This works was supported by funding from a Florida Health grant (Brain and spinal cord injury research fund) (KKW).

Tüm işlemler Florida Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından ve Laboratuvar Hayvanları Bakım ve Kullanım Sağlık Rehberi National Institutes göre onaylanan protokoller # 201207692 altında gerçekleştirildi. 1. Hayvan Bakımı <ol><li> 3-4 aylık erkek C57BL / 6J fareler kullanın. Yatak, yuvalama malzeme, yiyecek ve su ad libitum sağlamak. Sabit 12 saat ışık / 12 saat karanlık döngüsü ile 22 ° C - 20 kontrol edilen ortam sıcaklıklarında fareler tutun. </li></ol> 2. Ön impaksiyon Hazırlık <ol><li> Elektromanyetik stereotaksik darbe cihazına ısmarlama silikon kauçuk kaplı metal ucu takın. Ucunun düz tabanlı prob ucu (Şekil 1A) yüzeyine paralel olduğundan emin olun. </li><li> % 2.5 izofluran bakım anestezi ardından% 4 izofluran ile fare anestezisi. debimetre aracılığıyla anestezi kontrol edin. monitor hayvan kadar anestezi seviyesi pedalı geri çekme refleksi kaybı göstererek anestezi cerrahi düzeyine ulaşır. </li><li> bir ısıtma yastığı üzerinde yüzükoyun pozisyonda fare koyun. Anestezi altında fare tutmak için bir huni şeklinde bir burun konisi kullanın. Tamamen bir düzeltici kullanarak tıraş. anestezi altında iken kuruluğunu önlemek için farenin gözleri vazelin oftalmik merhem kullanın. </li></ol> 3. Etki Parametreleri Ayar NOT: Etki sistemi darbe parametrelerini ayarlamak için bir kontrol kutusu, impaction gerçekleştirmek için bir aktüatör ve 3-hareket eksenleri ile bir dijital stereotaktik çerçeve içerir. <ol><li> 4 m / sn darbe cihazın hızını önceden ayarlanmış ve kontrol kutusu üzerinde 240 msn bekleme süresi. </li></ol> 4. Konumlandırma Darbe Merkezi <ol><li> 39 ° C&#39;ye yakın vücut ısısını korumak için hayvanın vücudunun altında yumuşak bir ısıtma yastığı koyun. Bir pron bir stereotaktik çerçeve içinde fare montekünt uç kulak çubukları ile e konumu. </li><li> Z-sürücü hareket ettirerek farenin kafasına yakın darbe ucu indirin. Hedef sagital sütür üzerinde koordinatları ortada X- ve Y-sürücüleri hareket ettirerek düz darbe ucu (9 mm çapında) ayarlayın. </li><li> Dikey iki kulak (Şekil 1C) arasında çizilen hayali bir yatay çizgi paralel olan darbe ucunun emin bir kenarı olun. etkinin merkezi (0 mm, yanal 4.5 mm kulaklararası için interaural 9 mm) interfrontal ve lambdoidea&#39;nın sütürler arasındaki merkezi sagital sütür yarıda karşılık gelir. </li></ol> 5. Darbe Derinlik Ayarı <ol><li> doğru etki derinliğini ayarlamak için, yalıtımlı silikon kauçuk kaplı darbe ucu yerine ek prob ucunu kullanın. </li><li> ipuçları geçmeden önce sıfıra dijital stereotaksik kontrol panelindeki X ve Y kanalı ayarlamak anahtarlama ipuçları sonra darbe merkezinin herhangi bir kayma var olduğundan emin olmak için. </li><li> prob taşıEl X ve Y-sürücüleri hareket ettirerek darbe alanının merkezine e ucu. </li><li> farenin kuyruk klibi temas sensörü. </li><li> prob ucu darbe sitenin yüzeyine temas edene kadar aşağı impaktörü (Z sürücü) taşıyın. </li><li> sıfıra stereotaksik kontrol panelindeki Z kanalını ayarlayın. </li><li> elle X ve Y sürücüleri kadar X- ve Y-sürücüleri (NOT dijital stereotaksik kontrol panelinde sıfır düğmeleri) ayarlayarak geri etki alanının darbe ucu taşı (darbe ucu daha önce yerleştirildi) sıfırdır. </li><li> kontrol kutusu üzerindeki geri çekme anahtarı hareket ettirerek çalıştırıcıyı geri çekin. El ile 4 mm aşağı impaktörü (Z