Name: a surgical procedure for the administration of drugs to the inner ear in a non-human primate common marmoset (callithrix jacchus)
Uploaded: 2018-02-27T12:30:00.000+00:00
Duration: 6 min 55 s
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

仕事は日本政府文部科学省科研費 (24592560, 15 H 04991、15 K 15624) から m. f. と m. f. に武田科学財団、東京慈恵会医科大学戦略的な優先順位付け研究基金 H.J.O. の補助金によってサポートされて

すべての実験手順は、東京慈恵会医科大学、東京都で行われました。機関動物ケアおよび使用委員会東京慈恵会医科大学の動物取り扱いと実験プロシージャを承認 (承認なし: 26-060)、慈恵で実験動物の使用のための機関のガイドラインに沿って行われ、大学: 日本学術会議 (2006 年) で動物実験の適切な実施のためのガイドラインに同意します。ケアと実験動物の使用 (資源研究所の実験動物、1996) のガイドの推奨事項に従って動物のケアを行った。
1. 術前検査
<ol><li>少なくとも 1 ヶ月個々 の解剖学的構造の識別を容易にするため手術の前に動物 (コモンマーモ セット) の頭蓋の計算された断層レントゲン写真撮影 (CT) スキャンを取る。</li>
	<li>聴力検査手術の前に、少なくとも 1 ヶ月を実行します。 注: ここでは、聴覚の状態を評価するために、ABR を使用しました。簡単に言えば、肯定的な電極は、頂点と刺激耳背下部参照否定的な電極で皮下置かれました。接地電極は首の背の部分に皮下配置されました。17,18 Abr をトーン バーストと誘発した (1.0 ms トーン バースト 0.1 ms の上昇と持続時間と立下り時間)。音圧レベルは最大振幅から 10 dB ステップで、減った。各音のレベルで 512 応答を (交互刺激の極性) を平均しました。ABR 閾値はであらゆる波を検出できる最小の音圧レベル (SPL) として定義されました。</li>
</ol>2. 麻酔
<ol><li>全身麻酔下で手術前に、の晩から高速の動物。</li>
	<li>顔にマスク換気または低速誘導 (図 1 a) の誘導の商工会議所で 4% イソフルランと 5-6 mL/分で周囲の空気を管理します。動物はつま先のピンチなどの痛みを伴う刺激に応答しないことを確認します。</li>
	<li>Anesthestize コモンマーモ セット メデトミジン 0.04 mg/kg, ミダゾラム (0.4 mg/kg)、およびブトルファノール (0.4 mg/kg) の組み合わせ (大腿筋) に筋肉注射をします。</li>
	<li>アンピシリン (0.08 mg/g) を皮下注入術前抗生物質として皮膚切開前 30 分以上。</li>
	<li>上部で、その頭を仰臥位でマーモセットを置き、頭と首を拡張します。ナイロン糸をテーブルに上顎を修復良いビューを取得する舌を引くか、喉頭鏡を使用して、また、喉頭蓋、喉頭のエントリを視覚化 (図 1 b)。</li>
	<li>喉頭蓋は喉頭の表面に沿って気管内チューブを挿入します。 注: 16 ゲージの静脈のカテーテルを使用することがまたは 6 フランス語ゲージ栄養チューブの気管内チューブ (図 1)。</li>
	<li>気管に配置されて正しくいることを確認するために気管内チューブから EtCO2を確認してください。チューブが配置されている呼吸ガスの動きをチェックした後、テープでマーモセットの顔に気管内チューブを固定します。</li>
	<li>操作中に 1-3% イソフルランの麻酔を維持します。</li>
	<li>操作中に SpO2EtCO2体温、直腸の温度を監視します。体温の低下を防ぐために暖房のマットですべての手順を実行します。</li>
</ol>3. 術前準備
<ol><li>仰臥位でセットを準備し、頭を非手術側 (図 2 a) に 30-45 度の角度に傾斜します。</li>
