Name: positron emission tomography imaging for in vivo measuring of myelin content in the lysolecithin rat model of multiple sclerosis
Uploaded: 2021-02-28T14:30:00
Description: Watch this Scientific Journal Video about Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis at JoVE.com

&#946;-cube equipment (Molecubes NV, Belgium) was supported by the S&#227;o Paulo Research Foundation, FAPESP - Brazil (#2018/15167-1). LES has a PhD student scholarship from FAPESP - Brazil (#2019/15654-2).

All procedures were conducted in accordance with the guidelines of the National Council for the control of Animal Experimentation (CONCEA, Brazil) and were approved by the Ethics Committee for Animal Research of the Medical School of the University of Sao Paulo (CEUA-FMUSP, Brazil - protocol number: 25/15).
NOTE: In this protocol, we show how to induce a lysolecithin rat model of multiple sclerosis and how to acquire and analyze the myelin PET images.
1. Lysolecithin ...

<ol>
	<li>Oh, J., Vidal-Jordana, A., Montalban, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+sclerosis:+clinical+aspects.">Multiple sclerosis: clinical aspects.</a> Current Opinion in Neurology. 31 (6), 752-759 (2018).</li><li>Sand, I. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classification,+diagnosis,+and+differential+diagnosis+of+multiple+sclerosis.">Classification, diagnosis, and differential diagnosis of multiple sclerosis.</a> Current Opinion in Neurology. 28 (3), 193-205 (2015).</li><li>Thompson, A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+of+multiple+sclerosis:+2017+revisions+of+the+McDonald+criteria.">Diagnosis of multiple sclerosis: 2017 revisions of the McDonald criteria.</a> Lancet Neurology. 17 (2), 162-173 (2018).</li><li>Veronese, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+C-11+PIB+PET+for+imaging+myelin+in+the+human+brain:+a+test-retest+reproducibility+study+in+high-resolution+research+tomography.">Quantification of C-11 PIB PET for imaging myelin in the human brain: a test-retest reproducibility study in high-resolution research tomography.</a> Journal of Cerebral Blood Flow and Metabolism. 35 (11), 1771-1782 (2015).</li><li>Carvalho, R. H. 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The biggest advantage of using the lysolecithin model to study multiple sclerosis is the fast timeline for demyelination (about 1 week) and remyelination (about 4 weeks) to occur14. This model can also be induced in mice15, however, induction in rats is more advantageous for in vivo PET imaging due to the larger size of the rat brain compared to mice.
The first step of the induction model is to be extremely cautious. This model was valid...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/62094">Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis</a>. The citation&#160;was updated.
The citation was updated from:
de Paula Faria, D., Cristiano Real, C., Estessi de Souza, L., Teles Garcez, A., Navarro Marques, F. L., Buchpiguel, C. A. Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis. J. Vis. Exp. (168), e62094, doi:10.3791/62094 (2021).
to:
de Paula Faria, D., Real, C.C., Estessi de Souza, L., Teles Garcez, A., Navarro Marques, F. L., Buchpiguel, C. A. Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis. J. Vis. Exp. (168), e62094, doi:10.3791/62094 (2021).

An erratum was issued for:&#160;<a href="https://www.jove.com/t/62094">Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis</a>. The citation&#160;was updated.

