<ol style="padding:5px;"><li> Installation des équipements <ol style="padding:5px;"><li> Sélectionnez une zone dans le laboratoire où la température ambiante est égale ou supérieure à 20 ° C. </li><li> Éviter les endroits à proximité de bouches de climatisation et / ou un bruit fort. </li><li> Évitez d&#39;exposer l&#39;animal à des odeurs qui peuvent causer une irritation et / ou de stress, comme le parfum, eau de cologne, produits chimiques, et les nettoyeurs. </li><li> Mettez le contrôleur de l&#39;ADOC. </li><li> Tournez sur la plateforme de réchauffement et mis au niveau 3. Si la température ambiante est supérieure à 25 ° C, commencer le niveau 1 ou 2. </li><li> Diagnostics contrôleur de test <ol style="padding:5px;"><li> Ouvrez le logiciel CODA. </li><li> Sélectionnez le périphérique CODA en cliquant dessus. </li><li> Cliquez sur &quot;périphérique de test sélectionné&quot;. </li><li> Sélectionnez les menottes et les chaînes de test et cliquez sur &quot;Test&quot;. </li><li> Après le test est terminé, fermez la fenêtre de test de périphériques. </li></ol></li></ol></li><li> Logiciel Set-up <ol style="padding:5px;"><li> Gérer le personnel et les animaux <ol style="padding:5px;"><li> Gérer le personnel <ol style="padding:5px;"><li> Sélectionnez Outils&gt; Gérer le personnel. </li><li> Cliquez sur &quot;Chercheur 1&quot; pour changer le nom. </li><li> Cliquez sur les lignes suivantes à ajouter d&#39;autres chercheurs. </li><li> Cliquez sur &quot;Tab techniciens&quot; pour modifier ou ajouter des techniciens. </li><li> Cliquez sur &quot;Tab spécimens&quot; au nom de l&#39;animal pour faciliter l&#39;identification et de sélectionner le type d&#39;animal; rat ou la souris. </li><li> Cliquez sur &quot;Tab Animaux&quot; pour modifier le type d&#39;animal et de sensibilité. </li></ol></li></ol></li><li> Expérience Nouveau <ol style="padding:5px;"><li> Sélectionnez Fichier&gt; Nouveau&gt; Expérience </li><li> Commencez une nouvelle expérience <ol style="padding:5px;"><li> Entrez le nom d&#39;expérience. </li><li> Sélectionnez le chercheur clé. </li><li> Sélectionnez une date de début et cliquez sur &quot;Suivant&quot;. </li></ol></li><li> Infos session de base <ol style="padding:5px;"><li> Entrez le nom de session </li><li> Réglez &quot;Cycles Acclimatation&quot; à 5. Si vous préférez ne pas inclure les cycles d&#39;acclimatation, mis à zéro. Pour aider à acclimater l&#39;animal au titulaire et aux poignets, ajouter des cycles additionnels de votre expérience et d&#39;exécuter l&#39;expérience pendant 5 jours consécutifs. </li><li> Set &quot;Nombre de jeux&quot; à 1. </li><li> Réglez &quot;Temps entre les séries» à 30 secondes pour plusieurs jeux </li><li> Réglez &quot;Cycles par Set&quot; à 20 cycles. </li><li> Réglez &quot;temps entre les cycles&quot; à 5 secondes et cliquez sur &quot;Suivant&quot;. </li></ol></li></ol></li><li> Sélection des échantillons <ol style="padding:5px;"><li style="list-style-type:none;"><ol style="padding:5px;"><li> Sélectionnez le spécimen de la piscine échantillon et de l&#39;attribuer à la première chaîne disponible et cliquez sur &quot;Suivant&quot;. </li></ol></li><li> Paramètres de session <ol style="padding:5px;"><li> Réglez &quot;pression d&#39;occlusion maximale&quot; à 250mmHg. </li><li> Réglez &quot;Temps déflation&quot; à 20 secondes pour les souris, 15 secondes pour les rats. </li><li> Réglez «Volume minimum&quot; à 15ul. </li><li> Réglez &quot;Style d&#39;affichage&quot; pour un canal par le graphique. </li><li> Ajouter d&#39;autres chercheurs et techniciens et cliquez sur &quot;Suivant&quot;. </li></ol></li><li> Script de session <ol style="padding:5px;"><li> Lisez le script de session et cliquez sur &quot;Suivant&quot;. </li><li> Ne pas sélectionner Terminer jusqu&#39;à ce que vous remplir la section Préparation des animaux. </li></ol></li></ol></li></ol></li><li> Préparation des animaux <ol style="padding:5px;"><li> Mise en place des animaux <ol style="padding:5px;"><li> Titulaire d&#39;animaux <ol style="padding:5px;"><li> Pour former et améliorer l&#39;acclimatation des animaux, ajouter des cycles supplémentaires à votre expérience et exécuter