Name: repetitive transcranial magnetic stimulation to the unilateral hemisphere of rat brain
Uploaded: 2016-10-22T13:00:00
Description: Watch this Scientific Journal Video about Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain Video at JoVE.com

This work was supported by the Korea Research Foundation Grant funded by the Korean Government (KRF-2008-313-E00458). The authors thank Jin-Joo Lee for the technical assistance.

All of the procedures using animals were reviewed and approved by the Institutional Animal Care and Use Committee of Seoul National University Hospital.
1. Experimental Setup
<ol>
	<li>Animal preparation
	<ol>
		<li>Allow male Sprague-Dawley rats 1 week to adapt to their new environment before starting the experiment. 
		NOTE: Although 8-week old rats were used in the present study, a developing or adult brain can be chosen according to the research hypotheses.</li>
	</ol>
	</li>
	<li>Inhalation anesthesia ...

<ol>
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P., 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=Efficacy+and+safety+of+transcranial+magnetic+stimulation+in+the+acute+treatment+of+major+depression:+a+multisite+randomized+controlled+trial.">Efficacy and safety of transcranial magnetic stimulation in the acute treatment of major depression: a multisite randomized controlled trial.</a> Biol Psychiatry. 62 (11), 1208-1216 (2007).</li><li>Brighina, F., 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=Facilitatory+effects+of+1+Hz+rTMS+in+motor+cortex+of+patients+affected+by+migraine+with+aura.">Facilitatory effects of 1 Hz rTMS in motor cortex of patients affected by migraine with aura.</a> Exp Brain Res. 161 (1), 34-38 (2005).</li><li>Lefaucheur, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stroke+recovery+can+be+enhanced+by+using+repetitive+transcranial+magnetic+stimulation+(rTMS).">Stroke recovery can be enhanced by using repetitive transcranial magnetic stimulation (rTMS).</a> Neurophysiol Clin. 36 (3), 105-115 (2006).</li><li>Khedr, E. M., Ahmed, M. A., Fathy, N., Rothwell, J. 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R., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Opposite+impact+on+14C-2-deoxyglucose+brain+metabolism+following+patterns+of+high+and+low+frequency+repetitive+transcranial+magnetic+stimulation+in+the+posterior+parietal+cortex.">Opposite impact on 14C-2-deoxyglucose brain metabolism following patterns of high and low frequency repetitive transcranial magnetic stimulation in the posterior parietal cortex.</a> Exp Brain Res. 176 (4), 603-615 (2007).</li><li>Di Lazzaro, V., 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=The+physiological+basis+of+the+effects+of+intermittent+theta+burst+stimulation+of+the+human+motor+cortex.">The physiological basis of the effects of intermittent theta burst stimulation of the human motor cortex.</a> J Physiol. 586 (16), 3871-3879 (2008).</li><li>Post, A., Keck, M. 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The primary purpose of this study was to introduce an animal model of unilateral rTMS. Although unilateral stimulation is one of the most fundamental characteristics of human rTMS research, many studies have not adopted it in small animals. However, Rotenberg et al.15 recorded contralateral MEPs with stimulation of 100% MT using a figure-8 coil with an outside lobe diameter of 20 mm, whereas stimulation with 112.5% and 133.3% MT produced ipsilateral as well as contralateral MEPs. This might be because...

