Evaluation of spontaneous propulsive movement as a screening tool to detect rescue of Parkinsonism phenotypes in zebrafish models.
Evaluation of spontaneous propulsive movement as a screening tool to detect rescue of Parkinsonism phenotypes in zebrafish models.
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DOI：
10.1016/j.nbd.2011.05.016
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发表时间：
2011-10
影响因子：
6.1
通讯作者：
Burton, Edward A.
中科院分区：
文献类型：
--
作者：
Farrell, Thomas C.;Cario, Clinton L.;Milanese, Chiara;Vogt, Andreas;Jeong, Jong-Hyeon;Burton, Edward A.
关键词：
人类神经精神疾病的斑马鱼模型提供了通过基于表型的遗传或化学修饰筛选来确定新的治疗靶点和治疗方法的机会。为了建立一种检测帕金森病斑马鱼模型拯救的方法，我们以适合于筛选的微量试板的形式表征了受精后3~9天斑马鱼幼体的自发运动行为，并阐明了多巴胺能信号在其调节中的作用。在3~9dpf之间，幼虫运动的时间比例逐渐增加，而在5dpf至6dpf之间，幼虫的活动速度逐渐下降，因为零星的爆发式运动让位于更成熟的拍打和滑行模式。由于包括遗传背景在内的内在幼虫因素的影响，幼虫之间和记录过程中的自发活动有所不同。变异性随着年龄的增加而降低，因此暴露在不同实验条件下的幼虫组之间的微小差异可以在6到7个DPF中强烈地检测到。暴露于MPP+、氟哌啶醇或氯丙嗪可抑制内源性多巴胺能信号，通过降低自发运动的启动频率来降低平均运动速度，但不改变活动速度。通过分析每个数据点的幼虫组，得出可接受的筛选窗口系数，可以降低平均速度分析的变异性；每组所需的样本量由不同模型中运动表型的大小决定。对于氯丙嗪暴露，4只幼虫的样本允许牢固地分离处理和未处理的数据点(Z=0.42)，而MPP+引起的较轻微的损害需要8只幼虫组成的组，以便提供有用的发现分析(Z=0.13)。对幼虫自发活动的量化为确定运动系统的功能完整性提供了一种简单的方法，并可能成为分离帕金森症表型的新的分子调节剂的有用工具。
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.
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DOI：
10.1523/jneurosci.0714-09.2009
发表时间：
2009-07-01
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Peng J;Wagle M;Mueller T;Mathur P;Lockwood BL;Bretaud S;Guo S
通讯作者：
Guo S
影响因子：
4.7
作者：
Sallinen, V. .;Torkko, V. .;Panula, P.
通讯作者：
Panula, P.
影响因子：
2.9
作者：
Giacomini, NJ;Rose, B;Guo, S
通讯作者：
Guo, S
影响因子：
2.5
作者：
Vogt, Andreas;Cholewinski, Andrzej;Shen, Xiaoqiang;Nelson, Scott G.;Lazo, John S.;Tsang, Michael;Hukriede, Neil A.
通讯作者：
Hukriede, Neil A.
影响因子：
2.9
作者：
Bretaud, S;Lee, S;Guo, S
通讯作者：
Guo, S