Dopaminergic Neuronal Loss and Dopamine-Dependent Locomotor Defects in Fbxo7-Deficient Zebrafish 英文参考文献.docVIP

DopaminergicNeuronalLossandDopamine-Dependent LocomotorDefectsinFbxo7-DeficientZebrafish TiannaZhao,HermaZondervan-vanderLinde,Lies-AnneSeverijnen,BenA.Oostra,RobWillemsen, VincenzoBonifati* DepartmentofClinicalGenetics,ErasmusMC,Rotterdam,TheNetherlands Abstract Recessivemutations intheF-box onlyprotein 7gene(FBXO7)causePARK15,aMendelianformofearly-onset,levodopa- responsive parkinsonism with severe loss of nigrostriatal dopaminergic neurons. However, the function of the protein encodedbyFBXO7,andthepathogenesisofPARK15remainunknown.Noanimalmodelsofthisdiseaseexist.Here,we reportthegenerationofavertebratemodelofPARK15inzebrafish.WefirstshowthatthezebrafishFbxo7homologprotein (zFbxo7)isexpressedabundantlyinthenormalzebrafishbrain.Next,weusedtwozFbxo7-specificmorpholinos(targeting proteintranslationandmRNAsplicing,respectively),toknockdownthezFbxo7expression.Theinjectionofeitherofthese zFbxo7-specificmorpholinosinthefishembryosinducedamarkeddecreaseinthezFbxo7proteinexpression,andarange of developmental defects. Furthermore, whole-mount in situ mRNA hybridization showed abnormal patterning and significantdecreaseinthenumberofdiencephalictyrosinehydroxylase-expressingneurons,correspondingtothehuman nigrostriatal or ventral tegmental dopaminergic neurons. Of note, the number of the dopamine transporter-expressing neurons was much more severely depleted, suggesting dopaminergic dysfunctions earlier and larger than those due to neuronalloss.Last,thezFbxo7morphantsdisplayedseverelocomotordisturbances(bradykinesia),whichweredramatically improved by the dopaminergic agonist apomorphine. The severity of these morphological and behavioral abnormalities correlated with the severity of zFbxo7 protein deficiency. Moreover, the effects of the co-injection of zFbxo7- and p53- specific morpholinos were similar to those obtained with zFbxo7-specific morpholinos alone, supporting further the contention that the observed phenotypes were specifically due to the knock down of zFbxo7.