Bu çalışma Lundbeck Vakfı, Carlsberg Vakfı ve Dagmar Marshall Vakfı tarafından desteklenmiştir.

1. Ameliyat öncesinde von Frey başlangıç ​​ölçümleri Von Frey test prosedürü için, bölüm 5&#39;e bakınız. 2. Anestezi / hazırlık <ol><li> Hayvanların anestezi (ketamin ve 15 mg / kg Xylazine 100mg/kg bir karışımı intraperitoneal). </li><li> Tam anestezi (örneğin, bir kağıt havlu ile örtülü) kadar sakin bir yerde hayvan yerleştirin. </li><li> Bir cımbız ile kuyruk ucu ve pençeleri pinching refleksleri kontrol edin. Devam etmeden önce hayvanlar tepkisiz emin olun. </li><li> Deri altına (arka omuz bölgesi), antibiyotikler örneğin ampisilin (bu küçük bir cerrahi işlem olmasına rağmen, dehidratasyonu önlemek ve enfeksiyonu önlemek için) 0.5 ml izotonik tuzlu su enjekte. </li><li> Bir elektrikli tıraş makinesi kullanarak, diz, kalça bölgesine hafifçe altında ameliyat alanında tıraş (sol arka bacak sağ elini kullanan kişiler için tavsiye edilir). </li><li> Pamuk tomurcuk gözleri oftalmik merhem uygulayın. </li><li> Sağ tarafta hayvan yerleştirin ve sol hindlimb yüksekte tutmak için küçük bir platform üzerine yerleştirin. Bacak yapışkan bant ile sabitleyin. </li><li> , Cerrahi sitesini etanol ve betadin scrubs alternatif ile ameliyat alanında dezenfekte edin. </li></ol> 3. Cerrahi <ol><li> Sol elin başparmağı ile diz bulun ve bir uygulama yapmak için bir neşter kullanabilirsiniz. 1 cm&#39;lik kesi, diz proksimal boyuna. </li><li> Bir çift steril makas ucu künt diseksiyon cilt açın. </li><li> Künt diseksiyon tarafından açıkça görülebilir bir kan damarı yanında kas tabakası ayırın, uyluk kemiği (femur) yakın. Eğer doğru yapılırsa, kas tabakaları kolayca kasları sağ altında siyatik sinir açığa herhangi bir kanama olmadan ayrı olacaktır. Kanama örneğin diz yakın bir kan damarı hasar oluşursa, kan emmek için steril pamuk tomurcukları veya bakışın parçaları kullanın. Kanama durana kadar basın. </li><li> Stereo mikroskop altında fare yerleştirin ve dikkatli bir şekilde steril bir pens (nr. 2) bir çift ile siyatik sinir görselleştirmek için kasları ayrı. Retraktörler de uygulanabilir. </li><li> Siyatik sinir sural sinir dallarının alanı belirleyin. Sural sinir, sol bacak sağ dallanma, üç şube küçüğüdür. </li><li> Sütür (6-0 dikiş), hala, (tibial ve peroneal sinirler) paralel olarak çalışan sural şube dokunmak için çok dikkatli olmak diğer iki dal etrafında uygulayın. Sural sinir tamamen bozulmadan bırakılmalıdır Bu önemli bir adım. </li><li> Sıkı bir cerrahi düğüm olun. Ilk düğümü sıkı ise, bacak kasılmaları görülecektir. </li><li> Bir cımbız (nr. 5) ile dikiş altında kesilebilir sinirleri alın ve küçük bir çift makas ile cımbız, altında ve üstündeki sinirleri kesmek. Ilk bakışta, cımbız çifti üzerinde kesme, sinirlerin çekerek kaçınılmalıdır. </li><li> Mikro makas bir çift sütür uçları kesilmiş ve kas tabakası yavaşça kapatın. Lidokain, cerrahi knot ile yara ve dikiş için bir damla ekleyin. </li></ol> 4. Ameliyattan sonra <ol><li> Göz merhemi yine de yeterli olup olmadığını kontrol edin. </li><li> Fare, rahat bir duruş bir kağıt havlu altında temiz bir kafes yerleştirin. Oda soğuk ise, (sadece hayvan olarak kafesin parçası altında tercih eğer daha soğuk bir alana kaçmak gerekir) bir parçası kafesin altında bir ısı pedi yerleştirin. </li><li> Kolay erişilebilir su ve chow olun. </li></ol> 5. von Frey testi (bazal ve ameliyattan bir gün sonra) <ol><li> Bir tel örgü tablo, iki gün ve ameliyattan bir gün önce yerleştirilmiş kırmızı renkli plastik silindir fareler yerleştirin. 15 dakika önce silindir test için alıştırarak. Silindir kırmızı renk farelerin birbirlerini görmek ve daha karanlık bir alan olarak hızlı bir şekilde rahatlamak için yardımcı olur. </li><li> Fareler sakin olduğundan emin olun ve pençe lateral kısmı von Frey filamentler geçerlidir: 0.02 g filament ile başlayarak, ilk 30 saniye (uyaran başına yaklaşık 2 saniye) toplam süre içinde beş kez sol pençe kuvveti uygulamak ve Her uygulamadan sonra farenin tepki ölçmek. Aynı filament ve herhangi bir diğer farelerin sol pençe tekrarlayın ve sonra bir sonraki filament, örneğin geçmeden önce, ilk fare sağ pençe ile tekrar baştan başlamak: <ul><li> Fare 1: sol pençe filament 0,02 g 5 kez </li><li> Mouse 2: sol pençe filament 0,02 g vb 5 kez </li><li> Fare 1: sağ pençe filament 0,02 g ile 5 kez </li><li> Mouse 2: sağ pençe filament 0,02 g vb ile 5 kez </li><li> Fare 1: sol pençe filament 0,04 g 5 kez </li><li> Mouse 2: sol pençe filament 0,04 g vb 5 kez </li></ul></li><li> Üç beş uyaranlara yanıt olumlu bir tepki olarak kabul edilir. Threshol yukarıda filamentlerd eşik düzeyini doğrulamak için uygulanabilir. Tepki = ani pençe çekilmesi, yalama ani flinching, ani pençe. </li></ol> 6. Temsilcisi sonuçları <img src="/files/ftp_upload/3092/3092fig1.jpg" alt="Şekil 1" /> Şekil 1: 8-10 hafta eski C57BL / 6 farelerin yaş Von Frey test çalışması için 1 gün önce yapılmış ve tekrar tekrar ameliyat takip ediyordu . Her bir grupta 4-8 hayvanlar dahil edildi. Ameliyatlı tarafta Von Frey test ipsilateral tarafından temsil edilir ve non-kumanda tarafındaki kontralateral tarafından temsil edilmektedir. Olmayan çalışan sahte çalışan farelere göre sadece biraz etkilenen ise gün ameliyat sonrası hayvanlar çalışan pençe üzerinde önemli bir duyarlılık geliştirmesi. (* &lt;0.05, ** &lt;0.01, *** &lt;0.001, kontralateral için ipsilateral).

