この研究は、NIH / NIDDKからの助成金1K01DK079019A2（PT）によってサポートされていました。

1。マウス接種のための細菌の準備以下は無菌条件下で行わなければなりません。 <ol><li>キャップで1 17×100mmのポリプロピレンチューブを取り、新鮮なルリア培地（LB）培地3 mlを加える。オートクレーブしたチップを使用して、ひずみCP1 2の一部凍結細菌のグリセロールストックを取り、LB培地を含むチューブに培養液の少量を転送します。酸素がまだ管を入力することができるように、チューブのキャップを交換します - 文化は好気的条件下で成長する必要がある。で37箇所管℃で一晩220rpmで振盪インキュベーターのC。 </li><li>翌日、3 mlのLB培地で新しいチューブに一晩培養した培養液の5μlを転送し、37℃で一晩インキュベーター内の静的条件下で増殖</li><li> 40mlのLB培地それぞれを含むAA 50mlチューブに培養液40μlの3日目の転送で。一晩37℃のインキュベーター内で静的な場所。 </li><li>電源をオンにして、4℃に遠心分離機を設定一度遠心機が最終温度に達しており、40ミリリットルナルゲン遠心管にそれぞれ40 mlの培養液を移す。 </li><li> LB培地で音量を調整して、必要に応じて - 遠心分離機のバランスをとるためにチューブの重量を量る。 20分間6,000 rpmでチューブを軽くスピン。 </li><li>上清を慎重に除去し、穏やかに40ミリリットル滅菌PBSに細菌ペレットを中断します。 </li><li> PBSでステップ1.5を繰り返します。 </li><li>上清を吸引除去し、500μlの滅菌PBS中の細菌を一時停止。 1.5 mlのマイクロチューブに溶液を移します。懸濁液10μlを取り、氷冷したPBS 990μlに希釈します。接種に必要な量を決定するために、OD 420 nmを読み取るために分光光度計を使用しています。の適切な音量を決定するために、氷冷PBSでペレットを一時停止するには：外径420 nmの番号 （mlで）に乗り、細菌ペレット[元の推定量を減算します。 ODを420nmで = 0.545、ペレット約0.075ミリリットル。 0.545から0.075 = 0.470]。 </li><li>細菌懸濁液10μlを取り出して、ディ990μlの氷冷PBSでリュート。 OD 420 nmをお読みください。目標は1.000±0.010希釈懸濁液のOD 420 nmの値を達成することです。あなたの希釈の数がこの目標を上回っている場合、懸濁液に複数のボリュームを追加し、別の値を読み取ります。数が0.990を下回っている場合は、懸濁液をスピンダウンし、所望の読み取り値を取得するまで、外径420 nmの読みを繰り返す。 </li></ol> 2。動物の感染症マウス（5から7週齢、グループ当たり10）はジャクソン研究所（バーハーバー、メイン州）から購入した。 <ol><li>マウスはプレキシグラス室におけるイソフルラン1〜4％の吸入により麻酔し、リカンベントまで監視されています。 </li><li>一度マウスはガラスハミルトンシリンジ（1.5〜2.0センチメートルの長さ）の変更された30gの針に取り付けポリエチレン（PE10）カテーテルとカテーテル法によって細菌の点滴のために取られています。カテーテルは針のハブに挿入されます。マウスはplacですedは麻酔下で背側表面にノーズコーンを使用して維持。 </li><li>マウスの陰​​茎は陰茎尿道の入り口の潤滑に使用されている穏やかな圧力と綿棒で、潤滑剤のリベラルな量によって押し出される。 </li><li> 1×10 8個の細菌を含有するリン酸緩衝生理食塩水、10μlのカテーテル挿入に続いて麻酔したマウスの尿道内に導入される。 </li><li>細菌の付着を可能にするために、直ちに排尿を防ぐために、細菌の導入後マウスはプレキシグラス室中で15分間、麻酔状態に維持されている。 </li><li>マウスをケージに戻し、次の24時間監視されています。 </li></ol> 3。動作テストマウスは、感染前（ベースライン）へと3日、7日、14日、21日、感染後28日に試験した。呼ば痛覚過敏と触覚アロディニアは、腹部と足11,12 Rに適用von Freyフィラメントを用いて試験した後足13 egion。テストは固定時刻、​​標準的な方法論と、すべての動物の単一実験試験採用された時に施行した。グループの盲検試験は、動物モデルにおいて行動ベースの痛み試験の限界と戦うために利用された。グラム（Stoelting、米国）が撤退応答の腹部と周波数に適用されていた0.04、0.16、0.4、1.0、4.0の力を持つ5つの個々のvon Freyフィラメントを算出した。 <ol><li>順応の30分の期間の後、マウスはステンレススチールワイヤーグリッドの床が社内で行われた個々のプレキシグラス室（6×10×12 cm）で配置されます。 </li><li>呼ば痛覚過敏と触覚アロディニア5 von Freyフィラメントを用いて試験した。各フィラメントは合計10回に5秒の間刺激間隔で1〜2秒のために適用され、毛は力の昇順で試験した。 </li><li>各フィラメントは前立腺の一般付近で下腹部領域に適用されると注意が脱感作を回避したり、エフェクトを &quot;巻き上げる&quot;ために、この領域内の異なる領域を刺激するために取られた。フィラメントは、10回の合計のための刺激の間に少なくとも5秒の間隔をおいて1〜2秒のための力の低い順に適用された。 </li><li>行動は、3種類のフィラメント刺激に対する肯定的な反応であると考えています：腹部の1）鋭い引き込み、2）即時舐めるまたはフィラメント刺激の面積の傷、3）ジャンプ。 </li><li>応答周波数は、肯定応答の割合（10点、10 = 50％の例えば 5回答）として算出したとのデータが応答周波数の平均パーセンテージ±SEとして報告されました</li><li> 25以上の正の合計ベースライン応答を持つ動物は研究から除外されます。 </li><li>接触性アロディニアは0.04、0.16、0.4、1.0と4.0グラムの力とvon Freyフィラメントを用いて後肢の足の領域でテストされました。中央値50％引っ込め閾値（5）は、アップダウンを用いて評価したフィラメントが少し曲げるまでテストが0.04グラムのフィラメントで開始されたメソッドは、後肢の足底表面に垂直に印加。フィラメントは、肯定的な反応が観察されるまで、昇順で試験した。フィラメントへの積極的な応答は試験足の足や舐めるのシャープ撤退のいずれかとして定義されていました。肯定応答が記録されたときに次の弱いフィラメントが適用され、否定応答が観察された場合は、次の強いフィラメントを適用した。 </li><li>記載された実験と方法論は、ノースウェスタン大学の動物のケアと使用委員会によって審査され、承認されています。 </li></ol>