sürücüsü) hareket ettirin. </li></ol> 6. Darbe <ol><li> kontrol kutusu üzerindeki etkisi anahtarını tıklayarak etkiyi tetikleyebilir ve 4 mm deformasyon derinliği elde. </li></ol> 7. Sonrası impaksiyon <ol><li> farenin ilk nefes bir zamanlayıcı kullanarak kadar etkisinden zamanı ölçmek. </li><li&gt; Enstrüman hayvan çıkarın ve vücut ısısını korumak için sıcak bir ped üzerine koyun. sternal yatma korumak için yeterli bilinci yerine kadar sahipsiz hayvan bırakmayın. </li><li> Temiz bir kafes içine geri hayvan dönmeden önce kurtarma için izin verin. Tamamen iyileşene kadar diğer hayvanların şirkete bir hayvan iade etmeyin. </li><li> Gözlemleyin ve günlük fareler tartmak. Fareler ağrı belirtileri varsa, intraperitoneal 1 de Meloksikamın ile enjekte - 24 saat - 2 mg / her 12 kg. </li></ol> 8. Tekrarlanan Gömülü <ol><li> gün 4, 7 farelere ek yaralanmaları verin ve ilk yaralanma (etkileri arasındaki 72 saat aralığı) sonra 10. </li></ol> 9. İmmunohistokimya (İHK) <ol><li> transcardial perfüzyon <ol><li> 200 m / kg pentobarbital intraperitonal enjeksiyon yoluyla fareler anestezi. </li><li> Değerlendirmek ve bir ayak tutam cerrahi düzlem anestezi sağlamak. saniyeure hafifçe kimyasal davlumbaz içine plastik bir çalışma yüzeyine forepaws ve arka pençeleri bantlama ile yatar pozisyonda fare. </li><li> Sadece klavikulaya kılıç şeklinde bir süreç altından göğüs orta hat boyunca deri yoluyla bir kesi yapmak. kılıç şeklinde bir süreç iki ek cilt kesiler yapmak ve yanal ventral göğüs kafesi tabanı boyunca devam edin. </li><li> göğüs boşluğu açın ve göğüs kaslarının ve göğüs kafesi üzerinden keserek kalp maruz. </li><li> künt forseps ile kalpte sabitleyin ve 1 yapmak - sol ventrikül 2 mm kesi. </li><li> Hemen sağ atrium içine kelebek iğne takın. yavaş yavaş şırınga iterek 20ml tuzlu su infüzyon başlayın. </li><li> tuzlu dan% 4 paraformaldehid geçin. paraformaldehit 20ml ile perfüzyon devam edin. </li><li> fare başını kesmek ve makasla cilt kaldırmak. Bir kemik kesici kullanarak kafatası beyin izole edin. </li></ol></li><li> Ckesit ryostat <ol><li> -80 ° C&#39;de optimum kesme sıcaklığı (OKT) formülasyonu ve donma embed beyin dokuları. sagital yönde kriyostat beyin yerleştirin. Kes beyin kesitleri 5 mikron kalınlığında. </li></ol></li><li> boyanma <ol><li> 1 saat süre ile, oda sıcaklığında dondurulmuş bölümler kurutun. </li><li> Oda sıcaklığında 1 saat için fosfat tamponlu tuzlu suda 100% 2 keçi serumu ul ve% 0.1 Triton X-100 (PBS) ile slaytlar inkübe edin. </li><li> PBS 300 ul slaytlar 3 kere yıkayın. 4 ° C&#39;de gece boyunca ayrı (200 1) ya da anti-ferritin antikor sonra anti-GFAP (200 1) ile slaytlar kuluçkalayın. </li><li> PBS 300 ul slaytlar 3 kere yıkayın. Daha sonra biyotin-konjuge sekonder antikor ile oda sıcaklığında 2 st için slaytlar inkübe edin. </li><li> PBS 300 ul slaytlar 3 kere yıkayın. Daha sonra 30 dakika boyunca oda sıcaklığında avidin-biyotin kompleksi (ABC) solüsyonu (1:50) ile slaytlar kuluçkalayın. </li><li>PBS 300 ul slaytlar 3 kere yıkayın. 8 dakika - sonra 3,3&#39;-diaminobenzidin 5 için (DAB) alt-tabaka çözeltisi (50 ml PBS, 10 ul H2O 2, 10 mg, DAB hap, kullanımdan önce filtre) &#39;de inkübe edilir. pozitif hücreler ortaya çıkıncaya kadar mikroskop altında slaytlar uyun. </li><li> 5 dakika boyunca yavaş çalışan musluk suyunda slaytlar durulayın. laboratuar silin bir ile temiz slaytlar. Sonra montaj orta ve lamel ile bölümleri monte edin. </li></ol></li></ol>

