Name: surgical labyrinthectomy of the rat to study the vestibular system
Uploaded: 2018-05-19T13:00:00
Description: Watch this Scientific Journal Video about Surgical Labyrinthectomy of the Rat to Study the Vestibular System at JoVE.com

Denne forskning blev støttet af en bevilling fra Korea sundhed Technology R & D projekt gennem den Korea sundhed industri udvikling Institute (KHIDI), finansieret af Ministeriet for sundhed og velfærd, Republikken Korea (giver nummer: HI15C2651).

Denne undersøgelse blev udført i overensstemmelse med den institutionelle Animal Care og brug Udvalget af Seoul National University Hospital (14-0148-C1A1), som er akkrediteret af Association for evaluering og akkreditering af Laboratory Animal Care International.
Bemærk: Forsøgene blev udført på Sprague-Dawley (SD) mandlige rotter i 7-8 uger gamle (200/250 g). Hvert dyr blev akklimatiseret til laboratorieforhold for 1 uge før starten af forsøget. Dyrene har været opstaldet i en kontr...

<ol>
	<li>Curthoys, I. S., Halmagyi, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vestibular+compensation:+a+review+of+the+oculomotor,+neural,+and+clinical+consequences+of+unilateral+vestibular+loss.">Vestibular compensation: a review of the oculomotor, neural, and clinical consequences of unilateral vestibular loss.</a> Journal of Vestibular Research. 5 (2), 67-107 (1995).</li><li>Smith, P. F., Curthoys, I. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+recovery+following+unilateral+labyrinthectomy:+a+review.">Mechanisms of recovery following unilateral labyrinthectomy: a review.</a> Brain Research. Brain Research Reviews. 14 (2), 155-180 (1989).</li><li>Lacour, M., Helmchen, C., Vidal, P. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vestibular+compensation:+the+neuro-otologist's+best+friend.">Vestibular compensation: the neuro-otologist's best friend.</a> Journal of Neurology. 263, S54-S64 (2016).</li><li>Darlington, C. L., Smith, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+of+recovery+from+vestibular+damage+in+mammals:+recent+advances.">Molecular mechanisms of recovery from vestibular damage in mammals: recent advances.</a> Progress in Neurobiology. 62 (3), 313-325 (2000).</li><li>Shinder, M. E., Ramanathan, M., Kaufman, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymmetric+gene+expression+in+the+brain+during+acute+compensation+to+unilateral+vestibular+labyrinthectomy+in+the+Mongolian+gerbil.">Asymmetric gene expression in the brain during acute compensation to unilateral vestibular labyrinthectomy in the Mongolian gerbil.</a> Journal of Vestibular Research. 16 (4-5), 147-169 (2006).</li><li>Dutheil, S., Brezun, J. M., Leonard, J., Lacour, M., Tighilet, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurogenesis+and+astrogenesis+contribution+to+recovery+of+vestibular+functions+in+the+adult+cat+following+unilateral+vestibular+neurectomy:+cellular+and+behavioral+evidence.">Neurogenesis and astrogenesis contribution to recovery of vestibular functions in the adult cat following unilateral vestibular neurectomy: cellular and behavioral evidence.</a> Neuroscience. 164 (4), 1444-1456 (2009).</li><li>Gunther, L., 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=N-acetyl-L-leucine+accelerates+vestibular+compensation+after+unilateral+labyrinthectomy+by+action+in+the+cerebellum+and+thalamus.">N-acetyl-L-leucine accelerates vestibular compensation after unilateral labyrinthectomy by action in the cerebellum and thalamus.</a> PLoS One. 10 (3), e0122015 (2015).</li><li>Pericat, D., Farina, A., Agavnian-Couquiaud, E., Chabbert, C., Tighilet, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complete+and+irreversible+unilateral+vestibular+loss:+a+novel+rat+model+of+vestibular+pathology.">Complete and irreversible unilateral vestibular loss: a novel rat model of vestibular pathology.</a> Journal of Neuroscience Methods. 283, 83-91 (2017).</li><li>Cass, S. P., Goshgarian, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vestibular+compensation+after+labyrinthectomy+and+vestibular+neurectomy+in+cats.">Vestibular compensation after labyrinthectomy and vestibular neurectomy in cats.</a> Otolaryngology-Head and Neck Surgery. 104 (1), 14-19 (1991).</li><li>Vignaux, G., 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=Evaluation+of+the+chemical+model+of+vestibular+lesions+induced+by+arsanilate+in+rats.">Evaluation of the chemical model of vestibular lesions induced by arsanilate in rats.</a> Toxicology and Applied Pharmacology. 258 (1), 61-71 (2012).