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特別な利害関係は宣言されません。

UPECによるマウス前立腺の感染における生体内細菌性前立腺炎の病態に関与している可能性のある事象のモデリング、CP / CPPSまたは慢性炎症における素因イベントなどが可能になります。方法は女性の尿路感染症7,14でUPECモデルに文学の大きな体によって細菌準備や点滴の引き分けに記載。モデルは、潜在的なワクチン候補と免疫調節のメカニズムをテストする、病因を研究す?...

<table border="1"><tbody><tr><td> 試薬の名称 </td><td> 会社 </td><td> カタログ番号 </td></tr><tr><td>培養管</td><td> BDファルコン</td><td> 352059 </td></tr><tr><td> LBブロスミラー</td><td> EMD </td><td> EM1.10285.0500 </td></tr><tr><td>ナルゲン遠心管</td><td>サーモ</td><td> 3118-0050 </td></tr><tr><td>イソフルレン</td><td>バトラーシャイン</td><td> NDC 11695-6776-1 </td></tr><tr><td>カテーテルニードル30G </td><td>ハミルトン</td><td> 91030 </td></tr><tr><td> PE10チューブ</td><td> BD intramedic </td><td> 427400 </td></tr><tr><td> von Freyフィラメント</td><td> Stoelting </td><td> 58025から31 </td></tr></tbody></table>