	<li>手術側の後耳介領域を剃る。</li>
	<li>アルコール、ポビドン ヨード、後耳介や外耳道の両方の外科的領域を消毒します。</li>
	<li>マーモセット (図 2 b) に円形孔を有する手術用覆布シートを配置します。</li>
</ol>4. 手術
<ol><li>1% 塩酸リドカインを注入 (1: 100,000 エピネフリン 0.1 から 0.2 で mL) 皮下に皮膚切開部位。第 15 円形の刃を持つ、耳介後部の皮膚切開を行います。</li>
	<li>止血、1:5,000 希釈エピネフリンに浸したガーゼを使用して出血領域に圧力を適用します。バイポーラ熱傷も効果的です (図 2)。</li>
	<li>だからその耳介軟骨 (c) の後耳介領域の軟部組織に開くし、後耳介筋 (m) を認識することができます。外耳道骨軟骨 (図 3 a) の奥深くに表示すべきを公開します。</li>
	<li>鼓膜の深さを確認するため後方の外耳道の最深部で骨と皮の付着として認識されることができます鼓膜の縁の骨の一部を公開します。 注: この手順は、どのように深い中耳腔の外側の部分である (図 3 a) を認識することが出来ます。</li>
	<li>側頭骨から分離、尾の側面に向かって後耳介筋 (胸鎖乳突筋、頭板状筋、顎二腹筋の後部の腹) の皮をむきます。側頭骨 (図 3Bのアスタリスク) の外側表面は、公開します。</li>
	<li>1.0 mm ダイヤモンド bur. (図 3) と外耳道の後ろに約 5 mm の穴を作る。掘削サイトに応じてドリル回転数 1,000-8,000 rpm を使用します。 注: 低速の半規管や顔面神経の近くの繊細な操作に適しています。掘削サイト熱損傷を防止するために適切に生理食塩水を点滴します。</li>
	<li>ドリルで穴を拡大し、水平半規管 (HSC) では、(図 3 D) の乳様突起の空洞に向かって隆起の位置を確認します。</li>
	<li>垂直部分実行 HSC の前方顔面神経の位置を確認します。ドリルし、顔面神経と鼓索神経との間の三角形の面積の中心に位置している顔の凹部の明確なビューを取得する、存在する場合、すべての骨の隔壁を削除します。</li>
	<li>顔面神経と、鼓膜の垂直部分の間にドリルで小さな穴を作り 1,000 rpm で 0.6 mm 径ダイヤモンド バールとリム (図 4 aのアスタリスク)。</li>
	<li>薄くなった骨をピンクの線と見ることができる顔面神経を損傷しないように後部運河の壁と平行、バールに移動します。神経は白い線構造 (図 4 b) として公開する直前に、掘削を停止します。</li>
	<li>丸い窓のニッチが穴 (図 4) を視覚化できることを確認します。その後、25 ゲージまたは細い針でニッチに薬を管理します。丸い窓のニッチ市場で薬を保持するためにニッチ (図 4) 粘性ソリューションを配置します。</li>
	<li>その止血がしっかりと確認した後 6-0 吸収スレッドを使用して簡単な縫合と層 (表皮層と真皮層) を閉じる制御します。</li>
	<li>手術の傷口にガーゼを修正し、包帯を貼る。</li>
</ol>5. 術後のケア
<ol><li>メデトミジンを敵に回した大腿筋に筋肉内塩酸アチパメゾール (0.15 mg/kg) を管理します。</li>
	<li>自発呼吸の開始、気管内チューブを削除します。</li>
	<li>30% O2覚醒を完了するために麻酔が切れるまで 29 度を記録、集中治療室における共通のセットを維持します。</li>
	<li>眼振など前庭機能障害を示唆する所見がないことを確認し、飼育ケージに動物を返します。</li>
</ol>

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蝸牛の血流の量が非常に小さい 1/1,000,000 人間の総心臓出力の順番になっていると推定される、一般的な循環19 から内耳を隔てる血蝸牛バリアの存在によってアクセスを制限 ,20。これらの理由から、薬物や内耳へのウイルスのベクトルの管理は直接管理ではなく、全身投与ではなく、良好です。マウス、モルモット実験で様々 なアプローチが報告?...

