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Regulation of L-type voltage-dependent Ca2+ channels by hypoxia inducible factor-lα in hypoxia-enhanced cell death

Regulation of L-type voltage-dependent Ca2+ channels by hypo...

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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： Ran LI~1,Ling-Ling ZHU~2,Xiao-Li GONG~1,Fen YU~1,Fang SHI~1,Ming FAN~2,Xiao-Min WANG~1,Xuan WANG~(1,*) 1 Department of Physiology,Capital Medical University,Key Laboratory for Neurodegenerative Diseases of Education Ministry of China,Beijing 100069,China 2 Department of Brain Protection & Plasticity Research,Beijing Institute of Basic Medical Sciences,Beijing 100027, China

Purpose Though the oxygen concentration is 20%in atmosphere,it is around 3%in *** is the basic environment in vivo,moderate oxygen concentration can promote cell ***,if the oxygen concentration is too low or the hypoxia lasts too long,it will cause the cell *** phenomena are very clear now, but the mechanisms behind them are still *** we all know,HIF-1αis the main regulator during hypoxia. Our previous experimental results show that hypoxia has an effect on the expression of L-type voltage-dependent Ca *** this study,we have focused on the regulation of L-type voltage-dependent Ca channels by hypoxia inducible factor-1α（HIF-1α） in hypoxia-enhanced cell *** Detect the effect of HIF-1αinhibitor echinomycin on cell viability with *** the expression of L-type voltage-dependent Ca channel subtypes Ca1.2 and Ca1.3 by Western blot and real-time PCR ***（1） 3%O induced the death of PC12 cells.（2） After culturing PC12 cells in 3%O2 and 20%O for 24 h with HIF-1αinhibitor echinomycin in various concentrations, we detected the cell viability with *** found that 1 nM and 5 nM group had an acceptable mortality.（3） Western blot result showed that both Ca1.2 and Ca1.3 proteins were up-regulated during hypoxia,and down-regulated dosedependently when using HIF-1αinhibitor.（4） PCR data showed that the gene of Ca1.2 has no difference between hypoxia and normoxia treatment,while that of Ca 1.3 increased in hypoxia ***,both of them decreased dose-dependently when the culture was treated with the HIF-1α*** Ca1.2/Ca1.3 proteins are correlated positively with HIF-1αin protein level,however,the relationship between Ca1.2/Ca1.3 and HIF-1αis not very clear in gene level,which needs to be verified further.

关键词： HIF Regulation of L-type voltage-dependent Ca channels by hypoxia inducible factor-l in hypoxia-enhanced cell death

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A novel PLA2G6 gene mutation causes autosomal recessive early-onset parkinsonism with typical clinical phenotype of Parkinson's disease

A novel PLA2G6 gene mutation causes autosomal recessive earl...

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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： Ji-Feng GUO~(1,3),Chang-He SHI~1,Lei WANG~1,Zhan-Yun LV~1,Jian WANG~4,Lin-Zi LUO~1,Lu SHEN~(1,3),Hong JIANG~(1,3),Xin-Xiang YAN~(1,3),Qian PAN~2,Kun XIA~2,Bei-Sha TANG~(1,2,3,*) 1 Department of Neurology,Xiangya Hospital,Central South University,Changsha 410008,China 2 National Laboratory of Medical Genetics of China,Changsha 410008,China 3 Neurodegenerative Disorders Research Center,Central South University,Changsha 410008,China 4 Department of Neurology,Huashan Hospital,Fudan University,Shanghai 200040,China

Objective Mutations in the PLA2G6 gene at the PARK14 locus have been reported in complicated parkinsonism *** assess the prevalence of and phenotypes associated with PLA2G6 gene mutations,we screened PLA2G6 mutations in a cohort of patients with autosomal recessive early-onset parkinsonism(AREP). Methods We selected 12 families with AREP in which the Parkin,PINK1,DJ-1,ATP13A2,and FBX07 gene mutations had been previously *** patients came from the mainland of *** entire PLA2G6 coding region and exon-intron boundaries were sequenced from genomic DNA *** then performed PET studies on individuals in the pedigree with a homozygous PLA2G6 mutation,and investigated the enzyme activity level of the *** A homozygous missense mutation,C.G991T(p.D331Y) was identified in an autosomal recessive case.A younger sister of the p.D331Y-carrying patient was also homozygous for the mutation,but was with no extrapyramidal symptoms.A PET study showed a substantial reduction in dopamine transporter(DAT) binding in the p.D331Y patient,and a slight reduction in DAT binding in his *** vitro,we experimentally demonstrate that the D331Y mutation caused an approximately 70%reduction in enzyme *** We have confirmed that the PLA2G6 gene allocated PARK14 locus and is associated with AREP.