<ol>
	<li>Costigan, M., Scholz, J., Woolf, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropathic+pain:+a+maladaptive+response+of+the+nervous+system+to+damage.">Neuropathic pain: a maladaptive response of the nervous system to damage.</a> Annu. Rev. Neurosci. 32, 1-32 (2009).</li><li>Decosterd, I., Woolf, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spared+nerve+injury:+an+animal+model+of+persistent+peripheral+neuropathic+pain.">Spared nerve injury: an animal model of persistent peripheral neuropathic pain.</a> Pain. 87, 149-158 (2000).</li><li>Bourquin, A. -. F., S&#252;veges, M., Pertin, M., Gilliar, N., Sardy, S., Davidson, A. C., Spahn, D. R., Decosterd, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+and+analysis+of+mechanical+allodynia-like+behavior+induced+by+spared+nerve+injury+(SNI)+in+the+mouse.">Assessment and analysis of mechanical allodynia-like behavior induced by spared nerve injury (SNI) in the mouse.</a> Pain. 122, 14.e1-14.e14 (2006).</li><li>Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M., Yaksh, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+tactile+allodynia+in+the+rat+paw.">Quantitative assessment of tactile allodynia in the rat paw.</a> J. Neurosci. Methods. 53, 55-63 (1994).</li><li>Bennett, G. J., Xie, Y. -. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+peripheral+mononeuropathy+in+rat+that+produces+disorders+of+pain+sensation+like+those+seen+in+man.">A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man.</a> Pain. 33, 87-107 (1988).</li><li>Seltzer, Z., Dubner, R., Shir, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+behavioral+model+of+neuropathic+pain+disorders+produced+in+rats+by+partial+sciatic+nerve+injury.">A novel behavioral model of neuropathic pain disorders produced in rats by partial sciatic nerve injury.</a> Pain. 43, 205-218 (1990).</li><li>Hargreaves, K., Dubner, R., Brown, F., Flores, C., Joris, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+and+sensitive+method+for+measuring+thermal+nociception+in+cutanous+hyperalgesia.">A new and sensitive method for measuring thermal nociception in cutanous hyperalgesia.</a> Pain. 32, 77-88 (1988).</li><li>Erichsen, H. K., Blackburn-Munro, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+characterization+of+the+spared+nerve+injury+model+of+neuropathic+pain.">Pharmacological characterization of the spared nerve injury model of neuropathic pain.</a> Pain. 98, 151-161 (2002).</li></ol>