measurement of tactile allodynia in a murine model of bacterial prostatitis

尿路病原性大腸菌 （UPEC）は、尿路感染症や細菌性前立腺炎1に重要な役割を果たしている病原体である。我々は最近、UPECは、慢性骨盤痛2、慢性前立腺炎/慢性骨盤痛症候群（CP / CPPS）3,4の機能の開始に重要な役割を持っていることが示されている。臨床的に関連UPECによる前立腺の感染は、組織の損傷および自己免疫のメカニズム5の開始を含むことができるメカニズムを介して慢性痛を開始し、確立することができます。 前立腺内UPECの病因を理解するための課題は、操作に前立腺の相対的な到達不能です。我々は、それによって前立腺の上行性感染を確立雄マウスにUPECの臨床株を提供するために前述の尿道感染症法6を利用した 。ここでは、BAを標準化するための私達のプロトコルを記述cterial接種7と同様に細菌の点滴のために麻酔雄マウスをcatheterizingするための手順。 CP / CPPSは主に触覚アロディニア4の存在によって特徴付けられる。動作テストは呼ば内臓痛8-10起因する皮膚の痛覚過敏の概念に基づいていた。内臓組織における過敏焦点は通常非痛み刺激（アロディニア）に誇張された応答を可能にする皮膚の疼痛閾値を低減します。基盤となる内臓痛の強度に伴って増加する傾向がある異常反応で皮膚の結果に垂直力の応用。我々はUPEC細菌を持つ単一の感染に応答して時間をかけて接触性アロディニアを定量化するためのvon Frey繊維を利用したNOD / ShiLtJマウスでの方法論を記述する。

<html>我々は、細菌感染時に慢性的な痛みの有無をNOD / ShiLtjおよびC57BL/6J雄マウスを調べた。雄性マウス（5〜7週齢）尿道（ 図1）に生理食塩水や細菌を注入した。生理食塩水をたらしC57BL/6JマウスとUPEC感染C57BL/6Jマウスのvon Freyフィラメントと骨盤領域の機械的刺激は、実験の28日間コース（ 図1）の間に変更されなかった応答周波数が得られた。これとは対照的に、UPEC...

Watch this Scientific Journal Video about Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis at JoVE.com

Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis

前立腺の感染は、慢性前立腺炎における骨盤痛を媒介する要因かもしれません。我々は時間をかけて、マウスにおける接触性アロディニアを測定するための雄マウスと方法論の尿道への細菌の点滴、標準化された細菌の接種物の調製のための手順を記述している。

細菌性前立腺炎のマウスモデルにおける接触性アロディニアの測定

Uropathogenic Escherichia coli (UPEC) are pathogens that play an important role in urinary tract infections and bacterial prostatitis1. We have recently ...

measurement-tactile-allodynia-murine-model-bacterial

Research

JoVE Journal

Immunology and Infection

15.8K ビュー.  Northwestern University Feinberg School of Medicine. 前立腺の感染は、慢性前立腺炎における骨盤痛を媒介する要因かもしれません。我々は時間をかけて、マウスにおける接触性アロディニアを測定するための雄マウスと方法論の尿道への細菌の点滴、標準化された細菌の接種物の調製のための手順を記述している。

ビデオ: Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical condition. However, the underlying mechanisms of CRPS-I have yet to be revealed. It is known that ischemia/reperfusion is one of the leading factors that causes CRPS-I. By means of prolonged ischemia and reperfusion of the hind limb, the rat chronic post-ischemia pain (CPIP) model has been established to mimic CRPS-I. The CPIP model has become a well-recognized animal model for studying the mechanisms of CRPS-I. This protocol describes the detailed procedures involved in the establishment of the rat model of CPIP, including anesthesia, followed by ischemia/reperfusion of the hind limb. Characteristics of the rat CPIP model are further evaluated by measuring the mechanical and thermal hypersensitivities of the hind limb as well as the nocifensive responses to acute capsaicin injection. The rat CPIP model exhibits several CRPS-I-like manifestations, including hind limb edema and hyperemia in the early stage after establishment, persistent thermal and mechanical hypersensitivities, and increased nocifensive responses to acute capsaicin injection. These characteristics render it a suitable animal model for further investigation of the mechanisms involved in CRPS-I.

Provided here is a protocol that details steps to establish an animal model of chronic post-ischemia pain (CPIP). This is a well-recognized model mimicking human complex regional pain syndrome type-I. Mechanical and thermal hypersensitivities are further evaluated, as well as capsaicin-induced nocifensive behaviors observed in the CPIP rat model.

ラットにおける複雑な局所疼痛症候群タイプIに対する慢性虚血後疼痛モデル

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical ...

