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-synuclein amino terminus regulates mitochondrial membrane permeability in dopaminergic cells

-synuclein amino terminus regulates mitochondrial membrane p...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：杨慧高姗姗王雪张辉林申甲梅段春礼高歌首都医科大学基础医学院神经生物学系

Parkinson's disease(PD)is a common neurodegenerative movement disorder affecting an increasing number of *** studies have shown that mitochondrial dysfunction and abnormal protein aggregation are two major contributors to the progression of *** N terminus ofα-synuclein(α-Syn),which adopts aα-helical conformation upon lipid binding,is essential for membrane interaction;yet its role in mitochondria remains poorly defined.A functional characterization of theα-Syn N-terminal domain and investigation of its effect on mitochondrial membrane permeability were undertaken in this study.α-Syn/N andα-Syn/del N(amino acids 1–65 and 61–140,respectively)constructs were overexpressed in dopaminergic MN9D *** lactate dehydrogenase release and decreased cell viability was observed in cells transfected withα-Syn/N but notα-Syn/del *** addition,anα-Syn/N-induced increase in the level of intracellular reactive oxygen species,alteration in mitochondrial morphology,and decrease in mitochondrial membrane potential were accompanied by the activation of mitochondrial permeability transition *** changes were also associated with a decline in mitochondrial cardiolipin content and interaction with the voltage-dependent anion channel and adenine nucleotide translocator in the mitochondrial *** results suggest that the N terminus ofα-Syn is essential for the regulation of mitochondrial membrane permeability and is a likely factor in the neurodegeneration associated with PD.

关键词： Parkinson's disease α-synuclein mitochondrial membrane permeability

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GABAergic Actions Mediate Opposite Ethanol Effects on Dopaminergic Neurons in the Anterior and Posterior Ventral Tegmental Area

GABAergic Actions Mediate Opposite Ethanol Effects on Dopami...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：关艳中金秀东郭冉牡丹江医学院生理学教研室

It is known that the posterior ventral tegmental area(p-VTA)differs from the anterior VTA(a-VTA)in that rats learn to self-administer ethanol into the p-VTA,but not into the *** activation of VTA dopaminergic neurons by ethanol is a cellular mechanism underlying the reinforcement of ethanol consumption,we hypothesized that ethanol may exert different effects on dopaminergic neurons in the p-VTA and *** patch-clamp recordings in midbrain slices from young rats(postnatal days 22-32),we detected no significant difference in electrophysiological properties between p-VTA and a-VTA dopaminergic ***,acute exposure to ethanol(21–86 m M)stimulated p-VTA dopaminergic neurons but suppressed a-VTA dopaminergic ***,ethanol(>21 m M)dose-dependentlyreduced the frequency of the GABAergic spontaneous inhibitory postsynaptic currents(s IPSCs)generated by inhibitory neuronal firing but not miniature inhibitory postsynaptic currents(m IPSCs)in p-VTA dopaminergic *** contrast,ethanol increased the frequency and amplitude of both s IPSCs and m IPSCs in a-VTA dopaminergic *** of these effects of ethanol were abolished by a GABAA receptor *** was a strong negative correlation between ethanol-evoked modulation of s IPSCs and neuronal firing in VTA dopaminergic *** results indicate that GABAergic inputs play an important role in ethanol’s actions in the *** differential effects of ethanol on s IPSCs and neuronal firing in the p-VTA and a-VTA could be the basis for ethanol reinforcement via the p-VTA.

关键词： dopaminergic neuron GABAergic neuron ethanol VTA

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A mechanism of modulating dopamine release in the striatum by smoking-level nicotine

A mechanism of modulating dopamine release in the striatum b...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者： Li Wang Ruiying Jiao Wei Liu Huadong Xu Feipeng Zhu Xiaoxuan Sun Li Zhou Minli Li Qinglong Wang Sasa Teng Changhe Wang 周专 SKLM-CLS-Mc G-BKCMM Institute of Molecular MedicinePeking University 北京大学

Striatal dopamine(DA)release can be independently triggered by two pathways,the midbrain nigrostriatal ascending pathway and the cholinergic transmission-induced dopaminergic transmission(CTDT)***–level nicotine(SL-nicotine)causes addiction by modulating DA release,but with paradoxical ***,we investigated how SL-nicotine modulates DA release following optogenetics-induced CTDT in striatal *** found that(1)SL-nicotine abolished DA release via CTDT but not the ascending pathway;(2)an electrical-field stimulus(Estim)triggered DA release with 2phases of DA(2p DA),corresponding to ascending and CDTD pathways;(3)following a single Estim,SL-nicotine reduced the depletion of DA vesicle pool;(4)during a burst Estim,SL-nicotine inhibited the initial component but facilitated the 2*n ACh Rs wereβ6alater component of 2p DA due to reduced DA depletion;(5)responsible for striatal *** work resolves the paradoxical observations of 2*n ACh Rs and CTDT as targets to treatβ6aEstim-induced DA release and offer smoking addiction.

