This work was supported in part by the Lupus Research Institute, NCI grant CA138617, NIDDK grant DK090978, Charlotte Schmidlapp Award (to E.M.J.), and the Albert J. Ryan Fellowship (to J.K.). We are grateful for the support and instrumentation provided by the Research Flow Cytometry Core in the Division of Rheumatology at Cincinnati Children&#8217;s Hospital Medical Center, supported in part by NIH AR-47363, NIH DK78392 and NIH DK90971.

<ol>
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The authors have nothing to disclose.

Le modèle inductible de BM12 est un moyen relativement simple et efficace pour étudier les processus cellulaires et moléculaires de SLE. L&#39;activation chronique des cellules T CD4 par transfert adoptif dirigés contre les antigènes du soi conduit à l&#39;accumulation de Tfh, GC cellules B, les cellules plasmatiques et qui peut être mesurée par cytométrie en flux, comme décrit ici. Les futures études utilisant ce modèle peut rapidement et facilement interroger le rôle des gènes candidats et de nouvelles t...

Le lupus érythémateux disséminé (SLE) est une maladie auto-immune complexe caractérisé prototypiquement par l&#39;anticorps anti-nucléaire (ANA) et de la production glomérulonéphrite. De nombreux autres séquelles, y compris dermique, cardio-pulmonaire, et des lésions hépatiques sont associés à la maladie chez certains individus. Les estimations de prévalence aux États-Unis varient considérablement, de 1,2 150,000-1,500,000, avec incidence particulièrement élevée chez les femmes et les minorités 3. Bien que l&#39;étiologie de la SLE a été difficile à discerner, il est pensé pour résulter de l&#39;interaction de divers facteurs génétiques et environnementaux, qui culminent dans l&#39;auto-immunité systémique. De nombreux modèles animaux ont été utilisés pour étudier les facteurs menant à l&#39;apparition et la progression de la maladie. Des modèles de souris classiques de SLE comprennent des souches de souris génétiquement prédisposées, y compris le modèle NZB x NZW F1 et ses dérivés NZM, la souche / lpr MLR, et la souche BXSB / Yaa et systèmes inductibles, tels que le pristane et la maladie chronique du greffon contre l&#39;hôte (cGVHD) 4 modèles. Les premiers rapports de la production d&#39;auto-anticorps dans des modèles de GVHD utilisés souches de souris différentes souches ou de hamster pour parent dans les transferts de F1 5 - 8; méthodes les plus courantes utilisées pour étudier la maladie lupique comprennent actuellement les DBA / 2 → parent (C57BL / 6 x DBA / 2) F1, et le modèle de transfert de bm12 décrit ici. Chaque modèle a ses propres mises en garde, mais ils partagent en général un ensemble commun de caractéristiques qui sont en corrélation avec des caractéristiques cliniques de la maladie humaine. Les paramètres les plus souvent rapportés chez les souris modèles comprennent une splénomégalie, lymphadénopathie, la néphrite, la production ANA, et au niveau cellulaire, l&#39;expansion des cellules T folliculaires auxiliaires (TFH), germinales cellules centre (GC) de B et les cellules plasmatiques. Le modèle BM12 inductible est réalisé par le transfert adoptif de lymphocytes à partir de IA BM12 B6 (C) - H2 - Ab1 BM12 / KhEgJ (BM12) chez la souris, une souche identical de souris C57BL / 6 à l&#39;exception de 3 substitutions d&#39;acides aminés sur le CMH de classe II, en IA b / 6 (B6) des souris C57BL. Alloactivation des donateurs cellules T CD4 par bénéficiaire APC conduit à cGVHD avec des symptômes ressemblant étroitement SLE. Plus précisément, ils comprennent l&#39;expansion de Tfh donateurs dérivés, l&#39;expansion des cellules B GC bénéficiaires dérivés et les cellules plasmatiques, et la production d&#39;Anas y compris les anti-ADN double brin, anti-ADNsb, anti-chromatine, et des anticorps anti-RBC 9. Au fil du temps, les souris receveuses développent glomérulonéphrite associée à des dépôts d&#39;IgG dans le glomérulaire, interstitielle, et les régions vasculaires des reins 10. Nous avons récemment montré que, comme dans la maladie humaine, il y a également un rôle critique pour les IFN de type I dans 11 ce modèle. Notamment, les critères de définition des SLE humaine incluent le développement de la néphrite compatible avec SLE en présence d&#39;anticorps anti-ADN anticorps 12, qui sont tous deux traits saillants de ce modèle de souris. Il n&#39;y a en soisieurs avantages du modèle de bm12 sur les modèles spontanés. Modèles classiques qui développent des signes SLE spontanément reposent sur soit des souches de souris hybrides, des souches de souris consanguines pas sur le fond de B6, ou grande loci génétiques sur le fond de B6, qui font traversant au coup de grâce ou de souris autrement génétiquement modifié difficile et fastidieux. Avec le modèle inductible de bm12, des souris génétiquement modifiées peuvent servir soit donneur ou du receveur, qui permet une identification plus rapide du compartiment cellulaire dans lequel des gènes particuliers peuvent être importants pour la maladie. En outre, le développement de la maladie dans le modèle de bm12 est beaucoup plus rapide, ne nécessitant que deux semaines jusqu&#39;à l&#39;apparition d&#39;Anas, contre plusieurs mois pour les modèles les plus spontanées. En outre, à la différence des modèles spontanés qui développent la maladie à différents points dans le temps, le début et la progression de la maladie dans le modèle d&#39;→ B6 BM12 est fortement synchronisés. Cela permet la génération de cohortes de taille appropriée qui peut Be utilisé pour les stratégies d&#39;intervention ou thérapeutiques à tout stade de développement de la maladie. Ce qui suit est un protocole détaillé pour initier SLE auto-immunité par le transfert adoptif de lymphocytes BM12 dans souris C57BL / 6, ou des variantes génétiques sur le fond de B6. En outre, nous décrivons un protocole de coloration par cytométrie de flux pour dénombrer Tfh, GC cellules B, les cellules plasmatiques et les types de cellules associées à la maladie humaine. Fait important, ces protocoles peuvent également être utilisés pour caractériser la plupart des maladies dans les modèles murins de lupus et identifier Tfh, les cellules B GC et les cellules plasmatiques dans d&#39;autres modèles de maladie.