关键词： PLA2G6 PARK14 PET AREP

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The inhibitory effect of botulinum toxin type A on pyloric smooth muscle contractile response to substance P in rats in vitro

The inhibitory effect of botulinum toxin type A on pyloric s...

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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-Feng SHAO,Yan-Feng SONG,Yin-Xiang RENG,Xiang-Wen LIU,Yi-Ping HOU~* Department of Neuroscience,Anatomy,Histology and Embryology,Key Laboratory of Preclinical Study for New Drugs of Gansu Province,School of Basic Medical Sciences,Lanzhou University,Lanzhou 730000,China

Objective Our previous studies has demonstrated that botulinum toxin type A（BTX-A）intrasphincteric injection through guide cannula in free move rats dose-dependently caused an inhibition of slow wave in amplitude but not in frequency,a diminishment of spike activity in amplitude and spike burst of pyloric myoelectrical activity, and a reduction of substance P（SP）content and cell number of SP-immunoreactivity in *** data suggest that botulinum toxin may inhibit not only the acetylcholine（ACh）release,but also SP release from the autonomic and enteric nervous terminals in ***,a major excitatory non-cholinergic neurotransmitter,depolarizes the membrane potential and thus induces contraction in gastrointestinal smooth *** is an undecapeptide belonging to the tachykinin family and can induce strong contractions in pylorus via neurokinin 1 receptor（NK1R）,the preferred receptor for ***,no experimental evidence that BTX-A inhibits the pyloric contractile response induced by SP release from nervous terminals or SP dosed has been documented until *** aim of this study was to determine the inhibitory effect of BTX-A on pyloric smooth muscle contractile response to substance P in rats in vitro. Methods The effect of BTX-A on contractile tension and frequency of pyloric muscle strips incubated with Krebs bicarbonate buffer with electric field stimulation（EFS,100 V,4 Hz,0.5 ms pulse width duration）were detected after atropine（1 uM）abolishing cholinergic action and compared with the effect of[D-Arg,D-Phe,D-Trp,Leu]-SP （APTL-SP）,a NK1R *** pyloric smooth muscle contractile response to SP（1 uM）added at intervals of 30 min under a range dose of BTX-A（from 4 to 10 U/mL）incubation for 4 h was measured,and the distributions of SP and NK1R cells in pylorus were examined by *** BTX-A（10 U/mL）as APTLSP （1μM）further decreased EFS-induced pyloric contractile tension（from 1.524±0.029 to 1.362±0.027,P<0.01） and frequency（from

关键词： botulinum toxin type A substance P NK1-receptor antagonist pyloric smooth muscle electric field stimulation rat

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Comparative study on treatment of tail vein injection of bone marrow stromal cells and intraperitoneal injection of granulocyte colonystimulating factor after cerebral infarction in rats

Comparative study on treatment of tail vein injection of bon...

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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： Zhong-Xia ZHANG~1,Ping GU~(1,2,*),Mei-Gang ZHOU2,Yan-Yong WANG~(1,2),Zhen-Qing ZHANG~2,Dong-Sheng CUI~1, Yuan GENG~1,Ming-Wei WANG~(1,2) 1 Brain Aging and Cognitive Neuroscience Laboratory of Hebei Province,Shijiazhuang 050031,China 2 Department of Neurology,First Hospital of Hebei Medical University,Shijiazhuang 050031,China