Repetitive transcranial magnetic stimulation (rTMS), a tool for non-invasive brain stimulation and neuromodulation, has been applied in the treatment of various conditions such as central pain1,2, depression3, migraine4, and even stroke5-7. Rapidly changing electrical current through coils on the head induces an electrical field on the cerebral cortex and a resultant neuronal activation. The excitability of the cerebral cortex can be modulated by rTMS, which can last for more than 30 min after the stimulation is terminated.
Suggested mechanisms of the rTMS after-effect include long-term potentiation/depression-like effect8, transient shift in ionic balance9, and metabolic changes10. In addition, Di Lazzaro et al. suggest that intermittent theta-burst stimulation affects the excitatory synaptic inputs to pyramidal tract neurons, both in the stimulated and the contralateral hemisphere11.
Significant limitations, however, have hindered researchers from translating on-bench evidence to clinical situations. First, in previous animal studies, rTMS was used for whole-brain stimulation12. Whole-brain stimulation is quite different from the protocols used in human studies9. The other problem is related with the stimulation duration. This is at least partly attributable to the fact that an effective cooling system was unavailable for small coils in the past.
In recent years, seminal articles have been published suggesting ways for overcoming these difficulties in the rTMS experiment on the small animal brain. By these animal models, it was revealed that the rat brain also shows similar cortical excitability changes as in human in response to low-frequency rTMS13. More importantly, cellular and molecular mechanisms of rTMS are increasingly being investigated using animal models of rTMS. A case in point is that a distinct type of inhibitory interneuron is known to be most sensitive to intermittent theta burst stimulation14. Rodent models of rTMS, thus, offer new opportunities for exploring much-sought questions on the molecular underpinnings of rTMS-induced changes. If small animal models of rTMS can be used in more laboratories, it may greatly accelerate and strengthen research in this area.
We now describe how to apply rTMS to the unilateral hemisphere of rat brain, an extension of the previous work15. Stimulation-induced changes were evaluated by using micro-positron emission tomography (PET) and mRNA microarrays to study rTMS-induced changes in the stimulated cerebral cortex.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Homeothermic blanket with a rectal probe</td>
			<td>Harvard apparatus</td>
			<td>507222F</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane (Forane sol.)</td>
			<td>Choongwae</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Propofol (Provive Inj. 1% 20ml)</td>
			<td>Claris Lifesciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Repetitive magnetic stimulator (Magstim Rapid2)</td>
			<td>Magstim Company Ltd</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>25 mm figure-of-8 coil</td>
			<td>Magstim Company Ltd</td>
			<td>1165-00</td>
			<td></td>
		</tr>
		<tr>
			<td>PET-CT</td>
			<td>GE Healthcare</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>QIAzol Lysis Reagent</td>
			<td>Qiagen</td>
			<td>(US Patent No. 5,346,994)</td>
			<td></td>
		</tr>
		<tr>
			<td>RNeasy Lipid Tissue Mini Kit</td>
			<td>Qiagen</td>
			<td>74804</td>
			<td></td>
		</tr>
		<tr>
			<td>RNeasy Mini Spin Columns</td>
			<td>Qiagen</td>
			<td>(Mat No. 1011708)</td>
			<td></td>
		</tr>
		<tr>
			<td>Agilent 2100 Bioanalyzer</td>
			<td>Agilent Technologies</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ambion Illumina RNA amplification kit</td>
			<td>Ambion</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Nanodrop Spectrophotometer</td>
			<td>NanoDrop</td>
			<td>ND-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Illumina RatRef-12 Expression BeadChip</td>
			<td>Illumina, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amersham fluorolink streptavidin-Cy3</td>
			<td>GE Healthcare Bio-Sciences</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

repetitive transcranial magnetic stimulation to the unilateral hemisphere of rat brain

Previous rodent models of repetitive transcranial magnetic stimulation (rTMS) adopted whole-brain stimulation instead of unilateral hemispheric rTMS, which is unlike the protocols used for human subjects. We report a successful application of rTMS to the unilateral hemisphere of rat brain. The rTMS was delivered with a low-frequency (1 Hz), high-frequency (20 Hz), or sham stimulation protocol to one side of the brain by using a small 25-mm figure-8 coil. We placed the center of the coil 1 cm lateral to the vertex on the biauricular line and angulated the coil 45&#176; to the ground to minimize a potential direct effect of rTMS on the contralateral cortex. We also used an in-house water cooling system to enable repetitive magnetic stimulation for more than 20 min, even at a 20-Hz stimulation frequency. Increases in the transcriptions of immediate early genes (Arc, Junb, and Egr2) were greater after rTMS than after sham stimulation. After 5 consecutive days of 20-min 1-Hz rTMS, bdnf mRNA expression was significantly higher in stimulated cortex than in contralateral side. The model presented herein will elucidate the molecular mechanisms of rTMS by allowing analysis of the inter-hemispheric difference in its effect.

Fifteen 8-week old male Sprague-Dawley rats were used for a separate inter-rater reliability analysis of MT determination. Using palpation of muscle twitching, the MTs were obtainable in all rats and measured as 33.00 &#177; 4.21% maximal stimulator output (% MSO) and 33.93 &#177; 0.88% MSO, respectively, by two independent researchers. Bland-Altman bias was -0.93, and the 95% limits of agreement were -9.13 to 7.26%.
In the micr...

Watch this Scientific Journal Video about Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain Video at JoVE.com

Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain Video (Video) | JoVE

We applied repetitive transcranial magnetic stimulation (rTMS) to the unilateral hemisphere of rat brain, by placing a 25-mm figure-8 coil 1 cm lateral to the vertex on the biauricular line and angulating the coil by 45&#176;. An in-house water cooling system was used for rTMS for more than 20 min.

Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain

Previous rodent models of repetitive transcranial magnetic stimulation (rTMS) adopted whole-brain stimulation instead of unilateral hemispheric rTMS, ...

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