关键词： dopamine nicotine striatal slice

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Opposing dendritic spine plasticity of fear learning and unlearning in neocortex

Opposing dendritic spine plasticity of fear learning and unl...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者： Cora Sau Wan Lai Department of Physiology LKS Faculty of MedicineThe University of Hong Kong

Fear conditioning is an associative learning in which a conditioned neutral stimulus is paired with an aversive unconditioned stimulus to elicit fear *** exposures to conditioned stimulus diminish the expression of fear response,a process called *** it is generally believed that fear extinction is a form of new learning that inhibits rather than erases previously acquired fear memories,the hypothesis that extinction causes the partial erasure of fear memories remains *** investigate how neural circuits are modified by fear learning and extinction,we used transcranial two-photon microscopy to examine the formation and elimination of postsynaptic dendritic spines of layer V pyramidal neurons in mouse *** found that fear conditioning by pairing an auditory cue with a footshock increases the rate of spine elimination in mouse frontal association *** contrast,fear extinction by repeated presentation of the same auditory cue increases the rate of spine ***,spine elimination and formation induced by fear conditioning and extinction occur on the same dendritic branches in a cue-and location-specific ***,reconditioning preferentially induces elimination of dendritic spines that were formed after ***,within vastly complex neuronal networks,fear conditioning,extinction and reconditioning lead to opposing changes at the level of individual *** recently found that this opposing dendritic spine plasticity was also observed in primary auditory *** findings suggest that fear memory trace erasure could be a universal phenomenon in neocortex after fear extinction.

关键词： Fear conditioning Dendritic spine Plasticity Frontal association cortex Auditory cortex

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cPKCγ通过调节自噬降低氧-糖剥夺诱导的原代培养小鼠脑皮层神经元缺血损伤

cPKCγ通过调节自噬降低氧-糖剥夺诱导的原代培养小鼠脑皮层神经...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：陈璐勔罗艳琳朱弘倚于津泠张丹韩松李俊发首都医科大学神经生物学系北京脑重大疾病研究院

目的:在细胞水平,探讨经典型蛋白激酶C(c PKC)γ对原代培养小鼠脑皮层神经元氧-糖剥夺(OGD)损伤中作用及其与细胞自噬的关系。方法:利用C57BL/6J野生和c PKCγ基因敲除型小鼠,建立原代培养脑皮层神经元1h OGD/24h复糖复氧离体细胞缺血模... 详细信息

目的:在细胞水平,探讨经典型蛋白激酶C(c PKC)γ对原代培养小鼠脑皮层神经元氧-糖剥夺(OGD)损伤中作用及其与细胞自噬的关系。方法:利用C57BL/6J野生和c PKCγ基因敲除型小鼠,建立原代培养脑皮层神经元1h OGD/24h复糖复氧离体细胞缺血模型;借助3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法和乳酸脱氢酶(LDH)漏出率,检测OGD致神经元损伤情况;应用蛋白免疫印迹(Western blot),检测自噬相关蛋白Beclin-1表达水平和LC3Ⅱ/I蛋白水解情况;同时利用细胞自噬抑制剂3-甲基腺嘌呤(3-MA)和巴弗洛霉素A1(Baf A1),以及c PKCγ慢病毒表达载体,观察细胞自噬和c PKCγ在OGD致神经元损伤中作用。结果:c PKCγ基因敲除可显著加重OGD诱导的脑皮层神经元缺血损伤,即与OGD处理的野生型相比,存活率明显降低、死亡率显著升高(p<0.05,每组n=6);与野生型神经元相比,c PKCγ基因敲除显著降低OGD处理神经元自噬相关蛋白Beclin-1表达水平和LC3Ⅱ/I比值(p<0.05,每组n=6);自噬抑制剂3-MA和Baf A1可使OGD处理的野生型神经元存活率显著下降、死亡率明显升高(p<0.05,每组n=6),即与OGD处理的c PKCγ基因敲除型神经元相似;然而,用c PKCγ慢病毒载体转染后,OGD处理的c PKCγ基因敲除型神经元内自噬相关蛋白Beclin-1表达水平和LC3Ⅱ/I比值显著升高(p<0.05,每组n=6)的同时,神经元存活率明显增高、死亡率显著降低(p<0.05,每组n=6)。结论:c PKCγ可能通过调节细胞自噬水平,来减轻氧-糖剥夺诱发的原代培养小鼠脑皮层神经元的缺血损伤。

