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

这项工作得到了日本政府下个 KAKENHI (24592560、15H04991 和 15K15624) 和武田科学基金会中频和 Jikei 大学战略优先研究基金中频的资助, 以 H.J.O。

所有实验程序都是在日本东京的 Jikei 大学进行的。动物处理和实验程序经 Jikei 大学机构动物保育和使用委员会批准 (批准号: 26-060), 并按照《 Jikei 实验室动物使用机构指南》执行。大学, 同意由日本科学理事会正确进行动物实验的指南 (2006)。动物护理是按照《护理和使用实验动物指南》 (1996 实验室动物资源研究所) 的建议进行的。
1. 术前检查
<ol><li>采取颅内计算机断层扫描 (CT) 对动物 (常...

<ol>
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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+method+for+virally+mediated+gene+delivery+to+the+mouse+inner+ear+through+the+round+window+membrane.">Surgical method for virally mediated gene delivery to the mouse inner ear through the round window membrane.</a> J Vis Exp. (97), (2015).</li><li>Jero, J., Tseng, C. J., Mhatre, A. N., Lalwani, A. 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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H., Kanzaki, S., Brown, N., Raphael, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+functional+and+structural+outcome+of+inner+ear+gene+transfer+via+the+vestibular+and+cochlear+fluids+in+mice.">The functional and structural outcome of inner ear gene transfer via the vestibular and cochlear fluids in mice.</a> Mol Ther. 4 (6), 575-585 (2001).</li><li>Praetorius, M., Baker, K., Weich, C. M., Plinkert, P. 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耳蜗血流量非常小, 估计是在人类总心脏输出的 1/100万的顺序, 和访问是有限的血-耳蜗屏障的存在, 分离内耳从一般循环19 ,20。由于这些原因, 药物或病毒的媒介管理进入内耳是有利的, 不是通过系统性的管理, 而是由直接管理。在小鼠和豚鼠实验中, 报告了各种方法, 如采用耳后入路或腹侧方法的转圆窗、cochleostomy、淋巴囊传递和半圆管方法...

感音神经性听力丧失 (SNHL) 主要出现在耳蜗损伤或缺陷。SNHL 的常见原因包括老化 (例如聋)、遗传缺陷、暴露于噪音、感染和毒性药物1。世界卫生组织 (卫生组织) 估计, 占世界人口5.3% 的3.6亿多人患有听力损失2。据估计, 在900至1的2500新生儿中, 有轻度、严重和深刻的先天性永久性听力丧失, 而1/3 岁以上的成年人有一定程度的听力损失65。然而, 对于这些患者来说, 听力丧失没有有效的临床治疗方法。
听力研究一直使用啮齿动物或豚鼠模型, 并提出了许多方法, 如基因治疗和再生治疗, 被认为是一种新的治疗听力损失。然而, 人类和啮齿动物在听觉系统上有很大的区别, 很难将动物模型转化为人类的应用。狨 (jacchus) 是来自亚马逊的新一代世界猴, 由于多种原因, 它已成为各种基础研究研究的一种具有吸引力的非人类灵长类动物模型。首先, 它的解剖和生理学更类似于人类而不是啮齿动物。第二, 关于这个物种的基本生物学信息, 包括疾病、神经网络、行为和基因组, 都有很好的特点。听觉和声带处理, 周期性皮质编码, 音高的表示, 和常见的猴的听觉声音互动也被报告了4,5,6,7,8,9,10. 在最近的组织学研究中, 研究人员发现了20种耳聋基因和阴离子交换器在普通狨耳蜗中的独特表达模式, 发现五基因是渐进性耳聋的病因, 三阴离子交换器, 有不同的表达模式, 从啮齿动物11,12。这些概况使我们相信, 共同狨是听觉研究的有力工具。
作为实验动物的常见狨的最显著属性如下:
<ul><li>简单的处理与小体型相比, 一些老世界的猴子: 成年猕猴重 300-400 克, 身高约60厘米, 这是类似于大鼠。</li>
	<li>高度繁殖的灵长类动物: 在18月的年龄, 它能达到性成熟, 每年能承受两次, 每年产生4到6个子代。</li>
	<li>基因修改是可利用的: et 等人成功地创建了转基因的13和敲出14灵长类动物使用猴的基因修饰涉及神经系统疾病 (例如,帕金森的疾病和阿尔茨海默病)。</li>
	<li>胚胎干细胞和诱导的多能干细胞 (iPS) 细胞已经建立了15,16。虽然与啮齿目动物相比, 保持相同数量的绒猴是相当困难的, 但干细胞或 iPS 细胞的可用性提供了体外化验, 这将减少所需的体内化验的数量。</li>
</ul>为了促进耳聋及其潜在治疗领域的更好的翻译研究, 我们建立了一种利用听觉脑干反应 (ABRs) 进行 CT 和 MRI、全身麻醉和听力测试的成像研究协议。这些实验系统可以为我们提供更好的机会, 以获得临床前证明的概念研究, 这对于弥合啮齿类动物研究和临床试验之间的差距是必不可少的。在这里, 我们报告一个外科方法, 通过圆窗膜对普通狨的内耳进行药物管理。为了在圆形窗口周围获得清晰的视野, 在不操作可能导致传导性听力损失的鼓膜的情况下, 从乳突腔中接近是有用的。临床上, 这种方法, 称为 "后鼓膜置" 是很好的建立, 通常用于人工耳蜗植入和胆脂瘤手术。我们相信, 后鼓膜置允许我们进行准确的药物管理, 而不诱发听力损失。