Objective To explore the effects of intravenous injection of bone marrow stem cells（BMSCs） and intraperitoneal injection granulocyte colony stimulating factor（G-CSF） to mobilize stem cells on treatment of cerebral artery occlusion（MCAO） model in *** Modified Zea-longa suture method was applied on the SD rats to block the middle cerebral artery,thus establish the model of cerebral *** were injected via the tail vein and G-CSF was injected intraperitoneal 24 h after MCAO.7 and 14 days after the cell transplantation and stem cell mobilization,the neurological motor and conginitive function was evaluated by Morris water maze *** infarct volume,and the BrdU positive cell survival number,migration and distribution were detected by Paraffin a slice of the organization of the immunochemical *** therapeutic effect of MCAO rats was conducted in the two group ***（1） The swimming speed of both BMSCs group and G-CSF group at the 7 day and the 14 day after transplantation were higher than the PBS group,while the time for searching platform were shorter,and the performance of G-CSF group was better than the BMSCs group.（2） Compared with the model group,the infarction size of BMSCs group and G-CSF group decreased at the 7 day and the 14 day,and the size of G-CSF group was smaller than BMSC group.（3） The number of BrdU-positive cells of G-CSF group was more than BMSCs group at the 7 day and the 14 after the injection,and mobilized stem cells could be seen surrounding cerebral ischemia and at the corpus *** Intraperitoneal injection of G-CSF and tail vein injection of BMSCs could improve the motor and cognitive function,decrease the infarction size,and promote BMSCs to migrate to the surrounding cerebral ischemia and corpus *** the results suggest that the method of G-CSF by mobilized BMSCs may be better than transplantation BMSCs via vein injection for treatment of cerebral ischemia.

关键词： bone marrow stromal cells transplantation granulocyte colony-stimulating factor cerebral ischemic disease middle cerebral artery occlusion model

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Netrin-1 hyper-expression of in the brain promotes angiogenesis and improves neurological outcome in transient MCAO mice

Netrin-1 hyper-expression of in the brain promotes angiogene...

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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： Hai-Yan LU~1,Yong-Ting WANG~2,Xiao-Song HE~2,Fa-Lei YUAN~2,Xiao-Jie LING~2,Bo-Hua XIE~2,Guang-Hui TANG~2, Jun HUANG~2,Yao-Hui TANG~2,Sheng-Di CHEN~1,Guo-Yuan YANG~(1,2,*) 1 Departments of Neurology,Ruijin Hospital,Shanghai JiaoTong University School of Medicine,Shanghai 200025, China 2 Neuroscience and Neuroengineering Research Center,Med-X Research Institute,Shanghai Jiao Tong University, Shanghai 200030,China

Objective Netrin-1(NT-1) is one of the axon-guiding molecules that are critical for neuronal ***-1 stimulates proliferation and migration of human cerebral endothelial cells in vitro and promotes focal neovascularization in adult brain in *** is unknown whether NT-1 improves long-term functional outcome after ischemic *** aim of this study is to determine whether hyper-expression of NT-1 through adenoassociated virus(AAV) -mediated gene transfer enhances neurovascular remodeling and improves functional outcome in mouse model of focal cerebral *** AAV-NT-1 virus was stereotactic injected into the basal ganglia of male CD-1 mice(n=30).Seven days after injection,these mice underwent 60 minutes of transient middle cerebral artery occlusion(tMCAO) followed by 1 to 4 weeks of *** of NT-1 and its receptors was quantified by Western blot and *** volume and behavioral analyses were performed after tMCAO. Synchronic radiation angiography was used to detect the changes of cerebral *** NT-1 expression in the ischemic hemisphere was enhanced after AAV-NT-1 gene *** neurons and astrocytes but not endothelial cells expressed ***-1 receptors(DCC and UNC5H2) also expressed in neurons and ***-expression of NT-1 decreased infarct volume in the AAV-NT-1-transduced mice compared to the control(P<0.01).Long-term observation demonstrated that neurobehavioral outcomes were greatly improved in the AAV-NT-1-tranduced mice compared to the control(P<0.05).Further study demonstrated that the microvessel counts were significantly increased in the AAV-NT-1 transduced mice(P<0.05).Synchrotron radiation angiography showed that the brunches of MCA was increased in NT-1 treated *** Our data demonstrate that NT-1 hyper-expression reduced ischemic injury and promoted functional *** may due to enhancement of focal neovascularization.

