Questo lavoro è stato sostenuto dalla Fondazione Lundbeck, la Fondazione Carlsberg e Dagmar Marshall Foundation.

1. misurazioni di base von Frey prima dell&#39;intervento Per la procedura di von Frey test, fare riferimento alla sezione 5. 2. Anestesia / preparato <ol><li> Anestetizzare gli animali (iniezione intraperitoneale di una miscela di 100mg/kg ketamina e 15 mg / kg xilazina). </li><li> Posto l&#39;animale in un luogo tranquillo fino a quando completamente anestetizzato (ad esempio coperto da un tovagliolo di carta). </li><li> Controllare riflessi pizzicando la punta della coda e le zampe con un paio di pinzette. Assicurarsi che gli animali non rispondono prima di procedere. </li><li> Iniettare per via sottocutanea (zona delle spalle indietro) 0,5 ml soluzione salina isotonica con antibiotici, per esempio ampicillina (per evitare la disidratazione e prevenire l&#39;infezione, anche se questo è un piccolo intervento chirurgico). </li><li> L&#39;utilizzo di un rasoio elettrico, rasatura del campo operatorio da poco sotto l&#39;area ginocchio alla zona dell&#39;anca (per destrorsi persone l&#39;arto posteriore sinistra è consigliato). </li><li> Applicare una pomata oftalmica per gli occhi con un cotone-lana gemma. </li><li> Posto l&#39;animale sul suo lato destro e portare il arti posteriori a sinistra su una piccola piattaforma per mantenerla elevata. Fissare la gamba con nastro adesivo. </li><li> La disinfezione del campo operatorio con alternanza di macchie di etanolo e betadine da fuori del sito chirurgico. </li></ol> 3. Chirurgia <ol><li> Individuare il ginocchio con il pollice della mano sinistra e utilizzare un bisturi per fare un app. 1 incisione cm in senso longitudinale prossimale al ginocchio. </li><li> Aprire la pelle per via smussa con la punta di un paio di forbici sterili. </li><li> Separare lo strato muscolare per via smussa proprio accanto al vaso sanguigno ben visibile, vicino al femore (femore). Se fatto correttamente, gli strati muscolari facilmente separato senza alcun sanguinamento, rivelando il nervo sciatico destro sotto i muscoli. Se l&#39;emorragia si verifica ad esempio da un danno ad un vaso sanguigno vicino al ginocchio, l&#39;uso di cotone idrofilo sterile gemme o pezzi dello sguardo per assorbire il sangue. Premere finché l&#39;emorragia si ferma. </li><li> Posizionare il mouse al microscopio stereo e con attenzione separare i muscoli con un paio di pinzette sterili (nr. 2) per visualizzare il nervo sciatico. Retrattori può anche essere applicata. </li><li> Identificare l&#39;area in cui i rami del nervo surale dal nervo sciatico. Il nervo surale è il più piccolo dei tre rami, che si dirama a destra alla gamba sinistra. </li><li> Applicare sutura (6-0 sutura) intorno agli altri due rami, che sono ancora in esecuzione in parallelo (i nervi tibiale e peroneo comune), facendo molta attenzione a non toccare il ramo surale. Questo è un passo critico come il nervo surale deve essere lasciato completamente intatto. </li><li> Fare un nodo stretto chirurgico. Se il primo nodo è stretto, contrazioni degli arti sarà osservato. </li><li> Afferra i nervi da tagliare sotto la sutura con un paio di pinzette (nr. 5) e tagliare i nervi sopra e sotto le pinzette con un paio di piccole forbici. In primo luogo il taglio sopra il paio di pinzette, tirando dei nervi è evitato. </li><li> Tagliare le estremità di sutura con un paio di forbici micro e chiudere delicatamente lo strato muscolare. Aggiungere una goccia di lidocaina per la ferita e suturare con nodi chirurgici. </li></ol> 4. Dopo l&#39;intervento chirurgico <ol><li> Controllare se unguento occhio è ancora sufficiente. </li><li> Posizionare il mouse in una gabbia pulita sotto un tovagliolo di carta in una posizione comoda. Se la stanza è fredda, posto un pad termico sotto una parte della gabbia (solo in parte della gabbia come l&#39;animale deve essere in grado di sfuggire ad una zona più fredda se preferite). </li><li> Assicurare acqua facilmente accessibili e chow. </li></ol> 5. von Frey test (di base, e dal giorno dopo l&#39;intervento) <ol><li> Posizionare il mouse in cilindri di plastica di colore rosso posto su un tavolo rete metallica due giorni e un giorno prima dell&#39;intervento. Abituare per 15 minuti in cilindri prima del test. Il colore rosso dei cilindri aiuta i topi a rilassarsi in fretta perché non possono vedersi e si trovano in una zona più scura. </li><li> Verificare che i topi sono calmi e applicare von filamenti Frey alla parte laterale della zampa: partire con il filamento di 0,02 g, applicare forza alla zampa sinistra per cinque volte per un periodo totale di 30 secondi (circa 2 secondi per lo stimolo) e valutare la reazione del mouse dopo ogni applicazione. Ripetere con il filamento stesso e la zampa sinistra di altri topi, e poi ricominciare di nuovo con la zampa destra del primo mouse prima di passare al successivo filamento, ad esempio: <ul><li> Mouse 1: zampa sinistra 5 volte con filamento 0,02 g </li><li> Mouse 2: zampa sinistra 5 volte con filamento 0,02 g ecc </li><li> Mouse 1: zampa destra 5 volte con filamento 0,02 g </li><li> Mouse 2: zampa destra 5 volte con filamento 0,02 g ecc </li><li> Mouse 1: zampa sinistra 5 volte con filamento 0,04 g </li><li> Mouse 2: zampa sinistra 5 volte con filamento 0,04 g ecc </li></ul></li><li> Risposta a tre su cinque stimoli è considerato come una reazione positiva. Filamenti di sopra threshold può essere applicata per verificare il livello di soglia. = Risposta zampa ritiro improvviso, repentino battere ciglio, zampa improvvisa leccare. </li></ol> 6. Rappresentante risultati <img src="/files/ftp_upload/3092/3092fig1.jpg" alt="Figura 1" /> Figura 1. Test Von Frey su C57BL / 6 anni topi 8-10 settimane è stato eseguito 1 giorno prima dell&#39;operazione e ripetutamente dopo l&#39;intervento. 4-8 animali sono stati inclusi in ciascun gruppo. Von Frey test del lato operato è rappresentato da ipsilaterale e il non-operato lato il controllo è rappresentato da controlaterale. Il giorno dopo l&#39;intervento degli animali sviluppare ipersensibilità significativo sulla zampa utilizzati mentre il non-operato è solo leggermente influenzata rispetto ai topi sham operato. (* &lt;0,05, ** &lt;0.01, *** &lt;0,001, rispetto ipsilaterale al controlaterale).