行動学

The Sciatic Nerve Cuffing Model of Neuropathic Pain in Mice

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in injured sensory neurons and along the entire nociceptive pathway within the central nervous system. It is usually chronic and challenging to treat. In order to study neuropathic pain and its treatments, different models have been developed in rodents. These models derive from known etiologies, thus reproducing peripheral nerve injuries, central injuries, and metabolic-, infectious- or chemotherapy-related neuropathies. Murine models of peripheral nerve injury often target the sciatic nerve which is easy to access and allows nociceptive tests on the hind paw. These models rely on a compression and/or a section. Here, the detailed surgery procedure for the &#34;cuff model&#34; of neuropathic pain in mice is described. In this model, a cuff of PE-20 polyethylene tubing of standardized length (2 mm) is unilaterally implanted around the main branch of the sciatic nerve. It induces a long-lasting mechanical allodynia, i.e., a nociceptive response to a normally non-nociceptive stimulus that can be evaluated by using von Frey filaments. Besides the detailed surgery and testing procedures, the interest of this model for the study of neuropathic pain mechanism, for the study of neuropathic pain sensory and anxiodepressive aspects, and for the study of neuropathic pain treatments are also discussed.

Neuropathic pain is a consequence of a lesion or disease affecting the somatosensory system. The &#8220;cuff model&#8221; of neuropathic pain in mice consists of the implantation of a polyethylene cuff around the main branch of the sciatic nerve. Mechanical allodynia is tested using von Frey filaments.

マウスにおける神経因性疼痛の坐骨神経細胞浸潤モデル

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in ...

医学

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain perception in rodents. Withdrawal thresholds can be measured first at baseline and then following drug, venom, injury, allergen,&#160;or otherwise evoked inflammation by applying an accurate force on very specific areas of the skin. An electronic von Frey apparatus allows precise assessment of mouse hind paw withdrawal thresholds that are not limited by the available filament sizes in contrast to classical von Frey measurements. The ease and rapidity of measurements allow for incorporation of assessment of tactile sensitivity outcomes in diverse models of rapid-onset inflammatory and neuropathic pain as multiple measurements can be taken within a short time period. Experimental measurements for individual rodent subjects can be internally controlled against individual baseline responses and exclusion criteria easily established to standardize baseline responses within and across experimental groups. Thus, measurements using an electronic von Frey apparatus represent a useful modification of the well-established classical von Frey filament-based assays for rodent mechanical allodynia that may also be applied to other nonhuman mammalian models.

Electronic von Frey measurements are an effective and improved alternative to classical von Frey filaments, as they allow rapid and precise measurement of changes in rodent mechanical withdrawal thresholds to continuously applied pressure stimuli before and after an inflammatory event.

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain ...

神経科学

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila larvae, customized metal nickel-titanium alloy (nitinol) filaments. These mechanical probes are similar to the von Frey filaments used in vertebrates to measure mechanical nociception. Here, we demonstrate how to make and calibrate these mechanical probes and how to generate a full behavioral dose-response from subthreshold (innocuous or non-noxious range) to suprathreshold (low to high noxious range) stimuli. To demonstrate the utility of the probes, we investigated tissue damage-induced hypersensitivity in Drosophila larvae. Mechanical allodynia (hypersensitivity to a normally innocuous mechanical stimulus) and hyperalgesia (exaggerated responsiveness to a noxious mechanical stimulus) have not yet been established in Drosophila larvae. Using mechanical probes that are normally innocuous or probes that typically elicit an aversive behavior, we found that Drosophila larvae develop mechanical hypersensitization (both allodynia and hyperalgesia) after tissue damage. Thus, the mechanical probes and assay that we illustrate here will likely be important tools to dissect the fundamental molecular/genetic mechanisms of mechanical hypersensitivity.

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

The goal of this protocol is to show how to perform an improved assay for mechanical nociception in Drosophila larvae. We use the assay here to demonstrate that mechanical hypersensitivity (allodynia and hyperalgesia) exists in Drosophila larvae.

ショウジョウバエ幼虫における機械的な結知を測定するための改善されたアッセイとツール

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila ...

Assessment of Morphine-induced Hyperalgesia and Analgesic Tolerance in Mice Using Thermal and Mechanical Nociceptive Modalities

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular mechanisms underlying both phenomena are not well understood and their elucidation should benefit from the study of animal models and from the design of appropriate experimental protocols. 
We describe here a methodological approach for inducing, recording and quantifying morphine-induced hyperalgesia as well as for evidencing analgesic tolerance, using the tail-immersion and tail pressure tests in wild-type mice. As shown in the video, the protocol is divided into five sequential steps. Handling and habituation phases allow a safe determination of the basal nociceptive response of the animals. Chronic morphine administration induces significant hyperalgesia as shown by an increase in both thermal and mechanical sensitivity, whereas the comparison of analgesia time-courses after acute or repeated morphine treatment clearly indicates the development of tolerance manifested by a decline in analgesic response amplitude. This protocol may be similarly adapted to genetically modified mice in order to evaluate the role of individual genes in the modulation of nociception and morphine analgesia. It also provides a model system to investigate the effectiveness of potential therapeutic agents to improve opiate analgesic efficacy.

