Segna McCauley è supportato da un mentore-formazione NIH concedere 5T32HL066991-07 Xander Wehrens è supportato da NIH / NHLBI concede 1R01HL091947-01A209 e 3R01HL089598-03S109

1. Preparazione del Telemetro per impianto chirurgico <ol><li> Prima di inserire un telemetro ECG nel mouse, è importante assicurarsi che il telemetro ECG è sterile e in buone condizioni. Nuovi dispositivi di telemetria sono di norma forniti in confezione sterile da parte del fabbricante. Telemetri ECG possono essere riutilizzate a condizione che il dispositivo viene pulito con soluzione Tergazyme 1% per almeno 4 ore. Si può lavare il telemetro con acqua sterile dopo la pulizia con Tergazyme. Inoltre, l&#39;uso Wavicide-01 disinfettante durante la notte per sterilizzare il telemetro ECG. Assicurarsi di lavare disinfettante con acqua sterile 48 ore prima l&#39;impianto in topi. Conservare in un contenitore sterile. </li><li> Controllare il telemetro porta per l&#39;integrità di entrambi i fili conduttori e la guaina isolante. Accendere il trasmettitore telemetria con un magnete sventolato entro 5 cm dal telemetro, e testare il segnale con una radio AM, 530 frequenza. Il segnale deve essere forte e chiara e dovrebbe variare di intensità e tono basata sulla manipolazione dei fili. Registrare il numero di modello, il numero di telemetro di serie e il valore di calibrazione ECG. I produttori spesso raccomandano che il telemetro dovrebbe essere per 24 ore prima dell&#39;impianto, consultare il manuale di istruzioni del telemetro specifiche per i dettagli. In questo esempio, useremo un Data Scienze Internazionali (DSI) telemetro per l&#39;impianto. </li><li> Preparazione di piombo telemetro può essere raggiunto tagliando il negativo (bianco) portare a circa 3,5 cm e il positivo (rosso) a 2,5 cm. Queste sono le lunghezze ottimali per l&#39;impianto del polo negativo nel petto a destra, e portare in positivo l&#39;addome a sinistra (vedi figura 2). Quindi, rimuovere la guaina isolante di esporre 7 mm di filo. Dipingi la fine del piombo metallo con colla sterile, come Vet-Bond, e quindi collegare tappi portare le punte di metallo. Questi tappi eviterà l&#39;erosione della pelle a causa di guidare il posizionamento. Circa 2-3 millimetri filo metallico deve essere esposto per il rilevamento elettrico del ritmo cardiaco nativo. </li></ol> 2. L&#39;impianto chirurgico dei Telemetro <ol><li> Per poter utilizzare una tecnica sterile in chirurgia murini, avrete bisogno di guanti sterili, un telino sterile, e 6-0 punti di sutura Prolene. Si avrà bisogno anche di 2 paia di pinzette sterilizzate, 1 paio di blunt-ended forbici, un bisturi, e un driver ago. Gli strumenti chirurgici possono essere sterilizzati in uno sterilizzatore a sfere di vetro riscaldato a 250 gradi Celsius. Assicurati di sterilizzare e lavare il telemetro come descritto in precedenza prima dell&#39;uso. </li><li> Preparare il mouse per un intervento chirurgico per prima anestetizzare il mouse in una scatola di induzione utilizzando 3% isoflurano a 0,5 L / min 100% O2. Utilizzare una piastra elettrica, come una T-pompa che fa circolare acqua calda, per mantenere la temperatura del mouse è anestetizzato. Temperatura può essere controllata con sonda rettale, se lo si desidera. Quando il mouse è adeguatamente sedato, radersi completamente l&#39;addome e il torace del mouse con tagliaunghie elettrici. Re-anestetizzare il mouse con isoflurano prima di trasferire il mouse su un flusso costante tubo, supina, con la testa rivolta verso la coda verso di voi. Nastro giù le zampe del mouse sul tavolo riscaldato chirurgico. Pulire l&#39;addome con un tampone imbevuto di alcool per rimuovere i capelli rasati e per pulire il campo operatorio. Applicare tre strati separati di betadine con tamponi per disinfettare l&#39;addome e torace. Verificare il corretto livello di anestesia applicando pressione sul letto ungueale del mouse. Dopo il mouse è stato preparato per un intervento chirurgico, utilizzare guanti sterili e applicare la garza sterile al campo chirurgico. </li><li> Impianto telemetro intraperitoneale offre il vantaggio di sperimentazione fisiologia. Per l&#39;inserimento intraperitoneale, iniziare intervento chirurgico utilizzando un bisturi per creare una incisione verticale mediana della cute sovrastante l&#39;addome circa 2,5 cm di lunghezza. Cura della pelle separato dal sottostante tessuto connettivo con blunt-ended forbici. Quindi, creare una incisione verticale mediana in linea alba sovrastante il peritoneo, circa 1,5 cm di lunghezza. Inoltre, creare un piccolo foro nel peritoneo appena sopra (craniale) per l&#39;incisione peritoneale, che servirà come valvola di sfogo per via sottocutanea porta. Durante l&#39;esecuzione di interventi chirurgici, mantenere umido il sito chirurgico di tanto in tanto stillicidio soluzione salina sterile sul campo operatorio. </li><li> Inserire il telemetro nella cavità peritoneale a destra. Inserire pinza nel foro superiore al telemetro e tirare entrambi i cavi attraverso il foro di piombo in modo che siano sporgenti dal peritoneo. Uso continuo 6-0 punti di sutura per chiudere la Prolene cm 1.5 incisione peritoneale linea mediana verticale. </li></ol> 3. Piombo impianto dell&#39;ovulo e chiusura addominale <ol><li> Le derivazioni ECG sono posti nella configurazione di piombo II. Il piombo, con il bianco / trasparente guaina è negativo, ed è collocato in addome alto a sinistra. In primo luogo, creare una incisione cutanea 0,5 cm di petto in alto a destra del mouse.Quindi, usare il forbici per creare un tunnel indietro