<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>

Nessun conflitto di interessi dichiarati.

Passaggi critici Danni al nervo surale deve essere evitata al fine di studiare i cambiamenti patologici nel pregiudizio intatta a seguito del nervo surale ai nervi tibiale e peroneo comune. Danni collaterali al nervo surale può portare alla paralisi e può quindi essere visualizzato come il trascinamento dell&#39;arto operato posteriori. Solo la parte laterale della zampa è innervato dal nervo surale risparmiato e, quindi, solo questa zona si sviluppa neuropatia. Test altre zone, innervate dai nervi recisi, può fortemente influenzare la valutazione dei alterata soglia meccanica. Eventuali modifiche Altri tipi di legature possono essere complementari in termini di studio delle condizioni patologiche del dolore dopo un trauma del nervo periferico, come una lesione costrizione cronica [5] o la legatura parziale del nervo [6]. Ogni procedura di risultati sperimentali in diversi cambiamenti fenotipici che deve essere valutata prima di post-chirurgia test. Inoltre, altri test sensoriali come iperalgesia termica può essere applicato [7], anche se questo fenotipo è meno robusta SNI seguente. Applicazioni future Questa tecnica può essere utilizzata per la sperimentazione dei farmaci alterare lo sviluppo o la manutenzione di allodinia meccanica [8]. Analisi del nervo sciatico, gangli delle radici dorsali (DRG) e / o il midollo spinale lombare consentirà la ricerca dei meccanismi molecolari responsabili del fenotipo indotto. La sperimentazione animale è stata eseguita secondo buona prassi di laboratorio nel pieno rispetto della normativa danese ed europea. Tutti gli esperimenti sono stati approvati dal danese Animal Experiments Ispettorato del Ministero della Giustizia (autorizzazione numero 2006/561-1206).

