Name: increased recovery time and decreased lps administration to study the vagus nerve stimulation mechanisms in limited inflammatory responses
Uploaded: 2017-03-29T14:00:00.000+00:00
Duration: 6 min 43 s
Description: Watch this Scientific Journal Video about Increased Recovery Time and Decreased LPS Administration to Study the Vagus Nerve Stimulation Mechanisms in Limited Inflammatory Responses at JoVE.com

The study was supported by the Swedish Research Council, the Swedish Rheumatism Asociation, Karolinska Institute Foundations, Stockholm County Council, The Wallenberg Foundation, and the GV 80 Years' Foundation for research. The authors would also like to thank Hannah Aucott for proofreading the manuscript.

全ての動物実験は、カロリンスカ研究所、ストックホルムで地元の倫理委員会によって承認された動物の管理と使用のためのガイドラインに従って行いました。地元の倫理委員会は、動物のケアに関する欧州連合指令に従っています。 注：元のプロトコルからの主な変更点は、手術後の回復時間（1時間対 6時間）であり、LPSのレベルを（15ミリグラム/ kgの対 2mg / kgの）を注入しました。それ以外の場合は、手術自体に関連するさまざまな設定が変更されていません。 1.刺激のための材料を準備<ol><li>コンピュータ及び刺激電極（ 図1A）に連結されたデータ収集システムをオン。 </li><li>アクノリッジプログラムを入力します。 </li><li> 1×PBS中5mg / mlの濃度のLPSのストック溶液を調製し、アリコートにし、-20℃で保管します。実験の日に、アリコートを解凍し、ADEを準備重量に応じて、動物に約100μLを注入するために、LPSのquateサンプル（0.5 mg / mlと）。 </li></ol> 2.麻酔のための動物の準備<ol><li> C57BL / 6マウスを使用してください。 、12時間の明/暗サイクルで気候制御された条件下でそれらを維持し、それらを標準的なげっ歯類固形飼料を供給し、それらに水を自由に与えます。 </li><li>実験の日に、周りに25グラムの重量を量るマウスに手術を行います。 注：炎症反応が誘導されると、動物での臨床反応の定期検診や観察を行うことが特に重要です。動物の条件は倫理的な基準を満たしていない場合はCO 2吸入による早期の安楽死が必要とされています。 </li><li>麻酔マシンを設定します。 <ol><li>チューブが正しく接続されており、どのような方法で破損していないことを確認します。換気が正常に動作していることを確認してください。イソフルランボトルにキーフィルタを接続し、VAPOを埋めますイソフルランの十分な量とrizer。 </li><li>ガス供給（空気や酸素）を開き、ボトルは、実験のために十分なガスが含まれていることを確認してください。 3ウェイコネクタは、誘導室やマスクにイソフルランの流れを送信することができます。 </li></ol></li><li>誘導室に、三方コネクタを回します。