難聴 (SNHL) は、損傷や、蝸牛の欠乏から主に発生します。SNHL の一般的な原因は、高齢化 (例えば老人性難聴)、遺伝的欠陥、耳毒性薬物1感染症、騒音への暴露。世界保健機関 (WHO) は、以上 3 億 6000 万人、世界の人口の 5.3% を表すが損失2を聞いて苦しむ推定されます。1 で 2,500 新生児に 900 の 1 が中等度、重度と先天性重度の永続的な難聴、および 65 歳以上の約 3 つの大人はある程度聴力損失3とも推定されます。ただし、これらの患者の難聴の効果的な治療はありません。
聴覚研究はネズミやモルモット モデルを使用して実行されて長いが、難聴の新しい治療法として遺伝子治療や再生療法などさまざまな方法が提案されています。しかし、人間および齧歯動物聴覚システムの面で大きな差がある、人間のアプリケーションに動物モデルを変換する非常に困難であります。コモンマーモ (C. 用いた)、アマゾンから発信された新しい世界猿いくつかの理由のための様々 な基礎調査研究魅力的な非ひと霊長類モデルとなっています。まず、その解剖学と生理学よりこれらの齧歯動物ではなく人間に似ています。第二に、基本的な生物学的疾患、神経回路網、動作、およびゲノムを含むこの種についてはよく特徴付けられます。聴覚とボーカルの処理、周期、ピッチの表現、コモンマーモ セットの聴覚音声相互作用の皮質コーディングもされている報告4,5,6,7,8,9,10. 最近の組織学的研究で研究者 20 の難聴遺伝子と共通のマーモセットの蝸牛における陰イオン交換器の特徴的な発現パターンを識別・発見 5 進行性難聴と 3 つの原因である遺伝子陰イオン交換器、齧歯動物11,12の異なる発現パターンを持っていた。これらのプロファイルは、共通のマーモセットが聴覚研究の強力なツールであることを信じる私たちを導きます。
実験動物としてコモンマーモの最も顕著な属性は次のとおりです。
<ul><li>小さなボディサイズと簡単に処理に比べていくつかの種の古い世界のサル: アダルト マーモセット 300 から 400 グラムの重量を量る、ラットに似ている、高さ約 60 cm。</li>
	<li>高度生殖霊長類: マーモセット 18 ヶ月の頃に性的に成熟を達成してクマの年 2 回、毎年 4 に 6 の子孫を生産することができます。</li>
	<li>遺伝の修正が利用可能な: 佐々木 Eらは、トランスジェニック13とノックアウト14霊長類のマーモセットを用いた神経疾患 (例えばパーキンソンに関与する遺伝の修正の作成に成功病やアルツハイマー病)。</li>
	<li>胚性幹 (ES) 細胞と多能性幹 (iPS) 細胞の両方は、確立された15,16をされています。むしろげっ歯類に比べてするマーモセットの同じ番号を維持することは困難であるが、幹細胞や iPS 細胞の可用性は、生体内のアッセイに必要な数を削減するの in vitroアッセイを提供します。</li>
</ul>難聴とその潜在的な療法の分野でより良い橋渡し研究を容易にするために、CT と MRI、全身麻酔と聴性脳幹反応 (Abr) を使用して聴力検査を用いたイメージングの研究プロトコルを設立しました。これらの実験システムが齧歯動物の研究と臨床試験の間のギャップを埋めるために不可欠な概念研究前臨床証拠を取得するより多くの機会を提供します。ここでは、丸い窓の膜をコモンマーモの内耳に薬を管理する手術法を報告します。伝導性の聴力損失を引き起こす可能性があります鼓膜を操作することがなく丸い窓周りの明確なビューを取得するには、便利ですアプローチに乳様突起の空洞から。臨床的に、このアプローチに呼ばれます「後部鼓膜」の確立し、蝸牛注入と真珠手術に用い。その後鼓膜聴力損失を引き起こすことがなく薬の正確な管理を実行することができます考えています。

