黑质过表达α-突触核蛋白对小鼠运动行为的影响
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摘要
目的研究黑质(SN)区过表达α-突触核蛋白(α-syn)对小鼠运动行为的影响。方法将40只9周龄C57BL/6J雄性小鼠随机均分为2组,实验组小鼠SN区双侧注射200nL腺相关病毒(AAV)2/9-α-syn,对照组小鼠SN区注射等量的AAV2/9-EGFP。病毒注射4周后,采用爬杆实验和转棒实验观察小鼠的运动协调能力。结果 Western Blot检测显示,与对照组相比,实验组小鼠SN区α-syn表达明显增加(t=2.803,P<0.05)。爬杆实验显示,与对照组相比,实验组小鼠爬杆所用总时间和调头时间均明显增加(t=8.045、9.401,P<0.01)。转棒实验显示,实验组小鼠在转棒仪上停留的时间较对照组小鼠明显缩短(t=2.177,P<0.05)。结论 SN区过表达α-syn会导致小鼠运动功能障碍。
Objective To investigate the influence ofα-synuclein(α-syn)overexpression in the substantia nigra(SN)on motor function in mice. Methods A total of 40 male C57 BL/6 J mice aged 9 weeks were randomly divided into control group and experimental group,with 20 mice in each group.The mice in the experimental group were given injection of 200 nL AAV2/9-α-syn into the SN at both sides,and those in the control group were given injection of an equal volume of AAV2/9-EGFP into the SN.At 4 weeks after injection,the pole test and the rotarod test were used to observe motor function. Results Western blot showed that compared with the control group,the experimental group had a significant increase in the expression ofα-syn in the SN(t=2.803,P<0.05).The pole test showed that compared with the control group,the experimental group had significant increases in the total time to climb down the pole and the time to turn around(t=8.045 and 9.401,P<0.01).The rotarod test showed that the experimental group had a significantly shorter retention time on the rotarod than the control group(t=2.177,P<0.05).Conclusion Overexpression ofα-syn in the SN can lead to motor impairment in mice.
Objective To investigate the influence ofα-synuclein(α-syn)overexpression in the substantia nigra(SN)on motor function in mice. Methods A total of 40 male C57 BL/6 J mice aged 9 weeks were randomly divided into control group and experimental group,with 20 mice in each group.The mice in the experimental group were given injection of 200 nL AAV2/9-α-syn into the SN at both sides,and those in the control group were given injection of an equal volume of AAV2/9-EGFP into the SN.At 4 weeks after injection,the pole test and the rotarod test were used to observe motor function. Results Western blot showed that compared with the control group,the experimental group had a significant increase in the expression ofα-syn in the SN(t=2.803,P<0.05).The pole test showed that compared with the control group,the experimental group had significant increases in the total time to climb down the pole and the time to turn around(t=8.045 and 9.401,P<0.01).The rotarod test showed that the experimental group had a significantly shorter retention time on the rotarod than the control group(t=2.177,P<0.05).Conclusion Overexpression ofα-syn in the SN can lead to motor impairment in mice.
引文
[1]KANSARA S,TRIVEDI A,CHEN S,et al.Early diagnosis and therapy of Parkinson’s disease:can disease progression be curbed[J]?Journal of Neural Transmission(Vienna,Austria:1996),2013,120(1):197-210.
[2]BABA M,NAKAJO S,TU P H,et al.Aggregation of alphasynuclein in Lewy bodies of sporadic Parkinson’s disease and dementia with Lewy bodies[J].American Journal of Pathology,1998,152(4):879-884.
[3]DEHAY B,BOURDENX M,GORRY P A,et al.Targeting alpha-synuclein for treatment of Parkinson’s disease:mechanistic and therapeutic considerations[J].Lancet Neurology,2015,14(8):855-866.
[4]YU S,UEDA K,CHAN P.alpha-Synuclein and dopamine metabolism[J].Molecular Neurobiology,2005,31(1/3):243-254.
[5]BUTLER B,SAMBO D,KHOSHBOUEI H.Alpha-synuclein modulates dopamine neurotransmission[J].Journal of Chemical Neuroanatomy,2017,83(2):41-49.
[6]TOLMASOV M,DJALDETTI R,LEV N,et al.Pathological and clinical aspects of alpha/beta synuclein in Parkinson’s disease and related disorders[J].Expert Review of Neurotherapeutics,2016,16(5):505-513.
[7]FORTUNA J T,GRALLE M,BECKMAN D A,et al.Brain infusion of alpha-synuclein oligomers induces motor and nonmotor Parkinson’s disease-like symptoms in mice[J].Behavioural Brain Research,2017,333(4):150-160.
[8]BENDOR J T,LOGAN T P,EDWARDS R H.The function of alpha-synuclein[J].Neuron,2013,79(6):1044-1066.