We describe a protocol to examine the development of opioid-induced hyperalgesia and tolerance in mice. Based on the measurement of thermal and mechanical nociceptive responses of na&#239;ve and morphine-treated animals, it allows to quantify the increase in pain sensitivity (hyperalgesia) and decrease in analgesia (tolerance) associated with chronic opiate administration.

熱的·機械的侵害受容性モダリティを用いて、マウスにおけるモルヒネ誘発痛覚過敏および鎮痛耐性の評価

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular ...

Back Mechanical Sensitivity Assessment in the Rat for Mechanistic Investigation of Chronic Back Pain

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, animal models are needed to examine the mechanisms and effectiveness of novel therapies from a translational perspective. Several rodent models of back pain are used in current investigations. Surprisingly, however, no standardized behavioral test was validated to assess mechanical sensitivity in back pain models. This is critical to confirm that animals with presumed back pain present local hypersensitivity to nociceptive stimuli, and to monitor sensitivity during interventions designed to relieve back pain. The objective of this study is to lay down a simple and accessible test to assess mechanical sensitivity in the back of rats. A test cage was fabricated specifically for this method; length x width x height: 50 x 20 x 7 cm, having a stainless-steel mesh on the top. This test cage allows the application of mechanical stimuli to the back. To perform the test, the back of the animal is shaved in the region of interest, and the test area is marked to repeat the test on different days, as needed. The mechanical threshold is determined with Von Frey filaments applied to the paraspinal muscles, utilizing the up-down method described previously. The positive responses include (1) muscle twitching, (2) arching (back extension), (3) rotation of the neck (4) scratching or licking the back, and (5) escaping. This behavioral test (Back Mechanical Sensitivity (BMS) test) is useful for mechanistic research with rodent models of back pain for the development of therapeutic interventions for the prevention and management of back pain.

To develop novel therapeutic interventions for the prevention and management of back pain, animal models are required to examine the mechanisms and effectiveness of these therapies from a translational perspective. The present protocol describes the BMS test, a standardized method to assess back mechanical sensitivity in the rat.

慢性腰痛の機構的検討のためのラットの背部機械的感受性評価

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, ...

Urokinase-type Plasminogen Activator-induced Mouse Back Pain Model

A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid induction of a persisting back pain model in mice are provided here using an injection of urokinase-type plasminogen activator (urokinase), a serine protease present in humans and other animals. The methodology for induction of persisting lower back pain in mice involves a simple injection of urokinase along the ligamentous insertion region of the lumbar spine. The urokinase inflammatory agent activates plasminogen to plasmin. Typically, the model can be induced within 10 min and hypersensitivity persists for at least 8 weeks.
Hypersensitivity, gait disturbance, and other standard anxiety- and depression-like measures can be tested in the persisting model.&#160;Back pain is the most prevalent type of pain. To improve awareness of back pain, the International Association for the Study of Pain (IASP) named 2021 the &#34;Global Year about Back Pain&#34; and 2022 the &#34;Global Year for Translating Pain Knowledge to Practice.&#34; One limitation of the therapeutic advancement of pain therapeutics is the lack of suitable models for testing persistent and chronic pain. The&#160;features of this model&#160;are suitable for testing potential therapeutics aimed at the reduction of back pain and its ancillary characteristics, contributing to IASP&#39;s naming 2022 as the Global Year for Translating Pain Knowledge to Practice.

Methods for simple, rapid induction of a back pain model in mice are provided here using an intraligament injection of urinary plasminogen activator.

ウロキナーゼ型プラスミノーゲン活性化因子誘発マウス腰痛モデル

A model of persisting lower back pain can be induced in mice with the simple methodology described herein. Step-by-step methods for simple, rapid ...