l&#39;incisione addominale. Tirare il cavo attraverso il tunnel e usare un 6-0 sutura Prolene per ancorare il cavo al muscolo pettorale. Assicurarsi che la sutura è in cima alla parte esposta del piombo, e crea un buon contatto tra piombo e muscoli sottostanti. Utilizzare una sutura secondo prossimale al citato uno, per immobilizzare il cavo al muscolo. Chiudere l&#39;incisione cutanea con un 6-0 sutura Prolene. </li><li> Il cavo positivo (guaina rosso) viene inserito nell&#39;addome sinistra sotto il diaframma sinistra e sotto il cuore. Il cavo è ancorato al tessuto sottostante peritoneale da 6-0 sutura Prolene e deve avere un buon contatto con il tessuto peritoneale. Una incisione addizionale non è necessaria in questa fase perché il sito dell&#39;impianto piombo è vicino alla linea mediana incisione verticale, possono quindi essere facilmente accessibili con il campo esistente chirurgico. Si noti che il tessuto peritoneale deve essere sollevato per evitare la perforazione dell&#39;intestino sottostante. </li><li> Chiudere la fascia addominale e la pelle in modo sequenziale con 6-0 Prolene a strati. </li></ol> 4. Cure post-operatorie <ol><li> Dare il 0,1 mL del mouse su [1 mg / ml] buprenorfina per l&#39;analgesia immediatamente dopo la chirurgia. Lasciare che il mouse per riprendersi da un intervento chirurgico su un tappetino riscaldato. Se il mouse ha prolungato intervento chirurgico&gt; 30 min, o se il mouse appare secco, si può iniettare 0,2-0,3 ml di soluzione salina sterile nel peritoneo per la reidratazione. Per continuato analgesia post-operatoria è pratica standard per fornire topi con buprenorfina due volte al giorno per tre giorni, e poi, se necessario ogni otto ore. </li><li> Pulire betadine con un batuffolo di cotone imbevuto di alcol per evitare irritazioni post-chirurgica. Ricordatevi di registrare il post-chirurgico di peso per la successiva determinazione di post-chirurgica salute. </li><li> 8-24 ore dopo l&#39;intervento, dare il mouse analgesia supplementari con buprenorfina, quando necessario, in quanto riduce il dolore e ridurre l&#39;insuccesso di piombo di collocamento da topo artigliare a livello post-chirurgiche. </li></ol> 5. Rappresentante Risultati Quando eseguito correttamente, il mouse dovrebbe aver chiuso incisioni addominali con sottocutaneo porta sotto la pelle. Solitamente, il mouse prende 10-30 minuti per il recupero dall&#39;anestesia. Ci vogliono circa 7-10 giorni prima che i topi hanno completamente recuperato da un intervento chirurgico, come dimostra il recupero post-operatorio di perdita di peso e la normalizzazione della mobilità. La complicanza più comune dalla chirurgia è l&#39;erosione della porta degli arti attraverso la pelle nei primi 10 giorni dopo l&#39;intervento chirurgico. Questa complicanza comune possono essere evitate con fermezza aderendo sottocutaneo porta a dei tessuti sottocutanei o del peritoneo. Questa tecnica evita porta molle, che creano la pressione e l&#39;attrito sulla pelle quando il mouse ambulates. Anche se meno comuni, l&#39;infezione e sepsi può verificarsi da errori di tecnica sterile, o dalla pulizia incompleta telemetri utilizzati. Uscita ECG può essere registrato da una matrice ricevitore accoppiato con software di acquisizione dati, come dati Sciences International (DSI). Risultati della forma d&#39;onda dovrebbe includere un onda ben definito P, che denota depolarizzazione atriale, e anche un&#39;onda QRS che significa depolarizzazione ventricolare, come mostrato nella Figura 3. Una buona qualità del tracciato ECG dovrebbe avere un onda ben definita P che precede un ben definito QRS onda in un rapporto di 1:1. Ci dovrebbe essere segnale di fondo basso e gli intervalli RR dovrebbe essere regolare in topi wild-type. Una scarsa qualità tracciato ECG può essere indistinta o onde P, QRS, o segnale di fondo, che può rendere l&#39;analisi del computer successive difficile. Un povero tracciato ECG può essere fissato di re-impianto porta telemetro, di solito uno alla volta. Per re-impianto un cavo telemetro, posizionare il mouse in anestesia, e usando la tecnica chirurgica, fare un&#39;incisione nella pelle direttamente sovrastante il cavo in questione. Quando il cavo si accede, tagliare la sutura di ancoraggio del piombo, e spostare il cavo nella posizione desiderata prima di ancorare il cavo di nuovo. Infine, chiudere il sito chirurgico con 6-0 Prolene. <img src="/files/ftp_upload/1739/1739fig1.jpg" alt="Figura 1" /> Figura 1: Un esempio di un telemetro piccolo animale con i cavi positivi e negativi. In generale, il cavo rosso è il conduttore positivo, e la biacca è il cavo negativo. Ogni cavo deve avere una guaina isolante, 5-7 mm di filo a vista, e una punta in plastica con tappo, che impedisce l&#39;erosione del cavo attraverso la pelle. <img src="/files/ftp_upload/1739/1739fig2.jpg" alt="Figura 2" /> Figura 2: Cartoon mostrando adeguata siti telemetro impianto di un mouse. Il bianco (negativo) viene impiantato nel petto alto a destra del mouse, e la (positiva) rosso piombo è posizionato nell&#39;addome di sinistra, in modo tale che i sensi telemetro del &quot;portare due&quot; di configurazione. &lt;p class = &quot;jove_content&quot;&gt; <img src="/files/ftp_upload/1739/1739fig3.jpg" alt="Figura 3" /> Figura 3: forme d&#39;onda rappresentante di un topo nella normale ritmo sinusale. P onda rappresenta depolarizzazione atriale e complesso QRS rappresenta la depolarizzazione ventricolare. Una forma d&#39;onda di alta qualità deve avere un onda distinte P che precede ogni complesso QRS in un rapporto di 1:1.

