[1]段凌晗,吴永梅,李淑蓉,等.应用mycalolide B初步建立斑马鱼帕金森病模型[J].第三军医大学学报,2018,40(06):487-493.
 DUAN Linghan,WU Yongmei,LI Shurong,et al.A zebrafish model of Parkinson’s disease established using mycalolide B[J].J Third Mil Med Univ,2018,40(06):487-493.

应用mycalolide B初步建立斑马鱼帕金森病模型(/HTML )




A zebrafish model of Parkinson’s disease established using mycalolide B
DUAN Linghan WU Yongmei LI Shurong SU Bingyin

Department of Anatomy, Histology and Embryology, Sichuan Provincial Key Laboratory of Development and Regeneration, Chengdu Medical College, Chengdu, Sichuan Province, 610500, China

mycalolide B1-甲基-4-苯基-1236四氢吡啶帕金森病模型斑马鱼
mycalolide B1-methyl-4-phenyl-1236-tetrahydropyridineParkinson&rsquos disease modelszebrafish

目的    应用mycalolide B(MB)建立斑马鱼帕金森病(Parkison’s disease,PD)模型,观察斑马鱼行为学、形态学及多巴胺神经元的变化并进行系统评价。方法    同亲代斑马鱼单次产卵200~300枚,简单随机化分为7组:对照组(0.2% DMSO)、MB10组(10 μg/L MB)、MB20组(20 μg/L MB)、MB40组(40 μg/L MB)、MB80组(80 μg/L MB)、MB160组(160 μg/L MB)、1-甲基-4-苯基-1,2,3,6四氢吡啶(MPTP)组(200 μmol/L MPTP),每组30枚胚胎,于浓度梯度下培养;统计存活率,分析行为学变化,利用激光共聚焦显微镜观察多巴胺神经元生长情况,并与MPTP组进行比较分析。结果     MB效应浓度33.4~50.3 μg/L时,成斑马鱼出现发育迟缓、脊椎畸形、心包水肿、平衡失态、震颤等;运动速率降低明显(P<0.05);Vmat2-GFP标记的阳性细胞减少(P<0.01);synuclein蛋白沉积(P<0.05)。结论    建立斑马鱼PD模型的MB效应浓度为33.4~50.3 μg/L;MB诱导的斑马鱼PD模型与MPTP模型相比,行为学、多巴胺神经元数量减少方面相似,且较不产生synuclein的MPTP模型更具模式学意义。


Objective    To establish a zebrafish model of Parkinson’s disease (PD) using mycalolide B (MB), and evaluate the model by observing zebrafish behavior, morphology and dopamine neurons. Methods    A total of 200~300 parental zebrafish eggs from a single spawning were randomly divided into 7 groups: control group, MB10 group (10 μg/L), MB20 group (20 μg/L), MB40 group (40 μg/L), MB80 group (80 μg/L), MB160 group (160 μg/L) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) group (200 μmol/L), with 30 embryos in each group. The embryos were cultured under the concentration gradient, and the survival rate and behavioral changes were observed. Laser confocal microscopy was used to observe the growth of dopamine neurons. And all the results of MB treatment groups were compared with MPTP group. Results     The effective concentration of MB was 33.4~50.3 μg/L, which resulted in the abnormities such as growth retardation, spinal deformity, pericardial edema, balance loss and tremor, etc. The movement rate was significantly decreased (P<0.05). There were less Vmat2-GFP positive cells (P<0.01) and more synuclein protein deposition (P<0.05). Conclusion    MB at an effective concentration of 33.4~50.3 μg/L can be used to establish zebrafish PD model. The zebrafish model induced by MB are similar with the MPTP model in behaviors and fewer dopamine neurons, but better than the other one for synuclein production.


