Name: induction and assessment of levodopa-induced dyskinesias in a rat model of parkinson's disease
Uploaded: 2021-10-14T14:30:00
Description: Watch this Scientific Journal Video about Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease at JoVE.com

We want to acknowledge the struggles of all those with Parkinson&#39;s disease and the strength and resilience they show every day, especially the beloved father of KSC, Mark Steece. The work represented here was supported by the National Institute of Neurological Disorders and Stroke (NS090107, NS110398) and the Parkinson Disease Foundation International Research Grant Program, now the Parkinson Foundation.&#160;We also would like to acknowledge Molly VanderWerp for her excellent editorial assistance.

The animals presented here were maintained and handled in compliance with the institutional guidelines. All animal procedures were approved by the Michigan State University Institutional Animal Care and Use Committee (IACUC) in compliance with federal and state regulations.
1. Drug-free confirmation of 6-OHDA lesion status
<ol>
	<li>Postural Tail hang test11,12,13 
	NOTE: Asses...

Presented here are details for the reproducible induction and rating of LIDs in a parkinsonian rat model following unilateral 6-OHDA lesioning of the nigrostriatal DA system. While it was once thought that rodents did not develop LID and that rotational asymmetry may be the analog of LID in rats31, rat and mouse models have been characterized over the past two decades and are a well-accepted tool for LID research15,32,<sup class...

Although it has been more than 50 years since levodopa (L-DOPA) was first introduced as a treatment for individuals with PD1,2, it remarkably remains the most effective therapy for parkinsonian motor symptoms. The clinical motor symptoms associated with PD stem from the loss of dopamine (DA) neurons in the substantia nigra (SN) pars compacta, resulting in the dramatic decrease in available dopamine in the striatum. L-DOPA effectively restores striatal DA levels, resulting in motor benefit early in the disease3,4. Inopportunely, with long-term treatment, most individuals with PD will develop&#160;L-DOPA induced dyskinesias (LID), including chorea, dystonia, and athetosis, which often significantly impact activities of daily living5,6,7.
While several behavioral models of LID in rodents exist, differences in modeling and behavioral assessment of LIDs have called into question the reproducibility of results between labs as well as the reliability of these experimental tools for pre-clinical PD research. Developed in association with a clinical movement disorder specialist8, the current protocol is a straightforward method for LID induction and rating and is appropriate for use in a rat model of PD utilizing 6-hydroxydopamine (6-OHDA)-induced unilateral nigral lesioning9,10. The LID rating scale provided here includes scoring for both the intensity and frequency of dyskinetic behavior in various individual body parts. Pertinent information regarding workflow optimization of experiments and the appropriate care and handling of parkinsonian and dyskinetic animals is also provided.