Çıkar çatışması ilan etti.

Kritik adımlar Sural sinir hasar tibial ve peroneal sinirlerin sağlam sural sinir yaralanma sonrası patolojik değişiklikler araştırmak için kaçınılmalıdır. Kollateral hasar sural sinir felce yol açabilir ve böylece çalışan arka bacak sürükleyerek olarak görünür olabilir. Sadece pençe lateral tarafında ayrılan sural sinir tarafından innerve ve dolayısıyla, bu alanda sadece nöropati gelişir. Güçlü bir önyargı değişmiş mekanik eşik değerlendirme transeksiyon sinirler tarafından innerve diğer alanlarda Test. Olası değişiklikler [5] ligasyonu diğer türleri gibi kronik daralma yaralanma gibi periferik sinir hasarı sonrası patolojik ağrı durumları, eğitim açısından tamamlayıcı olabilir veya kısmi sinir ligasyonu [6]. Önce ameliyat sonrası test düşünülmelidir farklı fenotipik değişiklikler her deneysel işlemin sonuçları. Ayrıca, termal hiperaljezi gibi diğer duyusal testler, bu fenotip, daha sağlam aşağıdaki SNI olmasına rağmen [7] uygulanan olabilir. Gelecekteki uygulamalar Bu teknik, mekanik allodini geliştirme ya da bakım yapıcı ilaçların test edilmesi için kullanılabilir [8]. Analizi siyatik sinir, dorsal kök gangliyon (DRG) ve / veya lomber spinal kord kaynaklı fenotip moleküler mekanizmaları araştırma sorumlu izin verir. Iyi laboratuvar uygulamaları ile tam bir uyum içinde, Danimarka ve Avrupa yönetmeliklerine uygun olarak hayvan deneyleri yapıldı. Tüm deneyler kapsamında Danimarka Hayvan Deneyleri Müfettişliği, Adalet Bakanlığı (izin numarası 2006/561-1206) tarafından kabul edildi.