[9]PAUMIER K L,SUKOFF RIZZO S J,BERGER Z,et al.Behavioral characterization of A53T mice reveals early and late stage deficits related to Parkinson’s disease[J].PLoS One,2013,8(8):e70274.
[10]DAWSON T M,KO H S,DAWSON V L.Genetic animal models of Parkinson’s disease[J].Neuron,2010,66(5):646-661.
[11]CHESSELET M F.In vivo alpha-synuclein overexpression in rodents:a useful model of Parkinson’s disease[J]?Experimental Neurology,2008,209(1):22-27.
[12]CHESSELET M F,FLEMING S,MORTAZAVI F,et al.Strengths and limitations of genetic mouse models of Parkinson’s disease[J].Parkinsonism&Related Disorders,2008,14(Suppl 2):S84-S87.
[13]KOPRICH J B,JOHNSTON T H,REYES M G,et al.Expression of human A53Talpha-synuclein in the rat substantia nigra using a novel AAV1/2vector produces a rapidly evolving pathology with protein aggregation,dystrophic neurite architecture and nigrostriatal degeneration with potential to model the path[J].Molecular Neurodegeneration,2010,5(1):43.
[14]MARKS J,BARTUS R T,SIFFERT J A,et al.Gene delivery of AAV2-neurturin for Parkinson’s disease:a double-blind,randomised,controlled trial[J].Lancet Neurology,2010,9(12):1164-1172.
[15]ULUSOY A,DECRESSAC M,KIRIK D,et al.Viral vectormediated overexpression of alpha-synuclein as a progressive model of Parkinson’s disease[J].Progress in Brain Research,2010,184(3):89-111.
[16]WILLIAMS G P,SCHONHOFF A M,JURKUVENAITE AA,et al.Targeting of the classⅡtransactivator attenuates inflammation and neurodegeneration in an alpha-synuclein model of Parkinson’s disease[J].Journal of Neuroinflammation,2018,15(1):244.
[17]IP C W,KLAUS L C,KARIKARI A A,et al.AAV1/2-induced overexpression of A53T-alpha-synuclein in the substantia nigra Results in degeneration of the nigrostriatal system with Lewy-like pathology and motor impairment:a new mouse model for Parkinson’s disease[J].Acta Neuropathologica Communications,2017,5(1):11.
[18]FLEMING S M.Mechanisms of gene-environment interactions in Parkinson’s disease[J].Current Environmental Health Reports,2017,4(2):1-8.
[19]LO BIANCO C,RIDET J L,SCHNEIDER B L,et al.alphaSynucleinopathy and selective dopaminergic neuron loss in a rat lentiviral-based model of Parkinson’s disease[J].Proceedings of the National Academy of Sciences of the United States of America,2002,99(16):10813-10818.
[20]VAN ROMPUY A S,OLIVERAS S M,VAN D A,et al.Nigral overexpression of alpha-synuclein in the absence of parkin enhances alpha-synuclein phosphorylation but does not modulate dopaminergic neurodegeneration[J].Molecular Neurodegeneration,2015,10(1):23.
[21]KIRIK D,ROSENBLAD C,BURGER C,et al.Parkinsonlike neurodegeneration induced by targeted overexpression of alpha-synuclein in the nigrostriatal system[J].Journal of Neuroscience,2002,22(7):2780-2791.
[22]NEUMANN M,KAHLE P J,GIASSON B I,et al.Misfolded proteinase K-resistant hyperphosphorylated alpha-synuclein in aged transgenic mice with locomotor deterioration and in human alpha-synucleinopathies[J].Journal of Clinical Investigation,2002,110(10):1429-1439.
[23]MULCAHY P,O’DOHERTY A,PAUCARD A,et al.Development and characterisation of a novel rat model of Parkinson’s disease induced by sequential intranigral administration of AAV-alpha-synuclein and the pesticide,rotenone[J].Neuroscience,2012,203(11):170-179.
[24]DONG Z Z,FERGER B,FELDON J,et al.Overexpression of Parkinson’s disease-associated alpha-synucleinA53Tby recombinant adeno-associated virus in mice does not increase the vulnerability of dopaminergic neurons to MPTP[J].Journal of Neurobiology,2002,53(1):1-10.
[25]GOMBASH S E,MANFREDSSON F P,KEMP C J,et al.Morphological and behavioral impact of AAV2/5-mediated overexpression of human wildtype alpha-synuclein in the rat nigrostriatal system[J].PLoS One,2013,8(11):e81426.
[26]VAN DER PERREN A,TOELEN J,CASTEELS C,et al.Longitudinal follow-up and characterization of a robust rat model for Parkinson’s disease based on overexpression of alpha-synuclein with adeno-associated viral vectors[J].Neurobiology of Aging,2015,36(3):1543-1558.
