Evaluation of spontaneous propulsive movement as a screening tool to detect rescue of Parkinsonism phenotypes in zebrafish models

NEUROBIOLOGY OF DISEASE

卷:44

Article

Farrell, Thomas C.^1,2  Cario, Clinton L.^1,2  Milanese, Chiara^1,2,7  Vogt, Andreas^3,4  Jeong, Jong-Hyeon⁶  Burton, Edward A.^1,2,5,8,9 

[1] Univ Pittsburgh, Sch Med, Pittsburgh Inst Neurodegenerat Dis, Pittsburgh, PA USA

[2] Univ Pittsburgh, Sch Med, Dept Neurol, Pittsburgh, PA 15261 USA

[3] Univ Pittsburgh, Sch Med, Dept Computat & Syst Biol, Pittsburgh, PA USA

[4] Univ Pittsburgh, Sch Med, Drug Discovery Inst, Pittsburgh, PA USA

[5] Univ Pittsburgh, Sch Med, Dept Microbiol & Mol Genet, Pittsburgh, PA USA

[6] Univ Pittsburgh, Grad Sch Publ Hlth, Dept Biostat, Pittsburgh, PA 15261 USA

[7] RIMED Fdn, Palermo, Italy

[8] Pittsburgh VA Healthcare Syst, Geriatr Res Educ & Clin Ctr, Pittsburgh, PA USA

[9] Pittsburgh VA Healthcare Syst, Dept Neurol, Pittsburgh, PA USA

关键词: Zebrafish;    Parkinsonism;    Motor function;    Dopamine;    MPP+;    Chlorpromazine;    Haloperidol;    Apomorphine;    Ropnirole;    Spontaneous movement;    Automated behavior measurement;    Drug discovery;   

DOI  :  10.1016/j.nbd.2011.05.016

来源: Elsevier

【 摘 要 】

Zebrafish models of human neuropsychiatric diseases offer opportunities to identify novel therapeutic targets and treatments through phenotype-based genetic or chemical modifier screens. In order to develop an assay to detect rescue of zebrafish models of Parkinsonism, we characterized spontaneous zebrafish larval motor behavior from 3 to 9 days post fertilization in a microtiter plate format suitable for screening, and clarified the role of dopaminergic signaling in its regulation. The proportion of time that larvae spent moving increased progressively between 3 and 9 dpf, whereas their active velocity decreased between 5 and 6 dpf as sporadic burst movements gave way to a more mature beat-and-glide pattern. Spontaneous movement varied between larvae and during the course of recordings as a result of intrinsic larval factors including genetic background. Variability decreased with age, such that small differences between groups of larvae exposed to different experimental conditions could be detected robustly by 6 to 7 dpf. Suppression of endogenous dopaminergic signaling by exposure to MPP+, haloperidol or chlorpromazine reduced mean velocity by decreasing the frequency with which spontaneous movements were initiated, but did not alter active velocity. The variability of mean velocity assays could be reduced by analyzing groups of larvae for each data point, yielding acceptable screening window coefficients; the sample size required in each group was determined by the magnitude of the motor phenotype in different models. For chlorpromazine exposure, samples of four larvae allowed robust separation of treated and untreated data points (Z = 0.42). whereas the milder impairment provoked by MPP+ necessitated groups of eight larvae in order to provide a useful discovery assay (Z = 0.13). Quantification of spontaneous larval movement offers a simple method to determine functional integrity of motor systems, and may be a useful tool to isolate novel molecular modulators of Parkinsonism phenotypes. Published by Elsevier Inc.

【 授权许可】

Free