<table><tbody><tr><td>common marmoset</td><td>CLEA Japan</td><td>EDM:C.Marmoset(Jic)</td><td></td></tr><tr><td>isoflurane</td><td>Phizer</td><td></td><td></td></tr><tr><td>medetomidine</td><td>ZENOAQ</td><td></td><td></td></tr><tr><td>midazolam</td><td>astellas</td><td></td><td></td></tr><tr><td>butorphanol</td><td>Meiji Seika</td><td></td><td></td></tr><tr><td>ampicillin</td><td>Meiji Seika</td><td></td><td></td></tr><tr><td>lidocaine hydrochloride</td><td>AstraZeneca</td><td></td><td></td></tr><tr><td>&amp;nbsp;6-0 absorbent thread</td><td>ETHICON</td><td>RD-1</td><td></td></tr><tr><td>atipamezole hydrochloride</td><td>ZENOAQ</td><td></td><td></td></tr><tr><td>phosphate buffered saline</td><td>Wako</td><td>045-29795</td><td></td></tr><tr><td>Surge Wave</td><td>Morita</td><td>TR-900-OR</td><td></td></tr><tr><td>Diamond Bar 006</td><td>Morita</td><td>14070057</td><td></td></tr><tr><td>Diamond Bar 010</td><td>Morita</td><td>14070081</td><td></td></tr><tr><td>intensive care unit</td><td>Menix</td><td>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)

研究を聞いて長いによって促進されています齧歯動物モデルがいくつかの疾患では、ことはできませんですが、人間の症状が締めくくっています。コモンマーモ (用いた) が小さい、ハンドル簡単新しい世界猿と齧歯動物の比較よりも中耳の耳小骨連鎖と人間、内耳を含む頭骨のような解剖学を持っています。ここでは、一般的なマーモセットの内耳への薬物送達の蝸牛の丸い窓のニッチに再現可能な安全かつ合理的な手術アプローチを報告する.後部鼓膜、鼓膜伝導性の聴力損失を引き起こす可能性のある操作を避けるために人間の外科で臨床的に使用されるプロシージャを採用しました。この手術は、任意の重要な聴力損失には至らなかった。外側半規管、顔面神経の垂直部分を慎重に検討する必要がありますが、このアプローチは共通マーモセットの大きな嚢胞構造のために可能だった。この手術方法では、前臨床トランスレーショナルリサーチの概念実証を得ることに非常に重要なは、聴覚の損失、薬剤の安全で正確な管理を実行することが出来ます。

手術部位感染、血管の損傷や中耳の操作で発生する前庭障害と出血など合併症なく後部鼓膜を行った。7 歳コモンマーモの右の耳の後部の鼓膜を行い丸い窓のニッチに 1 μ L のリン酸緩衝生理食塩水を投与します。Abr は、前に、(図 5A B) 術後後 2 ヶ月を測定しました。波形と Abr のしきい値が以前研究17,<sup c...

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

丸い窓の膜を介してヒト以外の霊長類コモンマーモ (用いた) の内耳に薬を管理する手術法を報告する.

ヒト以外の霊長類コモンマーモ (用いた) で内耳に薬の管理術

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

a-surgical-procedure-for-administration-drugs-to-inner-ear-non-human

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)

8.6K ビュー.  Jikei University School of Medicine. 丸い窓の膜を介してヒト以外の霊長類コモンマーモ (用いた) の内耳に薬を管理する手術法を報告する.