关键词： adeno-associated virus angiogenesis gene transfer ischemia middle cerebral artery occlusion netrin-1

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Ser9 phosphorylation of GSK-3β attenuates calpain-mediated truncation of its N- and C-terminus

Ser9 phosphorylation of GSK-3β attenuates calpain-mediated ...

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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-Shan MA~(1,2,3),Shao-Jun LIU~(1,2,3),Qiao-Ying HUANG~(1,2,3),Bing-Quan LAI~(1,2,3),Chong WANG~(1,2,3),Ming-Tao LI~(1,2,3,*) 1 Department of Pharmacology,2Proteomics Center,and 3 Research center for Neurobiology,Zhongshan School of Medicine,Sun Yat-sen University,74 Zhongshan Road II,Guangzhou 510080,China

Glycogen synthase kinase-3β(GSK-3β),a key regulator of neuronal apoptosis,is inhibited via phosphorylation on Ser-9,and has also been recently shown to be cleaved by calpain at the N-terminus and subsequently *** the present study we show that calpain truncates GSK-3βat not only N-terminus but also C-terminus,which occurs following Ser9 dephosphorylation during neuronal *** found that,calpain-mediated cleavage was significantly attenuated by increasing Ser9 phosphorylation of GSK-3B via inhibition of phosphatase 1/2A in vivo or pretreatment with purified active Akt in *** serine 9 to alanine increased GSK- 3β's interaction with and sensitivity to *** results demonstrate that Ser9 phosphorylation protects GSK-3βfrom calpain-mediated truncation through interrupting their *** spectrometry analysis indicated GSK-3B cleavage occurred at residues Thr38-Thr39 and *** cleavage generated three truncated products all leaving the kinase domain intact,ΔN-GSK-3β(39-420aa),ΔC-GSK-3B(l-384aa) and AN/C-GSK-3B(39-384aa),each of them exhibiting increased kinase activity and pro-apoptotic *** results suggest that GSK-3βC-terminus, similar to N-terminus,serves as an auto-inhibitory *** agreement with this,C-terminal peptide inhibits GSK- 3βactivity and protects neurons from death in a Ser389 phosphorylation dependent *** therefore conclude that an important role of GSK-3βcleavage during apoptosis is the removal of its N-/C- terminal inhibitory domains to increase its activity,whereas an opposing role of Ser9 phosphorylation of GSK-3βduring survival is the direct suppression of its activity and the decrease of calpain-mediated cleavage.

关键词： GSK-3β calpain Ser9 phosphorylation cleavage neuronal apoptosis

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EA produces antidepressant effect by promoting the proliferation of both quiescent neural progenitors and amplifying neural progenitors in the dentate gyrus of hippocampus

EA produces antidepressant effect by promoting the prolifera...

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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： Liu YANG~1,Qiong LIU~2,Bing LI~1,Na YUE~1,Jin YU~1,Gen-Cheng WU~(1,3,*) 1 Department of Integrative Medicine and Neurobiology,State key lab of Medical Neurobiology,Shanghai Medical College,Fudan University,Shanghai 200032,China 2 Department of Anatomy,Histology and Embryology,Shanghai Medical College,Fudan University,Shanghai 200032, China 3 Institutes of Brain Science,Fudan University,Shanghai 200032,China