<table border="1"><tbody><tr><td> Anestesia: ketamina e xilazina </td><td> Bisturi </td></tr><tr><td> Pomata oftalmica </td><td> Paio di pinzette nr. 2 </td></tr><tr><td> Rasoio elettrico </td><td> Paio di pinzette nr. 5 </td></tr><tr><td> Guanti </td><td> Paio di forbici per dissezione </td></tr><tr><td> 70% (v / v) di etanolo </td><td> Micro forbici </td></tr><tr><td> Cotone-lana gemme / sguardo </td><td> 6,0 Prolene non riassorbibili sutura </td></tr><tr><td> Microscopio stereo </td><td> 2 x ago titolari </td></tr><tr><td> Nastro adesivo </td><td> Calore pad </td></tr><tr><td colspan="2"> filamenti di von Frey (0,02 - 2,0 g per i topi) (Stoelting) </td></tr></tbody></table>

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

Dolore neuropatico periferico è una condizione di dolore cronico grave che può derivare da traumi ai nervi sensoriali del sistema nervoso periferico. Il risparmiato lesioni nervose (SNI) modello induce sintomi del dolore neuropatico, come dolore allodinia meccanica cioè a causa di stimoli tattili che normalmente non provocano una risposta dolorosa [1]. Il modello murino SNI prevede la legatura di due delle tre branche del nervo sciatico (il nervo tibiale e il nervo peroneo comune), mentre il nervo surale è lasciata intatta [2]. I risultati lesione nei casi di ipersensibilità segnato nella zona laterale della zampa, che è innervato dal nervo surale risparmiato. La non-operato lato del mouse può essere utilizzato come controllo. I vantaggi del modello di SNI sono la robustezza della risposta e che non richiede competenze di microchirurgia esperti. La soglia del dolore per la risposta meccanica è determinata dal test con filamenti di Von Frey di crescente forza di piega, che vengono ripetutamente premuto contro la superficie laterale della zampa [3], [4]. Una reazione positiva dolore è definito come il ritiro zampa improvvisa, indietreggiare e / o leccarsi zampa indotto dal filamento. Una risposta positiva a tre su cinque stimoli ripetitivi è definita come la soglia del dolore. Come dimostrato nel protocollo video, topi C57BL / 6 allodinia profonda esperienza già a partire dal giorno seguente l&#39;intervento chirurgico e mantenere questo per diverse settimane.

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

Il Risparmiata lesioni nervose modello animale è descritto qui come un modello murino di dolore neuropatico periferico dopo denervazione parziale del nervo sciatico con lesione dei rami del nervo tibiale e peroneale comune, lasciando i rimanenti nervo surale intatta. Modificazione comportamentale che derivi dalla meccanica allodinia è quantificato da von filamenti Frey.

Risparmiato il Nerve pregiudizio (SNI) Modello di Allodinia meccanica indotta nei topi

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

62.8K Views.  Aarhus University. Il Risparmiata lesioni nervose modello animale è descritto qui come un modello murino di dolore neuropatico periferico dopo denervazione parziale del nervo sciatico con lesione dei rami del nervo tibiale e peroneale comune, lasciando i rimanenti nervo surale intatta. Modificazione comportamentale che derivi dalla meccanica allodinia è quantificato da von filamenti Frey.

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

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

Muscle strains are one of the most common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and physical exam alone, however the clinical presentation can vary greatly depending on the extent of injury, the patient's pain tolerance, etc. In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Biological markers, such as serum creatine kinase levels, are typically elevated with muscle injury, but their levels do not always correlate with the loss of force production. This is even true of histological findings from animals, which provide a "direct measure" of damage, but do not account for all the loss of function. Some have argued that the most comprehensive measure of the overall health of the muscle in contractile force. Because muscle injury is a random event that occurs under a variety of biomechanical conditions, it is difficult to study. Here, we describe an in vivo animal model to measure torque and to produce a reliable muscle injury. We also describe our model for measurement of force from an isolated muscle in situ. Furthermore, we describe our small animal MRI procedure.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo animal model of injury is described. The method takes advantage of the subcutaneous position of the fibular nerve. Velocity, timing of muscle activation, and arc of motion are all pre-determined and synchronized using commercial software. Post injury changes are monitored in vivo using MR imaging/spectroscopy.

Un In vivo Modello di roditore della contrazione indotta Lesioni e monitoraggio non invasivo di recupero

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

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Medicina

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.

Ischemia-riperfusione Modello di danno renale acuta e post Infortunio fibrosi nei topi

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;

Resezione del nervo tibiale - Un modello standardizzato per Denervation indotta scheletrico atrofia muscolare nei topi

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.

Il nervo sciatico ammanettare modello di dolore neuropatico nei topi

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.

Facial assotomia Nerve in topi: un modello per studiare motoneuroni risposta al danno

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.

Photothrombosis-indotta ischemia focale come un modello di lesioni del midollo spinale nei topi

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.

Un ripetitivo Concussive trauma cranico Modello nei topi

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.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

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.

Induzione di aterosclerosi accelerata nei topi: il modello "Wire-Injury"

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.

Un modello di lesioni del midollo spinale contusivo basato sullo spostamento tissutale nei topi

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.

Misurazione di contrattilità isometrica dell'arteria mesenterica Mouse con filo Myography

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