[1]GUSTAVSSON E K, TRINH J, GUELLA I, et al. DCTN1 p.K56R in progressive supranuclear palsy[J]. Parkinsonism Relat Disord, 2016, 28: 56-61. DOI: 10.1016/j.parkreldis.2016.04.025.
[2]CAVOLO S L, ZHOU C, KETCHAM S A, et al. Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of densecore vesicles[J]. Mol Biol Cell, 2015, 26(14): 2664-2672. DOI: 10.1091/mbc.e14-11-1564.
[3]ANICHTCHIK O V, KASLIN J, PEITSARO N, et al. Neurochemical and behavioural changes in zebrafish Danio rerio after systemic administration of 6-hydroxydopamine and 1-methyl-4-phenyl-1,2,3,6tetrahydropyridine[J]. J Neurochem, 2004, 88(2): 443-453. DOI: 10.1111/j.1471-4159.2004.02190.x. 
[4]KIM E K, CHOI E J. Pathological roles of MAPK signaling pathways in human diseases[J]. Biochim Biophys Acta, 2010, 1802(4): 396-405. DOI: 10.1016/j.bbadis.2009.12.009.
[5]ISHIKAWA K, SAIKI S, FURUYA N, et al. P150gluedassociated disorders are caused by activation of intrinsicapoptotic pathway[J]. PLoS ONE, 2014, 9(4): e94645. DOI: 10.1371/journal.pone.0094645.
[6]BEAL M F. Experimental models of Parkinson’s disease[J].Nat Rev Neurosci, 2001, 2(5): 325-334. DOI: 10.1038/35072550.
[7]SALLINEN V, TORKKO V, SUNDVIK M, et al. MPTP and MPP+ target specific aminergic cell populations in larval zebrafish[J]. J Neurochem, 2009, 108(3): 719-731. DOI: 10.1111/j.1471-4159.2008.05793.x.
[8]CHADDERTON L, KELLEHER S, BROW A, et al. Testing the efficacy of rotenone as a piscicide for New Zealand pest fish species[C]// Managing Invasive Freshwater Fish in New Zealand. Proceedings of A Workshop Hosted by Department of Conservation, Hamilton, New Zealand,  May 10-12, 2001.
[9]LAM C S, KORZH V, STRAHLE U. Zebrafish embryos are susceptible to the dopaminergic neurotoxin MPTP[J].Eur J Neurosci, 2005, 21(6): 1758-1762. DOI: 10.1111/j.1460-9568.2005.03988.x.
[10]BRETAUD S, LEE S, GUO S. Sensitivity of zebrafish to environmental toxins implicated in Parkinson’s disease[J].Neurotoxicol Teratol, 2004, 26(6): 857-864. DOI: 10.1016/j.ntt.2004.06.014.
[11]MCKINLEY E T, BARANOWSKI T C, BLAVO D O, et al. Neuroprotection of MPTP-induced toxicity in zebrafish dopaminergic neurons[J].Brain Res Mol Brain Res, 2005, 141(2): 128-137. DOI: 10.1016/j.molbrainres.2005.08.014.
[12]TAGLIAFERRO P, BURKE R E. Retrograde axonal degeneration in parkinson disease[J].J Parkinsons Dis, 2016, 6(1): 1-15. DOI: 10.3233/JPD-150769.
[13]POEWE W, SEPPI K, TANNER C M, et al. Parkinson disease[J]. Nat Rev Dis Primers, 2017, 3: 17013. DOI: 10.1038/nrdp.2017.13.
[14]ABOUD O, PARCON P A,DEWALL K M, et al. Aging, Alzheimer’s, and APOE genotype influence the expression and neuronal distribution patterns of microtubule motor protein dynactinP50[J]. Front Cell Neurosci, 2015, 9: 103. DOI: 10.3389/fncel.2015.00103.
[15]DEWEY E B, JOHNSTON C A. Diverse mitotic functions of the cytoskeletal?crosslinking protein Shortstop suggest a role in Dynein/Dynactin activity[J]. Mol Biol Cell, 2017, 28(19): 2555-2568. DOI: 10.1091/mbc.E17-04-0219.
[16]GIRI A, GUVEN G, HANAGASI H, et al. PLA2G6 mutations related to distinct phenotypes: A new case with earlyonset parkinsonism[J]. Tremor Other Hyperkinet Mov(NY), 2016, 6: 363. DOI: 10.7916/D81G0M12
[17]CRONIN M A, SCHWARZ T L. The CAP-Gly of p150: one domain, two diseases, and a function at the end[J]. Neuron, 2012, 74(2): 211-213. DOI: 10.1016/j.neuron.2012.04.003.
[18]LAZARUS J E, MOUGHAMIAN A J, TOKITO M K, et al. Dynactin subunit p150(Glued) is a neuronspecific anti-catastrophe factor[J]. PLoS Biol, 2013, 11(7): e1001611. DOI: 10.1371/journal.pbio.1001611.
[19]BRAUNSTEIN K E, ESCHBACH J, R NA-V R S K, et al. A point mutation in the dynein heavy chain gene leads to striatal atrophy and compromises neurite outgrowth of striatal neurons[J]. Hum Mol Genet, 2010, 19(22):4385-4398. DOI: 10.1093/hmg/ddq361.

更新日期/Last Update: 2018-03-23