In order to evaluate the antidepressant effect of electroacupuncture(EA) on the rat model of chronic unpredictable stress(CUS) and the proliferation promoting effect on stem / progenitor cells in the dentate gyrus (DG) of hippocampus,this study investigated the behavioral and brain section staining changes of 4 groups(normal, Model,Sham EA and EA).After 2 weeks of CUS procedure,EA,as well as Sham EA,was performed on Du-20 (Bai-Hui) and GB-34(Yang-Ling-Quan) for 15 d(8 treatments in all).Forced swimming test(FST),elevated plus-maze (EPM) test and open field test(OFT) suggested that EA had reversed the depressive like ***, Sham EA,as designed on the real acupoints without electrical current,also showed a little antidepressant *** shown by 5-bromo-2-deoxyuridine(BrdU) and SOX-2 double-labeling immunofluorescence,proliferation of DG stem cells was decreased in model *** with the depression model,the decrease of stem-cell proliferation in both Sham EA and EA groups had begun to be significantly blocked since 2 treatments were carried *** the subpopulations of stem cells into consideration,we found that EA could up-regulate both quiescent neural progenitors (QNPs,SOX-2 positive and GFAP positive) and amplifying neural progenitors(ANPs,SOX-2 positive but GFAP negtive) in DG while Sham EA could only promote the latter,through SOX-2 and GFAP double *** further demonstrated that both Sham EA and EA increased p-ERK and SOX-2 double positive cells number and the both groups had the increasing effect at the same *** the EA group had a lower ratio of p-ERK in SOX-2 in respect that the total stem cell number was *** results suggest that EA has the antidepressant effect mediated by promoting both QNPs and ANPs in DG and that acupuncture could treat depression by activating ANPs,ERK as a possible passway to *** other passways may be included in the mechanism of EA antidepressant effect.

关键词： EA produces antidepressant effect by promoting the proliferation of both quiescent neural progenitors and amplifying neural progenitors in the dentate gyrus of hippocampus

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Ibuprofen suppresses inflammation and ameliorates cognitive impairment in presenilin 1 and presenilin 2 conditional double knock-out mice

Ibuprofen suppresses inflammation and ameliorates cognitive ...

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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 YAN,Li LI,Meng-Dan LI,Ting-Ting JI,Bing MEI~* Shanghai Institute of Brain Functional Genomics,Key Lab of Brain Functional Genomics,MOE & STCSM,East China Normal University,3663 North Zhongshan Rd,Shanghai 200062,China

Objective Alzheimer's disease(AD)is one of the most common age-related neurodegenerative diseases. The brain of AD patients show a chronic inflammatory response characterized by activated glial cells and increased expression of cytokines,chemotactic factors and complement ***,whether the inflammatory response in AD pathologic process is a powerful pathogenetic force or a concomitant symptom remains *** studies have demonstrated that there is a reduced risk for AD patients using nonsteroidal anti-inflammatory drugs (NSAIDs).This present study is using ibuprofen,which is one of the most commonly used NSAIDs,to evaluate whether it could affect AD-like pathology in 3-month-old(mild inflammatory response and memory impairment) and 6-month-old(intensive inflammatory response and memory impairment)presenilin 1 and 2 conditional double knockout(cDKO)*** Conditional DKO mice were obtained by crossing the forebrain specific PS 1 heterozygous knockout mice with conventional PS2 heterozygous knockout mice on B6/CBAF1 genetic background. Mice with the Cre transgene,fPS1/fPS1,and PS2-/-served as cDKO,their littermates(no Cre,fPS1/+ and PS2+/+ or fPS1/+ and PS2+/-)served as *** were housed under 20-26℃temperature and 40%-70%humidity with 12 hrs light/dark cycles,and all the experiments were approved by the Animal Ethics Committee in ECNU, *** mice were employed a dose of dietary ibuprofen(375 ppm)at the early stage(3-month-old)and the middle stage(6-month-old),respectively,continuously treated for 6 months,while for the control group,the littermate cDKO mice received standard *** a series of behavioral tests were performed to assay the ibuprofen effects on cognitive impairment in cDKO ***(1)The 3-month-old cDKO mice showed improved recognition memory in 1-hour novel object recognition after the ibuprofen treatment.(2)The 6-month-old cDKO mice show improved memory in 1-hour novel object recognition,enhanced co

关键词： ibuprofen inflammation presenilin gene Alzheimer's disease

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The cellular localization of receptor-interacting protein 3 in adult rat retina

The cellular localization of receptor-interacting protein 3 ...