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The authors have no financial interest to disclose.

klinik durum morfolojik olarak benzer beyin yaralanmaları taklit etmek, post-sarsıntı belirtileri bekleniyor. Post-sarsıntı semptomlar genellikle baş ağrısı, baş dönmesi, baş dönmesi, yorgunluk, hafıza ve uyku problemleri, sorun konsantre yanı sıra anksiyete ve depresif ruh hali sayılabilir. somatik semptomlar henüz hayvan modellerinde ölçülebilir olabilir, bu motor ve bilişsel işlev ve duygusal davranış değişiklikleri mantıklı bir hayvan modellerinde beyin sarsıntısı değerlendirmek içi...

Sarsıntı da adlandırılan hafif travmatik beyin hasarı (HTBH), travmatik beyin hasarı (TBI) en sık görülen bir durumdur ve ABD&#39;de milyonlarca insanı etkilemektedir. Beyin sarsıntısı teşhis etmek zor olabilir ve sarsıntı için özel bir tedavisi yoktur. Büyüyen bir tanıma ve spor yaralanmaları, askeri mücadele ve diğer fiziksel ilgi çekici takipçiliği kaynaklanan hafif mekanik travma kümülatif ve kronik nörolojik sonuçları 1,2 olabileceği bazı kanıtlar vardır. Ancak, yine de sarsıntı ve onların etkilerine ilişkin bir bilgi eksikliği vardır. Sadece nörolojik değerlendirme ve görüntüleme değerlendirilmesi klinik tanı için kullanılabilir çünkü Güncel metodoloji insanlarda patoloji ve tedavi çalışmalarını kısıtlar. Hayvan modelleri, teşhis ve daha HTBH tedavisinde umut etkili, sıkı, ve kontrollü bir şekilde beyin sarsıntısı incelemek için bir araç sağlar. Çalışmalar geleneksel TBI adapte olmasıBöyle kontrollü kortikal etkisi (CCİ) olarak modeller, sıvı-perküsyon etkisi (FPI), ağırlık düşüşü yaralanma ve patlama hasarı HTBH gerçekleştirmek ve yaralanma parametreleri değiştirerek düşük yaralanma şiddet derecelerine uyarmak için. Bu modeller nedeniyle klinik durumuna morfolojik olarak benzer beyin travması çoğaltmak kabiliyetlerine için kullanmak yararlıdır; Ancak, aynı zamanda kendi sınırlamaları vardır. Bir ivme yaralanma (ağırlık damla) ile uyarılan yaralanma şiddeti genellikle oldukça değişkendir. Hafif CCI iki sonuçları - subaraknoid kanama ve fokal kontüzyon - tipik insan sarsıntı ile karşılaştırılabilir değildir. Blast yaralanma maruz 3-6 esnasında farklı pozlama pozisyon ve pik basınç ölçümleri açısından daha tartışmalı bir model yanı sıra değişken ikincil yaralanma ise CCİ ve FPI, klinik açıdan anlamlı olmayan bir kranyotomi gerektirir. Güncellenmiş sarsıcı hayvan modeli klinik Setti içine pre-klinik araştırma çevirebilirng araştırma gereklidir. Hafif TBI modelleme önemli bir konu en yakın bir klinik ortamda yaralanma çoğaltır deneysel yaralanma şiddetini, tanımlamaktır. Son zamanlarda, farklı araştırma grupları kapalı kafa travması veya Kafa travması (CHI) modeli 7-10 geliştirdi. CHI bir kraniotomi olmadan CCİ&#39;nin bir değişiklik, ama yine de bir kafa etkisini oluşturmak için geleneksel elektronik manyetik darbe sistemi kullanır. Bir CHI darbe parametrelerini ayarlayarak hafif ve orta şiddette değişen bir sarsıntı neden olabilir. bilinç (LOC) kaybı solunum hızı ya da solunum geçici fesih bir azalma tespit ederek bir darbeden sonra hemen görülebilir. LOC süresi yaralanmanın şiddetini belirlemek için kullanılır. Bu kağıt ayrıntılı adım-adım protokolü ve temsilcisi sonuçları ile birlikte farelerde tekrarlayan CHI (rCHI) modelin biraz geliştirilmiş ve güncelleştirilmiş bir sürümünü içerir. rCHI modeli araştırma stratejileri,sarsıntı kaynaklı patolojik değişikliklerin tümünü taklit edebilen hiçbir bireysel bir hayvan modeli var, özellikle de HTBH etkileri ve potansiyel tedaviler belirlenmesinde yararlı yeniden.