[27]FEBBRARO F,ANDERSEN K J,SANCHEZ-GUAJARDOV,et al.Chronic intranasal deferoxamine ameliorates motor defects and pathology in the alpha-synuclein rAAV Parkinson’s model[J].Experimental Neurology,2013,247(2):45-58.
[28]OLIVERAS-SALVA M,VAN DER PERREN A,CASADEIN,et al.rAAV2/7vector-mediated overexpression of alphasynuclein in mouse substantia nigra induces protein aggregation and progressive dose-dependent neurodegeneration[J].Molecular Neurodegeneration,2013,8(1):44.
[29]BOURDENX M,DOVERO S,ENGELN M,et al.Lack of additive role of ageing in nigrostriatal neurodegeneration triggered by alpha-synuclein overexpression[J].Acta Neuropathologica Communications,2015,3(1):46.
[30]DECRESSAC M,MATTSSON B,LUNDBLAD M,et al.Progressive neurodegenerative and behavioural changes induced by AAV-mediated overexpression of alpha-synuclein in midbrain dopamine neurons[J].Neurobiology of Disease,2012,45(3):939-953.
[31]POON H F,FRASIER M,SHREVE N,et al.Mitochondrial associated metabolic proteins are selectively oxidized in A30Palpha-synuclein transgenic mice-a model of familial Parkinson’s disease[J].Neurobiology of Disease,2005,18(3):492-498.
[32]DEVI L,RAGHAVENDRAN V,PRABHU B M,et al.Mitochondrial import and accumulation of alpha-synuclein impair complexⅠin human dopaminergic neuronal cultures and Parkinson disease brain[J].The Journal of Biological Chemistry,2008,283(14):9089-9100.
[2]BABA M,NAKAJO S,TU P H,et al.Aggregation of alphasynuclein in Lewy bodies of sporadic Parkinson’s disease and dementia with Lewy bodies[J].American Journal of Pathology,1998,152(4):879-884.
[3]DEHAY B,BOURDENX M,GORRY P A,et al.Targeting alpha-synuclein for treatment of Parkinson’s disease:mechanistic and therapeutic considerations[J].Lancet Neurology,2015,14(8):855-866.
[4]YU S,UEDA K,CHAN P.alpha-Synuclein and dopamine metabolism[J].Molecular Neurobiology,2005,31(1/3):243-254.
[5]BUTLER B,SAMBO D,KHOSHBOUEI H.Alpha-synuclein modulates dopamine neurotransmission[J].Journal of Chemical Neuroanatomy,2017,83(2):41-49.
[6]TOLMASOV M,DJALDETTI R,LEV N,et al.Pathological and clinical aspects of alpha/beta synuclein in Parkinson’s disease and related disorders[J].Expert Review of Neurotherapeutics,2016,16(5):505-513.
[7]FORTUNA J T,GRALLE M,BECKMAN D A,et al.Brain infusion of alpha-synuclein oligomers induces motor and nonmotor Parkinson’s disease-like symptoms in mice[J].Behavioural Brain Research,2017,333(4):150-160.
[8]BENDOR J T,LOGAN T P,EDWARDS R H.The function of alpha-synuclein[J].Neuron,2013,79(6):1044-1066.
[9]PAUMIER K L,SUKOFF RIZZO S J,BERGER Z,et al.Behavioral characterization of A53T mice reveals early and late stage deficits related to Parkinson’s disease[J].PLoS One,2013,8(8):e70274.
[10]DAWSON T M,KO H S,DAWSON V L.Genetic animal models of Parkinson’s disease[J].Neuron,2010,66(5):646-661.
[11]CHESSELET M F.In vivo alpha-synuclein overexpression in rodents:a useful model of Parkinson’s disease[J]?Experimental Neurology,2008,209(1):22-27.
[12]CHESSELET M F,FLEMING S,MORTAZAVI F,et al.Strengths and limitations of genetic mouse models of Parkinson’s disease[J].Parkinsonism&Related Disorders,2008,14(Suppl 2):S84-S87.
[13]KOPRICH J B,JOHNSTON T H,REYES M G,et al.Expression of human A53Talpha-synuclein in the rat substantia nigra using a novel AAV1/2vector produces a rapidly evolving pathology with protein aggregation,dystrophic neurite architecture and nigrostriatal degeneration with potential to model the path[J].Molecular Neurodegeneration,2010,5(1):43.
[14]MARKS J,BARTUS R T,SIFFERT J A,et al.Gene delivery of AAV2-neurturin for Parkinson’s disease:a double-blind,randomised,controlled trial[J].Lancet Neurology,2010,9(12):1164-1172.
[15]ULUSOY A,DECRESSAC M,KIRIK D,et al.Viral vectormediated overexpression of alpha-synuclein as a progressive model of Parkinson’s disease[J].Progress in Brain Research,2010,184(3):89-111.
