Name: measurement of tactile allodynia in a murine model of bacterial prostatitis
Uploaded: 2013-01-16T13:00:00.000+00:00
Duration: 6 min 46 s
Description: Watch this Scientific Journal Video about Measurement of Tactile Allodynia in a Murine Model of Bacterial Prostatitis at JoVE.com

Bu araştırma NIH / NIDDK hibe 1K01DK079019A2 (PT) tarafından desteklenmiştir.

1. Fare Aşılama için bakteri Hazırlık Aşağıdaki aseptik koşullar altında yapılmalıdır. <ol><li> Kapakları biri 17 x 100 mm polipropilen tüpler atın ve taze Luria broth (LB) medya 3 ml ekleyin. Otoklavlanmış ipuçlarını kullanarak, gerginlik CP1 2 bazı dondurulmuş bakteri gliserol stok almak ve LB ortamı içeren tüpe kültürünün küçük bir miktar transfer. Oksijen yine tüp girebilir, böylece tüp kapak değiştirin - kültür aerobik koşullar altında büyüyebilir gerekmektedir. 37 Sıra tüp ° gecede 220 rpm&#39;de bir sallayarak inkübatör C. </li><li> Ertesi gün, 3 ml LB ortamı ile yeni bir tüp vermek üzere gece boyunca kültür 5 ul aktarmak ve 37 ° C &#39;de bir gece boyunca kuluçka makinesi içinde statik koşullar altında büyüyebilir </li><li> Günde aa 50 ml tüp içinde 3 taşıma kültürün 40 ul, 40 ml LB ortamı her içeren. 37 ° C sabit inkübatör gecede yerleştirin. </li><li> Açın ve 4 ° C&#39;ye santrifüj ayarlayınBir kez santrifüj son sıcaklığa ulaştığını, 40 ml Nalgene santrifüj tüpüne her 40 ml kültürü aktarmak. </li><li> Gerektiğinde LB medya ile ses seviyesini ayarlamak - santrifüj dengelemek için tüpleri tartılır. 20 dakika boyunca 6000 rpm&#39;de tüpler Spin. </li><li> Dikkatlice süpernatant kaldırmak ve yavaşça 40 ml steril PBS bakteriyel pelet askıya. </li><li> PBS ile adım 1.5 tekrarlayın. </li><li> Süpernatantı kapalı aspire ve 500 ul steril PBS içinde bakteri askıya. 1.5 ml mikrofuge&#39;de tüpüne çözümü aktarın. Süspansiyonu 10 ul alın ve buz gibi soğuk PBS 990 ul seyreltin. Aşılama için gerekli hacim belirlemek için OD 420 nm okumak için bir spektrofotometre kullanın. Uygun miktarda belirlemek için buz gibi soğuk PBS içinde pelet askıya: OD 420 nm sayısı (ml) alın ve bakteriyel pelet [ex tahmini hacmi çıkarın. OD 420 nm = 0.545, pelet yaklaşık 0.075 ml. 0,545-0,075 = 0,470]. </li><li> 10. Bakteriyel süspansiyon ul ve di kaldırma990 ul buz gibi soğuk PBS içinde ud. OD 420 nm okuyun. Hedefi 1.000 seyreltilmiş süspansiyonu ± .010 bir OD 420 nm değeri elde etmektir. Sizin seyreltme sayısı bu hedefin üzerinde ise, süspansiyon daha fazla hacim kazandırmak ve başka bir okuma almak. Numarası altında ise 0,990 ardından istediğiniz okuma elde edilinceye kadar erteleme ve tekrar OD 420 nm okuma aşağı spin. </li></ol> 2. Hayvan Enfeksiyon Fareler (5-7 haftalık, her grupta on) Jackson Laboratory (Bar Harbor, ME) &#39;den satın alındı. <ol><li> Fare bir pleksiglas odasında izofluran% 1-4 solunması ile anestezi ve yatık kadar izlenir. </li><li> Bir seferde bir fare bir cam Hamilton şırınga (1.5-2.0 cm uzunluğunda) değiştirilmiş 30 G iğne takılı bir polietilen (PE10) kateter ile kateterizasyon ile bakterilerin instilasyonu için alınır. Kateter iğne merkeze yerleştirilir. Fare Plac olduğunued anestezi altında dorsal yüzeyi üzerinde bir burun konisi kullanılarak yapılmaktadır. </li><li> Fare penis penil üretra girişinde yağlanması