Name: a surgical procedure for the administration of drugs to the inner ear in a non-human primate common marmoset (callithrix jacchus)
Uploaded: 2018-02-27T12:30:00
Description: Watch this Scientific Journal Video about A Surgical Procedure for the Administration of Drugs to the Inner Ear in a Non-Human Primate Common Marmoset Callithrix jacchus at JoVE.com

Arbejdet er støttet af tilskud til MF fra den japanske regering MEXT KAKENHI (24592560, 15 H 04991 og 15 K 15624) og Takeda Science Foundation til MF og Jikei Universitet strategiske prioritering forskningsfonden til H.J.O.

Alle eksperimentelle procedurer blev udført på Jikei University, Tokyo Japan. Animalske håndtering og eksperimentelle procedurer blev godkendt af institutionelle Animal Care og bruge udvalg af det Jikei Universitet (godkendelse ingen: 26-060) og udført i overensstemmelse med de institutionelle retningslinjer for brugen af forsøgsdyr ved Jikei Universitet, som er enige med retningslinjerne for korrekt adfærd af dyr eksperimenter af videnskab Rådet af Japan (2006). Pasning af dyr blev gennemført i overensstemmelse med anbefalingerne i ...

<ol>
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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+surgical+approach+appropriate+for+targeted+cochlear+gene+therapy+in+the+mouse.">A surgical approach appropriate for targeted cochlear gene therapy in the mouse.</a> Hear Res. 151 (1-2), 106-114 (2001).</li><li>Iizuka, T., 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=Noninvasive+in+vivo+delivery+of+transgene+via+adeno-associated+virus+into+supporting+cells+of+the+neonatal+mouse+cochlea.">Noninvasive in vivo delivery of transgene via adeno-associated virus into supporting cells of the neonatal mouse cochlea.</a> Hum Gene Ther. 19 (4), 384-390 (2008).</li><li>Kilpatrick, L. 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Mængden af cochlear blodgennemstrømningen er ekstremt små, anslået til at være i størrelsesordenen 1/1.000.000 af de samlede minutvolumen i mennesker, og er begrænset af tilstedeværelsen af blod-cochlea barriere, der adskiller det indre øre fra den generelle cirkulation19 ,20. Af disse grunde er stof eller viral vektor administration i det indre øre gunstige, ikke via systemisk administration men snarere ved direkte administration. I mus og marsvin eks...

Perceptivt høretab (SNHL) opstår overvejende fra skader eller mangel i øresneglen. Almindelige årsager til SNHL omfatter aldring (fx presbycusis), genetiske defekter, eksponering for høj støj, infektion og ototoksiske narkotika1. Verdenssundhedsorganisationen (WHO) anslås, at over 360 mio mennesker, svarende til 5.3% af verdens befolkning lider af høre tab2. Det vurderes også, 1 i 900 til 1 i 2.500 nyfødte har moderat, svær, og dyb medfødt permanent høretab, og omkring en ud af tre voksne over 65 år har en vis grad af høring tab3. Der er imidlertid ingen effektiv klinisk behandling af høretab til disse patienter.
Hørelse forskning har længe udført ved hjælp af gnaver eller marsvin modeller, og mange tilgange, såsom genterapi og regenerativ terapi, er blevet foreslået som en ny behandling af høretab. Men der er stor forskel mellem mennesker og gnavere i form af det auditive system, og det er meget vanskeligt at oversætte dyremodeller for humane ansøgninger. Den fælles silkeabe (C. jacchus), en New World abe stammer fra Amazonas, er blevet et attraktivt primat model for forskellige grundlæggende undersøgelser af flere årsager. Først, dens anatomi og fysiologi er mere ligner dem af mennesker snarere end gnavere. Andet, de grundlæggende biologiske oplysninger om denne art, herunder sygdomme, neurale netværk, adfærd og genomet, er godt præget. Auditive og vocal forarbejdning, kortikal kodning af hyppighed, repræsentation af banen og auditive-vokal interaktioner af fælles silkeaber har også været rapporteret4,5,6,7 , 8 , 9 , 10. i de seneste histologiske undersøgelser, forskerne identificerede karakteristiske udtryk mønstre af 20 døvhed gener og anion vekslere i cochlea af den fælles silkeabe og fandt, at fem gener, som er sygdomsfremkaldende for progressiv døvhed og tre anion varmevekslere, havde forskellige udtryk mønstre fra dem af gnavere11,12. Disse profiler føre os til at tro, at den fælles silkeabe er et kraftfuldt værktøj til høring forskning.
De mest markante attributter af den fælles silkeabe som en eksperimentel dyr er som følger:
<ul><li>Nem håndtering med en lille kropsstørrelse i forhold til nogle arter af gamle verden aber: voksen silkeaber vejer 300-400 g og er ca 60 cm i højden, hvilket svarer til rotter.</li>
	<li>Stærkt reproduktive primat: silkeaber attain seksuelle modenhed i en alder af 18 måneder og er i stand til at bære to gange om året og producere 4 til 6 afkom årligt.</li>
	<li>Genetiske modifikationer er tilgængelige: Sasaki E et al. formået at skabe transgene13 og knock-out14 primater bruger silkeaber af genetiske modifikationer impliceret i neurologiske lidelser (fx Parkinson's sygdom og Alzheimers sygdom).</li>
	<li>Både embryonale stamceller (ES) og inducerede pluripotente stamceller (iPS) er blevet etableret15,16. Selvom det er temmelig svært at holde det samme antal silkeaber i forhold til gnavere, giver adgang til stamceller eller iPS-celler in vitro- assays, hvilket vil reducere antallet af i vivo assays kræves.</li>
</ul>For at lette bedre Translationel forskning i døvhed og dens potentielle terapi, etableret vi en billeddannelse undersøgelse protokollen ved hjælp af CT og Mr, generel anæstesi og høreprøver ved hjælp af auditive hjernestammen svar (ABRs). Disse eksperimentelle systemer kan give os bedre muligheder for at få præ-klinisk bevis koncept undersøgelser, som er afgørende for broer over kløfter mellem gnaver undersøgelser og kliniske forsøg. Her rapporterer vi en kirurgisk metode til at administrere medicin til det indre øre af den fælles silkeabe gennem det runde vindue membran. For at få et klart overblik omkring det runde vindue, uden at manipulere trommehinden, der kan forårsage et konduktivt høretab, er det nyttigt at tilgang fra mastoid hulrummet. Klinisk, omtales denne tilgang, som "posterior tympanotomy" er veletableret og normalt anvendes til cochlear implantation og cholesteatoma kirurgi. Vi mener, at bageste tympanotomy gør det muligt at udføre præcis administration af medicin uden inducerende høretab.