Assessment of Perigenital Sensitivity and Prostatic Mast Cell Activation in a Mouse Model of Neonatal Maternal Separation

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under the age of 50, yet it is the least understood and studied chronic pelvic pain disorder. A significant subset of patients with chronic pelvic pain report having experienced early life stress or abuse, which can markedly affect the functioning and regulation of the hypothalamic-pituitary-adrenal (HPA) axis. Mast cell activation, which has been shown to be increased in both urine and expressed prostatic secretions of CP/CPPS patients, is partially regulated by downstream activation of the HPA axis. Neonatal maternal separation (NMS) has been used for over two decades to study the outcomes of early life stress in rodent models, including changes in the HPA axis and visceral sensitivity. Here we provide a detailed protocol for using NMS as a preclinical model of CP/CPPS in male C57BL/6 mice. We describe the methodology for performing NMS, assessing perigenital mechanical allodynia, and histological evidence of mast cell activation. We also provide evidence that early psychological stress can have long-lasting effects on the male urogenital system in mice.

We are measuring perigenital mechanical sensitivity and mast cell activation in the prostate of male C57BL/6 mice that underwent an early life stress paradigm &#8211; neonatal maternal separation, in order to induce a preclinical model of chronic prostatitis/chronic pelvic pain syndrome.

新生児母子分離のマウスモデルにおけるPerigenital感度と前立腺肥満細胞の活性化の評価

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under the ...

Transurethral Instillation Procedure in Adult Male Mouse

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into the urinary bladder to either induce animal models of bladder pathologies or evaluate the effectiveness of intravesical treatments. Most rodent models of lower urinary tract (LUT) pathologies are induced in female mice due to ease of intravesical instillation of the substances via the female urethra. However, due to anatomical differences between the female and male LUT, transurethral instillation in a male mouse has been deemed a very challenging procedure, and it has not been previously described. In this manuscript, we provide a detailed description of how to prepare polyethylene (PE) tubing for subsequent insertion into the urethra of a male mouse. In addition, we discuss the ideal types of PE tubing to be used depending on the desired site of inoculation. Furthermore, we describe point by point how to prepare an animal for a successful transurethral instillation to avoid injury to the urethra and ensure the delivery of the solution to the desired location. The procedure is started by retracting the prepuce and the glans to expose the opening of the urethral meatus. Next, the glans are grasped by blunt non-crushing forceps to stabilize the penis and the PE tubing. The PE tubing is first inserted into the urethral meatus parallel to the animal body, then its angle is adjusted by tilting the catheter to maneuver it to follow the natural curvature of the urethra. This technique can be used to induced murine models of bladder pathologies and/or evaluate the effectiveness of intravesical treatments in male mice.

Transurethral instillation is a challenging procedure and has not been well described in the literature. The aim of this manuscript is to describe a technique for transurethral insertion of a catheter for intravesical delivery of liquids with active substances into the urinary bladder and/or prostate in adult male mice.

大人の雄のマウスに経尿道的注入手順

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into ...

免疫・感染学

T. evansi感染動物における疼痛および異痛症の評価

Electronic von Frey associated with the Mouse Grimace Scale to Assess Allodynia and Pain in Trypanosoma evansi-Infected Mice

Infectious disease pathogenesis is still a complex field to study. The course of several clinical signs, such as allodynia and pain, may be observed in domestic animals. However, the knowledge of their pathways and correct treatment need controlled experiments, many of them using laboratory animals. Measuring changes in mechanical thresholds of the hind paw and viscera is a useful technique to observe changes in pain perception in rodents. Withdrawal response can be measured first in baseline tests, which creates better control of experimental groups. Subsequent tests can be performed after inducing infection and adding drugs to the protocol. The use of an electronic von Frey apparatus associated with the use of a facial scale to observe pain-like changes allows a simple, precise, and consistent assessment to evaluate allodynia and pain in mice. Thus, experiments using the present methodology for Trypanosoma evansi infection represent a useful method to evaluate allodynia and pain in laboratory-infected animals, which can be applied to the conventional treatment for livestock animals.

著者スポットライト:トリパノソーマエバンシに感染した実験動物の痛みの評価と動物福祉の促進

Here, we present a protocol to evaluate nociception in mice infected with Trypanosoma evansi, using the electronic von Frey apparatus as a tool, measuring mechanical thresholds of the hind paw and viscera.

トリパノソーマ・エバンシ感染マウスにおけるアロディニアと疼痛を評価するためのマウスしかめっ面尺度に関連する電子フォン・フレイ

Infectious disease pathogenesis is still a complex field to study. The course of several clinical signs, such as allodynia and pain, may be observed in ...