için kullanılan hafif basınç ve pamuklu çubuk üzerine yağ bolca tarafından çekilmektedir. </li><li> 10 ul fosfat-tamponlu 1 x 10 8 bakteri içeren tuzlu kateterizasyon anestetize farelerin üretra içine katılmaktadır. </li><li> Bakteri bağlanma sağlamak için acil idrara çıkma ve önlemek için bakteri giriş sonra fareler pleksiglas bölme içinde 15 dakika süreyle bir anestezi altındaki bir durumda muhafaza edilir. </li><li> Fare onların tekrar kafeslerine yerleştirildi ve sonraki 24 saat boyunca izlenir. </li></ol> 3. Davranış Testi Fare öncesinde enfeksiyon (bazal) ve gün 3, 7, 14, 21 ve enfeksiyon sonrası 28 üzerinde test edildi. Referans hiperaljezi ve dokunsal allodini karın 11,12 ve plantar r uygulanan von Frey filamentleri kullanılarak test edildiarka pençe 13 egion. Test-saati günlük sabit bir de perfomed edildi, standart metodoloji ve tüm hayvanların tek deneyci testi kullanılmıştır. Grupların Kör test hayvan modellerinde davranış bazlı test ağrı ile ilgili sınırlamalar ile mücadele için kullanılmıştır. 0.04 arasında kuvvetler, 0.16, 0.4, 1.0 ve 4.0 g (Stoelting, ABD) ile beş ayrı von Frey filamentlerin çekilme yanıtların karın ve frekans uygulanmıştır hesaplanmıştır. <ol><li> Alışma sürecinden bir 30 dakika süre sonra, fareler bir paslanmaz çelik tel ızgara taban ile evde yapılan bireysel pleksiglas odacığı (6 x 10 x12 cm) yerleştirilmiştir. </li><li> Referans hiperaljezi ve dokunsal allodini beş von Frey filamentleri kullanılarak test edildi. Her filament toplam 10 kez 5 sn bir uyarıcılar arası süre ile 1-2 saniye uygulandı ve tüyleri kuvvet artan düzende test edildi. </li><li> Her bir fılaman, prostat Genel çevresinde alt karın bölgesine uygulanır vebakım duyarsızlaştırma önlemek veya etkilerini &quot;rüzgar&quot; Bu bölgedeki farklı alanlarda teşvik götürüldü. Filamentler 10 kez toplam uyarımlar arasında en az 5 saniye aralıklarla 1-2 saniye için kuvvet artan sırada uygulanmıştır. </li><li> Davranış üç tip filament stimülasyon olumlu bir yanıt olarak kabul edilir: karın 1) keskin retraksiyonu; 2) hemen yalama veya filament stimülasyon alan kazıma; 3) atlama. </li><li> Tepki frekansı olumlu yanıt yüzdesi (10 üzerinden, 10 =% 50 örneğin 5 karşılıklar) olarak hesaplandı ve data cevabı frekans ortalama yüzdesi ± SE olarak rapor edildi </li><li> 25&#39;den fazla pozitif toplam taban yanıtları Hayvanlar çalışmanın dışında tutulmuştur. </li><li> Dokunsal allodini 0.04 güçleri, 0.16, 0.4, 1.0 ve 4.0 g ile von Frey filamentleri kullanılarak arka pençenin plantar bölgeye üzerinde test edildi. Medyan% 50 geri çekme eşiği (5) yukarı-aşağı tuşlarını kullanarak değerlendirildifilaman hafif bir açı kadar test 0.04 g filaman ile başlatılan yöntem arka pençenin plantar yüzeye dik olarak uygulanır. Filament olumlu bir yanıt gözlenmiştir kadar artan sırayla test edildi. Filament pozitif tepki testi pençe pençe ya yalama keskin bir geri çekilme ya olarak tanımlandı. Olumlu bir yanıt kaydedildi zaman bir sonraki daha zayıf filaman uygulandı, ve bir negatif cevap gözlendi, sonra bir sonraki daha güçlü bir filaman uygulanmıştır. </li><li> Açıklanan deneyler ve metodoloji incelenen ve onaylanan Northwestern Üniversitesi hayvan bakımı ile ve komite kullanabilirsiniz edilmiştir. </li></ol>

