Yazarlar Bennett ve Xie (1988) tarafından CCİ modelin orijinal tanımı kabul etmek istiyorum. Bu çalışma Bilim ve GMT Tıbbi Araştırma Ulusal Sağlık ve Avustralya Tıbbi Araştırma Konseyi (ID # 568637) ve NSW hibe tarafından kısmen desteklenmiştir.

1. Siyatik Sinir Kronik daraltma Aseptik teknikler cerrahi işlem için kullanılmalıdır. % 70 etanol ile cerrahi iş yüzeyi dezenfekte edin ve otoklavlayarak önceden steril aletler (örn., ince makas, forseps ve retraktör), tüller, zımba ve bezlerden de hazırlar. Çoklu operasyonlar için, temiz ve% 70 etanol ya da sıçanlar arasında kuru bir boncuk sterilizanlar ile resterilise aletleri. Bir cerrahi maske, saç kaporta ve steril eldiven giyilmelidir. Sinir daralma için, kurumasını önlemek üzere, steril salin batırılır yaklaşık 3 cm uzunlukta küçük parçalar halinde kesilmesi ile kromik gut kenet hazırlanır. Yaranın kapatılması için, piyasada bulunan steril ipek dikişlerle kullanın. <ol><li> O 2 izofluran% 5 kullanan bir indüksiyon odasında sıçan Anaesthetise, sonra yüz maskesi yapılmış özel bir ile O 2 izofluran% 2 sunmak, ve işlem sırasında anestezi hayvan izlemek. Ortak alternatif anestezik halotan veya intraperitoneal sodyum pentobarbital teneffüs edilir. Bu reseptörler nöropatik ağrı geliştirilmesi önemlidir, çünkü ketamin, bir N-metil-D-aspartat (NMDA) reseptör antagonisti olarak, kaçınılmalıdır. </li><li> Sıçan sol arka bacak Tıraş. Sonra, 37 bir termo-regüle ısıtma hasır üzerine hayvan yere ° C ve gözlere yağlama oftalmik merhem uygulayın. </li><li> % 70 izopropil alkol ve iyot solüsyonu üç alternatif uygulamalarında traş alanı sterilize. </li><li> Sıçan onun göğüs / göğüs yatarken ile, sol arka bacak yükseltmek ve ayak maskeleme bandı kullanarak omurga 90 ° femor ile konumda tutun. </li><li> Bir deri paralel insizyon, ama femor altında 3-4 mm ve bağ dokusu ile keserek kesi çevreleyen kas ücretsiz cilt olun. </li><li> Gluteus superficialis ve pazı arasındaki bağ dokusu kesti künt makas, kullanmafemoris kasları. </li><li> Siyatik sinir açıkça görüntülenmesini sağlayan kaslar arasındaki farkın artırılmasına bir retraktörü kullanın. </li><li> Eğri künt uçlu forseps ve mikro-makas, çevresindeki bağ dokusu ile siyatik sinir yavaşça serbest yaklaşık 10 mm (siyatik trifurkasyon proksimalinde) kullanma. </li><li> Mikroskop ve iyi bir ışık kaynağı altında, bir çift düğüm ile siyatik sinir trifurkasyon proksimalinde dışında 1 mm, dört bitişik harfler (kromik katgüt 4.0, Ethicon, ABD) kravat. Her bağ, bir tek gevşek bir döngü başlar, sonra da döngü yakın iki ucu kavramak ve döngü sadece ancak rahat ve bağ sinir boyunca kaymıyorsa kadar sıkın. Kendi doğru pozisyonda döngü tutmak için, düğüm tamamlamak için ilk üstüne ikinci bir döngü koyun. Son olarak, 1mm civarına dikilen ve gevşek uçları kesilir. Sinir daralma en az olması ve kısa bir seğirme görülür ise derhal durdurulması, th tutuklamak önlemek için gerekene epinöral kan akımı. Ligatürleri sıkma fazla aksotomi yol açar ve autotomy (kendini yaralama), başarılı bir ağrı aşırı duyarlılık testlerine engel hem de istenmeyen yan etkiler. </li><li> Cildi sıkıştırmak için kas tabakası ve zımba (AutoCLIP, 9 mm) kapatmak için dikiş (Mersilk 