</li><li>Berryhill, W. E., Graham, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+and+physical+labyrinthectomy+for+Meniere's+disease.">Chemical and physical labyrinthectomy for Meniere's disease.</a> Otolaryngologic Clinics of North America. 35 (3), 675-682 (2002).</li><li>Morgenstern, C., Mori, N., Arnold, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+studies+on+the+effect+of+labyrinth+anesthesia.">Experimental studies on the effect of labyrinth anesthesia.</a> Archives of Oto-Rhino-Laryngology. 237 (3), 255-261 (1983).</li><li>Nadasy, G. L., Raffai, G., Feher, E., Schaming, G., Monos, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+standard+technique+for+labyrinthectomy+in+the+rat:+a+methodical+communication+with+a+detailed+description+of+the+surgical+process.">A simple standard technique for labyrinthectomy in the rat: a methodical communication with a detailed description of the surgical process.</a> Physiology International. 103 (3), 354-360 (2016).</li><li>Hitier, M., Besnard, S., Vignaux, G., Denise, P., Moreau, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ventrolateral+surgical+approach+to+labyrinthectomy+in+rats:+anatomical+description+and+clinical+consequences.">The ventrolateral surgical approach to labyrinthectomy in rats: anatomical description and clinical consequences.</a> Surgical and Radiologic Anatomy. 32 (9), 835-842 (2010).</li><li>Flourens, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Recherches experimentales sur les propri&#233;t&#233;s et les fonctions du syst&#232;me nerveux dans les animaux vert&#233;br&#233;s. , (1824).</li><li>T'Ang, Y., Wu, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+unilateral+labyrinthectomy+in+the+albino+rat.">The effects of unilateral labyrinthectomy in the albino rat.</a> Chinese Journal of Physiology. 10, 571-598 (1936).</li><li>Chang, M. Y., 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=MicroRNAs+218a-5p,+219a-5p,+and+221-3p+regulate+vestibular+compensation.">MicroRNAs 218a-5p, 219a-5p, and 221-3p regulate vestibular compensation.</a> Scientific Reports. 7 (1), 8701 (2017).</li><li>Bergquist, F., Ludwig, M., Dutia, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+the+commissural+inhibitory+system+in+vestibular+compensation+in+the+rat.">Role of the commissural inhibitory system in vestibular compensation in the rat.</a> The Journal of Physiology. 586 (18), 4441-4452 (2008).</li><li>Cameron, S. A., Dutia, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+basis+of+vestibular+compensation:+changes+in+intrinsic+excitability+of+MVN+neurones.">Cellular basis of vestibular compensation: changes in intrinsic excitability of MVN neurones.</a> NeuroReport. 8 (11), 2595-2599 (1997).</li><li>Park, M. K., Lee, B. D., Lee, J. D., Jung, H. H., Chae, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+profiles+during+vestibular+compensation+in+rats+after+unilateral+labyrinthectomy.">Gene profiles during vestibular compensation in rats after unilateral labyrinthectomy.</a> Annals of Otology, Rhinology &amp; Laryngology. 121 (11), 761-769 (2012).</li><li>Yamamoto, H., Tominaga, M., Sone, M., Nakashima, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+stapedial+artery+to+blood+flow+in+the+cochlea+and+its+surrounding+bone.">Contribution of stapedial artery to blood flow in the cochlea and its surrounding bone.</a> Hearing Research. 186 (1-2), 69-74 (2003).</li><li>Potegal, M., Abraham, L., Gilman, S., Copack, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technique+for+vestibular+neurotomy+in+the+rat.">Technique for vestibular neurotomy in the rat.</a> Physiology &amp; Behavior. 14 (2), 217-221 (1975).</li><li>Tung, V. W., Burton, T. J., Dababneh, E., Quail, S. L., Camp, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+assessment+of+the+aging+mouse+vestibular+system.">Behavioral assessment of the aging mouse vestibular system.</a> Journal of Visualized Experiments. (89), e51605 (2014).</li><li>de Jeu, M., De Zeeuw, C. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Video-oculography+in+mice.">Video-oculography in mice.</a> Journal of Visualized Experiments. 65 (65), e3971 (2012).</li></ol>

Denne teknik er en nyttig metode til at skabe pludselige, permanent og komplet vestibulære funktion tab. Dette kan bruges til at studere vestibulære patologier såsom vestibulær neuritis, akustisk tumor og Menières sygdom. Mange undersøgelser har brugt denne teknik til at studere den neuronale plasticitet af vestibulære kerner eller relaterede central proces5,17,18,19.
<p class="jove_...