ホームケージから1匹のマウスを選択し、チャンバー内に動物を挿入します。 1.0 L /分の酸素と1.0 L /分の空気を流量調整器を調整します。 5％ - 4にイソフルラン濃度を調整します。 </li><li>麻酔の所望のレベルに達したときに、麻酔の所望のレベルに達したとき、手術領域を剃るマスクにチャンバーから動物を移動させます。マスクの流れに、三方コネクタを回します。 0.25リットル/分の酸素および0.25リットル/分の空気を流量調整器を調整します。 <ol><li> 2.5％ - 1.5イソフルラン濃度を調整します。外科procedurを開始する前に、反射制御および呼吸数によって麻酔のレベルをチェックしてください電子。 </li></ol></li><li>粘着テープを使用した作業台にマウスの足を固定してください。動物の鼻はまだ慎重にマスク内に配置されていることを確認してください。 </li></ol>迷走神経の3手術と刺激<ol><li> 70％エタノールで手術領域を消毒します。 </li><li>メスを使用して、慎重に首（約1〜1.5センチの切開部）のレベルで皮膚を切開。 注意：プロトコルでは、手術の手順は、偽手術動物のためにここで終了。確かに、それだけで金属製の工具で神経に触れることは、すでにある程度までそれを刺激することが示されています。したがって、LPSの少量を使用した場合、より正確な手術コントロール動物を取得するには、この段階で手術を停止します。 </li><li>顕微鏡（12.5Xの目的）の助けを借りて、解剖ピンセットを用いて頸動脈から左迷走神経を分離します。まず皮膚や脂肪の層を除去することにより、胸鎖乳突筋を見つけて、それを撤回神経と動脈の両方の後ろに鉗子を置くため。 注：神経および動脈が互いに密接に付着される次のステップは、非常に厄介です。したがって、血管を切断して、動物を殺すことは非常に簡単です。しかし、神経と動脈の間で非常に慎重にピンセットを置くことによって、彼らは最終的に分離し、神経を単離することが可能です。 </li><li>迷走神経の下電極（ 図1B）を置きます。針電極はかなり長いので、神経が刺激中に、わずかに移動しても、それは常に、電極と接触することになります。 </li><li> （サイトカインレベルに対する読み出しのために、 すなわち、ダウンレギュレーションの効果を測定するために）注射器の助けを借りて、LPSの腹腔内（ip）注射（2mg / kgの）を実行します。 </li><li>刺激を開始する前に、5分待ってください。 </li><li>アクノリッジプログラムのスタートボタンを押して5 V及び1Hzで5分間、迷走神経を刺激します。 </li><li>再電極を移動させ、手術用縫合糸を用いて動物の創傷を縫合。 </li><li>治癒を向上させ、感染から保護するために傷口にはありませんスティングバリアフィルム（NSBF）をスプレーしてください。 </li></ol>動物の4.回復<ol><li>手術後、目覚めと回復のために戻ってそのホームケージに動物を移動します。体温を維持するために、赤外光の下では、完全な意識が回復されるまで、動物を監視するようにしてください。 </li><li>動物は、分析のために犠牲にする前に6時間、そのケージに回復しましょう。 注意：迷走神経刺激の効果は非常に高速であり、また、（48時間まで）持続長いことが示されているので、回復時間は、研究のニーズに応じて、実験者によって設定することができます。 </li></ol>さらなる分析のための5サクリファイス<ol><li> CO 2投与装置にリンクされているケージに動物を置きます。 </li><li> 5上のデバイスを設定します。CO 2吸入の-minサイクル。 </li><li>安楽死が行われると、関心のある器官を収集し、直接、さらなる分析（ 例えば、マウスTH1 / TH2 9プレックスアッセイを用いて脾臓抽出物中のサイトカインレベルの測定）のためにドライアイス上でそれらを凍結3。 </li></ol>