<ol>
	<li>Cerrone, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bidirectional+ventricular+tachycardia+and+fibrillation+elicited+in+a+knock-in+mouse+model+carrier+of+a+mutation+in+the+cardiac+ryanodine+receptor.">Bidirectional ventricular tachycardia and fibrillation elicited in a knock-in mouse model carrier of a mutation in the cardiac ryanodine receptor.</a> Circ Res. 96 (10), e77-e77 (2005).</li><li>Chelu, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calmodulin+kinase+II-mediated+sarcoplasmic+reticulum+Ca2++leak+promotes+atrial+fibrillation+in+mice.">Calmodulin kinase II-mediated sarcoplasmic reticulum Ca2+ leak promotes atrial fibrillation in mice.</a> J Clin Invest. 119 (7), 1940-1940 (1940).</li><li>Sood, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+calcium+leak+due+to+FKBP12.6+deficiency+in+mice+facilitates+the+inducibility+of+atrial+fibrillation.">Intracellular calcium leak due to FKBP12.6 deficiency in mice facilitates the inducibility of atrial fibrillation.</a> Heart Rhythm. 5 (7), 1047-1047 (2008).</li><li>Swoap, S. J., Gutilla, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+changes+during+daily+torpor+in+the+laboratory+mouse.">Cardiovascular changes during daily torpor in the laboratory mouse.</a> Am J Physiol Regul Integr Comp Physiol. 297 (3), R769-R769 (2009).</li><li>Costa-Goncalves, d. a., C, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+the+multidomain+protein+spinophilin+in+blood+pressure+and+cardiac+function+regulation.">Role of the multidomain protein spinophilin in blood pressure and cardiac function regulation.</a> Hypertension. 52 (4), 702-702 (2008).</li><li>Wehrens, X. H., Kirchhoff, S., Doevendans, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+electrocardiography:+an+interval+of+thirty+years.">Mouse electrocardiography: an interval of thirty years.</a> Cardiovasc Res. 45 (1), 231-231 (2000).</li></ol>

