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 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.

जानवरों का उपयोग प्रक्रियाओं की समीक्षा और सियोल नेशनल यूनिवर्सिटी अस्पताल के संस्थागत पशु की देखभाल और उपयोग समिति द्वारा अनुमोदित किया गया। 1. प्रायोगिक सेटअप <ol><li> पशु तैयारी <ol><li> प्रयोग शुरू करन...

<ol>
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<html> इस अध्ययन के प्राथमिक उद्देश्य एकतरफा rTMS के एक पशु मॉडल पेश करने के लिए किया गया था। हालांकि एकतरफा उत्तेजना मानव rTMS अनुसंधान के सबसे मौलिक विशेषताओं में से एक है, कई अध्ययनों छोटे जानवरों में इसे नही?...

दोहराव transcranial चुंबकीय उत्तेजना (rTMS), गैर इनवेसिव मस्तिष्क उत्तेजना और neuromodulation के लिए एक उपकरण है, इस तरह के केंद्रीय दर्द 1,2, अवसाद 3, माइग्रेन 4, और यहां तक कि स्ट्रोक 5-7 के रूप में विभिन्न शर्तों के उपचार में लागू किया गया है। तेजी से कॉयल के माध्यम से विद्युत धारा को बदलने के सिर सेरेब्रल कॉर्टेक्स और एक परिणामी न्यूरोनल सक्रियण पर एक बिजली के क्षेत्र लाती है। सेरेब्रल कॉर्टेक्स के excitability rTMS, जो अधिक से अधिक 30 मिनट के लिए पिछले के बाद उत्तेजना समाप्त होता है सकते द्वारा संग्राहक जा सकता है। बाद प्रभाव rTMS का सुझाव दिया तंत्र दीर्घकालिक potentiation / अवसाद की तरह प्रभाव 8, आयनिक संतुलन 9 में क्षणिक पारी शामिल है, और चयापचय परिवर्तनों 10। इसके अलावा, Di Lazzaro एट अल। सुझाव है कि रुक-रुक कर थीटा-फट उत्तेजना दोनों प्रेरित में, पथ न्यूरॉन्स पिरामिड को उत्तेजक synaptic आदानों को प्रभावित करता हैऔर contralateral गोलार्द्ध 11। महत्वपूर्ण सीमाओं, हालांकि, नैदानिक ​​स्थितियों पर बेंच सबूत अनुवाद से शोधकर्ताओं रुकावट है। सबसे पहले, पिछले जानवरों के अध्ययन में, rTMS पूरे मस्तिष्क की उत्तेजना 12 के लिए इस्तेमाल किया गया था। पूरे मस्तिष्क उत्तेजना मानव अध्ययन 9 में इस्तेमाल किया प्रोटोकॉल से काफी अलग है। अन्य समस्या उत्तेजना की अवधि के साथ संबंधित है। यह कम से कम आंशिक रूप से तथ्य यह है कि एक प्रभावी शीतलन प्रणाली अतीत में छोटे कॉयल के लिए उपलब्ध नहीं था के कारण है। हाल के वर्षों में, मौलिक लेख छोटे जानवर मस्तिष्क पर rTMS प्रयोग में इन कठिनाइयों पर काबू पाने के लिए तरीके का सुझाव प्रकाशित किया गया है। इन पशु मॉडल के द्वारा, यह पता चला था कि चूहे के मस्तिष्क भी कम आवृत्ति rTMS 13 के जवाब में मानव के रूप में समान cortical excitability परिवर्तन दिखाता है। इससे भी महत्वपूर्ण बात, rTMS के सेलुलर और आणविक तंत्र तेजी से bei हैंएनजी rTMS के पशु मॉडल का उपयोग कर जांच की। बिंदु में एक मामला है कि निरोधात्मक interneuron की एक अलग प्रकार रुक-रुक कर थीटा फट उत्तेजना से 14 सबसे संवेदनशील हो जाना जाता है। rTMS के कृंतक मॉडल, इस प्रकार, rTMS प्रेरित परिवर्तन के आणविक आधार पर बहुत समय से मांग सवालों की खोज के लिए नए अवसर प्रदान करते हैं। rTMS के छोटे पशु मॉडल और अधिक प्रयोगशालाओं में इस्तेमाल किया जा सकता है, यह बहुत तेजी लाने और इस क्षेत्र में अनुसंधान को मजबूत कर सकता है। अब हम वर्णन कैसे चूहे के मस्तिष्क, पिछले काम 15 का एक विस्तार के एकतरफा गोलार्द्ध के लिए rTMS लागू करने के लिए। उत्तेजना प्रेरित परिवर्तन प्रेरित सेरेब्रल कॉर्टेक्स में rTMS प्रेरित परिवर्तन का अध्ययन करने के लिए माइक्रो-पोजीट्रान एमिशन टोमोग्राफी (पीईटी) और mRNA प्रोटीन का उपयोग करके मूल्यांकन किया गया।

<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.