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

Canalostomy As a Surgical Approach to Local Drug Delivery into the Inner Ears of Adult and Neonatal Mice

Local delivery of therapeutic drugs into the inner ear is a promising therapy for inner ear diseases. Injection through semicircular canals (canalostomy) has been shown to be a useful approach to local drug delivery into the inner ear. The goal of this article is to describe, in detail, the surgical techniques involved in canalostomy in both adult and neonatal mice. As indicated by fast-green dye and adeno-associated virus serotype 8 with the green fluorescent protein gene, the canalostomy facilitated broad distribution of injected reagents in the cochlea and vestibular end-organs with minimal damage to hearing and vestibular function. The surgery was successfully implemented in both adult and neonatal mice; indeed, multiple surgeries could be performed if required. In conclusion, canalostomy is an effective and safe approach to drug delivery into the inner ears of adult and neonatal mice and may be used to treat human inner ear diseases in the future.

Here we describe canalostomy procedure which allows local drug delivery into the inner ears of adult and neonatal mice through the semicircular canal with minimum damage to hearing and vestibular function. This method can be used to inoculate viral vectors, pharmaceuticals, and small molecules into the mouse inner ear.

成人および新生児マウス内耳への薬剤の局所投与への外科的アプローチとしての Canalostomy

Local delivery of therapeutic drugs into the inner ear is a promising therapy for inner ear diseases. Injection through semicircular canals (canalostomy) ...

遺伝学

Surgical Method for Virally Mediated Gene Delivery to the Mouse Inner Ear through the Round Window Membrane

Gene therapy, used to achieve functional recovery from sensorineural deafness, promises to grant better understanding of the underlying molecular and genetic mechanisms that contribute to hearing loss. Introduction of vectors into the inner ear must be done in a way that widely distributes the agent throughout the cochlea while minimizing injury to the existing structures. This manuscript describes a post-auricular surgical approach that can be used for mouse cochlear therapy using molecular, pharmacologic, and viral delivery to mice postnatal day 10 and older via the round window membrane (RWM). This surgical approach enables rapid and direct delivery into the scala tympani while minimizing blood loss and avoiding animal mortality. This technique involves negligible or no damage to essential structures of the inner and middle ear as well as neck muscles while wholly preserving hearing. To demonstrate the efficacy of this surgical technique, the vesicular glutamate transporter 3 knockout (VGLUT3 KO) mice will be used as an example of a mouse model of congenital deafness that recovers hearing after delivery of VGLUT3 to the inner ear using an adeno-associated virus (AAV-1).

The described post-auricular surgical approach allows rapid and direct delivery into the mouse cochlear scala tympani while minimizing blood loss and animal mortality. This method can be used for cochlear therapy using molecular, pharmacologic and viral delivery to postnatal mice through the round window membrane.

円窓膜を介してマウスインナーイヤーにウイルスにより媒介遺伝子送達のための手術方法

Gene therapy, used to achieve functional recovery from sensorineural deafness, promises to grant better understanding of the underlying molecular and ...

神経科学

The Mouse Round-window Approach for Ototoxic Agent Delivery: A Rapid and Reliable Technique for Inducing Cochlear Cell Degeneration

Investigators have utilized a wide array of animal models and investigative techniques to study the mammalian auditory system. Much of the basic research involving the cochlea and its associated neural pathways entails exposure of model cochleae to a variety of ototoxic agents. This allows investigators to study the effects of targeted damage to cochlear structures, and in some cases, the self-repair or regeneration of those structures. Various techniques exist for delivery of ototoxic agents to the cochlea. When selecting a particular technique, investigators must consider a number of factors, including the induction of inadvertent systemic toxicity, the amount of cochlear damage produced by the surgical procedure itself, the type of lesion desired, animal survivability, and reproducibility/reliability of results. Currently established techniques include parenteral injection, intra-peritoneal injection, trans-tympanic injection, endolymphatic sac injection, and cochleostomy with perilymphatic perfusion. Each of these methods has been successfully utilized and is well described in the literature; yet, each has various shortcomings. Here, we present a technique for topical application of ototoxic agents directly to the round window niche. This technique is non-invasive to inner ear structures, produces rapid onset of reliably targeted lesions, avoids systemic toxicity, and allows for an intra-animal control (the contra-lateral ear). Results stemming from this approach have helped deeper understanding of auditory pathophysiology, cochlear cell degeneration, and regenerative capacity in response to an acute injury. Future investigations may use this method to conduct interventional studies involving gene therapy and stem cell transplantation to combat hearing loss.

