Rating L-DOPA-Induced Dyskinesias in the Unilaterally 6-OHDA-Lesioned Rat Model of Parkinson's Disease

Keila Bariotto-dos-Santos; Danilo Leandro Ribeiro; Rayanne Poletti Guimar&#227;es; Fernando  Eduardo Padovan-Neto

doi:10.3791/62924

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Biology

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Rating L-DOPA-Induced Dyskinesias in the Unilaterally 6-OHDA-Lesioned Rat Model of Parkinson’s Disease

评价L-DOPA诱导的运动障碍在帕金森病单侧6-OHDA病变大鼠模型中

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06:45 min

October 4, 2021

DOI: 10.3791/62924-v

Keila Bariotto-dos-Santos*¹, Danilo Leandro Ribeiro*¹, Rayanne Poletti Guimarães¹, Fernando Eduardo Padovan-Neto¹

¹Department of Psychology, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto,University of São Paulo

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study utilizes the 6-hydroxydopamine (6-OHDA) rat model of Parkinson's disease to investigate L-DOPA-induced dyskinesias, aiming to identify therapeutic interventions. The chronic treatment with L-DOPA allows for the assessment of abnormal involuntary movements, providing insights into potential antidyskinetic strategies.

Key Study Components

Research Area

Neuroscience
Pharmacology
Pediatric Medicine

Background

L-DOPA treatment is commonly associated with dyskinesias in Parkinson's disease patients.
Rodent models enable the study of therapeutic effects on dyskinesias.
The 6-OHDA lesion mimics pathophysiological changes seen in human Parkinson's disease.

Methods Used

Stereotaxic administration of 6-OHDA in Sprague-Dawley male rats.
Chronic L-DOPA treatment with benserazide.
Video analysis of abnormal involuntary movements post-treatment.

Main Results

L-DOPA treatment resulted in peak-dose dyskinesias between 30 to 90 minutes.
Patterns of axial, limb, and orolingual movements were systematically scored.
The model effectively mimicked clinical observations of dyskinesias.

Conclusions

This study demonstrates a reliable method to assess L-DOPA-induced dyskinesias.
It underscores the significance of rodent models in developing antidyskinetic therapies.

Frequently Asked Questions

What is L-DOPA-induced dyskinesia?

L-DOPA-induced dyskinesia refers to abnormal involuntary movements that occur as a side effect of long-term treatment with L-DOPA in Parkinson's disease patients.

How is the 6-OHDA model relevant to Parkinson's research?

The 6-OHDA model replicates dopamine depletion seen in Parkinson's disease, allowing for the study of treatments aimed at managing symptoms.

What are the key observations in this study?

Key observations include the timing of dyskinetic movements relative to L-DOPA administration and the assessment of these movements in a controlled environment.

Why is video analysis used in this research?

Video analysis allows for accurate, qualitative scoring of abnormal movements from multiple angles, enhancing the reliability of the observations.

How does this study contribute to antidyskinetic therapy research?

The findings provide insights into the mechanisms and timing of dyskinesias, informing the development of targeted therapeutic interventions.

What implications does this research have for clinical practice?

Understanding dyskinesia onset and severity can guide more effective treatment strategies for Parkinson's patients, improving their quality of life.

Are the animal models used in this study relevant for humans?

Yes, rodent models like the 6-OHDA model have significant predictive validity for human conditions, making them valuable for preclinical research.

左旋多巴诱导的运动障碍的啮齿动物模型是确定治疗干预措施的宝贵工具，以减轻发展或减轻由于反复施用左旋多巴而出现的表现.该协议演示了如何在帕金森病的单侧6-OHDA病变大鼠模型中诱导和分析运动障碍样运动。

在帕金森病的6-羟基多巴胺大鼠模型中评估左旋多巴诱导的运动障碍是确定有效抗运动障碍干预措施的重要临床前工具。这种模式相对简单，成本低，并且与临床上发生的事情有相似之处。该程序将由实验室的博士生Danilo Leandro Ribeiro演示。

用重200至250克的Sprague-Dawley雄性大鼠开始实验。在标准实验室条件下，每个笼子饲养两到三只动物，随意提供食物和水。手术前，腹膜内给予大鼠去甲肾上腺素转运抑制剂丙咪嗪。

在通过对脚趾捏缺乏反应确认麻醉后，将大鼠置于加热垫顶部的立体定位装置中的俯卧位置。一旦大鼠被定位，使用手术刀在将发生显微注射的区域做一个一厘米长的切口。然后，用棉签清洁颅骨区域，并确保前膛和λ暴露在外。

取前脑内侧束或MFB，距前胸4.3毫米，右侧外侧1.6毫米，距硬度计腹侧8.3毫米处的内侧前脑束或MFB，立体定位坐标。确定坐标后，在右侧MFB中以每分钟0.4微升的速率单方面施用6-羟基多巴胺或6-OHDA，使用50微升汉密尔顿玻璃注射器。在手术结束时，缝合头皮切口，并以每公斤10毫升的剂量用温热，无菌的0.9%盐水溶液皮下给动物补充水分。

将动物从立体定位框架中取出，并将其放入温暖的恢复笼中，同时监测直到恢复意识。在病变后四周，通过评估病变对侧前肢的运动不能，使用阶梯试验测量多巴胺能病变的有效性。对侧前肢出现三个或更少调整步骤的大鼠应作为推定的严重 6-OHDA 病变大鼠包括在研究中。

6-OHDA病变四周后，周一开始慢性治疗，周三记录异常的不自主运动。为此，将大鼠放在直径为20厘米，高度为40厘米的透明圆柱体内，并使其适应至少15分钟。确保地板上覆盖着垫料，镜子位于圆柱体后面，以便从所有可能的角度观察动物。

放置高分辨率摄像机，以便查看轴向、肢体和口舌异常不自主运动。气缸后面的后视镜将允许以 360 度角跟踪异常的不自主运动。相机可以与动物成15度角放置在平面下方，以观察口语异常的不自主运动。

确定位置后，将相机直接固定在工作台上。一旦动物适应了 15 分钟，将动物从圆筒中取出，每公斤 5 毫克新鲜制备的左旋多巴胺或左旋多巴联合 12.5 毫克每公斤苄丝肼皮下注射.将动物放回圆筒中并启动计时器以跟踪异常的不自主运动。

使用摄像机记录注射后180分钟的异常不自主运动，每隔30分钟进行一次，以进行离线评分。在六个观察期之后，分数必须在一到两分钟的时期内给出，并分为轴向、肢体或口语。确保不包含正常行为。

左旋多巴和苄丝肼的治疗应持续三周，每天一次，从周一到周五.在代表性的时间历程分析中，显示了应用于轴向，肢体，和口舌异常不自主运动的评分三周慢性左旋多巴给药， .注意到左旋多巴诱导的峰值剂量运动障碍发生在 30 至 90 分钟之间，注射后 120 分钟后逐渐减少.

此外，轴向，肢体，和口舌异常不自主运动的总和呈现在个体，评分天数和每周三周的慢性左旋多巴给药.异常不自主运动方案通常用于研究具有抗运动障碍平移潜力的新治疗工具。

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