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			<td height="21" style="height:21px;width:441px;">&#160;100 Minutes Digital Timer</td>
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			<td height="21" style="height:21px;width:441px;">&#160;Compass CX Compact Scale</td>
			<td style="width:193px;">Ohaus</td>
			<td style="width:182px;">30428202</td>
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			<td height="42" style="height:42px;">5-(2-aminoethyl)-1,2,4-benzenetriol, monohydrobromide</td>
			<td style="width:193px;">Cayman Chemicals</td>
			<td style="width:182px;">25330</td>
			<td style="width:559px;">6-OHDA is a catecholaminergic neurotoxin that is used to induce dopaminergic lesions and parkinsonian symptoms in rodents.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:441px;">Allentown cages</td>
			<td style="width:193px;">Allentown, LLC</td>
			<td style="width:182px;">Rat900</td>
			<td style="width:559px;">Allentown cages provide the ability to view the rats from all sides.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:441px;">BD Allergist Trays with Permanently Attached Needle</td>
			<td style="width:193px;">BD</td>
			<td style="width:182px;">BD 305540</td>
			<td style="width:559px;">For subcutaneous L-DOPA injections</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:441px;">Benserazide hydrochloride</td>
			<td style="width:193px;">Sigma-Aldrich</td>
			<td style="width:182px;">B7283</td>
			<td style="width:559px;">Benserazide is a peripheral decarboxylase inhibitor used with L-DOPA to to induce dyskinesia in rodent models of PD.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:441px;">Glass amber scintillation vials</td>
			<td style="width:193px;">Thermo Scientific</td>
			<td style="width:182px;">B7921</td>
			<td style="width:559px;">Used for storage of L-DOPA/benserazide at -20 &#176;C until mixed with sterile saline.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">L-3,4-Dihydroxyphenylalanine methyl ester hydrochloride</td>
			<td style="width:193px;">Sigma-Aldrich</td>
			<td style="width:182px;">D1507</td>
			<td style="width:559px;">L-3,4-Dihydroxyphenylalanine methyl ester is a precursor to&#160;L-DOPA that crosses the blood-brain barrierand use to treat parkinsonian symptoms in rodents.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:441px;">Paper Mate Sharpwriter Mechanical Pencils</td>
			<td style="width:193px;">Staples</td>
			<td style="width:182px;">107250</td>
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			<td height="21" style="height:21px;width:441px;">Rodent nutritionally complete enrichment treats</td>
			<td style="width:193px;">Bio-Serv</td>
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			<td height="21" style="height:21px;width:441px;">Round Ice Bucket with Lid, 2.5 L</td>
			<td style="width:193px;">Corning</td>
			<td style="width:182px;">432129</td>
			<td style="width:559px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:441px;">Standard Plastic Clipboard</td>
			<td style="width:193px;">Staples</td>
			<td style="width:182px;">1227770</td>
			<td style="width:559px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:441px;">Steel wired 6&#39; long movable shelving units</td>
			<td style="width:193px;">Uline</td>
			<td style="width:182px;">H9488</td>
			<td style="width:559px;">Width/Height can be adjusted to need/number of rats per experiment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:441px;">Sterile Saline 0.9%</td>
			<td style="width:193px;">Covidien/Argyle</td>
			<td style="width:182px;">1020</td>
			<td style="width:559px;">For mixing with L-DOPA/benserazide prior to subcutaneous injections.</td>
		</tr>
	</tbody>
</table>

induction and assessment of levodopa-induced dyskinesias in a rat model of parkinson's disease

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements known as L-DOPA-induced dyskinesias (LIDs) develop with prolonged use of this dopamine precursor.&#160;It is estimated that the incidence of LIDs escalates to approximately 90% of individuals with PD within 10&#8211;15 years of treatment. Understanding the mechanisms of this malady and developing both novel and effective anti-dyskinesia treatments requires consistent and accurate modeling for pre-clinical testing of therapeutic interventions. A detailed method for reliable induction and comprehensive rating of LIDs following 6-OHDA-induced nigral lesioning in a rat model of PD is presented here. Dependable LID assessment in rats provides a powerful tool that can be readily utilized across laboratories to test emerging therapies focused on reducing or eliminating this common treatment-induced burden for individuals with PD.

LIDs in parkinsonian rats can manifest as a range of abnormal involuntary movements (AIMs), including dystonic, hyperkinetic, and stereotypic behaviors. LID rating criteria for such behaviors are presented here to include both intensity (Table 1) and frequency (Table 2). This provides an overall LID severity score for each rat that reflects both the quality (intensity) and quantity of time spent engaging (frequency) in these behaviors at each rating timepoint. The final LID severity scor...

Watch this Scientific Journal Video about Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease at JoVE.com

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease

This article describes methods to induce and evaluate levodopa-induced dyskinesias in a rat model of Parkinson&#39;s disease. The protocol offers detailed information regarding the intensity and frequency of a range of dyskinetic behaviors, both dystonic and hyperkinetic, providing a reliable tool to test treatments targeting this unmet medical need.

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements ...

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