le test pendant cinq jours consécutifs. Vous pouvez également aider à l&#39;acclimatation en plaçant l&#39;animal dans le support pendant 15 minutes pendant 3 jours consécutifs avant l&#39;étude réelle. </li><li> Placer chaque animal dans un support en ramassant l&#39;animal par la queue et plaçant délicatement l&#39;animal dans l&#39;arrière de la porte qui fait face à l&#39;extrémité ouverte du cône de nez. Note: la taille de l&#39;animal par rapport à la taille du support sélectionné. L&#39;animal doit être en mesure d&#39;entrer librement sur le support. </li><li> Soigneusement sécuriser le hayon arrière au titulaire en tournant le bouchon à vis rouge sur le hayon arrière. Des précautions doivent être prises pour éviter de pincer la queue ou de toute autre partie du corps tout en sécurisant le hayon arrière. </li><li> Glissez le cône de nez vers le hayon arrière, limitant le mouvement de l&#39;animal. Le cône de nez devrait être en mesure de limiter l&#39;animal de se retourner tout l&#39;intérieur du support. </li><li> Placez le support sur la plateforme du réchauffement de la position désignée. Si les tests du même groupe d&#39;animaux sur une période de temps, assurez-vous de tourner la position de titulaire sur la plateforme réchauffement d&#39;améliorer la cohérence et la précision de vos mesures de pression artérielle. Laisser l&#39;animal au moins 5 minutes pour s&#39;acclimater à son titulaire. </li><li> Ne pas toucher ou manipuler l&#39;animal tandis que l&#39;intérieur du support. La multiplication des contacts pourraient irriter l&#39;animal. </li><li> Ne jamais laisser l&#39;animal dans la détenteurs sans surveillance. </li></ol></li><li> Placement manchette <ol style = &quot;padding: 5px;&quot;&gt; <li> Discussion de la queue à travers la &quot;manchette d&#39;occlusion.&quot; Placer ce brassard à proximité de la base de la queue que possible sans la force. </li><li> Fixez le &quot;Tuyau d&#39;occlusion du brassard&quot; dans l&#39;encoche à l&#39;arrière haut de la porte. </li><li> Discussion de la queue à travers la &quot;manchette VPR», le plaçant au sein de 2mm de la &quot;manchette d&#39;occlusion». </li><li> Fixez le &quot;Tube Cuff VPR» dans l&#39;encoche à l&#39;arrière haut de la porte. </li><li> Attacher les poignets au contrôleur CODA. </li></ol></li><li> Température des animaux <ol style="padding:5px;"><li> Laisser l&#39;animal au moins 5 minutes de thermorégulation. </li><li> Enregistrer la température de l&#39;animal fréquemment avec un thermomètre infrarouge. La température des animaux doit être de 32 à 35 ° C. </li></ol></li></ol></li></ol></li><li> Expérience <ol style="padding:5px;"><li> Commencez l&#39;expérience <ol style="padding:5px;"><li> Cliquez sur «Finish» et l&#39;expérience va commencer immédiatement. </li></ol></li><li> Expérience Fin <ol style="padding:5px;"><li> Des animaux <ol style="padding:5px;"><li> Immédiatement enlever l&#39;animal de les poignets et titulaire. </li><li> Retour à l&#39;animal de leur cage après le «Rapport sommaire de la session Simple&quot; est affiché. </li></ol></li><li> Collecte des données <ol style="padding:5px;"><li> Après l&#39;achèvement du dernier cycle des cycles acceptés seront automatiquement affichées sous forme de tableur dans l&#39;application CODA, cliquez sur &quot;Exporter vers Excel&quot; bouton. </li><li> Sélectionnez le dossier à placer le fichier dans, le nom du fichier et cliquez sur &quot;Enregistrer&quot;. </li><li> Ouvrez l&#39;application Excel et cliquez sur «Ouvrir». </li><li> Voir tous les types de fichiers à afficher le &quot;. Csv sauvé&quot;, cliquez sur le fichier. </li></ol></li><li> Traitement des données <ol style="padding:5px;"><li> Il ya plusieurs façons de traiter les données dans le fichier Excel. Une pratique courante est d&#39;obtenir l&#39;écart moyen et standard. </li><li> Supprimer les mesures si l&#39;écart type est supérieur à 30. </li><li> En outre, les cycles de rejet peut être consulté et toute la base peuvent être facilement exportées vers Excel. Consultez le manuel d&#39;utilisation pour des informations supplémentaires. </li></ol></li></ol></li></ol></li></ol>

