alkaline phosphatase（AP）,were injected into the lateral ventricles of postnatal day 4 Wistar rats to label dividing cells in the *** days after injection,AP labeled cells had migrated into the *** weeks after injection,AP/NeuN double labeled cells were found not only in the GCL and GL, but also in the EPL of the *** the EPL,most AP labeled cells were calcium-binding protein parvalbumin-（PV） immunoreactive （+） interneurons.A subset of these cells were calcium-binding protein calretinin（CR） + *** to their structural features,these AP/PV double labeled cells were Van Gehuchten cells,multipolar cells and superficial shortaxon ***,the neural stem/progenitor cells in the postnatal SVZ give rise not only to granular and periglomerular interneurons,but also to interneurons in the EPL of the ***,these data suggest that that SVZ-derived progenitors give rise to virtually all subpopulations of interneurons in the OB.

关键词： subventricular zone olfactory bulb external plexiform layer neurogenesis parvalbumin calretinin

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Role of KCC2 in seizure induction and intervention

Role of KCC2 in seizure induction and intervention

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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： Yun WANG institutes of brain science and state key laboratory for medical neurobiology,Fudan University,Shanghai 200032, China

The neuron specific K-Cl cotransporter KCC2 in adult plays an important role to maintain intracellular Cl homeostasis,hence renders GABA receptor inhibitory ***,KCC2 expression levels are found to be reduced in various pathological conditions,including *** current study,we analyzed KCC2 expression and function and its involvement in GABA receptor mediated inhibition change during acute seizure *** using cyclothiazide(CTZ) induced both in vivo and in vitro acute seizure model,we discovered that the hippocampal neuron somatic membrane KCC2 expression was down regulated,the GABA reversal potential(E) was up- shifted and a profound reduction in Cl extrusion *** molecular manipulation of KCC2 expression in cultured hippocampal neurons,CTZ treatment induced epileptic neurons was significantly reduced in association with KCC2 over expression,and KCC2 down regulated by shKCC2 resulted a E up-shift but without effect on CTZ induction of epileptiform *** addition,furosemide,a KCC2 inhibitor,while applied continuously mimicked CTZ effect to cause the E up-shift and facilitate the CTZ induced epileptiform ***,furosemide co-treated with CTZ prevented CTZ induced KCC2 down regulation from the somatic membrane and in turn rescued acute convulsant stimulation induced GABA inhibitory function *** conclusion,our data suggest that KCC2 plays an important role in seizure induction and prevent KCC2 down regulation may be a novel target for anticonvulsant development.

关键词： CTZ epilepsy hippocampus KCC2 GABAA receptor

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Involvement of the anterior cingulate cortex in the affective pain

Involvement of the anterior cingulate cortex in the affectiv...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Yu-Qiu ZHANG Institute of neurobiology,institutes of brain science and state key laboratory of medical neurobiology,Fudan University,Shanghai 200032,China.

Pain is a multidimensional conscious experience including sensory and affective *** former has been extensively studied,while the latter is poorly *** evidence from morphological, electrophysiological,neuroimaging,and behavioral studies as well as clinical observations indicate that the anterior cingulate cortex(ACC) is a key structure that contributes to pain negative affect or *** formalin-induced conditioned place avoidance(F-CPA) in rats,which is believed to reflect the pain-related negative emotion,we found that lesions of the bilateral ACC markedly blocked the painful stimulation-induced CPA,but not non-painful fear stimulation-induced CPA(Gao et al.,2004).Intracerebroventricular or intra-rostral ACC(rACC) administration of NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid(APV),but not AMPA/KA receptor antagonist 6,7- dinitroquinoxaline-2,3-dione(DNQX),significantly decreased both conditioning scores of F-CPA(Lei et al.,2004).Similarly, intra-rACC injection of 7-Chlorokynurenate(7C1-KYNA),an antagonist of the glycine site of NMDA receptors,can also effectively eliminate the induction of ***,degradation of endogenous D-serine,an endogenous ligand for the glycine site of NMDA receptors,by DAAO(D-amino acid oxidase),an endogenous D-serine degrading enzyme, effectively inhibited NMDA-evoked currents and attenuated F-CPA scores,but did not affect formalin-induced acute nociceptive behaviors,suggesting that endogenous D-serine in the rACC is necessary for NMDA receptor-mediated pain-related aversion(Ren et al.,2006).Extracellular signal-regulated kinase(ERK),a family member of mitogen-activated protein kinases(MAPKs),has been implicated in NMDA-dependent synaptic *** of NMD A receptor markedly increased expression levels of phosphor-ERK(pERK) through a Ca-dependent mechanism,and PKA might mediate the NMDA-dependent ERK *** ERK translocates from the cytosol to the