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2000年代初頭におけるその発見以来、CAPのメカニズムは、徹底的に研究されてきました。私たちは今、ダウンレギュ炎症性メディエーター2に経路の良い写真を持って、特に、標的臓器、脾臓、非常に効率的なチームとしてNE、メモリーT細胞、のAch、およびマクロファージの仕事。また、最近明らかにCAP 3の活性化後の脾臓におけるACh放出のための必須の成?...

先天性免疫は、生物の広い範囲の感染および疾患に対する防御の即時の最初のラインを提供します。それは脅威を排除するための一次免疫応答を開始するだけでなく、それはまた、病原体特異的に二次免疫応答を行い、適応免疫を活性化し、教育において重要な役割を果たしています。炎症は順番に感染部位に他の免疫細胞を誘引する能力を持っていると、このような赤み、腫れ、痛み、機能の喪失、および発熱などの炎症の基本的兆候を誘導するサイトカインおよびケモカインの過多によって画策されました。期間や炎症の強さは、いくつかの要因に依存するが、炎症を解決し、恒常性を回復することは慢性炎症性疾患の発症を回避するための重要なステップです。神経科学および免疫学の分野における最近の進歩はINFLを制御するために巨大な治療的可能性を持つ特定の神経機構を解明してきました中枢神経系における及び末梢の両方でammation。これらのメカニズムのいずれかはまた、自律神経系4、5により駆動される炎症性の反射として知られているコリン作動性抗炎症経路（CAP）、です。 現在、炎症性メディエーターは、感覚神経を活性化し、中枢神経系に炎症の状態に関するシグナルを送ることを考えています。反射反応は、その後、遠心性迷走神経を介して活性化されます。 CAPの解剖学的詳細について鋭意研究がそれぞれ二つの神経、迷走神経および脾臓神経、6からなる副交感神経、交感神経モデルを明らかにしました。 CAPにおいて、活性化されたコリン作動性遠心性迷走神経は、まだ探求される機構によってアドレナリン脾臓神経の活性化をもたらす、セリアック病、腸間膜神経節で終わります。このようにして活性化脾臓神経は、内側にはよく知られています白色パルプ、マージナルゾーン、および脾臓の赤脾髄、CAP 7,8の主及び必須の器官における免疫細胞に近接しvate。脾臓神経終末からのノルエピネフリン（NE）は、脾臓Tリンパ球上に発現される、対応するβ2アドレナリン受容体に結合します。これは、今度は、それによりサイトカイン産生および炎症2を限定マクロファージ上のα7ニコチン性アセチルコリン受容体（α7nACh）を、活性化コリンアセチルトランスフェラーゼ（ChAT）媒介性アセチルコリン（ACH）放出を誘導します。したがって、神経系は、末梢組織に炎症を調節すると、局所免疫恒常性を復元することが可能であることが明らかです。 経路の名前が示すように、AChのシステムでは、この神経免疫調節経路の機能に中心的な重要性です。興味深いことに、メカニズムは、の活性化に関与しますCAPは、周囲の中枢神経系で異なるように見えます。脾臓中のニコチン性受容体（α7nAChR）の重要性は以前9が実証されてきたが、ムスカリン受容体（のmAChR）は経路10、11の中心活性のために必須です。致死マウス内毒素血症時より最近、中枢作用性M1ムスカリン性アゴニストの末梢投与が有意に抑制血清および脾臓腫瘍壊死因子α（TNFα）、12シグナリング無傷の迷走神経及び脾臓神経を必要なアクション。我々はまた、プロスタグランジンE 2（PGE 2）を欠損したマウスは、迷走神経刺激に応答し、血清および脾臓3におけるサイトカインのLPS誘導性の解放をダウンレギュレートしませんでしたができなかったことが最近示されています。したがって、CAPは、メインアセチルコリンpathw以外のシステムにより調節されるかもしれませんAY。 迷走神経は、理由は体内での放浪もちろん、肝臓、肺、脾臓、腎臓、および腸13を含む主要臓器を支配などの名前が挙がっています。この大規模な神経支配と迷走神経の非常に強力な免疫抑制効果を考慮すると、CAPの治療可能性は、炎症状態の広い範囲をカバーできます。迷走神経は、体に加えない薬剤と、電気的に（または機械的に）活性化され、電圧および周波数の制御と、従来の治療法とは逆であってもよいです。試験は、慢性炎症14を治療する際のVNSの臨床的意義をテストするために、例えば、リウマチ患者では、現在進行中です。要するに、炎症の神経免疫通信および調節は、従来の治療に対する可能な代替治療を提供しており、現在調査中です。そのため、迷走神経の分析stimulation個の異なる神経支配器官における効果が、慢性炎症の動物モデルでの潜在的な治療行為のも特徴づけは、間違いなく洞察を与え、新しい潜在的な治療標的のために望んでいるだろう。 トレイシーと同僚4によって開発されたオリジナルの方法論が原因（LPSの致死量によって）炎症反応の過剰刺激とCAPの活性化と読み出しの間にあまりにも短い時間帯に研究の私たちのフィールドに置き換えることができませんでした。本論文では、我々は、元のプロトコルに対する変更を提示サイトカインレベル上の2つの異なる方法を比較し、標的臓器（脾臓）に新しい反対観察を強調表示します。