The authors have nothing to disclose.

measuring blood pressure in mice using volume pressure recording, a tail-cuff method

Le CODA 8-canaux à haut débit système non invasif de pression artérielle mesure la pression sanguine dans jusqu&#39;à 8 simultanément souris ou les rats. Le CODA-queue coiffe système utilise enregistrement de la pression du volume (VPR) pour mesurer la pression artérielle en déterminant le volume de sang de la queue. Un capteur différentiel de pression spécialement conçu et une occlusion de la queue-coiffe mesurer le volume total de sang dans la queue, sans la nécessité d&#39;obtenir le signal d&#39;impulsion individuelle. Une attention particulière est accordée à la longueur de la manchette d&#39;occlusion afin d&#39;en tirer les lectures les plus précises pression sanguine. VPR peuvent facilement obtenir des lectures sur la peau foncée rongeurs, comme les souris C57BL6 et est compatible avec l&#39;IRM. Le système ADOC vous fournit des mesures de six (6) différents paramètres de pression artérielle; pression artérielle systolique et diastolique, fréquence cardiaque, pression artérielle moyenne, le flux sanguin queue, et le volume sanguin de la queue. Les mesures peuvent être faites sur des souris soit éveillé ou anesthésié ou des rats. Le système comprend un contrôleur CODA, ordinateur portable, logiciel, poignets, les détenteurs d&#39;animaux, plaquettes réchauffement infrarouge et un thermomètre infrarouge. Il ya sept tailles de supports différents pour les souris aussi petits que 8 grammes pour les rats aussi gros que 900 grammes.

Watch this Scientific Journal Video about Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method at JoVE.com

Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

Le CODA 8-canaux à haut débit système non invasif de pression artérielle mesure la pression sanguine dans jusqu&#39;à 8 simultanément souris ou les rats. Ce système utilise la queue-coiffe enregistrement de la pression du volume (VPR) pour mesurer la pression artérielle en déterminant le volume de sang de la queue.

Mesurer la pression sanguine chez des souris en utilisant enregistrement de la pression du volume, une méthode Queue-coiffe

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. The CODA ...

measuring-blood-pressure-mice-using-volume-pressure-recording-tail

Research

JoVE Journal

Biology

51.3K vues.  University of Kentucky. Le CODA 8-canaux à haut débit système non invasif de pression artérielle mesure la pression sanguine dans jusqu'à 8 simultanément souris ou les rats. Ce système utilise la queue-coiffe enregistrement de la pression du volume (VPR) pour mesurer la pression artérielle en déterminant le volume de sang de la queue.

Vidéo: Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

Pressure-polishing Pipettes for Improved Patch-clamp Recording

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording. We first developed this method for fabricating pipettes suitable for recording from small (&lt;3 m) neuronal cell bodies. The basic principal is similar to glass-blowing and combines air pressure and heat to modify the shape of patch pipettes prepared by a conventional micropipette puller. It can be applied to so-called soft (soda lime) and hard (borosilicate) glasses. Generally speaking, pressure polishing can reduce pipette resistance by 25% without decreasing the diameter of the tip opening (Goodman and Lockery, 2000). It can be applied to virtually any type of glass and requires only the addition of a high-pressure valve and fitting to a microforge. This technique is essential for recording from ultrasmall cells (&lt;5 m) and can also improve single-channel recording by minimizing pipette resistance. The blunt shape is also useful for perforated-patch clamp recording since this tip shape results in a larger membrane bleb available for perforation.

This is a guide to modifying the shape of glass micropipettes. Specifically, by using heat and air pressure the taper is widened without increasing the tip opening, leading to lower pipette resistance. This is critical to obtain low noise recordings of small cells but is useful in many applications.

Pression de polissage Pipettes Amélioration patch-clamp

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording.  We first developed this method for fabricating pipettes suitable for ...

