Pan-Montojo Francisco J, Funk Richard H W
Institute of Anatomy, Technische Universität Dresden.
J Vis Exp. 2010 Oct 26(44):2123. doi: 10.3791/2123.
In Parkinson's disease (PD) patients, the associated pathology follows a characteristic pattern involving inter alia the enteric nervous system (ENS) (1,2), the olfactory bulb (OB), the dorsal motor nucleus of the vagus (DMV)(3), the intermediolateral nucleus of the spinal cord (4) and the substantia nigra, providing the basis for the neuropathological staging of the disease(4,5). The ENS and the OB are the most exposed nervous structures and the first ones to be affected. Interestingly, PD has been related to pesticide exposure(6-8). Here we show in detail two methods used in our previous study (9). In order to analyze the effects of rotenone acting locally on the ENS, we administered rotenone using a gavage to one-year old C57/BL6 mice. Rotenone is a widely used pesticide that strongly inhibits mitochondrial Complex I (10). It is highly lipophylic and poorly absorbed in the gastrointestinal tract (11). Our results showed that the administration of 5 mg/kg of rotenone did not inhibit mitochondrial Complex I activity in the muscle or the brain. Thus, suggesting that using our administration method rotenone did not cross the hepatoportal system and was acting solely on the ENS. Here we show a method to administer pesticides using a gavage and the image analysis protocol used to analyze the effects of the pesticide in alpha-synuclein accumulation in the ENS. The first part shows a method that allows intragastric administration of pesticides (rotenone) at a desired precise concentration. The second method shows a semi-automatic image analysis protocol to analyze alpha-synuclein accumulation in the ENS using an image analysis software.
在帕金森病（PD）患者中，相关病理变化呈现出一种特征性模式，尤其涉及肠神经系统（ENS）（1,2）、嗅球（OB）、迷走神经背运动核（DMV）（3）、脊髓中间外侧核（4）和黑质，为该疾病的神经病理学分期提供了依据（4,5）。ENS和OB是最易暴露且最先受到影响的神经结构。有趣的是，PD与接触杀虫剂有关（6 - 8）。在此我们详细展示了我们先前研究中使用的两种方法（9）。为了分析鱼藤酮局部作用于ENS的效果，我们通过灌胃给一岁的C57/BL6小鼠施用鱼藤酮。鱼藤酮是一种广泛使用的杀虫剂，能强烈抑制线粒体复合物I（10）。它具有高度脂溶性，在胃肠道中吸收不良（11）。我们的结果表明，施用5 mg/kg的鱼藤酮并未抑制肌肉或大脑中的线粒体复合物I活性。因此，这表明使用我们的给药方法，鱼藤酮未穿过肝门静脉系统，仅作用于ENS。在此我们展示了一种通过灌胃施用杀虫剂的方法以及用于分析杀虫剂对ENS中α-突触核蛋白积累影响的图像分析方案。第一部分展示了一种允许以所需精确浓度进行胃内施用杀虫剂（鱼藤酮）的方法。第二种方法展示了一种半自动图像分析方案，用于使用图像分析软件分析ENS中α-突触核蛋白的积累情况。