<table><tbody><tr><td>Computer</td><td>Toshiba</td><td>-</td><td>Any computer is actually compatible</td></tr><tr><td>MP-150 data acquisition system</td><td>Biopac Systems</td><td>MP150WSW</td><td></td></tr><tr><td>Acknowledge software</td><td>Biopac Systems</td><td></td><td></td></tr><tr><td>Mice C57Bl/6</td><td>Charles River</td><td></td><td></td></tr><tr><td>Anesthetic machine</td><td>Simtec Engineering</td><td></td><td></td></tr><tr><td>Medical oxygen bottle</td><td>AGA</td><td>107563</td><td></td></tr><tr><td>Medical air bottle&amp;nbsp;</td><td>AGA</td><td>108639</td><td></td></tr><tr><td>Vetflurane (1,000 mg/g)</td><td>Virbac</td><td>137317</td><td></td></tr><tr><td>LPS</td><td>Sigma-Aldrich</td><td>L2630</td><td></td></tr><tr><td>Saline</td><td>Merck Millipore</td><td>1024060080</td><td></td></tr><tr><td>PBS 10x</td><td>Sigma-Aldrich</td><td>P5493</td><td>Diluted 10 times for used concentration</td></tr><tr><td>Syringe (1 ml)</td><td>BD Plastipak</td><td>303172</td><td></td></tr><tr><td>Needles 23 G</td><td>KD-FINE</td><td>900284</td><td>0.6 x 30 mm (blue)</td></tr><tr><td>Microdissecting forceps (curved)</td><td>Sigma-Aldrich</td><td>F4142</td><td></td></tr><tr><td>Dissecting scissors</td><td>Sigma-Aldrich</td><td>Z265969</td><td></td></tr><tr><td>Surgical suture 4-0</td><td>Ethicon</td><td>G667G</td><td></td></tr><tr><td>Euthanasia unit</td><td>Euthanex Smartbox</td><td>EA-32000</td><td></td></tr><tr><td>Cavilon No Sting Barrier Film</td><td>3M Health Care</td><td>3346N</td><td></td></tr><tr><td>TH1/TH2 9-Plex assay, ultrasensitive kit</td><td>MesoScale Discovery</td><td>K15013C-1</td><td></td></tr><tr><td>Stimulating electrode device</td><td>Biopac Systems</td><td>STIMSOC</td><td></td></tr><tr><td>Aesculap Isis shaver</td><td>Agnthos</td><td>GT420</td><td></td></tr><tr><td>R70</td><td></td><td></td><td>Rodent diet from Lantmannen, Stockholm, Sweden</td></tr></tbody></table>

increased recovery time and decreased lps administration to study the vagus nerve stimulation mechanisms in limited inflammatory responses

Inflammation is a local response to infection and tissue damage mediated by activated macrophages, monocytes, and other immune cells that release cytokines and other mediators of inflammation. For a long time, humoral and cellular mechanisms have been studied for their role in regulating the immune response, but recent advances in the field of immunology and neuroscience have also unraveled specific neural mechanisms with interesting therapeutic potential. The so-called cholinergic anti-inflammatory pathway (CAP) has been described to control innate immune responses and inflammation in a very potent manner. In the early 2000s, Tracey and collaborators developed a technique that stimulates the vagus nerve and mimics the effect of the pathway. The methodology is based on the electrical stimulation of the vagus nerve at low voltage and frequency, in order to avoid any side effects of overstimulation, such as deregulation of heart rate variability. Electrical devices for stimulation are now available, making it easy to set up the methodology in the laboratory. The goal of this research was to investigate the potential involvement of prostaglandins in the CAP. Unfortunately, based on earlier attempts, we failed to use the original protocol, as the induced inflammatory response either was too high or was not suitable for enzymatic metabolism properties. The different settings of the original surgery protocol remained mostly unchanged, but the conditions regarding inflammatory induction and the time point before sacrifice were improved to fit our purposes (i.e., to investigate the involvement of the CAP in more limited inflammatory responses). 
The modified version of the original protocol, presented here, includes a longer time range between vagus nerve stimulation and analysis, which is associated with a lower induction of inflammatory responses. Additionally, while decreasing the level of lipopolysaccharides (LPS) to inject, we also came across new observations regarding mechanistic properties in the spleen.