5.0, Ethicon) kullanın. Sonra, yara sterilize etmek (Riodine) iyot çözeltisi kullanın. </li><li> Boğulma bilinçsiz hayvan önlemek için, anestezi iyileşme döneminde yakından sıçan dikkat edin, düz kağıt yataklar (değil standart hayvancılık yatak) ile ayrı bir kafese kurtarmak için izin verir. Herhangi bir post-operatif analjezi araştırmacılar vermeden önce bu ağrı aşırı duyarlılık testleri etkileyebilir gerçeğini göz önünde bulundurmalıdır. Aslında, yaralanma sırasında anestezi, lidokain, lokal uygulama nöropatik ağrı gelişimi, değiştirebilir ve önceki hafta 7 hiperaljezi derecesini azaltabilir dair kanıtlar vardır. Bu m daha fazla tartışma içinBennett ve arkadaşları 8 tarafından gözden bakın ardıl. </li></ol> 2. Ağrı Davranışları ölçümleri <ol><li> Deney başlamadan önce birkaç gün süreyle sıçanlar Günlük kullanım, onların işleyicisi sıçan alıştırmak stres ve ısırma riskini azaltmak için önerilmektedir. </li><li> Davranış testinden önce, sıçanlarda test prosedürü ve hayvan muhafaza etmek habitüe gerekir. Test ortamı sessiz kaldı ve iyi sabit sıcaklık ve nem düzeyleri ile kontrol ve gün benzer bir zaman her test oturumunda yapılmalıdır. </li><li> Ağrı aşırı duyarlılık testi istediğiniz gibi, CCI (bazal) öncesi ve CCİ sonra farklı zaman noktalarında birkaç kez yapılmalıdır. </li></ol> 3. Mekanik Çekme Eşik <ol><li> Test gününde, yeri fareler artmış örgü katlı test kafesin içine çekilmesi eşikleri ölçülerek 15-30 dakika önce, kafes arama ve groo izinming faaliyetleri durdurma ve sıçanların rahat ve sabit olmak. </li><li> Doğrudan sıçanın sol arka ayağını orta plantar yüzeyinin altında bir dinamik taban von Frey aesthesiometer (Ugo Basile, İtalya), pozisyon filamanlı dokunmatik stimülatör ünitesi kullanarak metal örgü kaçınarak, ve start tuşuna basın. Bu mekanik olarak artan bir kuvvet ile sıçanın sol arka ayağını plantar uyarmak için filament kaldırır. </li></ol> Manuel von Frey tüyleri cilde uygulandığında belirli bir kuvveti teşvik için kalibre çapında filamentler artan bir dizi vardır. Bunlar pençe çekme eşiği, daha önce tarif 4 ölçülmesi için bir elektronik dinamik taban von Frey aesthesiometer alternatif olarak kullanılabilir. Elektronik von Frey aesthesiometers (örneğin IITC, ABD ve Somedic, İsveç) birkaç alternatif üreticileri de vardır. <ol start="3"><li> Yaralı ve elin hem hindpa gelen mekanik çekme eşiği kaydetmekws, bir denge için. Bu cihaz, pençe için basınç uygulayan ve geri çekilmesi üzerine reaksiyon sistemi otomatik kaydeder ve (gram cinsinden) uygulanan maksimum kuvvet gösterir. Belirsiz bir yanıt (veya ambulasyon tüylerini temizleme durumunda örn) oluşursa, uyaran tekrar tekrar eşik kaydedebilir. Yalama veya sıçanın sol arka ayağını aşağıdaki stimülasyon sallayarak ağrı ile ilişkili bir davranış işaretidir. </li><li> Mekanik stimülasyonu uyaranlara arasında yaklaşık 5 dakika arayla 3 kez tekrarlayın, eşikler kaydetmek ve sonra pençe çekme eşikleri ortalamasını hesaplamak. </li></ol> 4. Termal Çekilme Gecikme Plantar test cihazı ile kızılötesi (IR) ısı yoğunluğu deney başlamadan önce ayarlanmalıdır. Laboratuarımızda, biz naif sıçanlarda yaklaşık 10 saniye ortalama bir pençe çekme gecikme vermek