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Çıkar çatışması ilan etti.

UPEC ile fare prostat enfeksiyonu bakteriyel prostatit, CP / CPPS patogenezinde veya kronik inflamasyon bir predispozan olay olarak dahil olabilir olayların in vivo modelleme için sağlar. Yöntemler kadın idrar yolu enfeksiyonu 7,14 in UPEC modellerde edebiyat büyük bir gövde üzerine bakteriyel hazırlanması ve instillasyon beraberlik için nitelendirdi. Modeli potansiyel aşı adayları ve immün modülasyon mekanizmalarının test patogenezinde çalışmak için geniş uygulanabilirliği v...

<table border="1"><tbody><tr><td> Reaktif Adı </td><td> Şirket </td><td> Katalog numarası </td></tr><tr><td> Kültür tüpleri </td><td> BD Falcon </td><td> 352059 </td></tr><tr><td> LB Broth Miller </td><td> EMD </td><td> EM1.10285.0500 </td></tr><tr><td> Nalgene Santrifüj Tüpleri </td><td> Thermo </td><td> 3118-0050 </td></tr><tr><td> İzofluran </td><td> Butler Schein </td><td> NDC 11695-6776-1 </td></tr><tr><td> Kateter iğne 30G </td><td> Hamilton </td><td> 91030 </td></tr><tr><td> PE10 boru </td><td> BD intramedic </td><td> 427400 </td></tr><tr><td> Von Frey filamentler </td><td> Stoelting </td><td> 58025-31 </td></tr></tbody></table>

measurement of tactile allodynia in a murine model of bacterial prostatitis

Üropatojenik Escherichia coli (UPEC) idrar yolu enfeksiyonları ve bakteriyel prostatit 1 önemli bir rol oynamaktadır patojenlerdir. Biz son zamanlarda UPEC kronik pelvik ağrı 2, Kronik prostatit / kronik pelvik ağrı sendromu (CP / CPPS) 3,4 bir özelliği başlatılmasında önemli bir role sahip olduğunu göstermiştir. Klinik UPEC ile prostat enfeksiyonu doku hasarı ve otoimmünite 5 mekanizmalarının başlatılması içerebilir mekanizmalar yoluyla kronik ağrı başlatmak ve kurabilir. Prostat UPEC patogenezini anlamak için bir meydan okuma manipülasyon prostat bezinin nispi erişilememesi olduğunu. Böylece de prostat bir artan enfeksiyon oluşturma erkek farelere UPEC bir klinik suşu vermek üzere, daha önce anlatılan intraüretral enfeksiyon yöntemiyle 6 kullanılmıştır. Burada, biz ba standartlaştırılması için protokoller tarifgibi bakteri perfüze anestetize erkek fareler catheterizing için işlem olarak cterial inokulum 7. CP / CPPS öncelikle dokunsal allodini 4 varlığı ile karakterizedir. Davranış testleri ifade viseral ağrı 8-10 kaynaklanan kutanöz hiperaljezi kavramına dayanmaktadır. Visseral dokularda bir irritabl odak normal olmayan ağrılı uyaranlar (allodini) için abartılı bir tepki veren Kutanöz ağrı eşikleri azaltır. Yatan visseral ağrının şiddeti ile artış eğilimindedir anormal yanıtları cilt sonucu normal kuvvet uygulanması. Bu UPEC bakteri ile bir tek enfeksiyonuna yanıt olarak zaman içinde dokunsal allodini ölçmek için von Frey lifler kullanmak NOD / ShiLtJ farelerde yöntemi açıklanmaktadır.