手術後の経過時間を増加し、LPSの投与量を減少した後、TNFαおよびインターロイキン-1β（IL-1β）のレベル先に示したように、元のプロトコルを使用して、VNSは、TNFαのレベルおよびIL-1β（360.0±40.21 PG / MGに（39.7±10.8 PG / VNSのMg、P &lt;0.001対SHAMで169.3±24.9 PG / mg）を減少しました腹腔内LPS注射（15ミリグラム/ kg）を（ 図2A?...

Watch this Scientific Journal Video about Increased Recovery Time and Decreased LPS Administration to Study the Vagus Nerve Stimulation Mechanisms in Limited Inflammatory Responses at JoVE.com

Increased Recovery Time and Decreased LPS Administration to Study the Vagus Nerve Stimulation Mechanisms in Limited Inflammatory Responses

Vagus nerve stimulation has proven to have a strong efficacy for decreasing peripheral inflammation. Here, we present a modified vagus nerve stimulation protocol that allows for further examinations of the cholinergic anti-inflammatory mechanisms in limited inflammatory responses.

回復時間の増加とリミテッド炎症反応における迷走神経刺激メカニズムを研究するためにLPSの管理を減少

Inflammation is a local response to infection and tissue damage mediated by activated macrophages, monocytes, and other immune cells that release ...

increased-recovery-time-decreased-lps-administration-to-study-vagus

Universidad de Cartagena UDEC

Research

JoVE Journal

Immunology and Infection

9.8K ビュー.  Karolinska University Hospital. Vagus nerve stimulation has proven to have a strong efficacy for decreasing peripheral inflammation. Here, we present a modified vagus nerve stimulation protocol that allows for further examinations of the cholinergic anti-inflammatory mechanisms in limited inflammatory responses.

ビデオ: Increased Recovery Time and Decreased LPS Administration to Study the Vagus Nerve Stimulation Mechanisms in Limited Inflammatory Responses

Vagus Nerve Stimulation as a Tool to Induce Plasticity in Pathways Relevant for Extinction Learning

Extinction describes the process of attenuating behavioral responses to neutral stimuli when they no longer provide the reinforcement that has been maintaining the behavior. There is close correspondence between fear and human anxiety, and therefore studies of extinction learning might provide insight into the biological nature of anxiety-related disorders such as post-traumatic stress disorder, and they might help to develop strategies to treat them. Preclinical research aims to aid extinction learning and to induce targeted plasticity in extinction circuits to consolidate the newly formed memory. Vagus nerve stimulation (VNS) is a powerful approach that provides tight temporal and circuit-specific release of neurotransmitters, resulting in modulation of neuronal networks engaged in an ongoing task. VNS enhances memory consolidation in both rats and humans, and pairing VNS with exposure to conditioned cues enhances the consolidation of extinction learning in rats. Here, we provide a detailed protocol for the preparation of custom-made parts and the surgical procedures required for VNS in rats. Using this protocol we show how VNS can facilitate the extinction of conditioned fear responses in an auditory fear conditioning task. In addition, we provide evidence that VNS modulates synaptic plasticity in the pathway between the infralimbic (IL) medial prefrontal cortex and the basolateral complex of the amygdala (BLA), which is involved in the expression and modulation of extinction memory.

Vagus nerve stimulation (VNS) has emerged as a tool to induce targeted synaptic plasticity in the forebrain to modify a range of behaviors. This protocol describes how to implement VNS to facilitate the consolidation of fear extinction memory.

ツールとしての迷走神経刺激は、絶滅の学習のための関連経路の可塑性を誘導するために

Extinction describes the process of attenuating behavioral responses to neutral stimuli when they no longer provide the reinforcement that has been ...