पंद्रह 8 सप्ताह पुराने पुरुष Sprague-Dawley चूहों मीट्रिक टन दृढ़ संकल्प का एक अलग अंतर करदाता विश्वसनीयता विश्लेषण के लिए इस्तेमाल किया गया। मांसपेशियों में ऐंठन की टटोलने का कार्य का उपयोग करना,...

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

Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain

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.

चूहा मस्तिष्क के एकतरफा गोलार्ध के लिए दोहराव transcranial चुंबकीय उत्तेजना

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

The overall goal of this experiment is to conduct a unilateral repetitive transcranial magnetic stimulation, or rTMS, in a small animal model. This method can help answer key questions regarding molecular mechanisms of rTMS, by enabling unilateral hemispheric stimulation to the rat brain. The main advantage of this technique is that it provides an opportunity to explore the interhemispheric differences of molecular events, in response to unilateral rTMS.Individuals new to this method may struggle because the rTMS coil can be so large that they stimulate the whole brain, and because the timing of the motor threshold is time consuming and laborious. An experienced technician from our laboratory, Jin-Joo Lee, will be demonstrating the procedure. Place a rectal probe in an anesthetized adult male rat to monitor the body temperature, using a homeothermic blanket to maintain the temperature at 37 degrees Celsius.Apply ointment to the animal's eyes. Then, 10 minutes after the complete transition to IV anesthesia, mark a point 0.5 centimeters lateral to the vertex on the biauricular line, and fix the coil holder firmly to the apparatus. Then, locate the center of a 25 millimeter figure of eight coil at the point, and angulate the coil 45 degrees to the ground.Holding the coil surface flat against the calvaria, gradually increase the stimulation intensity to determine the minimum stimulus intensity required to evoke five or more palpable contralateral forepaw contractions, by ten consecutive biphasic stimuli, through the coil. Verify that the stimulation is primarily causing the contralateral muscle contraction, without any ipsilateral contraction, to ensure a unilateral stimulation. Use a water-cooling system to cool the coil for 10 minutes.Maintaining the coil temperature with the cooling system, use the software console to set the stimulation intensity at 100 to 110%of the motor threshold. Then, to apply the rTMS at a one hertz stimulation, first set the input to 1200 shots, for 20 minutes. Then, move the center of the coil to the target site on one cerebral cortex, and tilt the coil, to ensure the direct contact between the coil center and the surface of the skull at the point of stimulation.For a 20 hertz stimulation, set the input to 1600 shots over 20 minutes, and conduct two seconds of stimulation, followed by 28 seconds of rest. For a sham stimulation, tilt the coil at a 90 degree rotation to the calvaria, and place the edge of the coil two centimeters from the head surface. At the end of the stimulation, allow the animal to recover with full monitoring.In this representative experiment, six eight-week-old rats were imaged by micro-positron emission tomography to evaluate the uptake of a radio isotope in the cerebral cortices regions of interest. The radioactivity in the contralateral area was used as a reference to normalize the data obtained in the ipsilateral area. The mean differential uptake ratios obtained from three consecutive transverse images, were then averaged to calculate the differential uptake ratio for the rats, demonstrating a focal increase in the glucose metabolism in the stimulated left cortical area, in the one hertz group.Supporting the unilaterality of the rTMS. In this mRNA microarray study, the expression levels of several immediate early genes were observed to be significantly higher in the rTMS animals than in the sham group. In addition, after a one hertz stimulation, brain-derived neurotrophic factor mRNA expression was also significantly higher in the stimulated cortex, revealing differential rTMS-induced changes in the stimulated versus the contralateral cerberal cortices.Once administered, the rTMS technique can be completed in one hour, if it's performed properly. While attempting this procedure, it's important to take care to properly position the rTMS coil, and to prepare the contra and ipsilateral forepaw contractions to confirm the unilateral stimulation of the hemispheres. This technique will be useful in the field of translational neurorehabilitation research, for exploring the molecular mechanisms of rTMS, through the analysis of interhemispheric differences, and the various pathologic conditions.After watching this video, you should have a good understanding of how to conduct unilateral rTMS, in a small animal model.

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