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		<col />
		<col />
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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">common marmoset</td>
			<td style="width:92px;">CLEA Japan</td>
			<td style="width:160px;">EDM:C.Marmoset(Jic)</td>
			<td style="width:207px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">isoflurane</td>
			<td style="width:92px;">Phizer</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">medetomidine</td>
			<td style="width:92px;">ZENOAQ</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">midazolam</td>
			<td style="width:92px;">astellas</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">butorphanol</td>
			<td style="width:92px;">Meiji Seika</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:216px;">ampicillin</td>
			<td style="width:92px;">Meiji Seika</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">lidocaine hydrochloride</td>
			<td style="width:92px;">AstraZeneca</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">&#160;6-0 absorbent thread</td>
			<td style="width:92px;">ETHICON</td>
			<td style="width:160px;">RD-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">atipamezole hydrochloride</td>
			<td style="width:92px;">ZENOAQ</td>
			<td style="width:160px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">phosphate buffered saline</td>
			<td style="width:92px;">Wako</td>
			<td style="width:160px;">045-29795</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surge Wave</td>
			<td style="width:92px;">Morita</td>
			<td style="width:160px;">TR-900-OR</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Diamond Bar 006</td>
			<td style="width:92px;">Morita</td>
			<td style="width:160px;">14070057</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Diamond Bar 010</td>
			<td style="width:92px;">Morita</td>
			<td style="width:160px;">14070081</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">intensive care unit</td>
			<td style="width:92px;">Menix</td>
			<td style="width:160px;">P-100</td>
			<td></td>
		</tr>
	</tbody>
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a surgical procedure for the administration of drugs to the inner ear in a non-human primate common marmoset (callithrix jacchus)

Høre forskning er længe blevet fremmet af gnavere modeller, selv om nogle sygdomme, menneskelige symptomer ikke kan blive sammenfattet. Den fælles silkeabe (Callithrix jacchus) er en lille, let at håndtere New World abe, som har en lignende anatomi af den tidsmæssige knoglen, herunder mellemøret ossicular kæder og indre øre til mennesker, end i forhold til gnavere. Her, rapportere vi en reproducerbar, sikker og rationel kirurgisk tilgang til cochlear runde vinduet niche for medicinafgivelse til det indre øre af den fælles silkeabe. Vi vedtog bageste tympanotomy, en procedure, der anvendes klinisk i menneskelige kirurgi, for at undgå manipulation af trommehinden, der kan forårsage konduktivt høretab. Denne kirurgiske procedure førte ikke til nogen betydelig høretab. Denne tilgang var mulig på grund af den store bulla struktur af den fælles silkeabe, selv om den laterale halvrunde kanalen og lodrette del af ansigtets nerve bør overvejes nøje. Denne kirurgiske metode gør det muligt for os at udføre sikker og præcis administration af medicin uden tab af hørelse, som er af stor betydning at få præ-klinisk påvisning i id for Translationel forskning.

Bageste tympanotomy blev udført uden komplikationer såsom operationsstedet infektion, blødning med vaskulære skade eller vestibulær dysfunktion, som undertiden opstår med mellemøret manipulation. Vi udførte bageste tympanotomy på 7-årige fælles silkeabe højre øre og administreres 1 µL af fosfatbufferet saltopløsning til runde vinduet niche. ABRs blev målt før og to måneder efter operationen (figur 5A-B). Waveforms og tærskl...

Watch this Scientific Journal Video about A Surgical Procedure for the Administration of Drugs to the Inner Ear in a Non-Human Primate Common Marmoset Callithrix jacchus at JoVE.com

A Surgical Procedure for the Administration of Drugs to the Inner Ear in a Non-Human Primate Common Marmoset Callithrix jacchus

Vi rapporterer en kirurgisk metode til at administrere medicin til det indre øre af en ikke-menneskelige primater, den fælles silkeabe (Callithrix jacchus), via det runde vindue membran.

En kirurgisk Procedure for Administration af medicin til det indre øre i en ikke-menneskelige primater fælles silkeabe (Callithrix jacchus)

Hearing research has long been facilitated by rodent models, although in some diseases, human symptoms cannot be recapitulated. The common marmoset ...

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A Surgical Procedure for the Administration of Drugs to the Inner Ear in a Non-Human Primate Common Marmoset (Callithrix jacchus)

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