<table border="0" cellpadding="0" cellspacing="0" style="width:675px;" width="674">
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		<col />
		<col />
	</colgroup>
	<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>
</table>

a surgical procedure for the administration of drugs to the inner ear in a non-human primate common marmoset (callithrix jacchus)

虽然在某些疾病中, 人类的症状不能概括, 但长期以来, 啮齿类动物模型已经促进了听觉研究。常见的狨 (Callithrix jacchus) 是一种小型的、易于处理的新世界猴, 它具有类似于颞骨的解剖, 包括中耳听骨链和内耳对人类, 比与啮齿动物的比较。在这里, 我们报告一个可重现的, 安全的, 合理的手术方法的耳蜗圆形窗口的利基为药物传递到内耳的共同狨。我们采用后鼓膜置, 这是一个在人体外科临床应用的程序, 以避免对鼓膜的操纵, 可能导致传导性听力丧失。这个手术过程并没有导致任何重大的听力损失。这种方法是可能的, 由于共同狨的大泡结构, 虽然侧半圆管和面神经的垂直部分应仔细考虑。这种手术方法使我们能够在无听力损失的情况下对药物进行安全、准确的管理, 这对于获得临床翻译研究概念的前期证明具有重要意义。

术后鼓膜置无并发症, 如手术部位感染, 血管损伤出血, 前庭功能障碍, 有时发生中耳操纵。我们在7岁的普通狨的右耳上进行后鼓膜置, 并将1µL 磷酸盐缓冲盐水用于圆窗利基。ABRs 在手术前和两个月后进行测量 (图 5A-B)。ABRs 的波形和阈值与以前的研究17、18相同, 在手术前后这些参数没有变化 (<st...

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

我们报告一种外科方法, 以管理药物的内耳的非人类灵长类, 常见的狨 (Callithrix jacchus), 通过圆窗膜。

一种非人类灵长类常见狨中内耳药物的外科手术方法 (Callithrix jacchus)

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

Research

JoVE Journal

Medicine

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

A Strategy to Identify de Novo Mutations in Common Disorders such as Autism and Schizophrenia

一项战略，以确定在常见的疾病，如自闭症和精神分裂症的新突变

There are several lines of evidence supporting the role of de novo mutations as a mechanism for common disorders, such as autism and schizophrenia. First, ...

科研

医学

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

一种新的手术方法气管内注射生物活性剂在胎鼠模型

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs

外科技术治疗脊髓交付：在哥廷根小型猪演示程序

This is a  compact visual description of a combination of surgical technique and device  for the  delivery of (gene and cell) therapies into the spinal ...

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

基于音频的虚拟游戏环境的发展，以帮助盲人的导航技巧

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice

异位黏膜移植程序直接给药到小鼠的大脑

Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high ...

A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair

外科手术切除为鼠标肋骨：模型进行大规模的长骨修复

This protocol introduces researchers to a new model for large-scale bone repair utilizing the mouse rib. The procedure details the following: preparation ...

Quantification of the Immunosuppressant Tacrolimus on Dried Blood Spots Using LC-MS/MS

在免疫抑制剂他克莫司对干血斑使用LC-MS / MS定量

The calcineurin inhibitor tacrolimus is the cornerstone of most immunosuppressive treatment protocols after solid organ transplantation in the United ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

执行永久远端大脑中与老年大鼠颈总动脉闭塞研究皮层缺血具有持续残疾

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

动静脉内瘘失败的新型小鼠模型：在详细介绍手术过程

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

使用Saccadometry与深部脑刺激来研究正常和病理脑功能

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...