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</table>

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.

Bu modelde (Şekil 1 AC) ise, nefes nefese ve sığ respirations kısa dönemler vardı. (Bilinçsiz) bilinç kaybı solunum hızı veya normal solunumu devam etmeden önce nefes geçici fesih bir azalma olarak tanımlanır. Başın ortasına bir darbe kısa süreli bilinç kaybı nedeniyle (7.5 ± 4.7, ± 5.5 7.8, ± 8.8 10.2, ayrı ayrı etkisine 9.5 ± 8.0 sn, Şekil 1D). Fare beyinleri etkisi (Şekil 2A) bir sonucu olarak ortaya beli...

Watch this Scientific Journal Video about A Repetitive Concussive Head Injury Model in Mice at JoVE.com

A Repetitive Concussive Head Injury Model in Mice

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 ...

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Research

JoVE Journal

Medicine

11.8K Görüntüleme.  University of Florida. 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.

Video: A Repetitive Concussive Head Injury Model in Mice

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

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared nerve injury (SNI) model induces symptoms of neuropathic pain such as mechanical allodynia i.e. pain due to tactile stimuli that do not normally provoke a painful response [1]. 
The SNI mouse model involves ligation of two of the three branches of the sciatic nerve (the tibial nerve and the common peroneal nerve), while the sural nerve is left intact [2]. The lesion results in marked hypersensitivity in the lateral area of the paw, which is innervated by the spared sural nerve. The non-operated side of the mouse can be used as a control. The advantages of the SNI model are the robustness of the response and that it doesn&#x2019;t require expert microsurgical skills.
The threshold for mechanical pain response is determined by testing with von Frey filaments of increasing bending force, which are repetitively pressed against the lateral area of the paw [3], [4]. A positive pain reaction is defined as sudden paw withdrawal, flinching and/or paw licking induced by the filament. A positive response in three out of five repetitive stimuli is defined as the pain threshold. 
As demonstrated in the video protocol, C57BL/6 mice experience profound allodynia as early as the day following surgery and maintain this for several weeks.

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

The Spared Nerve Injury animal model is described here as a mouse model of peripheral neuropathic pain following partial denervation of the sciatic nerve by lesioning the tibial and common peroneal nerve branches, leaving the remaining sural nerve intact. Behavioral modification resulting from mechanical allodynia is quantified by von Frey filaments.

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 ...

Tıp

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 ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

Sıçanlarda Şiddetli Spinal Kord Yaralanması bir bere modeli

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

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 ...

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable during renal transplantation. Experimental models that accurately and reproducibly recapitulate renal IRI are crucial in dissecting the pathophysiology of AKI and the development of novel therapeutic agents. Presented here is a mouse model of renal IRI that results in reproducible AKI. This is achieved by a midline laparotomy approach for the surgery with one incision allowing both a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia of the left kidney. By careful monitoring of the clamp position and body temperature during the period of ischaemia this model achieves reproducible functional and structural injury. Mice sacrificed 24 hr&#160;following surgery demonstrate loss of renal function with elevation of the serum or plasma creatinine level as well as structural kidney damage with acute tubular necrosis evident. Renal function improves and the acute tissue injury resolves during the course of 7 days following renal IRI such that this model may be used to study renal regeneration. This model of renal IRI has been utilized to study the molecular and cellular pathophysiology of AKI as well as analysis of the subsequent renal regeneration.

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

Renal İskemi Reperfüzyon Hasarı: Yaralanma Fare Modeli ve Rejenerasyon

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

Farelerde Basit Kritik ölçekli Femoral Hata Modeli

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

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 ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

Farelerde Tekrarlanan Hipoksik Önkoşullanma ve Geçici Orta Serebral Arter Tıkanıklığı ardından Nörovasküler Koruma Kantitasyonu

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

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) ...

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. ...