行動学

Characterization of Inflammatory Responses During Intranasal Colonization with Streptococcus pneumoniae

Nasopharyngeal colonization by Streptococcus pneumoniae is a prerequisite to invasion to the lungs or bloodstream1. This organism is capable of colonizing the mucosal surface of the nasopharynx, where it can reside, multiply and eventually overcome host defences to invade to other tissues of the host. Establishment of an infection in the normally lower respiratory tract results in pneumonia. Alternatively, the bacteria can disseminate into the bloodstream causing bacteraemia, which is associated with high mortality rates2, or else lead directly to the development of pneumococcal meningitis. Understanding the kinetics of, and immune responses to, nasopharyngeal colonization is an important aspect of S. pneumoniae infection models.
Our mouse model of intranasal colonization is adapted from human models3 and has been used by multiple research groups in the study of host-pathogen responses in the nasopharynx4-7. In the first part of the model, we use a clinical isolate of S. pneumoniae to establish a self-limiting bacterial colonization that is similar to carriage events in human adults. The procedure detailed herein involves preparation of a bacterial inoculum, followed by the establishment of a colonization event through delivery of the inoculum via an intranasal route of administration. Resident macrophages are the predominant cell type in the nasopharynx during the steady state. Typically, there are few lymphocytes present in uninfected mice8, however mucosal colonization will lead to low- to high-grade inflammation (depending on the virulence of the bacterial species and strain) that will result in an immune response and the subsequent recruitment of host immune cells. These cells can be isolated by a lavage of the tracheal contents through the nares, and correlated to the density of colonization bacteria to better understand the kinetics of the infection.

Characterization of Inflammatory Responses During Intranasal Colonization with Streptococcus pneumoniae

Colonization of the murine nasopharynx with Streptococcus pneumoniae and the subsequent extraction of adherent or recruited cells is described. This technique involves flushing the nasopharynx and collection of the fluid through the nares and is adaptable for various readouts, including differential cell quantification and analysis of mRNA expression in situ.

肺炎球菌による鼻腔内コロニー形成時の炎症反応の特徴

Nasopharyngeal colonization by Streptococcus pneumoniae is a prerequisite to invasion to the lungs or bloodstream1. This organism is capable of colonizing ...

免疫・感染学

A Method of Nodose Ganglia Injection in Sprague-Dawley Rat

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located in the organs of the abdomen and thorax. The vagus nerve communicates information from stimuli such as heart rate, blood pressure, bronchopulmonary irritation, and gastrointestinal distension to the nucleus of solitary tract of the medulla. The cell bodies of the vagus nerve are located in the nodose and petrosal ganglia, of which the majority are located in the former. The nodose ganglia contain a wealth of receptors for amino acids, monoamines, neuropeptides, and other neurochemicals that can modify afferent vagus nerve activity. Modifying vagal afferents through systemic peripheral drug treatments targeted at the receptors on nodose ganglia has the potential of treating diseases such as sleep apnea, gastroesophageal reflux disease, or chronic cough. The protocol here describes a method of injection neurochemicals directly into the nodose ganglion. Injecting neurochemicals directly into the nodose ganglia allows study of effects solely on cell bodies that modulate afferent nerve activity, and prevents the complication of involving the central nervous system as seen in systemic neurochemical treatment. Using readily available and inexpensive equipment, intranodose ganglia injections are easily done in anesthetized Sprague-Dawley rats.

Afferent vagal signaling transmits important information to central nervous system from receptors located in organs of the abdomen and thorax. The nodose ganglia of vagus nerves contain many types of receptors that modulate vagal activity. This protocol describes a method of local injections of neurochemicals into the nodose ganglia.

スプラーグ - ドーリーラットのメソッド節状神経節の注入

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located ...

神経科学

Laboratory Administration of Transcutaneous Auricular Vagus Nerve Stimulation (taVNS): Technique, Targeting, and Considerations

Non-invasive vagus nerve stimulation (VNS) may be administered via a novel, emerging neuromodulatory technique known as transcutaneous auricular vagus nerve stimulation (taVNS). Unlike cervically-implanted VNS, taVNS is an inexpensive and non-surgical method used to modulate the vagus system. taVNS is appealing as it allows for rapid translation of basic VNS research and serves as a safe, inexpensive, and portable neurostimulation system for the future treatment of central and peripheral disease. The background and rationale for taVNS is described, along with electrical and parametric considerations, proper ear targeting and attachment of stimulation electrodes, individual dosing via determination of perception threshold (PT), and safe administration of taVNS.