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Cation-chloride co-transportors involved in cyclothiazide induced depolarizing shift of EGABA in CA1 pyramid neurons during seizure

Cation-chloride co-transportors involved in cyclothiazide in...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Zheng WU Yun WANG~* institutes of brain science and state key laboratory for medical neurobiology Fudan University Shanghai 200032 China

＜正＞Imbalance between inhibitory and excitatory in the brain is one of the basic mechanisms for epileptogenesis. Previous study from our lab has shown cyclothiazide（CTZ） is a potant convulsant in inducing epilepsy both in vitro and in vivo by activating glutamatergic and suppressing GABAergic ***-R-induced fast response depends on the concentration gradient of intracellular and extracellular chloridion,which is regulated partly by cation-chloride cotransportors （CCCs）.The aim of current study is to investigate the functional role of CCCs in CTZ induced epileptogenesis. Rat hippocampal slice（P21-P30） were incubated with 50μM CTZ for 2 hours before CA1 pyramid neurons were recorded using whole cell patch *** induced robust spontaneous discharges to epileptiform bursts in CA1 pyramid neurons and a depolarizing shift in EGABA（control = -62.56±1.06 mV,n = 14 vs CTZ = -56.15±0.93 mV,n=l 1;P＜0.05）.To determine the contributions of CCCs in setting EGABA under these conditions,furosemide,a CCCs inhibitor,was tested on its effect on *** with previous reports,furosemide（100μM） also induced a depolarizing shift in EGABA（Furosemide = -57.58±1.51 mV,n = 5）.In conclusion,our results demonstrate that down-regulation of CCCs activity may contribute to the CTZ-induced epileptiform activity in CA1 pyramid neurons.

关键词： epilepsy EGABA shift cyclothiazide hippocampus KCC2

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Kir andⅠh channels regulate the excitability of rat retinal ganglion cells

Kir andⅠh channels regulate the excitability of rat retinal...

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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： Shu-Xia JIANG,Zhongfeng WANG~* institutes of brain science,Institute of neurobiology and state key laboratory of medical neurobiology,Fudan University,Shanghai 200032,China

Objective Retinal ganglion cells,output neurons of the retina,receive excitatory inputs from bipolar cells and inhibitory inputs from amacrine *** change of RGC excitability directly influences visual processing in the *** both inwardly rectifying potassium channels(Kir) and hyperpolarization-activated cation channel(I) were reported to be involved in the regulation of cell excitability in the CNS,the effects of these channels in regulating the excitability of RGCs are largely *** Whole-cell patch-clamp recordings were performed on ganglion cells in retinal slice *** activities were recorded under current-clamp *** (1) Blocking synaptic inputs almost completely eliminated spontaneous firing both in ON- and OFF-types of RGCs. Blocking the inhibitory synaptic inputs alone enhanced the firing frequency,shortened interspike interval of the RGCs, while inhibition of excitatory inputs alone terminated the spontaneous firing in almost all RGCs tested.(2) In the presence of synaptic blockers(NBQX,D-APV,bicuculline and strychnine),extracellular application of Ba,a Kir channel blocker,significantly increased the firing frequency both in ON- and OFF-types of RGCs,as well as induced a membrane ***,the firing pattern of the cells became much more regular.(3) In the presence of synaptic blockers,Cs,a nonspecific I blocker,also remarkably increased the firing frequency and induced a membrane depolarization both in ON- and OFF-types RGCs,while the firing pattern of the cells became much more irregular after Cs ***,ZD7288,a specific I blocker,induced an increase of the frequency of spontaneous firing in most,if not all,of *** These results suggest that both Kir and I channels are functionally expressed in rat RGCs,and these channels are involved in the regulation of RGC excitability.