IR yoğunluğunu ayarlayın ve doku zedelenmesine önlemek için 20 saniye kesme gecikme setBir pençe kaldırmak için arıza durumunda yaşı. <ol><li> Test gününde, bir yerde 15-30 dakika gecikme ölçümünden önce cam tabanlı test kafesin içine sıçan, kafes arama izin ve durdurma faaliyetleri damat ve sıçanların rahat ve sabit olmak. </li><li> Doğrudan sıçanın sol arka ayağını orta plantar yüzeyinin altında bir termal plantar analjezi cihazı (Ugo Basile, İtalya), pozisyon kızılötesi kaynağı kullanarak, ve start tuşuna basın. Bu şeffaf bir cam yüzeyi sayesinde mükemmel bir ısı ışın sıçanın sol arka ayağını plantar ortaya koyar. Termal plantar analjezi aletler (örn. IITC, ABD ve Stoelting, ABD) alternatif üreticileri de vardır. </li><li> Bir denge için, yaralı ve zarar görmemiş hem hindpaws çekilme gecikme kaydedin. Bu cihaz otomatik olarak ısı kaynağından pençe kesilmesine termal uyaranın başlangıcından (saniyede) geçen süreyi kaydeder. Belirsiz bir yanıtı ortaya çıkarsa (ambula durumunda örnvermekte veya damat), uyarıcı tekrar ve tekrar gecikme kaydedin. Yalama veya sıçanın sol arka ayağını aşağıdaki stimülasyon sallayarak ağrı ile ilişkili bir davranış işaretidir. </li><li> Isı stimülasyonu uyaranlara arasında yaklaşık 5 dakika ara ile en az 3 kez tekrarlayın, gecikmeleri kaydetmek ve sonra pençe çekme gecikmeleri ortalamasını hesaplamak. </li></ol> 4. Temsilcisi Sonuçlar Wistar sıçanların (yaralanmamış) ipsilateral (yaralı) ve kontralateral pençeleri mekanik çekme eşiği ve termal çekilme gecikme bir örnek, daha önce gösterilmeyen (bazal) ve 2-12 gün Şekil 3A CCİ ve Şekil 3B sonra. Edilir Bu rakamlara bakıldığında CCİ, mekanik ve termal ağrı hipersensitivite varlığını gösteren, mekanik çekme eşikleri ve ipsilateral sıçanın sol arka ayağını termal çekilmesi gecikmeleri hem de önemli bir düşüş neden olduğu görülebilir kronik nöropatik ağrı hem de özellikleri<html> . <img alt="Şekil 1" src="/files/ftp_upload/3393/3393fig1.jpg" /> Şekil 1. CC modelinde ligatürü konumlandırma açıklayan şematik bir resimdir. Nöropatik ağrı, bu hayvan modelinde orta uyluk düzeyinde siyatik sinir dört gevşek kromik-bağırsak bitişik harfler, yerleştirme içerir. Bu sıçanın sol arka ayağını ayağı ve bunu takip eden ağrı aşırı duyarlılık olacak afferentleri sadece bir kısmını yaralanmasına neden olur. <img alt="Şekil 2" src="/files/ftp_upload/3393/3393fig2.jpg" /> Şekil 2. Ağrı testi için kullanılan sıçanın sol arka ayağını alanının Diyagramı. Olarak kırmızı noktalı bir daire ile gösterilen ayak yastıklarından sadece posterior orta plantar, ağrı, test için kullanılır. Bu bölge siyatik sinirin terminal dalları çoğunlukla innerve edilir. Topuk ve ayak pedleri diferansiyel cilt hassasiyeti nedeniyle kaçınılmalıdır. &quot;3.jpg /&gt; Sıçanlarda siyatik sinirin kronik daralma sonrasında Şekil 3. Ağrı aşırı duyarlılık. CC sonra mekanik uyarıcılar (A) ve (B) yaralı termal uyarılara pençe çekme gecikmesi (ipsilateral) ile pençe çekme eşiği önemli bir azalma elin (taraf) sıçanın sol arka ayağını (P &lt;0.01 tekrar ölçümlü 2 ile karşılaştırıldığında vardır sıçanın sol arka ayağını olduğu yönlü ANOVA). Veriler ortalama göstermek ± sem, n = 7 Wistar sıçan.