Erratum: Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis

Multiple sclerosis (MS) is a neuroinflammatory disease that affects the central nervous system, characterized by inflammation, demyelination, and axonal loss1. The prognosis of this disease is variable even with advances in treatment, and it is one of the most common causes of neurological deficits in young people1. The diagnosis of MS is based on the criteria of clinical manifestation and visualization of characteristic lesions by magnetic resonance imaging (MRI)2,3.
Positron emission tomography (PET) can be a useful tool for in vivo monitoring of MS progression and therapeutic effects. The Pittsburgh compound B radiotracer (PIB) labeled with carbon-11 (11C-PIB) is widely used to quantify &#946;-amyloid plaques; however, in the last decade, it has been studied to quantify myelin content and show dynamic demyelination and remyelination4,5,6.
Different amyloid PET tracers (11C-PIB, 18F-florbetaben,18F-florbetapir, 18F-flutemetamol) can be used to quantify myelin and provide important information about disease progression and therapeutic response, allowing identification of demyelination and remyelination processes, without the interference of neuroinflammation, which can occur with conventional magnetic resonance images (MRI)7. Amyloid PET imaging showed decreased tracer uptake in active MS patients compared to non-active patients which could be explained by early white matter damage in the active patients8. Lower amyloid tracer uptake was also associated with cognitive decline in a follow-up study, showing this technique to be a valuable tool for studying the pathophysiology of the disease and clinical outcomes9.
The lysolecithin (LPC) rat model is a chemical induced model of multiple sclerosis, where the injected toxin, LPC, induces a high response of macrophages that results in increased inflammation and, consequently, demyelination10,11. The demyelination is rapidly reversed, in approximately 4 weeks, which makes this a good model for evaluating demyelination and remyelination processes in rodents. This model has already been evaluated using PET imaging, with good results and correlation with post-mortem essays12.
Here we present the protocol for myelin PET imaging with 11C-PIB in the lysolecithin rat model, showing this imaging technique to be a useful tool for in vivo measurement of myelin content.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Analytical Balance</td>
			<td>Marte</td>
			<td>AUWZZOD</td>
			<td>max: 220 g- min: 1 mg</td>
		</tr>
		<tr>
			<td>Anestesia vaporizer</td>
			<td>Nanitech</td>
			<td>15800</td>
			<td></td>
		</tr>
		<tr>
			<td>Beta-cube</td>
			<td>Molecubes</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bulldog clamp</td>
			<td>Stoelting</td>
			<td>5212043P</td>
			<td></td>
		</tr>
		<tr>
			<td>clorexidine</td>
			<td>Rioquimica</td>
			<td></td>
			<td>0.5%/100 mL</td>
		</tr>
		<tr>
			<td>Cotton swabs</td>
			<td>johnson e johnson</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dose calibrator</td>
			<td>Capintech</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Drill</td>
			<td>Kinzo powertools</td>
			<td>352901</td>
			<td>Model Q0M-DC3C</td>
		</tr>
		<tr>
			<td>Eppendorf tube</td>
			<td>Eppendorf</td>
			<td>30125150</td>
			<td>1.5 mL</td>
		</tr>
		<tr>
			<td>Eye lubricant</td>
			<td>ADVFARMA</td>
			<td>30049099</td>
			<td>&#160;vaseline 15 g (pharmaceutical purity)</td>
		</tr>
		<tr>
			<td>Fine forceps</td>
			<td>Stoelting</td>
			<td>52102-38P</td>
			<td></td>
		</tr>
		<tr>
			<td>Gloves</td>
			<td>Descarpack</td>
			<td>212101</td>
			<td>&#160;6.5 size</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Softhear</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Injection Syringe</td>
			<td>Hamilton</td>
			<td>80314</td>
			<td>10&#181;, 32ga, model 701</td>
		</tr>
		<tr>
			<td>Insuline syringe</td>
			<td>BD</td>
			<td>328328</td>
			<td>1 mL insulin syringes with needle</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Crist&#225;lia</td>
			<td>410525</td>
			<td>100 mL , concentration 1 mL/1 mL</td>
		</tr>
		<tr>
			<td>Ketoprofen or other analgesic</td>
			<td>Sanofi</td>
			<td></td>
			<td>100 mg/2 mL</td>
		</tr>
		<tr>
			<td>lidocaine</td>
			<td>Hipolabor</td>
			<td>1.1343.0102.001-5</td>
			<td>2%/20mL</td>
		</tr>
		<tr>
			<td>L-&#945;-Lysophosphatidylcholine from egg yolk</td>
			<td>Sigma-aldrich</td>
			<td>L-4129</td>
			<td>25 mg - &#8805;99%, Type I, powder</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Stoelting</td>
			<td>5212290P</td>
			<td></td>
		</tr>
		<tr>
			<td>Oxygen</td>
			<td>White Martins</td>
			<td>7782-44-7</td>
			<td>Compressed gas</td>
		</tr>
		<tr>
			<td>PMOD software</td>
			<td>PMOD technologies</td>
			<td>Version 4.1</td>
			<td>module fuse it</td>
		</tr>
		<tr>
			<td>Rat anesthesia mask</td>
			<td>KOPF</td>
			<td>Model 906</td>
			<td></td>
		</tr>
		<tr>
			<td>Saline</td>
			<td>Farmace</td>
			<td>0543325/ 14-8</td>
			<td>0.9% sodium chloride for injection, 10 mL</td>
		</tr>
		<tr>
			<td>Scapel blades</td>
			<td>Stoelting</td>
			<td>52173-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Scapel handles</td>
			<td>Stoelting</td>
			<td>52171P</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissor</td>
			<td>Stoelting</td>
			<td>52136-50P</td>
			<td></td>
		</tr>
		<tr>
			<td>Semi-analytical Balance</td>
			<td>Quimis</td>
			<td>BK-3000</td>
			<td>max:3,100 g; min:0.2 g</td>
		</tr>
		<tr>
			<td>shaver</td>
			<td>Mega profissional</td>
			<td></td>
			<td>AT200 model</td>
		</tr>
		<tr>
			<td>Stereotactic Apparatus</td>
			<td>KOPF</td>
			<td>Nodel 900</td>
			<td></td>
		</tr>
		<tr>
			<td>Universal holder with needle support</td>
			<td>KOPF</td>
			<td>Model 1772-F1</td>
			<td>Hamilton support for 5 and 10 &#181;L</td>
		</tr>
	</tbody>
</table>