I passi fondamentali di questa procedura sono una corretta pulizia e preparazione del mouse prima dell&#39;intervento sopravvivenza. Quando un telemetro e la sede operativa del mouse sono adeguatamente puliti, c&#39;è un molto migliore post-chirurgica risultato. Inoltre, attenzione a guidare il posizionamento post-chirurgica riduce l&#39;erosione della pelle. Eventuali modifiche alla tecnica comprendono configurazioni alternative per valutare la funzione della parete laterale LV (piombo I configurazione), o l&#39;uso d...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Steri 250 Bead Sterilizer</td>
<td>&nbsp;</td>
<td>Inotech</td>
<td>IS-250</td>
<td>&nbsp;</td>
<tr>
<td>Blunt-Ended Scissors</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5980</td>
<td>&nbsp;</td>
<tr>
<td>Blunt-Ended Forceps (x2)</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5138</td>
<td>&nbsp;</td>
<tr>
<td>Veterinary Anesthesia Vaporizer</td>
<td>&nbsp;</td>
<td>Surgivet</td>
<td>Isotec 4</td>
<td>&nbsp;</td>
<tr>
<td>T/Pump</td>
<td>&nbsp;</td>
<td>Gaymar</td>
<td>P/N11184-000</td>
<td>&nbsp;</td>
<tr>
<td>Propylene Blue Monofilament Suture</td>
<td>&nbsp;</td>
<td>Ethicon</td>
<td>8718</td>
<td>&nbsp;</td>
<tr>
<td>Povodone Iodine, 10%</td>
<td>&nbsp;</td>
<td>Equate</td>
<td>N/A</td>
<td>&nbsp;</td>
<tr>
<td>Alcohol Swabs</td>
<td>&nbsp;</td>
<td>PDI</td>
<td>B339</td>
<td>&nbsp;</td>
<tr>
<td>Tergazyme</td>
<td>&nbsp;</td>
<td>Alconox</td>
<td>21837-118</td>
<td>&nbsp;</td>
<tr>
<td>Wavidicde-01</td>
<td>&nbsp;</td>
<td>Medical Chemical Corporation</td>
<td>Wavicide-01</td>
<td>&nbsp;</td>
<tr>
<td>ECG Telemeter</td>
<td>&nbsp;</td>
<td>DataSciences International</td>
<td>EA-F20</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

ambulatory ecg recording in mice

Telemetrici registrazione ECG nei topi è essenziale per comprendere i meccanismi alla base aritmie, disturbi della conduzione, e morte cardiaca improvvisa. Anche se la superficie ECG è utilizzato per misure a breve termine degli intervalli di forma d&#39;onda, non è pratico di studi a lungo termine della variabilità della frequenza cardiaca o la cattura di rari episodi di aritmie. Impiantabili telemetri ECG offrire i vantaggi di semplice impianto chirurgico, a lungo termine la registrazione di elettrogrammi nei topi ambulatoriale, e la scalabilità con registrazioni simultanee di animali diversi. Qui vi presentiamo uno step-by-step per l&#39;impianto di telemetri per la registrazione ECG ambulatoriale nei topi. Stretta aderenza alla tecnica asettica è necessario per la sopravvivenza risultati favorevoli con la possibilità di impianto e la registrazione da settimane a mesi. Così, impiantabile telemetria ECG è uno strumento prezioso per la rilevazione di informazioni critiche sui elettrofisiologia cardiaca nei modelli animali ambulatoriale come il mouse.

Watch this Scientific Journal Video about Ambulatory ECG Recording in Mice at JoVE.com

Ambulatory ECG Recording in Mice

Telemetrico ECG è emerso come uno strumento essenziale per la valutazione dei modelli animali per aritmie cardiache e morte cardiaca improvvisa. Qui vi presentiamo una guida graduale per telemetrici registrazioni ECG per l&#39;applicazione nel monitoraggio a lungo termine ECG ambulatoriale nei topi.