关键词： retinal ganglion cell Kir Ih excitability spontaneous firing

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The transcription factor Sp8 characterizes the configuration of the subventricular zone,rostral migratory stream and olfactory bulb in the adult rhesus monkeys

The transcription factor Sp8 characterizes the configuration...

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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： Rui CHEN,Zhen-Gang YANG~* institutes of brain science and state key laboratory of medical neurobiology,Fudan University,Shanghai 200032, China

＜正＞Neurogenesis in the primate subventricular zone（SVZ） of the lateral ventricle（LV） persisted throughout adulthood and the existence of rostral migratory stream（RMS） pathway in the adult primates has been intensely ***,the cellular organization and neurogenesis process in vertebrates,especially in monkeys remains *** the present study,we described the general configuration of the SVZ and RMS in adult monkeys via the expression of zinc finger transcription factor *** we detected the expression of Sp8 in the amphibians,reptiles, birds,rodents and *** of the neuroblasts in SVZ,RMS,olfactory tract and olfactory bulb are Sp8 ***,this transcription factor is highly expressed in the interneurons（CR,CB,TH and PV） of adult monkey olfactory *** with immunohistochemical detection of a marker for DNA replication（BrdU） and several cell type-specific markers,newly generated intemeurons are detected in the monkey olfactory *** together,our data suggest that Sp8 could be used as a marker to characterize neurogenesis of adult SVZ and RMS in rhesus monkeys.

关键词： Sp8 rhesus monkey neurogenesis neuroblasts subventricularzone rostral migratory stream olfactory bulb

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CB1 receptor-mediated modulation of synaptic transmission in rat retinal ganglion cells

CB1 receptor-mediated modulation of synaptic transmission in...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Xiao-Han WANG,Yi WU,Zhong-Feng WANG institutes of brain science,Institute of neurobiology and state key laboratory of medical neurobiology,Fudan University, Shanghai 200032,China

Endocannabinoid serves as a retrograde signaling to regulate neurotransmitter release via activating presynaptic CB1 receptors in the CNS,thus playing a critical role in neural development,synaptic plasticity,learning and memory,*** CB1 receptors are known to be widely distributed in the retina,the functions of endocannabinoid are still largely *** the present study,we tried to explore modulation by endocannabinoid signaling of glycine receptor-mediated miniature inhibitory postsynaptic currents(mIPSCs) and AMPA receptor-mediated miniature excitatory postsynaptic currents(mEPSCs) in retinal ganglion cells(RGCs) by patch-clamp techniques in rat retinal *** room temperature(22℃),when 5μmol/L WIN55212,a CB1 receptor agonist, was supplied to the normal 2 mmol/L Ca-containing bath solution,the frequency of mIPSCs was significantly decreased and the effect was reversed by co-application of 10μmol/L AM251,a CB1 receptor ***, the activation of CB1 receptor did not affect the frequency of mEPSCs both in room and in higher temperature(32℃).When extracellular Ca([Ca]) was elevated from 2 mmol/L to 5 mmol/L,the frequency of both mIPSCs and mEPSCs increased significantly,which could be inhibited by ***,under([Ca])-free condition, the frequency of mIPSCs was reduced to a extremely low level,whereas the frequency of mEPSCs was hardly changed as compared with normal 2 mmol/L[Ca],and not be affected by *** of the slices with thapsigargin,an inhibitor of sarco-endoplasmic reticulum Ca-ATPases,significantly increased the frequency of mEPSCs,indicating that Ca released from intracellular Ca stores plays an important role in triggering glutamate release from terminal of the retinal bipolar *** these results suggest that activation of presynaptic CB1 recep- tors modulates synaptic transmission in rat RGCs in a Ca-dependent *** regulation of inhibitory and excitatory synaptic transmission by cannabinoi