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Hayvanlar üzerinde deneyler New South Wales, Avustralya Üniversitesi Hayvan Bakım ve Etik Komitesi tarafından onaylandı ve Ağrı Çalışmaları Uluslararası Birliği tarafından yayınlanan izledi.

CCİ kronik nöropatik ağrı yaygın olarak kullanılan periferik sinir yaralanması modelidir. Bunu gerçekleştirmek için oldukça basit ve yaralanma sonrası en az bir ay için sağlam ve kararlı bir ağrı aşırı duyarlılık üretir. CCİ takiben sıçan yaralı sıçanın sol arka ayağını (parmakları bir arada tutan ve plantar fleksiyon ve pençe everte) yanı sıra, sallayarak koruma ve spontan ağrı 9 varlığını düşündüren yaralı sıçanın sol arka ayağını yalamaya tekrarlanan an...

<table border="1"><tbody><tr><td> Reaktif / ekipman Adı </td><td> Şirket </td><td> Katalog numarası </td><td> Yorumlar </td></tr><tr><td> Kromik gut </td><td> Ethicon </td><td> G-211 </td><td> 4-0 kalınlığı </td></tr><tr><td> Izofluran </td><td> Delvet Pty Ltd, Seven Hills, NSW </td><td> n / a </td><td></td></tr><tr><td> Mersilk </td><td> Ethicon </td><td> W-580 </td><td> 5-0 kalınlığı </td></tr><tr><td> AutoCLIP </td><td> Becton Dickinson </td><td> 427631 </td><td> 9 mm paslanmaz çelik </td></tr><tr><td> Riodine </td><td> Orion </td><td> R1000802F </td><td> % 1 w / v iyot </td></tr><tr><td> Termal plantar analjezi cihazı </td><td> Ugo Basile </td><td> 37370 </td><td></td></tr><tr><td> Dinamik plantar aesthesiometer </td><td> Ugo Basile </td><td> 37400 </td><td></td></tr></tbody></table>

chronic constriction of the sciatic nerve and pain hypersensitivity testing in rats

Kronik nöropatik ağrı, merkezi ya da periferal sinir sisteminin hasar görmesi sonucu, nüfusun 1,2% 7-18 etkileyen, yaygın bir ve zayıflatıcı bir durumdur. Belirtileri spontan (gibi karıncalanma, yanma, elektrik çarpması) ağrı, disestezi, parestezi, allodini (ağrı normal olmayan ağrılı uyaranlar kaynaklanan) ve hiperaljezi (ağrılı uyaranlara artmış yanıt) içerir. Duysal semptomlar uykusuzluk ve depresyon gibi davranışsal özürlü ile komorbid bulunmaktadır. Kronik nöropatik ağrı çalışmak için periferik sinir yaralanması taklit eden birçok hayvan modellerinde geliştirilmiş olup, en yaygın olarak kullanılan bir Bennett ve Xie (1988) tek taraflı siyatik sinir kronik daralma yaralanma (CCI) 3 (Şekil 1). Burada CC ve test ağrısı aşırı duyarlılık gerçekleştirmek için bir yöntem sunulmaktadır. CCİ olarak yaparak maruz kalan bir tarafta siyatik sinir ile, anestezi altında yapılırkin insizyon ve gluteus süperfisiyalis ve biseps femoris kaslar arasındaki bağ dokusu ile kesme. Dört kromik katgüt Ligatürleri sadece kapamak için, 1 mm aralıklarla siyatik sinir etrafındaki gevşek bağlanmış ama epinöral kan akışını tutuklama değildir. Yara deri kas ve mahsullerindeki dikişler ile kapatılır. Hayvan daha sonra ağrı aşırı duyarlılık testleri başlamadan önce 24 saat için ameliyatın izin verilir. Davranış testi için, sıçanlarda test cihazları yerleştirilir ve test prosedürü alışmakta izin verilir. Test alanı siyatik sinir dağılımında içinde düşer sıçanın sol arka ayağını orta plantar (Şekil 2) &#39;dir. Mekanik çekme eşiği mekanik elektronik bir dinamik taban von Frey aesthesiometer veya manuel von Frey kıllar 4 kullanarak yaralı ve yaralanmamış hindpaws hem de uyararak değerlendirilir. Mekanik bir geri çekme eşiği uygulanan yüksek basınç(Gram cinsinden) pençe çekme tetikler. İlk Hargreaves ve ark (1988) tarafından açıklanan ısıl çekmede gecikme, ölçülmesi için, sıçanın sol arka ayağını bir plantar analjezi metre 5,6 kullanarak şeffaf bir cam yüzeyi sayesinde mükemmel bir ısı ışınına maruz kalmaktadır. Isı uyarana çekilme gecikme yaralı ve yaralanmamış hindpaws hem pençe çekme için zaman olarak kaydedilir. Aşağıdaki CC, yaralı pençede mekanik çekme eşiği, hem de ısı geri çekme gecikmesi bazal ölçümler ve elin pençe (Şekil 3) ile karşılaştırıldığında, gerek önemli ölçüde azalır. Ağrı duyarlılık testi ile birlikte periferik sinir hasarı CCİ modeli kronik nöropatik ağrı değiştirmek için potansiyel terapötik ajanların etkinliğini araştırmak için bir model sistemi sağlar. Laboratuarımızda, biz patogenezinde nöro-immün etkileşimlerin rolünü araştırmak için hindpaws termal ve mekanik hassasiyet yanında CCİ kullanmaknöropatik ağrı ve tedavisi.