Recherche

Biologie

Making Gynogenetic Diploid Zebrafish by Early Pressure

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from heterozygous females allow F1 mutagenized females to be screened directly for deleterious mutations in an accelerated forward genetic screen. Streisinger et al.1,2 described methods for making gynogenetic (homozygous) diploid zebrafish by activating zebrafish eggs with ultraviolet light-inactivated sperm and preventing either the second meiotic or the first zygotic cell division using physical treatments (heat or pressure) that deploymerize microtubules. The "early pressure" (EP) method blocks the meiosis II, which occurs shortly after fertilization. The EP method produces a high percentage of viable embryos that can develop to fertile adults of either sex. The method generates embryos that are homozygous at all loci except those that were separated from their centromere by recombination during meiosis I. Homozygous mutations are detected in EP clutches at between 50% for centromeric loci and less than 1% for telomeric loci. This method is reproduced verbatim from the Zebrafish Book3.

This is a method for generating gynogenetic diploid zebrafish embryos (embryos whose only genetic contribution comes from the mother) by blocking the second meiotic division immediately after fertilization with ultraviolet light-inactivated sperm. EP embryos are not fully homozygous due to recombination during the first meiotic division, however they are homozygous at all loci that have not been separated from their centromere by recombination.

Faire Zebrafish diploïdes gynogénétiques par Pression Early

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from ...

Ambulatory ECG Recording in Mice

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although the surface ECG is utilized for short-term measurements of waveform intervals, it is not practical for long-term studies of heart rate variability or the capture of rare episodes of arrhythmias. Implantable ECG telemeters offer the advantages of simple surgical implantation, long-term recording of electrograms in ambulatory mice, and scalability with simultaneous recordings of multiple animals. Here, we present a step-by-step guide to the implantation of telemeters for ambulatory ECG recording in mice. Careful adherence to aseptic technique is required for favorable survival results with the possibility of implantation and recording from weeks to months. Thus, implantable ECG telemetry is a valuable tool for detection of critical information on cardiac electrophysiology in ambulatory animal models such as the mouse. 

Telemetric ECG has emerged as an essential tool in evaluating animal models for cardiac arrhythmias and sudden cardiac death. Here, we present a stepwise guide to telemetric ECG recordings for application in long-term ambulatory ECG monitoring in mice.

Ambulatoires enregistrement de l&#39;ECG chez des souris

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although ...

Isolation and Culture of Pulmonary Endothelial Cells from Neonatal Mice

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells (HUVECs) have shown to be convenient, easy to obtain and culture, and thus are the most widely studied endothelial cells. However, for research focused on processes like angiogenesis, permeability or many others, microvascular endothelial cells (ECs) are a much more physiologically relevant model to study 2. Furthermore, ECs isolated from knockout mice provide a useful tool for analysis of protein function ex vivo. Several approaches to isolate and culture microvascular ECs of different origin have been reported to date 3-7, but consistent isolation and culture of pure ECs is still a major technical problem in many laboratories. Here, we provide a step-by-step protocol on a reliable and relatively simple method of isolating and culturing mouse lung endothelial cells (MLECs). In this approach, lung tissue obtained from 6- to 8-day old pups is first cut into pieces, digested with collagenase/dispase (C/D) solution and dispersed mechanically into single-cell suspension. MLECS are purified from cell suspension using positive selection with anti-PECAM-1 antibody conjugated to Dynabeads using a Magnetic Particle Concentrator (MPC). Such purified cells are cultured on gelatin-coated tissue culture (TC) dishes until they become confluent. At that point, cells are further purified using Dynabeads coupled to anti-ICAM-2 antibody. MLECs obtained with this protocol exhibit a cobblestone phenotype, as visualized by phase-contrast light microscopy, and their endothelial phenotype has been confirmed using FACS analysis with anti-VE-cadherin 8 and anti-VEGFR2 9 antibodies and immunofluorescent staining of VE-cadherin. In our hands, this two-step isolation procedure consistently and reliably yields a pure population of MLECs, which can be further cultured. This method will enable researchers to take advantage of the growing number of knockout and transgenic mice to directly correlate in vivo studies with results of in vitro experiments performed on isolated MLECs and thus help to reveal molecular mechanisms of vascular phenotypes observed in vivo.

Here, we describe a protocol for isolation and culture of murine pulmonary endothelial cells. This method comprises mechanic and enzymatic lung tissue dissociation as well as a 2-step purification process using anti-PECAM-1 and anti-ICAM-2 antibodies conjugated to magnetic beads, which produces a pure endothelial cell population of mostly microvascular origin.