[16]WILLIAMS G P,SCHONHOFF A M,JURKUVENAITE AA,et al.Targeting of the classⅡtransactivator attenuates inflammation and neurodegeneration in an alpha-synuclein model of Parkinson’s disease[J].Journal of Neuroinflammation,2018,15(1):244.
[17]IP C W,KLAUS L C,KARIKARI A A,et al.AAV1/2-induced overexpression of A53T-alpha-synuclein in the substantia nigra Results in degeneration of the nigrostriatal system with Lewy-like pathology and motor impairment:a new mouse model for Parkinson’s disease[J].Acta Neuropathologica Communications,2017,5(1):11.
[18]FLEMING S M.Mechanisms of gene-environment interactions in Parkinson’s disease[J].Current Environmental Health Reports,2017,4(2):1-8.
[19]LO BIANCO C,RIDET J L,SCHNEIDER B L,et al.alphaSynucleinopathy and selective dopaminergic neuron loss in a rat lentiviral-based model of Parkinson’s disease[J].Proceedings of the National Academy of Sciences of the United States of America,2002,99(16):10813-10818.
[20]VAN ROMPUY A S,OLIVERAS S M,VAN D A,et al.Nigral overexpression of alpha-synuclein in the absence of parkin enhances alpha-synuclein phosphorylation but does not modulate dopaminergic neurodegeneration[J].Molecular Neurodegeneration,2015,10(1):23.
[21]KIRIK D,ROSENBLAD C,BURGER C,et al.Parkinsonlike neurodegeneration induced by targeted overexpression of alpha-synuclein in the nigrostriatal system[J].Journal of Neuroscience,2002,22(7):2780-2791.
[22]NEUMANN M,KAHLE P J,GIASSON B I,et al.Misfolded proteinase K-resistant hyperphosphorylated alpha-synuclein in aged transgenic mice with locomotor deterioration and in human alpha-synucleinopathies[J].Journal of Clinical Investigation,2002,110(10):1429-1439.
[23]MULCAHY P,O’DOHERTY A,PAUCARD A,et al.Development and characterisation of a novel rat model of Parkinson’s disease induced by sequential intranigral administration of AAV-alpha-synuclein and the pesticide,rotenone[J].Neuroscience,2012,203(11):170-179.
[24]DONG Z Z,FERGER B,FELDON J,et al.Overexpression of Parkinson’s disease-associated alpha-synucleinA53Tby recombinant adeno-associated virus in mice does not increase the vulnerability of dopaminergic neurons to MPTP[J].Journal of Neurobiology,2002,53(1):1-10.
[25]GOMBASH S E,MANFREDSSON F P,KEMP C J,et al.Morphological and behavioral impact of AAV2/5-mediated overexpression of human wildtype alpha-synuclein in the rat nigrostriatal system[J].PLoS One,2013,8(11):e81426.
[26]VAN DER PERREN A,TOELEN J,CASTEELS C,et al.Longitudinal follow-up and characterization of a robust rat model for Parkinson’s disease based on overexpression of alpha-synuclein with adeno-associated viral vectors[J].Neurobiology of Aging,2015,36(3):1543-1558.
[27]FEBBRARO F,ANDERSEN K J,SANCHEZ-GUAJARDOV,et al.Chronic intranasal deferoxamine ameliorates motor defects and pathology in the alpha-synuclein rAAV Parkinson’s model[J].Experimental Neurology,2013,247(2):45-58.
[28]OLIVERAS-SALVA M,VAN DER PERREN A,CASADEIN,et al.rAAV2/7vector-mediated overexpression of alphasynuclein in mouse substantia nigra induces protein aggregation and progressive dose-dependent neurodegeneration[J].Molecular Neurodegeneration,2013,8(1):44.
[29]BOURDENX M,DOVERO S,ENGELN M,et al.Lack of additive role of ageing in nigrostriatal neurodegeneration triggered by alpha-synuclein overexpression[J].Acta Neuropathologica Communications,2015,3(1):46.
[30]DECRESSAC M,MATTSSON B,LUNDBLAD M,et al.Progressive neurodegenerative and behavioural changes induced by AAV-mediated overexpression of alpha-synuclein in midbrain dopamine neurons[J].Neurobiology of Disease,2012,45(3):939-953.
[31]POON H F,FRASIER M,SHREVE N,et al.Mitochondrial associated metabolic proteins are selectively oxidized in A30Palpha-synuclein transgenic mice-a model of familial Parkinson’s disease[J].Neurobiology of Disease,2005,18(3):492-498.
[32]DEVI L,RAGHAVENDRAN V,PRABHU B M,et al.Mitochondrial import and accumulation of alpha-synuclein impair complexⅠin human dopaminergic neuronal cultures and Parkinson disease brain[J].The Journal of Biological Chemistry,2008,283(14):9089-9100.