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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： Lei SHANG,Kun XIONG,Ju-Fang HUANG~* Department of Anatomy and Neurobiology,Xiangya School of Medicine,Central South University,Changsha 410013, China

Objective RIP family includes a group of Ser/Thr kinases that modulate cell death as essential sensors to various cellular ***-interacting protein 3(RIP3),a member of the RIP family,has been found to induce necroptosis(programmed necrosis).Till now,studies in RIP3 were focus on its biology functions and general distribution in peripheral ***,nothing has been known about which cell types express RIP3 in normal adult rat retina,which is the important animal model in neuroscience ***,our purpose is to identify the cellular localization of RIP3 in rat *** The profile expression of RIP3 in rat retina was detected by immunohistochemistry,the double labeling between RIP3 and retina cell markers were identified by immunofluorescence staining,then laser con-focal microscopy(Nikon CZ-01) were performed to analyze the presence of double-fluorescent label in all types of retina ***(1) The immunohistochemistry shows RIP3 express in nerve fiber layer(NFL),ganglion cell layer(GCL) and inner nuclear layer(INL) in normal rat retina;(2) The double labeling of RIP3 and NeuN(ganglion cell specific marker),Parvalbumin(amacrines cell specific marker),Calbindin (horizontal cell specific marker) suggests a preferential localization of RIP3 in ganglion cell,amacrines cell and horizontal cell;(3) The double labeling of RIP3 and GS(M(u|¨)ller cell specific marker),GFAP(microglial cell specific marker),CD11b(macroglial cell specific marker) indicates a localization of RIP3 in Muller cell,microglial cell and marcoglial cell in the NFL,GCL and INL;(4) There is no distinctly double staining between RIP3 and PKC-α(bipolar cell specific marker),Rhodopsin(rod and cone cell specific marker),which suggests RIP3 does not express in bipolar cell and rod and cone *** RIP3 is expressed in NFL,GCL and INL,furthermore,it has present in amacrine cells,horizontal cells,ganglion cells,M(u|¨)ller cells,macroglial cells and microglial cells

关键词： RIP3 immunolocalization rat retina

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Experimental research on effects and mechanisms of ginsenoside Rd on the allodynia in rats with SNI

Experimental research on effects and mechanisms of ginsenosi...

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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： Ling-Feng TU~#,Rong CAO~#,Li DUAN,Ying-Fei XIONG,Zhi-Ren RAO~* Institute of Neurosciences of PLA,the Fourth Military Medical University,Xi'an 710032,China

Objective To investigate the analgesis and mechanisms of ginsenoside Rd(GR-d)on the allodynia in rats with *** Thirty male SD rats were randomly divided into five groups:Blank control,SNI,Sham operation,SNI + Saline(1 mL/kg body weight,intra-peritoneal injection,i.p.),and SNI + G-Rd(10 mg/1 mL/kg body weight,i.p.)*** measured paw withdrawal mechanical thresholds(PWMT)using calibrated von Frey filaments to stimulate the glabrous surface of the lateral plantar of the hindpaw on SNI *** staining was used to measure the mean fluorescent intensity(MFI)of glutamate immunoreactive substance and N-methyl-D-aspartale receptor-2-A/B subunit(NR2A/B)immunoreactive substances in the spinal cord dorsal horn (L4-6)in five groups,so was SP immunoreactive substance and NK-1 immunoreactive *** At 10 d after SNI,the mean PWMT value on SNI side revealed clearly lower than that of control and sham operation groups, at 20 d PWMT value dropped to the *** SNI + G-Rd group,the PWMT value significantly ascend than that of SNI + saline group(P<0.05).The immunofluorescent staining revealed that the MFI of glutamate-like,NR2A/B subunit-like and SP-like immunoreactive substances together with the number of NK-1 immunoreactive substance in the dorsal horn of the SNI side in spinal cord L4-6 segments showed significantly higher compared with that of control and sham operation *** SNI + G-Rd group,the MFI of glutamate-like,SP-like immunoreactive substances and the number of NK-1 immunoreactive substance revealed clearly lower than that of SNI and SNI + saline groups(P< 0.05),However,the MFI of anti-NR2A/B subunit-like immunoreactive substance did not showed significant changes in three groups mentioned *** GR-d revealed obvious analgesic effects on neuropathic pain induced by *** descent of the glutamate and SP content together with the number of NK-1 in the dorsal horn of spinal cord L4-6 segment was a possible mechanism

关键词： neuropathic pain ginsenoside-Rd analgesis spinal cord glutamate NR2A/B SP NK-1 receptor

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