Den vestibulære orgel er afgørende for balance og øjet kontrol. En normal vestibulære funktion afhænger af symmetriske afferente signaler fra de vestibulære organer i de to indre ører. Vestibulære hypofunction eller tab inducerer svimmelhed, nystagmus og postural ubalance. Efter en akut skade genopretter den vestibulære funktion spontant inden for flere dage, en proces kendt som vestibulære erstatning1,2. Den vestibulære erstatning af statisk underskud er en proces af recovery relateret til ubalance i spontan hvilende aktivitet mellem de ipsilaterale og kontralaterale vestibulære kerner. Den vestibulære kompensation i dynamisk underskud opnås hovedsagelig via sensorisk og adfærdsmæssige udskiftninger (ved hjælp af visuelle eller somatosensoriske input)3. Disse processer er attraktive for neuronal plasticitet undersøgelser4,5.
En række metoder har været udviklet til at studere det vestibulære system og mekanismerne bag neuronal plasticitet under vestibulære kompensation, som kirurgisk og kemiske labyrinthectomy og vestibulære neurectomy5,6 ,7,8. Vestibulære neurectomy er en bestemt måde at fremkalde komplet vestibulære tab, men det er en mere vanskelig og invasiv procedure og kan medføre hjerneskade8,9. Denne metode kræver større kirurgiske færdigheder og tager længere tid end labyrinthectomy. Kemiske labyrinthectomy herunder phosphatbufferet, arsanilate og tetracain, er lettere og kan give pålidelige resultater10,11,12. Men øresneglen kan også være beskadiget og vestibulære tab kan udvikle sig over tid11. Derudover er virkningerne af kemikalier på hjernen, som bør bevares for nøjagtig evaluering, uklart. Kirurgisk labyrinthectomy blev først indført i dyreforsøg i 184215 og blev først rapporteret i rotte i 193616. Denne teknik er siden blevet brugt i mange dyre undersøgelser5,17,18,19. Kirurgisk labyrinthectomy er en specifik, pålidelig og relativt enkel metode. 13 , 14 Derudover symptomerne på vestibulære skader ses umiddelbart efter operationen. Her beskriver vi vores kirurgisk teknik til rotte labyrinthectomy.