关键词：氧-糖剥夺(OGD) 细胞自噬经典型蛋白激酶C(cPKC)γ 3-甲基腺嘌呤(3-MA) 巴弗洛霉素A1(Baf A1)

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Mitochondrial Transplantation Attenuates Hypoxic Pulmonary Hypertension

Mitochondrial Transplantation Attenuates Hypoxic Pulmonary H...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：胡清华华中科技大学同济医学院

Aims:Mitochondria are essential for the onset of hypoxia-induced pulmonary vasoconstriction and pulmonary vascular-remodeling,two major aspects underlying the development of pulmonary ***,hypoxia induces relaxation of systemic *** heterogeneity appears critically involved in the initiation of hypoxia-induced constriction of pulmonary artery and relaxation of systemic *** remains unknown whether mitochondria control the distinct responses of pulmonary versus systemic artery smooth muscle cells(SMCs)to hypoxia,and whether mitochondrialheterogeneity can be experimentally exploited for a potential therapy against pulmonary ***:We showed that the transplantation of mitochondria derived from femoral artery smooth muscle cells inhibited acute hypoxia-induced membrane depolarization,[Ca2+]i elevation and constriction of pulmonary artery SMCs,and attenuated pulmonary hypertension by modulating pulmonary vasoconstriction and vascular-remodeling in rats exposed to chronic ***,the low or high amount of reactive oxygen species generation from mitochondria in SMCs was found to determine the relaxation of femoral artery versus constriction of pulmonary artery in response to ***:Mitochondria can be transplanted into SMCs in cell culture preparations,in isolated arteries in vitro and in *** behaviors can be manipulated by transplantation of exogenous mitochondria and the strategy of mitochondrial transplantation can be employed to determine the specific role(s)of mitochondria in any cellular signaling cascades and in pathophysiological processes underlying a ***:Our findings suggest that mitochondria necessarily determine the responses of vascular SMCs to hypoxia and mitochondrial transplantation have potential implications for exploring a novel therapeutic and preventive strategy against pulmonary hypertension.

关键词： Hypoxia Mitochondria Pulmonary hyertension

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Analysis of skin and secretions of Rana Dybowskii exposed to Staphylococcus aureus or Escherichia coli identifies immune response proteins

Analysis of skin and secretions of Rana Dybowskii exposed to...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者： Xiang-Hong Xiao Jing-Yu Zhang Long-Hui Chai Hui-Min Miao Xue-Can Shi Northeast Forestry University

The aim of the present study was to investigate responses in Dybowski’s frogs（Rana dybowskii）exposed to bacteria,using proteomic and transcriptomic *** aureus and Escherichia coli were used as representative Gram-positive and Gram-negative bacteria,respectively,in an infectiouschallenge *** skin and skin secretions were collected and protein expression in infected frogs compared to control frogs by two-dimensional gel electrophoresis,silver staining,and image *** that demonstrated differential expression were analysed by mass spectrometry and identified by searching protein *** than 180 protein spots demonstrated differential expression in ***-or ***-challenged groups and,of these,more than 55 spots were up-or down-regulated at least sixfold,*** with a potential function in the immune response were identified,such as stathmin 1a,annexin A1,superoxide dismutase A,C-type lectin,lysozyme,antimicrobial peptides,cofilin-1-B,mannose receptor,histone H4,prohormone convertase 1,carbonyl reductase 1 and some components of the Toll-like receptor（TLR）signalling *** molecules are potential candidates for further investigation of immune mechanisms in ***;in particular,TLR-mediated responses,which might be activated in frogs exposed to pathogenic bacteria as part of innate immune defence,but which might also impact on adaptive immunity to infection.