<table border="1"><tbody><tr><td> Anestezi: Ketamin ve Xylazine </td><td> Skalpel </td></tr><tr><td> Oftalmik merhem </td><td> Cımbız nr eşleştirme. 2 </td></tr><tr><td> Tıraş makinesi </td><td> Cımbız nr eşleştirme. 5 </td></tr><tr><td> Eldiven </td><td> Künt diseksiyon makas </td></tr><tr><td> % 70 (v / v) etanol </td><td> Mikro makas </td></tr><tr><td> Pamuk-yün tomurcukları / bakışları </td><td> 6.0 Prolen emilmeyen dikiş </td></tr><tr><td> Stereo mikroskop </td><td> 2 x iğnelikler </td></tr><tr><td> Yapışkan bant </td><td> Isı pedi </td></tr><tr><td colspan="2"> von Frey filamentler (0,02 - 2,0 gr, fareler için) (Stoelting) </td></tr></tbody></table>

the spared nerve injury (sni) model of induced mechanical allodynia in mice

Periferik nöropatik ağrı, travma, periferik sinir sisteminin duyu sinirleri neden olabilir ağır bir kronik ağrı bir durumdur. Ayrılan sinir hasarı (SNI) modeli, nöropatik ağrı, mekanik allodini yani ağrı gibi belirtileri nedeniyle normalde acı bir cevabı provoke dokunsal uyaranlara yol açar [1]. Sural sinir değişmeden kalır ise SNI fare modelinde, siyatik sinir (tibial sinir ve peroneal sinir) üç şube iki ligasyonu içerir [2]. Lezyon sonuçları ayrılan sural sinir tarafından innerve pençe, lateral alanda belirgin aşırı duyarlılık. Fare olmayan çalışan yan bir kontrol olarak kullanılabilir. SNI modelinin avantajları yanıt sağlamlık ve uzman mikrocerrahi beceri gerektirmez. Tekrarlayan pençe lateral alanına karşı bastırdı artan bükme kuvveti von Frey filamentler, test, mekanik ağrı yanıtı için eşik tarafından belirlenir [3] [4]. Filament tarafından uyarılan yalamaya ve / veya pençe flinching olumlu bir ağrı reaksiyonu, ani bir pençe geri çekilme olarak tanımlanır. Üç beş tekrarlayan uyaranlara olumlu bir yanıt ağrı eşiği olarak tanımlanır. Video protokolü gösterdiği gibi, C57BL / 6 fareler cerrahi sonrası gün olarak en erken zamanda derin allodini deneyim ve birkaç hafta boyunca bu korumak.

Watch this Scientific Journal Video about The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice at JoVE.com

The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

Ayrılan Sinir Sakatlık hayvan modeli kalan sural sinir zedelemeden, siyatik sinirin kısmi denervasyon aşağıdaki tibial ve peroneal sinir dallarının lezyon periferik nöropatik ağrı bir fare modeli olarak burada açıklanmıştır. Mekanik allodini kaynaklanan davranış değişikliği von Frey filamentler miktarı.

Fare İndüklenmiş Mekanik allodini ayrılan sinir hasarı (SNI) Model

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared ...

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Research

JoVE Journal

Medicine

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice

63.1K Görüntüleme.  Aarhus University. Ayrılan Sinir Sakatlık hayvan modeli kalan sural sinir zedelemeden, siyatik sinirin kısmi denervasyon aşağıdaki tibial ve peroneal sinir dallarının lezyon periferik nöropatik ağrı bir fare modeli olarak burada açıklanmıştır. Mekanik allodini kaynaklanan davranış değişikliği von Frey filamentler miktarı.