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Localization of dopamine Dl receptor in carp retina

Localization of dopamine Dl receptor in carp retina

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中国神经科学学会第四次会员代表大会暨第七届全国学术会议

作者： Xiao-Min ZHANG,Yong-Mei ZHONG,Xiong-Li YANG Institute of neurobiology,institutes of brain science and state key laboratory of medical neurobiology,Fudan University,Shanghai 200032,China

＜正＞Dopamine,as an important neuromodulator in the vertebrate retina,exerts its actions through two classes of receptors,known as the D1 and D2 *** this work,localization of D1 receptors in carp retina was studied using immunofluorescence double labeling and confocal laser scanning microscopy.D1-immunoreactivity（-IR） was observed in a lot of cells in the inner nuclear layer（INL） and some cells in the ganglion cell layer（GCL）.Anti-protein kinase C（PKC）, tyrosine hydroxylase（TH） and choline acetyltransferase（ChAT） antibodies were used to label rod-ON bipolar cells, dopaminergic amacrine cells（ACs） and cholinergic ACs,*** labeling experiments showed that D1 was strongly expressed in the somata of rod-ON bipolar cells,whereas the dendrites in the outer plexiform layer（OPL） and the terminals in the inner plexiform（IPL） of these cells were weakly ***-labeled dopaminergic ACs were located in the inner part of the INL and sent processes mainly into the outer sublayer of the IPL.D1 -IR was clearly observed in the somata but not seen in the processes of dopaminergic *** addition,D1 was also expressed in the somata of two types of cholinergic ACs（ACh-a,ACh-b）,with their cell bodies located in INL and GCL,respectively,but not in their processes in the ***,labeling for D1 was also observed in the ganglion cells which were retrogradely labelled by fluorogold. The wide expression profile of D1 suggests that the retinal dopaminergic system is organized in a way that favors its working through both wiring and volume transmission.

关键词： dopamine D1 receptor immunofluorescence double labeling bipolar cell amacrine cell ganglion cell

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Single Emxl-expressing neural stem cells give rise to both projection neurons and interneurons in vitro

Single Emxl-expressing neural stem cells give rise to both p...

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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： Yu-Qun CAI,Yue ZHANG,Zhen-Gang YANG~* institutes of brain science and state key laboratory of medical neurobiology,Fudan University,Shanghai 200032, China

＜正＞There are two major subtypes of neurons in the brain:projection neurons and *** Drosophila nervous system development,a subpopulation of single neuroblasts（neural stem cells） can give rise to both projection neurons and ***,in mammalian brains,whether single neuroepithelial cells or radial glias（primary neural stem cells） can produce both projection neurons and interneurons remains largely unknown. Using fluorescence-activated cell sorting（FACS）,we cultured single Emxl-expressing neural stem cells derived from E10.5-11.5 mouse *** weeks after culturing in defined serum-free medium,multiple-immunostaining revealed that a subpopulation of single Emxl-expressing neural stem cells can produce both projection neurons and *** projection neurons in a single cell-derived clone express typical projection neuronal markers,such as Tbr1,Cux1,Ctip2,Foxp1 and Satb2,while interneurons in the same clone co-express Sp8 and ***, our preliminary results suggest that,during marine forebrain development,at least some projection neurons and interneurons share single multipotent progenitors.

关键词： projection neurons interneurons share single neural stem cells

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