Various methods exist for introducing ototoxic agents to the cochleae of animal models. Presented is a surgical protocol for delivery of ototoxic agents to the round window niche. The procedure is reliable, creates targeted intra-cochlear lesions, and avoids mechanical damage to the microarchitecture. Examination of cochlear self-repair/regeneration is possible.

蝸牛細胞の変性を誘導するための迅速かつ信頼性の高いテクニック：耳毒性剤送達のためのマウスラウンドウィンドウアプローチ

Investigators have utilized a wide array of animal models and investigative techniques to study the mammalian auditory system. Much of the basic research ...

医学

Posterior Semicircular Canal Approach for Inner Ear Gene Delivery in Neonatal Mouse

Inner ear gene therapy offers great promise as a potential treatment for hearing loss and dizziness. One of the critical determinants of the success of inner ear gene therapy is to find a delivery method which results in consistent transduction efficiency of targeted cell types while minimizing hearing loss. In this study, we describe the posterior semicircular canal approach as a viable method for inner ear gene delivery in neonatal mice. We show that gene delivery through the posterior semicircular canal is able to perfuse the entire inner ear. The easy anatomic identification of the posterior semicircular canal, as well as minimal manipulation of the temporal bone required, make this surgical approach an attractive option for inner ear gene delivery.

In this study, we describe the posterior semicircular canal approach as a reliable method for inner ear gene delivery in neonatal mice. We show that gene delivery through the posterior semicircular canal is able to perfuse the entire inner ear.

新生児マウス内耳遺伝子デリバリーの後半規管アプローチ

Inner ear gene therapy offers great promise as a potential treatment for hearing loss and dizziness. One of the critical determinants of the success of ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

一般的なセットの全皮質のシェットランドシープドッグ配列の慢性注入

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Stereotaxic Surgery for Implantation of Microelectrode Arrays in the Common Marmoset (Callithrix jacchus)

Marmosets (Callithrix jacchus) are small non-human primates that are gaining popularity in biomedical and preclinical research, including the neurosciences. Phylogenetically, these animals are much closer to humans than rodents. They also display complex behaviors, including a wide range of vocalizations and social interactions. Here, an effective stereotaxic neurosurgical procedure for implantation of recording electrode arrays in the common marmoset is described. This protocol also details the pre- and postoperative steps of animal care that are required to successfully perform such a surgery. Finally, this protocol shows an example of local field potential and spike activity recordings in a freely behaving marmoset 1 week after the surgical procedure. Overall, this method provides an opportunity to study the brain function in awake and freely behaving marmosets. The same protocol can be readily used by researchers working with other small primates. In addition, it can be easily modified to allow other studies requiring implants, such as stimulating electrodes, microinjections, implantation of optrodes or guide cannulas, or ablation of discrete tissue regions.

Stereotaxic Surgery for Implantation of Microelectrode Arrays in the Common Marmoset Callithrix jacchus

This work presents a protocol to perform a stereotaxic, neurosurgical implantation of&#160;microelectrode arrays in the common marmoset. This method specifically enables electrophysiological recordings in freely behaving animals but can be easily adapted to any other similar neurosurgical intervention in this species (e.g., cannula for drug administration or electrodes for brain stimulation).

共通マーモセットにおけるマイクロ電極アレイの移植のための立体外科手術(カリスリックス・ジャカス)

Marmosets (Callithrix jacchus) are small non-human primates that are gaining popularity in biomedical and preclinical research, including the ...