Video: The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

Farelerde Kontrollü Servikal yırtığı Sakatlık

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Tıp

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

Farelerde Akut Böbrek Hasarı ve Post Yaralanma Fibrozis iskemi-reperfüzyon Model

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. Although originally developed and used extensively in the rat due to its larger size, the tibial nerve in mice is big enough that it can be easily manipulated with either crush or transection, leaving the peroneal and sural nerve branches of the sciatic nerve intact and thereby preserving their target muscles. Thus, this model offers the advantages of inducing less morbidity and impediment of ambulation than the sciatic nerve transection model and also allows investigators to study the physiologic, cellular and molecular biologic mechanisms regulating the process of muscle atrophy in genetically engineered mice. The tibial nerve supplies the gastrocnemius, soleus and plantaris muscles, so its transection permits the study of denervated skeletal muscle composed of fast twitch type II fibers and/or slow twitch type I fibers. Here we demonstrate the tibial nerve transection model in the C57Black6 mouse. We assess the atrophy of the gastrocnemius muscle, as a representative muscle, at 1, 2, and 4 weeks post-denervation by measuring muscle weights and fiber type specific cross-sectional area on paraffin-embedded histologic sections immunostained for fast twitch myosin.

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of skeletal muscle atrophy. The model surgical protocol is described and demonstrated in C57Black6 mice. &#160;

Tibial Sinir transeksiyonu - Farelerde denervasyon bağlı İskelet Kas Atrofi için bir Standart Model

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

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 ...

Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and either cut or crush it to induce peripheral nerve injury. Advantages of this surgery are its simplicity, high reproducibility, and the lack of effect on vital functions or mobility from the subsequent facial paralysis, thus resulting in a relatively mild surgical outcome compared to other nerve injury models. A major advantage of using a cranial nerve injury model is that the motoneurons reside in a relatively homogenous population in the facial motor nucleus in the pons, simplifying the study of the motoneuron cell bodies. Because of the symmetrical nature of facial nerve innervation and the lack of crosstalk between the facial motor nuclei, the operation can be performed unilaterally with the unaxotomized side serving as a paired internal control. A variety of analyses can be performed postoperatively to assess the physiologic response, details of which are beyond the scope of this article. For example, recovery of muscle function can serve as a behavioral marker for reinnervation, or the motoneurons can be quantified to measure cell survival. Additionally, the motoneurons can be accurately captured using laser microdissection for molecular analysis. Because the facial nerve axotomy is minimally invasive and well tolerated, it can be utilized on a wide variety of genetically modified mice. Also, this surgery model can be used to analyze the effectiveness of peripheral nerve injury treatments. Facial nerve injury provides a means for investigating not only motoneurons, but also the responses of the central and peripheral glial microenvironment, immune system, and target musculature. The facial nerve injury model is a widely accepted peripheral nerve injury model that serves as a powerful tool for studying nerve injury and regeneration.

We present a surgical protocol detailing how to perform a cut or crush axotomy on the facial nerve in the mouse. The facial nerve axotomy can be employed to study the physiological response to nerve injury and test therapeutic techniques.

Farelerde Yüz Sinir Aksotomi: Sakatlık Motonöron Yanıtı Eğitim için Bir Model

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Farelerde Omurilik Yaralanma Modeli Olarak Odak iskemisi Fototromboz kaynaklı

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

A Repetitive Concussive Head Injury Model in Mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

Farelerde Bir Tekrarlanan Concussive Kafa Travması Modeli

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

Farelerde Hızlandırılmış Ateroskleroz Indüksiyonu: "Tel-Yaralanma" Modeli

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between experimental animals. Several contusive SCI models have been developed to produce injuries using different mechanisms. The severity of the SCI is based on the height that a given weight is dropped, the injury force, or the spinal cord displacement. In the current study, we introduce a novel mouse contusive SCI device, the Louisville Injury System Apparatus (LISA) impactor, which can create a displacement-based SCI with high injury velocity and accuracy. This system utilizes laser distance sensors combined with advanced software to produce graded and highly-reproducible injuries. We performed a contusive SCI at the 10th thoracic vertebral (T10) level in mice to demonstrate the step-by-step procedure. The model can also be applied to the cervical and lumbar spinal levels.

We introduce a tissue displacement-based contusive spinal cord injury model that can produce a consistent contusive spinal cord injury in adult mice.

Farelerde Doku Yerinden Olma Esaslı Taşan Spinal Kord Yaralanması Modeli

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

İzometrik Contractility ölçüm tel Myography kullanarak fare mezenterik arter

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...