关键词： Immune responses Microbial infection Proteomic analysis Rana dybowskii

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Isolation of synaptic and nonsynaptic mitochondria from brain by iodixanol density gradient centrifugation

Isolation of synaptic and nonsynaptic mitochondria from brai...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：林元喜 Wan Tian-Lang Xi'an Jiaotong University School of Medicine

Isolation of synaptic and nonsynaptic mitochondria from brain by iodixanol density gradient centrifugation Lin Yuan-Xi,Wan Tian-Lang(Xi'an Jiaotong University School of Medicine,Xi'an,710061)The investigation of mitochondrial abnormalities in brain commonly requires isolation of these *** for isolating brain mitochondria rarely rely solely on differential centrifugation as a purification process,because of the mitochondrial fractions were heavily contaminated with synaptosomes and *** this report,differential centrifugation separation crude mitochondria from homogenates of brain tissues and purification by iodixanol density gradient centrifugation are *** of non-synaptic mitochondria and synaptososme from brain by using Percoll gradient *** digitonin,osmotic lysis,nitrogen cavitation technique disrupt the synaptosomal plasma membrane to release synaptic *** synaptosomal disruption,synaptic mitochondria can be *** key procedure as following:1 Continuous gradient:homogenate ordisrupted synaptosome(or crude mitochondria)adjust to 8%optiprep and loaded on top of 12-40%iodixanol gradient,and centrifuged at 100000g at 4°C for 90min.2 Discontinuous gradient:the crude mitochondria adjusted to 40%optiprep and was overlain sequentially by 25%and 20%optiprep,the tube was spun at 170 000 g for 3 h at *** were collected from the 44/25%and 25/20%at the *** techniques allowed isolation of mitochondria with relatively good purity,with acceptable metabolic functions and respiratory ***(60%w/v,Opti Prep)is appropriated for the separation of mitochondria from brain and for their subsequent use.

关键词： Brain Mitochondria Synaptososme Iodixanol Centrifugation

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A in vivo Study on the Neuroprotective Effects of Rapamycin Against Neurotoxic Aβ in Rats

A in vivo Study on the Neuroprotective Effects of Rapamycin ...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：郝明 Jia-Qing Tong Sheng-Xiao Zhang Jin-Shun Qi Department of physiology key Laboratory of Cellular PhysiologyMinistry of EducationShanxi Medical University Second Affiliated Hospital Shanxi Medical University

Extensive research has shown that the aggregation and toxicity of amyloid-beta protein(Aβ)are closely related to the cognitive deficits in Alzheimer's disease(AD).Inhibition of mammalian target of rapamycin(m TOR)by rapamycin can clear Aβ,reduce inflammation and improve cognitive ***,there is little evidence that rapamycin protects cognitive function and synaptic plastiscity of rats against neurotoxic Aβ*** present study investigated the effects of rapamycin on Aβ1-42-induced deficits in cognitive function and synaptic plasticity by using Morris water maze test and in vivo hippocampal field potentials *** results showed that:(1)Hippocampal injection of Aβ1-42 alone impaired the spatial learning and memory of rats;(2)Rapamycin did not affect cognitive function of rats,but alleviated Aβ1-42-induced the spatial learning and memory deficits,campared with Aβ1-42 alone group;(3)Aβ1-42remarkably suppressed in vivo hippocampal long-term potentiation(LTP)off EPSPs in the CA1region;(4)Rapamycin alone did not affect LTP,but prevented Aβ1-42-induced suppression of *** data supports the notion that rapamycin could protect against Aβ1-42-induced impairments in spatial cognition and synaptic plasticity in rats and suggests that rapamycin probably has a potential clinical value in the prevention and treatment of AD in the future.

关键词： Rapamycin Aβ1-42 Morris water maze Long-term potentiation

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Neuroprotective effects of rosmarinic acid in the MPTP mouse model of Parkinson's disease

Neuroprotective effects of rosmarinic acid in the MPTP mouse...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：贾文婷 Hua-Min XU Hong JIANG Jun-Xia XIE Department of Physiology Shandong Provincial Key Laboratory of Pathogenesis and Prevention of Neurological Disorders and State Key Disciplines:PhysiologyQingdao University

Parkinson’s disease(PD)is a common neurodegenerative disease characterized by the selective loss of dopaminergic neurons in substantia nigra(SN).Rosmarinic acid(RA)is a naturally occurring polyphenolic compound found in herbs,and has multiple biological activities such as antioxidative,anti-inflammatory and antiviral *** this study,we investigated the neuroprotective effects of RA on dopaminergic neurons in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced C57BL/6 mouse model of *** showed that the levels of dopamine and its metabolites in the striatum decreased in MPTP-induced PD *** mice received oral administration of RA before MPTP injection elevated the striatal levels of dopamine and its ***,the tyrosine hydroxylase expression in the SN decreased in MPTP intoxicated PD *** could be significantly inhibited by pretreatment with *** addition,RA could restore the decrease of SOD induced by *** study suggests that RA could suppress MPTP-induced degeneration of the nigrostriatal dopaminergic system by antioxidative ***,RA should be clinically evaluated for the prevention of neurodegenerative diseases.

关键词： Rosmarinic acid Parkinson's disease MPTP Neuroprotective

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