Laboratory Administration of Transcutaneous Auricular Vagus Nerve Stimulation taVNS: Technique, Targeting, and Considerations

A methodological description of the technique, potential targets, and proper administration of transcutaneous auricular vagus nerve stimulation (taVNS) on the human ear is described.

研究室管理の経皮的心房迷走神経刺激 (taVNS): 技術、ターゲット、および考慮事項

Non-invasive vagus nerve stimulation (VNS) may be administered via a novel, emerging neuromodulatory technique known as transcutaneous auricular vagus ...

Nerve Stimulator-guided Injection of Autologous Stem Cells Near the Equine Left Recurrent Laryngeal Nerve

Recurrent laryngeal neuropathy (RLN) commonly affects horses and is characterized by abnormal respiratory sounds and exercise intolerance. The recurrent laryngeal nerve shows lesions of demyelination. The benefit of applying stem cells to demyelinated nerves has been demonstrated in various animal models. The aim of the study was to test the feasibility and safety of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to the left recurrent laryngeal nerve in healthy horses by using an electrical nerve stimulator.
Muscle-derived stems cell are obtained from five healthy Standardbred horses by sampling 20 mg of muscle tissue with a semi-automatic 14 G biopsy needle from the triceps muscle. Movements of the larynx are monitored via upper-airway video endoscopy. The left recurrent laryngeal nerve is approached with an insulated nerve block needle. Nerve stimulation is applied, starting at 2 mA, and the successful abduction of the left arytenoid is monitored. The stimulation intensity is reduced progressively. When a loss of the motor response is observed at 0.5 mA, 107 autologous muscle-derived stem cells are injected. Two examiners, who are blinded to the time point, score the laryngeal function of the horses prior to the treatment and at day 1, day 7, and day 28 after the injection of the cells. In a sixth horse, 1 mL of 2% lidocaine is injected to further confirm the correct positioning of the needle. This leads to a temporary paralysis of the left arytenoid cartilage.
This study proves that the recurrent laryngeal nerve can be approached with the help of an electrical nerve stimulator and that the electrical stimulation of the nerve is well tolerated by the horses. No modification of the laryngeal function was observed in any of the horses after the injection of the stem cells. Further studies should be conducted to describe the effects of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to horses suffering from RLN.

Here, we present a protocol to inject autologous muscle-derived stem cells in the proximity to the recurrent laryngeal nerve with the help of an electrical nerve stimulator. This novel technique may become useful for the treatment of equine recurrent laryngeal neuropathy.

神経刺激誘導馬の左反回神経に近い自家幹細胞注入

Recurrent laryngeal neuropathy (RLN) commonly affects horses and is characterized by abnormal respiratory sounds and exercise intolerance. The recurrent ...

医学

Vagus Nerve Stimulation As an Adjunctive Neurostimulation Tool in Treatment-resistant Depression

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression (TRD). VNS exhibits antidepressive and stabilizing effects. This method is particularly useful as a long-term treatment, in which up to two-thirds of patients respond. The vagus nerve stimulator is positioned on the left vagus nerve during a surgical procedure and is activated telemetrically by a wand connected to a handheld computerized device. The treating physician can perform various adjustments of the vagus nerve stimulator during in-office visits (e.g., by modifying stimulation intensity or stimulation frequency) to achieve maximum therapeutic effects with low side effects. Set-up of the device usually takes several months. Typical side effects include wound infection, temporary salivation, coughing, paralysis of the vocal cords, bradycardia, or even asystole. The patient can stop the VNS by placing a magnet over the generator. The current protocol describes delivery of the specific stimulation tool and methods for adjusting the tuning parameters to achieve the best remission rates in patients with TRD.

Vagus nerve stimulation (VNS) has been shown to be effective as an adjunct treatment for treatment-resistant depression (TRD). VNS leads to antidepressive and antisuicidal effects and quality of life improvements. This protocol offers a step-by-step guide to managing and adjusting a vagus nerve stimulator for efficient treatment of TRD.