マーモセットの精密神経記録のためのX-Y電極ステージの開発

Electrophysiology of Laminar Cortical Activity in the Common Marmoset

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with linear arrays. The marmoset has recently grown in popularity due to its similar neural functional organization to other primates and its technical advantages for recording and imaging. However, neurophysiology in this model poses some unique challenges due to the small size and lack of gyri as anatomical landmarks. Using custom-built micro-drives, researchers can manipulate linear array placement to sub-millimeter precision and reliably record at the same retinotopically targeted location across recording days. This protocol describes the step-by-step construction of the micro-drive positioning system and the neurophysiological recording technique with silicon linear electrode arrays. With precise control of electrode placement across recording sessions, researchers can easily traverse the cortex to identify areas of interest based on their retinotopic organization and the tuning properties of the recorded neurons. Further, using this laminar array electrode system, it is possible to apply a current source density analysis (CSD) to determine the recording depth of individual neurons. This protocol also demonstrates examples of laminar recordings, including spike waveforms isolated in Kilosort, which span multiple channels on the arrays.

著者スポットライト:マーモセットでの正確な電極位置決めのためのマイクロドライブシステムの最適化

Custom-built micro-drives enable the sub-millimeter targeting of cortical recording sites with linear silicon arrays.

コモンマーモセットにおける層状皮質活動の電気生理学

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with ...

A Comparative Study of Drug Delivery Methods Targeted to the Mouse Inner Ear: Bullostomy Versus Transtympanic Injection

We present two minimally invasive microsurgical techniques in rodents for specific drug delivery into the middle ear so that it may reach the inner ear. The first procedure consists of perforation of the tympanic bulla, termed bullostomy; the second one is a transtympanic injection. Both emulate human clinical intratympanic procedures.
Chitosan-glycerophosphate (CGP) and Ringer&#180;s Lactate buffer (RL) were used as biocompatible vehicles for local drug delivery. CGP is a nontoxic biodegradable polymer widely used in pharmaceutical applications. It is a viscous liquid at RT&#160;but it congeals to a semi solid phase at body temperature. RL is an isotonic solution used for intravenous administrations in humans. A small volume of this vehicle is precisely placed on the Round Window (RW) niche by means of a bullostomy. A transtympanic injection fills the middle ear and allows less control but broader access to the inner ear.
The safety profiles of both techniques were studied and compared by using functional and morphological tests. Hearing was evaluated by registering the Auditory Brainstem Response (ABR) before and several times after microsurgery. The cytoarchitecture and preservation level of cochlear structures were studied by conventional histological techniques in paraformaldehyde-fixed and decalcified cochlear samples. In parallel, unfixed cochlear samples were taken and immediately frozen to analyze gene expression profiles of inflammatory markers by quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR).
Both procedures are suitable as drug delivery methods into the mouse middle ear, although transtympanic injection proved to be less invasive compared to bullostomy.

A Comparative Study of Drug Delivery Methods Targeted to the Mouse Inner Ear: Bullostomy Versus Transtympanic Injection

Two microsurgery approaches for local drug delivery to the inner ear are described here and compared in terms of impact on hearing parameters, cochlear cytoarchitecture and expression of inflammatory markers.

マウス内耳に標的薬の配布方法の比較研究：Bullostomy<em&gt;対</em&gt; Transtympanicインジェクション

We present two minimally invasive microsurgical techniques in rodents for specific drug delivery into the middle ear so that it may reach the inner ear. ...

ヒト以外の霊長類コモンマーモ (用いた) で内耳に薬の管理術

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成人および新生児マウス内耳への薬剤の局所投与への外科的アプローチとしての Canalostomy

円窓膜を介してマウスインナーイヤーにウイルスにより媒介遺伝子送達のための手術方法

蝸牛細胞の変性を誘導するための迅速かつ信頼性の高いテクニック：耳毒性剤送達のためのマウスラウンドウィンドウアプローチ

新生児マウス内耳遺伝子デリバリーの後半規管アプローチ

一般的なセットの全皮質のシェットランドシープドッグ配列の慢性注入

共通マーモセットにおけるマイクロ電極アレイの移植のための立体外科手術(カリスリックス・ジャカス)

コモンマーモセットにおける層状皮質活動の電気生理学

コモンマーモセットにおける層状皮質活動の電気生理学

コモンマーモセットにおける層状皮質活動の電気生理学

マウス内耳に標的薬の配布方法の比較研究：Bullostomy