Watch this Scientific Journal Video about Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats at JoVE.com

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Cerrahi basitlik ve sağlam davranışsal sonuçları nedeniyle, siyatik sinirin kronik daralma nöropatik ağrı seçkin hayvan modelleri biridir. Ameliyat sonrası 24 saat içinde, ağrı aşırı duyarlılık kurulan ve kantitatif bir von Frey aesthesiometer (mekanik test) ve plantar analjezi metre (termal testi) kullanılarak ölçülebilir.

Siyatik Sinir Sıçanlarda Ağrı aşırı duyarlılık Test Kronik daraltma

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

chronic-constriction-sciatic-nerve-pain-hypersensitivity-testing

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

58.6K Görüntüleme.  University of New South Wales. Cerrahi basitlik ve sağlam davranışsal sonuçları nedeniyle, siyatik sinirin kronik daralma nöropatik ağrı seçkin hayvan modelleri biridir. Ameliyat sonrası 24 saat içinde, ağrı aşırı duyarlılık kurulan ve kantitatif bir von Frey aesthesiometer (mekanik test) ve plantar analjezi metre (termal testi) kullanılarak ölçülebilir.

Video: Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

Sıçanlarda Spinal Kord Yaralanma sonrası Akut ve Kronik Dokunsal Duyu Testi

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

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Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart failure. Here, we describe a detailed surgical procedure for creating pressure overload and cardiac hypertrophy in rats by constriction of the ascending aorta using a small metallic clip. After anesthesia, the trachea is intubated by inserting a cannula through a half way incision made between two cartilage rings of trachea. Then a skin incision is made at the level of the second intercostal space on the left chest wall and muscle layers are cleared to locate the ascending portion of aorta. The ascending aorta is constricted to 50&#8211;60% of its original diameter by application of a small sized titanium clip. Following aortic constriction, the second and third ribs are approximated with prolene sutures. The tracheal cannula is removed once spontaneous breathing was re-established. The animal is allowed to recover on the heating pad by gradually lowering anesthesia. The intensity of pressure overload created by constriction of the ascending aorta is determined by recording the pressure gradient using trans-thoracic two dimensional Doppler-echocardiography. Overall this protocol is useful to study the remodeling events and contractile properties of the heart during the gradual onset and progression from compensated cardiac hypertrophy to heart failure stage.

We describe a stepwise procedure for creating pressure overload and left ventricular hypertrophy in Wistar rats by constriction of the ascending aorta using a small metallic clip. This model is extensively used for studying remodeling changes during cardiac hypertrophy and for identifying and evaluating strategies for regression of such changes.