治療抵抗性うつ病の補助神経ツールとして迷走神経刺激

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression ...

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. Several recent studies have demonstrated the effectiveness of chronic VNS in enhancing central nervous system plasticity to improve motor rehabilitation, extinction learning, and sensory discrimination. Construction of chronically implantable devices for use in such studies is challenging due to rats&#8217; small size, and typical protocols require extensive training of personnel and time-consuming microfabrication methods. Alternatively, commercially available implantable cuff electrodes can be purchased at a significantly higher cost. In this protocol, we present a simple, low-cost method for construction of small, chronically implantable peripheral nerve cuff electrodes for use in rats. We validate the short and long-term reliability of our cuff electrodes by demonstrating that VNS in ketamine/xylazine anesthetized rats produces decreases in breathing rate consistent with activation of the Hering-Breuer reflex, both at the time of implantation and up to 10 weeks after device implantation. We further demonstrate the suitability of the cuff electrodes for use in chronic stimulation studies by pairing VNS with skilled lever press performance to induce motor cortical map plasticity.

Existing approaches for constructing chronically implantable peripheral nerve cuff electrodes for use in small rodents often require specialized equipment and/or highly trained personnel. In this protocol we demonstrate a simple, low-cost approach for fabricating chronically implantable cuff electrodes, and demonstrate their effectiveness for vagus nerve stimulation (VNS) in rats.

ラットの慢性移植のための末梢神経刺激電極の調製

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...

経皮的耳介迷走神経刺激による神経調節の測定

Understanding the Effects of Non&#45;Invasive Transauricular Vagus Nerve Stimulation on EEG and HRV

Several studies have demonstrated promising results of transcutaneous auricular vagus nerve stimulation (taVNS) in treating various disorders; however, no mechanistic studies have investigated this technique's neural network and autonomic nervous system effects. This study aims to describe how taVNS can affect EEG metrics, HRV, and pain levels. Healthy subjects were randomly allocated into two groups: the active taVNS group and the sham taVNS group. Electroencephalography (EEG) and Heart Rate Variability (HRV) were recorded at baseline, 30 min, and after 60 min of 30 Hz, 200-250 &#181;s taVNS, or sham stimulation, and the differences between the metrics were calculated. Regarding vagal projections, some studies have demonstrated the role of the vagus nerve in modulating brain activity, the autonomic system, and pain pathways. However, more data is still needed to understand the mechanisms of taVNS on these systems. In this context, this study presents methods to provide data for a deeper discussion about the physiological impacts of this technique, which can help future therapeutic investigations in various conditions.

著者スポットライト:経耳介性迷走神経刺激の影響を探る

This protocol provides information on how to apply transcutaneous auricular vagus nerve stimulation (taVNS) in a clinical trial, including potential biomarkers such as EEG metrics and heart rate variability (HRV) to measure the effect of this treatment on the autonomic nervous system.

非侵襲的経auricurgus神経刺激がEEGおよびHRVに及ぼす影響の理解

Several studies have demonstrated promising results of transcutaneous auricular vagus nerve stimulation (taVNS) in treating various disorders; however, no ...

回復時間の増加とリミテッド炎症反応における迷走神経刺激メカニズムを研究するためにLPSの管理を減少

要約

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この動画の章

ツールとしての迷走神経刺激は、絶滅の学習のための関連経路の可塑性を誘導するために

肺炎球菌による鼻腔内コロニー形成時の炎症反応の特徴

スプラーグ - ドーリーラットのメソッド節状神経節の注入

研究室管理の経皮的心房迷走神経刺激 (taVNS): 技術、ターゲット、および考慮事項

神経刺激誘導馬の左反回神経に近い自家幹細胞注入

治療抵抗性うつ病の補助神経ツールとして迷走神経刺激

ラットの慢性移植のための末梢神経刺激電極の調製

非侵襲的経auricurgus神経刺激がEEGおよびHRVに及ぼす影響の理解

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