Isolement et culture de cellules endothéliales pulmonaires des souris néonatales

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells ...

Assessing Murine Resistance Artery Function Using Pressure Myography

Pressure myograph systems are exquisitely useful in the functional assessment of small arteries, pressurized to a suitable transmural pressure. The near physiological condition achieved in pressure myography permits in-depth characterization of intrinsic responses to pharmacological and physiological stimuli, which can be extrapolated to the in vivo behavior of the vascular bed. Pressure myograph has several advantages over conventional wire myographs. For example, smaller resistance vessels can be studied at tightly controlled and physiologically relevant intraluminal pressures. Here, we study the ability of 3rd order mesenteric arteries (3-4 mm long), preconstricted with phenylephrine, to vaso-relax in response to acetylcholine. Mesenteric arteries are mounted on two cannulas connected to a pressurized and sealed system that is maintained at constant pressure of 60 mmHg. The lumen and outer diameter of the vessel are continuously recorded using a video camera, allowing real time quantification of the vasoconstriction and vasorelaxation in response to phenylephrine and acetylcholine, respectively.
	To demonstrate the applicability of pressure myography to study the etiology of cardiovascular disease, we assessed endothelium-dependent vascular function in a murine model of systemic hypertension. Mice deficient in the &alpha;1 subunit of soluble guanylate cyclase (sGC&alpha;1-/-) are hypertensive when on a 129S6 (S6) background (sGC&alpha;1-/-S6) but not when on a C57BL/6 (B6) background (sGC&alpha;1-/-B6). Using pressure myography, we demonstrate that sGC&alpha;1-deficiency results in impaired endothelium-dependent vasorelaxation. The vascular dysfunction is more pronounced in sGC&alpha;1-/-S6 than in sGC&alpha;1-/-B6 mice, likely contributing to the higher blood pressure in sGC&alpha;1-/-S6 than in sGC&alpha;1-/-B6 mice. 
	Pressure myography is a relatively simple, but sensitive and mechanistically useful technique that can be used to assess the effect of various stimuli on vascular contraction and relaxation, thereby augmenting our insight into the mechanisms underlying cardiovascular disease.

In pressure myography, an intact small segment of a vessel is mounted onto two small cannulas and pressurized to a suitable luminal pressure. Here, we describe the method to measure vasorelaxation response of the mouse 3rd order mesenteric arteries in c57 and sGC&alpha;1-/- mice using pressure myography.

Évaluer murin Fonction Artère résistance à l&#39;aide myographie de pression

Pressure myograph systems are  exquisitely useful in the functional assessment of small arteries, pressurized  to a suitable transmural pressure. The  ...

One-channel Cell-attached Patch-clamp Recording

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate depends on properties intrinsic to each channel protein as well as the mechanism by which it is generated and controlled and represents an important area of current research. Information about the operational dynamics of ion channel proteins can be obtained by observing long stretches of current produced by a single molecule. Described here is a protocol for obtaining one-channel cell-attached patch-clamp current recordings for a ligand gated ion channel, the NMDA receptor, expressed heterologously in HEK293 cells or natively in cortical neurons. Also provided are instructions on how to adapt the method to other ion channels of interest by presenting the example of the mechano-sensitive channel PIEZO1. This method can provide data regarding the channel&#8217;s conductance properties and the temporal sequence of open-closed conformations that make up the channel&#8217;s activation mechanism, thus helping to understand their functions in health and disease.

Described here is a procedure for obtaining long stretches of current recording from one ion channel with the cell-attached patch-clamp technique. This method allows for observing, in real time, the pattern of open-close channel conformations that underlie the biological signal. These data inform about channel properties in undisturbed biological membranes.

Un canal cellulaire attaché patch-clamp enregistrement

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate ...

Measuring Respiratory Function in Mice Using Unrestrained Whole-body Plethysmography

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative assessment of lung function in rodent models is an important tool in the development of future therapies. Commonly used techniques for assessing respiratory function including invasive plethysmography and forced oscillation. While these techniques provide valuable information, data collection can be fraught with artefacts and experimental variability due to the need for anesthesia and/or invasive instrumentation of the animal. In contrast, unrestrained whole-body plethysmography (UWBP) offers a precise, non-invasive, quantitative way by which to analyze respiratory parameters. This technique avoids the use of anesthesia and restraints, which is common to traditional plethysmography techniques. This video will demonstrate the UWBP procedure including the equipment set up, calibration and lung function recording. It will explain how to analyze the collected data, as well as identify experimental outliers and artefacts that results from animal movement. The respiratory parameters obtained using this technique include tidal volume, minute volume, inspiratory duty cycle, inspiratory flow rate and the ratio of inspiration time to expiration time. UWBP does not rely on specialized skills and is inexpensive to perform. A key feature of UWBP, and most appealing to potential users, is the ability to perform repeated measures of lung function on the same animal.