Basınç Aşırı Kardiyak hipertrofi Modeli Oluşturma Ratlarda Aort daralma, Artan

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

Farelerde Mesane Ağrı için bir model olarak Mesane Distansiyonunun Uyarılmış Visceromotor Cevapları

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

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 ...

Chronic Constriction Injury of the Rat's Infraorbital Nerve (IoN-CCI) to Study Trigeminal Neuropathic Pain

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral procedures to study trigeminal neuropathic pain in rats. To meet the specificity of trigeminal neuropathic pain syndromes, the infraorbital nerve (IoN) is subjected to a chronic constriction injury (CCI) by loosely ligating the nerve. An intra-orbital approach is presented here to expose and ligate the IoN in the orbital cavity. After IoN ligation, rats exhibit changes in spontaneous behavior and in response to von Frey hair stimulation that are indicative of persistent pain and mechanical allodynia. Two phases can be defined in the development of the behavioral changes. During the first week following IoN-CCI (phase 1), rats show an increased and asymmetric face grooming activity, i.e., with face wash strokes primarily directed to the nerve-injured IoN territory. A distinction is made between face grooming behavior that is part of a more general body grooming behavior, which remains largely unaffected by IoN-CCI, and face grooming that is neither preceded nor followed by body grooming, which is significantly increased after IoN-CCI. During this period, responsiveness to mechanical stimulation of the IoN territory is reduced. This hyporesponsiveness is abruptly replaced by an extreme hyperresponsiveness whereby even very weak stimulus intensities provoke nocifensive behavior (phase 2). The phenomenological similarities between these behavioral alterations and reported signs of facial pain (i.e., responses to noxious stimulation of the face) suggest the presence of dysesthesia/paresthesia and mechanical allodynia in the ligated IoN territory.

Chronic Constriction Injury of the Rat's Infraorbital Nerve IoN-CCI to Study Trigeminal Neuropathic Pain

This manuscript describes a rodent model to study trigeminal neuropathic pain. The methods described include surgical procedures to perform a chronic constriction injury of the rat&#8217;s infraorbital nerve and the post-surgical behavioral tests to measure the changes in spontaneous and evoked behavior that are indicative of persistent pain and allodynia.

Trigeminal Nöropatik Ağrı Eğitim için Rat adlı infraorbital Sinir (ION-CCI) kronik Daralma Yaralanma

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

Sıçanlarda Karın Aort Konstrüksiyon sayesinde Kardiyak Yaptırma Bir Model

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

Histological Quantification of Chronic Myocardial Infarct in Rats

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ischemic injury. Following the initial acute phase, chronic remodeling of the left ventricle (LV) modifies the structure and function of the heart. Permanent coronary ligation in small animals has been widely used as a reference model for a chronic model of MI. Thinning of the infarcted wall progressively develops to transmural fibrosis. Histological assessment of infarct size is commonly performed; nevertheless, a standardization of the methods for quantification is missing. Indeed, important methodological aspects, such as the number of sections analyzed and the sampling and quantification methods, are usually not described and therefore preclude comparison across investigations. Too often, quantification is performed on a single section obtained at the level of the papillary muscles. Because novel strategies aimed at reducing infarct expansion and remodeling are under investigation, there is an important need for the standardization of accurate heart sampling protocols. We describe an accurate method to quantify the infarct size using a systematic sampling of harvested rat heart and image analyses of trichromatic stained histological sections obtained from base to apex. We also provide evidence that calculating the expansion index (EI) allowed for infarct size assessment, taking into account changes of the left ventricle throughout the remodeling.

The post-mortem assessment of myocardial infarction (MI) in rodents is based on quantification of the infarct on stained heart sections. We describe an accurate method to quantify the infarct size using systematic sampling of harvested rat hearts from base to apex and image analyses of trichrome-stained histological sections.

Sıçanlar Kronik Kalp Enfarktüsü histolojik Niceleme

Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ...