<html> Biz bakteriyel enfeksiyon üzerine kronik ağrı varlığı için NOD / ShiLtj ve C57BL/6J erkek farelerde incelenmiştir. Erkek fareler (5-7 haftalık) üretra (Şekil 1) içine salin ya da bakteri ile damlatıldı edildi. Salin-coşturup C57BL/6J fareler ve UPEC ile enfekte C57BL/6J farelerin von Frey filamentleri ile pelvik bölgeye mekanik uyarılması deney 28 günlük kurs (Şekil 1) sırasında değişmedi bir tepki frekansı sonuçlandı. Buna karşılık, UPEC-enfekte NOD...

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

Prostat enfeksiyonu kronik prostatit pelvik ağrı arabuluculuk katkıda bulunan bir faktör olabilir. Biz standardize edilmiş bakteriyel inokulum, zamanla farelerde dokunsal allodini ölçmek için erkek farelerde ve metodoloji üretra içine bakteri instilasyon hazırlama prosedürü açıklanmaktadır.

Bakteriyel Prostatit bir fare modelinde Dokunsal allodini Ölçümü

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 Görüntüleme.  Northwestern University Feinberg School of Medicine. Prostat enfeksiyonu kronik prostatit pelvik ağrı arabuluculuk katkıda bulunan bir faktör olabilir. Biz standardize edilmiş bakteriyel inokulum, zamanla farelerde dokunsal allodini ölçmek için erkek farelerde ve metodoloji üretra içine bakteri instilasyon hazırlama prosedürü açıklanmaktadır.

Video: 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.

Sıçanlarda Kompleks Bölgesel Ağrı Sendromu Tip-I kronik Post-Iskemi Ağrı Modeli

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

Davranış

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.

Farelerde Nöropatik Ağrı Siyatik Sinir cuffing Modeli

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

Tıp

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 ...

Nörobilim

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.

Drosophila Larvalarında Mekanik Nosiception Ölçümü için Geliştirilmiş Bir Tahlil ve Araçlar

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.

Termal ve Mekanik nosiseptik Usuller kullanma Farelerde Morfin kaynaklı Hiperaljezi ve Analjezik Hoşgörü Değerlendirilmesi

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.

Kronik Sırt Ağrısının Mekanik Araştırılması için Sıçanda Geri Mekanik Duyarlılık Değerlendirmesi

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.

Ürokinaz tipi Plazminojen Aktivatörü ile İndüklenen Fare Sırt Ağrısı Modeli

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.

Yenidoğan Anneden Ayrılma Fare Modelinde perigenital Duyarlılık ve Prostat Mast Hücre aktivasyonu Değerlendirilmesi

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.

Yetişkin erkek fare yordamda transüretral korumak

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

İmmünoloji ve Enfeksiyon

T. evansi ile Enfekte Hayvanlarda Ağrı ve Allodininin Değerlendirilmesi

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.

Yazar Spotlight: Trypanosoma evansi ile Enfekte Laboratuvar Hayvanlarında Ağrının Değerlendirilmesi ve Hayvan Refahının Teşvik Edilmesi

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.

Elektronik von Frey, Trypanosoma evansi ile Enfekte Farelerde Allodini ve Ağrıyı Değerlendirmek için Fare Yüz Buruşturma Ölçeği ile ilişkili

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 ...