<table border="0">
	<tbody>
		<tr>
			<td>B6.SJL-Ptprca Pepcb/BoyJ</td>
			<td>The Jackson Laboratory</td>
			<td>001162</td>
			<td>CD45.1+ BoyJ mouse strain</td>
		</tr>
		<tr>
			<td>B6(C)-H2-Ab1bm12/KhEgJ</td>
			<td>The Jackson Laboratory</td>
			<td>001162</td>
			<td>Bm12 mouse strain</td>
		</tr>
		<tr>
			<td>FastDigest PsuI</td>
			<td>Life Technologies</td>
			<td>FD1554</td>
			<td>Restriction digest enzyme for genotyping</td>
		</tr>
		<tr>
			<td>1X RBC Lysis Buffer</td>
			<td>eBioscience</td>
			<td>00-4333-57</td>
			<td></td>
		</tr>
		<tr>
			<td>IMDM</td>
			<td>GE Healthcare</td>
			<td>SH30228.01</td>
			<td></td>
		</tr>
		<tr>
			<td>Plasma Separation Tube (PST)</td>
			<td>BD</td>
			<td>365974</td>
			<td>Blood collection tube with Dipotassium EDTA</td>
		</tr>
		<tr>
			<td>Serum Separation Tube (SST)</td>
			<td>BD</td>
			<td>365967</td>
			<td>Blood collection tube with Clot activator / SST Gel</td>
		</tr>
		<tr>
			<td>Ficoll</td>
			<td>GE Healthcare</td>
			<td>17-1440-02&#160;</td>
			<td>High density cell separation solution</td>
		</tr>
		<tr>
			<td>Lympholyte-M</td>
			<td>Cedarlane</td>
			<td>CL5030</td>
			<td>High density cell separation solution</td>
		</tr>
		<tr>
			<td>GL-7-biotin</td>
			<td>eBioscience</td>
			<td>13-5902-82&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptavidin-BUV395</td>
			<td>BD</td>
			<td>564176</td>
			<td></td>
		</tr>
		<tr>
			<td>CD138-BV421</td>
			<td>BioLegend</td>
			<td>142508</td>
			<td></td>
		</tr>
		<tr>
			<td>CD4-BV510</td>
			<td>BioLegend</td>
			<td>100559</td>
			<td></td>
		</tr>
		<tr>
			<td>TCR&#946;-BV605</td>
			<td>BD</td>
			<td>562840</td>
			<td></td>
		</tr>
		<tr>
			<td>CD45.1-BV711</td>
			<td>BioLegend</td>
			<td>110739</td>
			<td></td>
		</tr>
		<tr>
			<td>CD45.2-FITC</td>
			<td>BioLegend</td>
			<td>109806</td>
			<td></td>
		</tr>
		<tr>
			<td>PD-1-PE</td>
			<td>BioLegend</td>
			<td>135206</td>
			<td></td>
		</tr>
		<tr>
			<td>CD19-PerCP</td>
			<td>BioLegend</td>
			<td>115532</td>
			<td></td>
		</tr>
		<tr>
			<td>Fas-PE-Cy7</td>
			<td>BD</td>
			<td>557653</td>
			<td></td>
		</tr>
		<tr>
			<td>CXCR5-APC</td>
			<td>BioLegend</td>
			<td>145506</td>
			<td></td>
		</tr>
		<tr>
			<td>Fixable Viability Dye ef780</td>
			<td>eBioscience</td>
			<td>65-0865-18</td>
			<td></td>
		</tr>
		<tr>
			<td>CD4-BV421</td>
			<td>BioLegend</td>
			<td>100443</td>
			<td></td>
		</tr>
		<tr>
			<td>1.2 ml FACS tube inserts, racked</td>
			<td>USA Scientific</td>
			<td>1412-1400</td>
			<td></td>
		</tr>
		<tr>
			<td>BD Falcon&#8482; Round-Bottom Tubes</td>
			<td>BD</td>
			<td>352017</td>
			<td></td>
		</tr>
	</tbody>
</table>