The assessment of respiratory physiology has traditionally relied upon techniques, which require restraint or sedation of the animal. Unrestrained whole-body plethysmography, however, provides precise, non-invasive, quantitative analysis of respiratory physiology in animal models. In addition, the technique allows repeated respiratory assessment of mice allowing for longitudinal studies.

Mesure de la fonction respiratoire chez la souris utilisant effrénée pléthysmographie corps entier

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative ...

A Method for Screening and Validation of Resistant Mutations Against Kinase Inhibitors

The discovery of BCR/ABL as a driver oncogene in chronic myeloid leukemia (CML) resulted in the development of Imatinib, which, in fact, demonstrated the potential of targeting the kinase in cancers by effectively treating the CML patients. This observation revolutionized drug development to target the oncogenic kinases implicated in various other malignancies, such as, EGFR, B-RAF, KIT and PDGFRs. However, one major drawback of anti-kinase therapies is the emergence of drug resistance mutations rendering the target to have reduced or lost affinity for the drug. Understanding the mechanisms employed by resistant variants not only helps in developing the next generation inhibitors but also gives impetus to clinical management using personalized medicine. We reported a retroviral vector based screening strategy to identify the spectrum of resistance conferring mutations in BCR/ABL, which has helped in developing the next generation BCR/ABL inhibitors. Using Ruxolitinib and JAK2 as a drug target pair, here we describe in vitro screening methods that utilizes the mouse BAF3 cells expressing the random mutation library of JAK2 kinase.

Emergence of genetic resistance against kinase inhibitor therapy poses significant challenge for effective cancer therapy. Identification and characterization of resistant mutations against a newly developed drug helps in better clinical management and next generation drug design. Here, we describe our protocol for in vitro screening and validation of resistant mutations.

Un procédé de criblage et de validation des mutations de résistance aux inhibiteurs de kinase

The discovery of BCR/ABL as a driver oncogene in chronic myeloid leukemia (CML) resulted in the development of Imatinib, which, in fact, demonstrated the ...

Cardiac Pressure-Volume Loop Analysis Using Conductance Catheters in Mice

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular systolic and diastolic function. Measures of ventricular contractility and compliance are obtained through examination of cardiac response to changes in afterload and preload. These techniques were originally developed nearly three decades ago to measure cardiac function in large mammals and humans. The application of these analyses to small mammals, such as mice, has been accomplished through the optimization of microsurgical techniques and creation of conductance catheters. Conductance catheters allow for estimation of the blood pool by exploiting the relationship between electrical conductance and volume. When properly performed, these techniques allow for testing of cardiac function in genetic mutant mouse models or in drug treatment studies. The accuracy and precision of these studies are dependent on careful attention to the calibration of instruments, systematic conduct of hemodynamic measurements and data analyses. We will review the methods of conducting pressure-volume loop experiments using a conductance catheter in mice.

Cardiac pressure-volume loop analysis is the most comprehensive way to measure cardiac function in the intact heart. We describe a technique to perform and analyze cardiac pressure volume loops, using conductance catheters.

Cardiac pression-volume Analyse boucle aide de la conductance cathéters chez la souris

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular ...

Measuring Pressure Volume Loops in the Mouse

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the mouse provides great potential to explore cardiac function at the molecular level. The mouse&#39;s small size does present some challenges in regards to performing detailed cardiac phenotyping. Miniaturization and other advancements in technology have made many methods of cardiac assessment possible in the mouse. Of these, the simultaneous collection of pressure and volume data provides a detailed picture of cardiac function that is not available through any other modality. Here a detailed procedure for the collection of pressure-volume loop data is described. Included is a discussion of the principles underlying the measurements and the potential sources of error. Anesthetic management and surgical approaches are discussed in great detail as they are both critical to obtaining high quality hemodynamic measurements. The principles of hemodynamic protocol development and relevant aspects of data analysis are also addressed.

This manuscript describes a detailed protocol for the collection of pressure-volume data from the mouse.

Mesure de boucles de volume de pression chez la souris

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the ...