<table>
	<tbody>
		<tr>
			<td>ASPIRATOR KB-012</td>
			<td>KOH BONG &#38; CO., LTD.</td>
			<td>KB-012</td>
			<td>Medical aspirator</td>
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		<tr>
			<td>Blade: #15</td>
			<td>Fine Science Tools</td>
			<td>#10015-00</td>
			<td>Blades for #7 Scalpel Handles, #15</td>
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			<td>Carbon Steel Burrs</td>
			<td>Fine Science Tools</td>
			<td>#19007-05</td>
			<td>shaft diameter: 2.3 mm, length: 44 mm, package of 10 burrs</td>
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			<td>Carl Zeiss Surgical GmbH</td>
			<td>Carl Zeiss</td>
			<td>#6627100863</td>
			<td>Surgical microscope</td>
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		<tr>
			<td>Dumont #3c</td>
			<td>Fine Science Tools</td>
			<td>#11231-20</td>
			<td>Standard tip 0.17 x 0.10 mm, 11 cm</td>
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			<td>Dumont #5SF</td>
			<td>Fine Science Tools</td>
			<td>#11252-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #7B</td>
			<td>Fine Science Tools</td>
			<td>#11270-20</td>
			<td>Serrated 0.17 x 0.10 mm, 11 cm</td>
		</tr>
		<tr>
			<td>Extra Fine Bonn: straight</td>
			<td>Fine Science Tools</td>
			<td>#14084-08</td>
			<td>Iris scissors, best suited for microdissection under high magnification</td>
		</tr>
		<tr>
			<td>Fine Iris Scissors: straight</td>
			<td>Fine Science Tools</td>
			<td>#14094-11</td>
			<td>Made from martensitic stainless steel, combined with molybdenum and vanadium</td>
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			<td>Finger Loop Ear Punch</td>
			<td>Fine Science Tools</td>
			<td>#24212-01</td>
			<td>1 mm. Provides stability and control for researchers using the numbering system</td>
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			<td>Hartman</td>
			<td>Fine Science Tools</td>
			<td>#13002-10</td>
			<td>Tip width: 1 cm, serrated, 10 cm</td>
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			<td>Short Scalpel Handle #7 Solid</td>
			<td>Fine Science Tools</td>
			<td>#10003-12</td>
			<td>#7 short, 12 cm</td>
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			<td>Small Vessel Cauterizer</td>
			<td>Fine Science Tools</td>
			<td>#18000-03</td>
			<td>Replacement tip, straight knife, keeps bleeding to a minimum and therefore provides a surgical field clear of clamps and hemostats</td>
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			<td>Strong 207S</td>
			<td>SAESHIN</td>
			<td>207S</td>
			<td>Powerful torque at low speed, available with speed or on/off foot controller</td>
		</tr>
		<tr>
			<td>Suction Tubes</td>
			<td>JEUNGDO B&#38;P CO., LTD.</td>
			<td>H-1927-8</td>
			<td>Frazier, 18 cm</td>
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			<td>VICRYL</td>
			<td>ETHICON</td>
			<td>W9570T</td>
			<td>Synthetic absorbable sterile surgical suture</td>
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			<td>Weitlaner-Locktite</td>
			<td>Fine Science Tools</td>
			<td>#17012-13</td>
			<td>Maximum spread: 4.5 cm, 2 x 3 blunt teeth, 11 cm</td>
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		<tr>
			<td>Zoletil</td>
			<td>Virbac, France</td>
			<td></td>
			<td>Tiletamine-zolazepam</td>
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			<td>Rompun</td>
			<td>Bayer</td>
			<td></td>
			<td>Xylazine</td>
		</tr>
		<tr>
			<td>Rimadyl</td>
			<td>Pfizer</td>
			<td></td>
			<td>Carprofen</td>
		</tr>
		<tr>
			<td>Septra</td>
			<td>Pfizer</td>
			<td></td>
			<td>Trimethoprim-sulfonamide&#160;</td>
		</tr>
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surgical labyrinthectomy of the rat to study the vestibular system

For at studere det vestibulære system eller det vestibulære kompensationsprocessen, er blevet udviklet en række metoder til at forårsage vestibulære skade, herunder kirurgisk eller kemiske labyrinthectomy og vestibulære neurectomy. Kirurgisk labyrinthectomy er en relativt enkel, pålidelig og hurtig metode. Her beskrives den kirurgiske teknik til rotte labyrinthectomy. Et postauricular snit er lavet under generel anæstesi til at afsløre den ekstern auditive kanal og trommehinden, hvorefter trommehinden og øreknoglerne er fjernet uden stapes. Stapes arterie, som er beliggende mellem stapes og ovale vindue, er en sårbar struktur og skal bevares for at opnå et klart kirurgiske felt. Et hul til fenestrate forhallen er lavet med en 2.1-mm drill bur overlegen i forhold til stapes. Så, 100% ethanol er injiceres gennem dette hul og indsugning flere gange. Omhyggelig dissektion under et mikroskop og omhyggelig blødende kontrol er afgørende for at opnå pålidelige resultater. Symptomer af vestibulære tab, nystagmus, hoved vippe og en rullende bevægelse, ses umiddelbart efter operationen. Rotarod eller rotation stol test kan anvendes til objektivt og kvantitativt evaluere den vestibulære funktion.

Succesen af operationen blev valideret af adfærdsmæssige test. Alle dyr udstillet den typiske adfærd af en ensidig tab af vestibulære funktion. Spontan tønde-rullende var tydeligt umiddelbart efter operationen, at være fremkaldt af en luft pust over hovedet eller en let berøring til kroppen i den tidlige opvågningsfasen (Video 1). 3 d efter kirurgi, dyrene flyttes rundt hælder mod den lesioned side med lejlighedsvis falder til venstre side. Spontan nystagmus blev...

Watch this Scientific Journal Video about Surgical Labyrinthectomy of the Rat to Study the Vestibular System at JoVE.com

Surgical Labyrinthectomy of the Rat to Study the Vestibular System

Denne protokol beskriver den kirurgiske labyrinthectomy af en rotte, som er en nyttig metode til at studere det vestibulære system.

Kirurgisk Labyrinthectomy af rotte at studere det vestibulære System

To study the vestibular system or the vestibular compensation process, a number of methods have been developed to cause vestibular damage, including ...

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