Yuan Hong, Sarre Sophie, Ebinger Guy, Michotte Yvette
Department of Pharmaceutical Chemistry and Drug Analysis, Research Group Experimental Pharmacology, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium.
J Neurosci Methods. 2005 May 15;144(1):35-45. doi: 10.1016/j.jneumeth.2004.10.004. Epub 2004 Dec 8.
We compared the effect of an injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB) and into the striatum on different parameters for evaluation of motor dysfunction and dopamine denervation in rats, as a function of time. A combination of behavioural, neurochemical and histological techniques was employed. Amphetamine-induced rotation is shown to provide a first rough estimation of motor impairment. Indeed, the number of rotations observed after amphetamine administration can distinguish between a partial and a near complete (>90%) denervation in the substantia nigra. However, lesion sizes of 50-80% resulted in similar rotational behaviour. Similarly, the elevated body swing test (EBST) can determine severe lesions, but is not sensitive enough in the partial model. In both models, determination of the dopamine tissue content with HPLC is a more precise measure of striatal dopamine innervation than striatal TH-immunostaining. The number of cells estimated by TH- and Nissl-staining correlated well in the striatal model, but there was a discrepancy between both measures in the MFB-lesioned animals. Therefore, additional Nissl-staining is necessary for better estimation of the size of the lesion at the level of the substantia nigra or ventral tegmental area in the severely lesioned animals. The MFB lesion model mimics end-stage Parkinson's disease. The striatal injection of 6-OHDA described here cannot be considered a progressive model, since there was no change in the number of TH-immunoreactive cells in the substantia nigra up to 8 weeks post-lesioning. However, the partial denervation renders its quite suitable for mimicking early stage Parkinson's disease, and is thus suitable for testing possible neuroprotective and neurotrophic drugs.
我们比较了向大鼠内侧前脑束（MFB）和纹状体注射6-羟基多巴胺（6-OHDA）对评估运动功能障碍和多巴胺去神经支配的不同参数的影响，并将其作为时间的函数进行研究。采用了行为学、神经化学和组织学技术相结合的方法。安非他明诱导的旋转被证明可对运动损伤提供初步的粗略估计。实际上，安非他明给药后观察到的旋转次数可区分黑质中的部分去神经支配和接近完全（>90%）的去神经支配。然而，50-80%的损伤大小会导致相似的旋转行为。同样，抬高身体摆动试验（EBST）可确定严重损伤，但在部分模型中不够敏感。在这两种模型中，用高效液相色谱法测定多巴胺组织含量比纹状体酪氨酸羟化酶免疫染色更能精确测量纹状体多巴胺神经支配。在纹状体模型中，通过酪氨酸羟化酶和尼氏染色估计的细胞数量相关性良好，但在MFB损伤的动物中，这两种测量方法之间存在差异。因此，对于严重损伤动物，在黑质或腹侧被盖区水平更好地估计损伤大小需要额外进行尼氏染色。MFB损伤模型模拟终末期帕金森病。本文所述的纹状体注射6-OHDA不能被视为一种进行性模型，因为在损伤后8周内黑质中酪氨酸羟化酶免疫反应性细胞的数量没有变化。然而，部分去神经支配使其非常适合模拟早期帕金森病，因此适合测试可能的神经保护和神经营养药物。
