title: Dopaminergic neuronal loss and dopamine-dependent locomotor defects in Fbxo7-deficient zebrafish.
abstract: Recessive mutations in the F-box only protein 7 gene (FBXO7) cause PARK15, a mendelian form of early-onset, levodopa-responsive parkinsonism with severe loss of nigrostriatal dopaminergic neurons. However, the function of the protein encoded by FBXO7, and the pathogenesis of PARK15 remain unknown. No animal models of this disease exist. Here, we report the generation of a vertebrate model of PARK15 in zebrafish. We first show that the zebrafish Fbxo7 homolog protein (zFbxo7) is expressed abundantly in the normal zebrafish brain. Next, we used two zFbxo7-specific morpholinos (targeting protein translation and mRNA splicing, respectively), to knock down the zFbxo7 expression. The injection of either of these zFbxo7-specific morpholinos in the fish embryos induced a marked decrease in the zFbxo7 protein expression, and a range of developmental defects. Furthermore, whole-mount in situ mRNA hybridization showed abnormal patterning and significant decrease in the number of diencephalic tyrosine hydroxylase-expressing neurons, corresponding to the human 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 neuronal loss. Last, the zFbxo7 morphants displayed severe locomotor disturbances (bradykinesia), which were dramatically 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. In conclusion, this novel vertebrate model reproduces pathologic and behavioral hallmarks of human parkinsonism (dopaminergic neuronal loss and dopamine-dependent bradykinesia), representing therefore a valid tool for investigating the mechanisms of selective dopaminergic neuronal death, and screening for modifier genes and therapeutic compounds.
authors.affiliation: Department of Clinical Genetics, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Tianna
authors.initials: T
authors.lastname: Zhao
authors.forename: Herma
authors.initials: H
authors.lastname: Zondervan-van der Linde
authors.forename: Lies-Anne
authors.initials: LA
authors.lastname: Severijnen
authors.forename: Ben A
authors.initials: BA
authors.lastname: Oostra
authors.forename: Rob
authors.initials: R
authors.lastname: Willemsen
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Bonifati
mesh_terms: D000818:Animals; D001058:Apomorphine / Q000494:pharmacology; D019521:Body Patterning; D001921:Brain / Q000378:metabolism; D004195:Disease Models, Animal; D004294:Domperidone / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D018492:Dopamine Antagonists / Q000494:pharmacology; D005091:Exons; D020224:Expressed Sequence Tags; D044783:F-Box Proteins / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization; D007438:Introns; D008124:Locomotion; D009474:Neurons / Q000378:metabolism*; D020734:Parkinsonian Disorders / Q000378:metabolism; D010641:Phenotype; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D015027:Zebrafish
chemical_list: D018492:Dopamine Antagonists; D044783:F-Box Proteins; C489149:FBXO7 protein, human; D012333:RNA, Messenger; D004294:Domperidone; D001058:Apomorphine; D004298:Dopamine