the bm12 inducible model of systemic lupus erythematosus (sle) in c57bl/6 mice

Systemic lupus erythematosus (SLE) is an autoimmune disease with diverse clinical and immunological manifestations. Several spontaneous and inducible animal models mirror common components of human disease, including the bm12 transfer model. Upon transfer of bm12 splenocytes or purified CD4 T cells, C57BL/6 mice rapidly develop large frequencies of T follicular helper cells (Tfh), germinal center (GC) B cells, and plasma cells followed by high levels of circulating anti-nuclear antibodies. Since this model utilizes mice on a pure C57BL/6 background, researchers can quickly and easily study disease progression in transgenic or knockout mouse strains in a relatively short period of time. Here we describe protocols for the induction of the model and the quantitation Tfh, GC B cells, and plasma cells by multi-color flow cytometry. Importantly, these protocols can also be used to characterize disease in most mouse models of SLE and identify Tfh, GC B cells, and plasma cells in other disease models.

Souris malades développent une splénomégalie en aussi peu que 14 jours, présentant les rates 2-3 fois la taille de souris saines en termes de masse et de la cellularité (Figure 2). Les splénocytes sont séquentiellement débloqué diffusion de la lumière (FSC-A par SSC-A), l&#39;élimination des doublets (FSC-W ou -H par le FSC-A), les cellules viables (faible coloration de la viabilité colorant) et CD4 + TCRβ + (Figure 3A). Le...