Minimally Invasive Transverse Aortic Constriction in Mice

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard open-chest transverse aortic constriction (TAC). Although transverse aortic constriction is a highly functional method for the induction of high pressure in the left ventricle, it is a more difficult and lengthy procedure due to its use of artificial ventilation with tracheal intubation. TAC is oftentimes also less survivable, as the newer method, MTAC, neither requires the cutting of the ribs and intercostal muscles nor tracheal intubation with a ventilation setup. In MTAC, as opposed to a thoracotomy to access to the chest cavity, the aortic arch is reached through a midline incision in the anterior neck. The thyroid is pulled back to reveal the sternal notch. The sternum is subsequently cut down to the second rib level, and the aortic arch is reached simply by separating the connective tissues and thymus. From there, a suture can be wrapped around the arch and tied with a spacer, and then the sternal cut and skin can be closed. MTAC is a much faster and less invasive way to induce left ventricular hypertension and enables the possibility for high-throughput studies. The success of the constriction can be verified using high-frequency trans-thoracic echocardiography, particularly color Doppler and pulsed-wave Doppler, to determine the flow velocities of the aortic arch and left and right carotid arteries, the dimension of the blood vessels, and the left ventricular function and morphology. A successful constriction will also trigger significant histopathological changes, such as cardiac muscle cell hypertrophy with interstitial and perivascular fibrosis. Here, the procedure of MTAC is described, demonstrating how the resulting flow changes in the carotid arteries can be examined with echocardiography, gross morphology, and histopathological changes in the heart.

Minimally invasive transverse aortic constriction (MTAC) conserves the essentials of regular transverse aortic constriction (TAC) while eliminating the use of a ventilator with tracheal intubation. It proves to be a highly desirable method for high-throughput studies on left ventricular overload, particularly in translational studies.

Farelerde Minimal İnvaziv Enine Aort Konstrüksiyon

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

Subcutaneous Trigeminal Nerve Field Stimulation for Refractory Facial Pain

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. Neuromodulation techniques for chronic facial pain include deep brain stimulation and motor cortex stimulation, which are complex to perform. Subcutaneous nerve field stimulation is certified for chronic back pain and is the least invasive form of neuromodulation. We applied this technique to treat chronic and neuropathic trigeminal pain as an individual therapy concept. First, trial stimulation is performed. Subcutaneous leads are placed in the painful trigeminal dermatome under local anesthesia. The leads are connected to an external neurostimulator that applies constant stimulation. Patients undergo a 12 day outpatient trial to assess the effect of the stimulation. Electrodes are removed after the trial. If the patient reports pain reduction of at least 50% in intensity and/or attack frequency, a reduction in medication or increase in quality of life, permanent implantation is scheduled. New electrodes are implanted under general anesthesia and are subcutaneously tunneled to an infraclavicular internal pulse generator. Patients are able to turn stimulation on and off and to increase or decrease the stimulation amplitude as needed.
This technique represents a minimal invasive alternative to other more invasive means of neuromodulation for trigeminal pain such as motor cortex stimulation or deep brain stimulation.

Treating chronic or neuropathic facial pain can be challenging when medical or standard treatment fails. Subcutaneous nerve field stimulation is the least invasive form of neuromodulation and is used for chronic back pain. We applied this technology to treat chronic and neuropathic trigeminal facial pain.

Refrakter Yüz Ağrısı İçin Subkutan Trigeminal Sinir Sahası Stimülasyonu

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. ...

Development of Recombinant Proteins to Treat Chronic Pain

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates for treating debilitating pain conditions due to their capacity to regulate aberrant neuro-immune interactions. However, physiologically they work in a network of various cytokines, and therefore their therapeutic effect may not be optimal when used as stand-alone drugs. To overcome this limitation, we developed a fusion protein of the anti-inflammatory cytokines IL4 and IL10. Here, we describe the methods for production and quality control of IL4-10 recombinant fusion protein and we test the effectiveness of the IL4-10 fusion protein to resolve pain in a mouse model of persistent inflammatory pain.

In this paper we provide details for the methods of production and quality control of an IL4-10 recombinant fusion protein. We also show how to test the effectiveness of this protein to resolve pain in a mouse model of inflammatory pain.

Kronik ağrı tedavisinde rekombinant proteinler gelişimi

Chronic pain is difficult to treat and new approaches to resolve persistent pain are urgently needed. Anti-inflammatory cytokines are promising candidates ...