positron emission tomography imaging for in vivo measuring of myelin content in the lysolecithin rat model of multiple sclerosis

Multiple sclerosis (MS) is a neuroinflammatory disease with expanding axonal and neuronal degeneration and demyelination in the central nervous system, leading to motor dysfunctions, psychical disability, and cognitive impairment during MS progression. Positron emission tomography (PET) is an imaging technique able to quantify in vivo cellular and molecular alterations.
Radiotracers with affinity to intact myelin can be used for in vivo imaging of myelin content changes over time. It is possible to detect either an increase or decrease in myelin content, what means this imaging technique can detect demyelination and remyelination processes of the central nervous system. In this protocol we demonstrate how to use PET imaging to detect myelin changes in the lysolecithin rat model, which is a model of focal demyelination lesion (induced by stereotactic injection) (i.e., a model of multiple sclerosis disease). 11C-PIB PET imaging was performed at baseline, and 1 week and 4 weeks after stereotaxic injection of lysolecithin 1% in the right striatum (4 &#181;L) and corpus callosum (3 &#181;L) of the rat brain, allowing quantification of focal demyelination (injection site after 1 week) and the remyelination process (injection site at 4 weeks).
Myelin PET imaging is an interesting tool for monitoring in vivo changes in myelin content which could be useful for monitoring demyelinating disease progression and therapeutic response.

Figure 1&#160;shows illustrative 11C-PIB PET images with myelin changes over time. In the baseline scan, no differences can be seen in myelin content (i.e., no demyelination is present). In the 1-week time-point image, it is possible to see the focal demyelinated lesion (in the right hemisphere) as indicated by the white arrow. Images are presented in the 3 anatomical planes (coronal, axial, and sagittal) and it is possible to identify the demyelinated lesion in all of them. The 1...

Watch this Scientific Journal Video about Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis at JoVE.com

Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis

This protocol has the aim of monitoring in vivo myelin changes (demyelination and remyelination) by positron emission tomography (PET) imaging in an animal model of multiple sclerosis.

Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis

Multiple sclerosis (MS) is a neuroinflammatory disease with expanding axonal and neuronal degeneration and demyelination in the central nervous system, ...

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