Watch this Scientific Journal Video about The bm12 Inducible Model of Systemic Lupus Erythematosus SLE in C57BL/6 Mice at JoVE.com

The bm12 Inducible Model of Systemic Lupus Erythematosus SLE in C57BL/6 Mice

The transfer of bm12 lymphocytes into a C57BL/6 recipient is an established model of systemic lupus erythematosus. Here we describe how to initiate disease using this model and how to characterize T follicular helper cells, germinal center B cells and plasma cells by flow cytometry.

Le bm12 inductible modèle du lupus érythémateux disséminé (SLE) dans C57BL / 6

Systemic lupus erythematosus (SLE) is an autoimmune disease with diverse clinical and immunological manifestations. Several spontaneous and inducible ...

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Research

JoVE Journal

Medicine

The bm12 Inducible Model of Systemic Lupus Erythematosus (SLE) in C57BL/6 Mice

17.1K vues.  Cincinnati Children's Hospital Medical Center and University of Cincinnati College of Medicine. The transfer of bm12 lymphocytes into a C57BL/6 recipient is an established model of systemic lupus erythematosus. Here we describe how to initiate disease using this model and how to characterize T follicular helper cells, germinal center B cells and plasma cells by flow cytometry.

Vidéo: The bm12 Inducible Model of Systemic Lupus Erythematosus SLE in C57BL/6 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.

Les blessures du nerf Epargnée (SNI) Modèle d&#39;allodynie mécanique induite chez la souris

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

Recherche

Médecine

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 Nerve Transection - un modèle standardisé pour dénervation induit l&#39;atrophie musculaire chez les souris

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

Assessing Changes in Volatile General Anesthetic Sensitivity of Mice after Local or Systemic Pharmacological Intervention

One desirable endpoint of general anesthesia is the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is less straightforward than it is in human patients. A widely used behavioral surrogate for hypnosis in rodents is the loss of righting reflex (LORR), or the point at which the animal no longer responds to their innate instinct to avoid the vulnerability of dorsal recumbency. We have developed a system to assess LORR in 24 mice simultaneously while carefully controlling for potential confounds, including temperature fluctuations and varying gas flows. These chambers permit reliable assessment of anesthetic sensitivity as measured by latency to return of the righting reflex (RORR) following a fixed anesthetic exposure. Alternatively, using stepwise increases (or decreases) in anesthetic concentration, the chambers also enable determination of a population's sensitivity to induction (or emergence) as measured by EC50 and Hill slope. Finally, the controlled environmental chambers described here can be adapted for a variety of alternative uses, including inhaled delivery of other drugs, toxicology studies, and simultaneous real-time monitoring of vital signs.

Loss of the righting reflex has long served as a standard behavioral surrogate for unconsciousness, also called hypnosis, in laboratory animals. Alterations in volatile anesthetic sensitivity caused by pharmacological interventions can be detected with a carefully controlled high-throughput assessment system, which may be adapted for delivery of any inhaled therapeutic.

Évaluer les changements dans volatils anesthésie générale sensibilité des souris après l&#39;intervention pharmacologique local ou systémique

One desirable endpoint of general anesthesia is  the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is  less ...

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.

Le nerf sciatique menottes modèle de douleur neuropathique chez les souris

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

Monitoring of Systemic and Hepatic Hemodynamic Parameters in Mice

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. However, due to the small size of the mouse, technical details of the intraoperative monitoring procedure suitable for the mouse were rarely described. Previously we have reported a monitoring procedure to obtain hemodynamic parameters for rats. Now, we adapted the procedure to acquire systemic and hepatic hemodynamic parameters in mice, a species ten-fold smaller than rats. This film demonstrates the instrumentation of the animals as well as the data acquisition process needed to assess systemic and hepatic hemodynamics in mice. Vital parameters, including body temperature, respiratory&#160;rate&#160;and heart&#160;rate were recorded throughout the whole procedure. Systemic hemodynamic parameters consist of carotid artery pressure (CAP) and central venous pressure (CVP). Hepatic perfusion parameters include portal vein pressure (PVP), portal flow rate as well as the flow rate of the common hepatic artery (table 1). Instrumentation and data acquisition to record the normal values was completed within 1.5 h. Systemic and hepatic hemodynamic parameters remained within normal ranges during this procedure.
This procedure is challenging but feasible. We have already applied this procedure to assess hepatic hemodynamics in normal mice as well as during 70% partial hepatectomy and in liver lobe clamping experiments. Mean PVP after resection (n= 20), was 11.41&#177;2.94 cmH2O which was significantly higher (P&#60;0.05) than before resection (6.87&#177;2.39 cmH2O). The results of liver lobe clamping experiment indicated that this monitoring procedure is sensitive and suitable for detecting small changes in portal pressure and portal flow rate. In conclusion, this procedure is reliable in the hands of an experienced micro-surgeon but should be limited to experiments where mice are absolutely needed.

This film demonstrates how to acquire systemic and hepatic hemodynamics in mice. The whole monitoring includes acquisition of vital parameters, systemic blood pressure, central venous pressure, common hepatic artery flow rate, and portal vein pressure as well as the portal flow rate in mice.

Surveillance des paramètres hémodynamiques systémiques et hépatique chez la souris

The use of mouse models in experimental research is of enormous importance for the study of hepatic physiology and pathophysiological disturbances. ...

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 PtenL/L p53L/L LSL-mTERTL/L and thymic T-cell lymphoma model G4 Atm-/- mTERTER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.

Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

Utilisant murin inductible télomérase allèles dans les études de la dégénérescence des tissus / Régénération et cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

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.

Induction de l’athérosclérose accélérée chez les souris : le modèle « 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. ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

Le modèle Pilocarpine d’épilepsie du Lobe Temporal et EEG surveillance d’un système de télémétrie chez la souris

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a diverting enterostomy. The etiology of this disease remains ill-defined but appears to differ from that of classical inflammatory bowel diseases such as Crohn's disease and ulcerative colitis. Research aimed to decipher the pathophysiological mechanisms leading to the development of this disease has been severely hampered by the lack of an appropriate murine model. This protocol generates a murine model of DC that facilitates the study of the immune system's role and its interaction with the microbiome in the development of DC. In this model using C57BL/6 animals, distal parts of the colon are excluded from the fecal stream by creating a distal colostomy, triggering the development of mild to moderate inflammation in the excluded bowel segments and reproducing the hallmark lesions of human DC with a moderate systemic inflammatory response. In contrast to the rat model, a large number of genetically-modified murine models on the C57BL/6 background are available. The combination of these animals with our model allows the potential roles of individual cytokines, chemokines, or receptors of bioactive molecules (e.g., interleukin (IL)-17; IL-10, chemokine CXCL13, chemokine receptors CXCR5 and CCR7, and the sphingosine-1-phosphate receptor 4) to be assessed in the pathogenesis of DC. The availability of congenic mouse strains on the C57BL/6 background largely facilitates transfer experiments to establish the roles of distinct cell types involved in the etiology of DC. Finally, the model offers the opportunity to assess the influences of local interventions (e.g., modification of the local microbiome or local anti-inflammatory therapy) on mucosal immunity in affected and non-affected bowel segments and the on systemic immune homeostasis.

Murine distal colostomy provides a murine model for human diversion colitis, a predominantly lymphocytic colitis in colon segment excluded from the fecal stream.

Murine colostomie distale, un nouveau modèle de colite détournement chez les souris C57BL/6

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a ...

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.

Mesure de la contractilité isométrique de l’artère mésentérique de souris à l’aide de fil myographie

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