La registrazione ECG ambulatoriale nei topi

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

Research

JoVE Journal

Biology

23.5K Views.  Baylor College of Medicine (BCM). Telemetrico ECG è emerso come uno strumento essenziale per la valutazione dei modelli animali per aritmie cardiache e morte cardiaca improvvisa. Qui vi presentiamo una guida graduale per telemetrici registrazioni ECG per l'applicazione nel monitoraggio a lungo termine ECG ambulatoriale nei topi.

Video: Ambulatory ECG Recording in Mice

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable for studying rejection and immune response now that newer transgenic and gene knockout mice are available, and a large number of immunologic reagents have been developed. The heart transplant model is less stringent than the skin transplant models, although technically more challenging. We have developed a modified technique and have completed over 1000 successful cases of heterotopic heart transplantation in mice. When making anastomosis of the ascending aorta and abdominal aorta, two stay sutures are placed at the proximal and distal apexes of recipient abdominal aorta with the donor s ascending aorta, then using 11-0 suture for anastomosis on both side of aorta with continuing sutures. The stay sutures make the anastomosis easier and 11-0 is an ideal suture size to avoid bleeding and thrombosis.
When making anastomosis of pulmonary artery and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s pulmonary artery. The left wall of the inferior vena cava and donor s pulmonary artery is closed with continuing sutures in the inside of the inferior vena cava after, one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s pulmonary artery are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

Il trapianto cardiaco eterotopico nel Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

Ricerca

Biologia

Small Bowel Transplantation In Mice

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive medications, infection prophylaxis, and suitable patient selection helped improve actuarial graft and patient survival rates for all types of intestine transplantation. Patients with irreversible intestinal failure and complications of parenteral nutrition should now be routinely considered for small intestine transplantation. However, Survival rates for small intestinal transplantation have been slow to improve compares increasingly favorably with renal, liver, heart and lung. The small bowel transplantation is still unsatisfactory compared with other organs. Further progress may depend on better understanding of immunology and physiology of the graft and can be greatly facilitated by animal models. A wider use of mouse small bowel transplantation model is needed in the study of immunology and physiology of the transplantation gut as well as efficient methods in diagnosing early rejection. However, this model is limited to use because the techniques involved is an extremely technically challenging. We have developed a modified technique. When making anastomosis of portal vein and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s portal vein. The left wall of the inferior vena cava and donor s portal vein is closed with continuing sutures in the inside of the inferior vena cava after, after one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s portal vein are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis.  In this article, we provide details of the technique to supplement the video.

The mouse small bowel transplantation model has been recognized as an important tool to study mechanismes of immune rejection and screen new immunosuppressive drugs. However, this model is limited to use because the techniques involved is an extremely technically challenge. Now we introduce the modified technique.

Trapianto di intestino piccolo Nei topi

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive ...

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

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 tail-cuff system uses Volume Pressure Recording (VPR) to measure the blood pressure by determining the tail blood volume. A specially designed differential pressure transducer and an occlusion tail-cuff measure the total blood volume in the tail without the need to obtain the individual pulse signal. Special attention is afforded to the length of the occlusion cuff in order to derive the most accurate blood pressure readings. VPR can easily obtain readings on dark-skinned rodents, such as C57BL6 mice and is MRI compatible. The CODA system provides you with measurements of six (6) different blood pressure parameters; systolic and diastolic blood pressure, heart rate, mean blood pressure, tail blood flow, and tail blood volume. Measurements can be made on either awake or anesthetized mice or rats. The CODA system includes a controller, laptop computer, software, cuffs, animal holders, infrared warming pads, and an infrared thermometer. There are seven different holder sizes for mice as small as 8 grams to rats as large as 900 grams.

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. This tail-cuff system uses Volume Pressure Recording (VPR) to measure the blood pressure by determining the tail blood volume.

Misurazione della pressione sanguigna nei topi mediante pressione del volume di registrazione, una coda-bracciale Metodo

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 ...

Electrophysiological Recording in the Drosophila Embryo

Drosophila is a premier genetic model for the study of both embryonic development and functional neuroscience. Traditionally, these fields are quite isolated from each other, with largely independent histories and scientific communities. However, the interface between these usually disparate fields is the developmental programs underlying acquisition of functional electrical signaling properties and differentiation of functional chemical synapses during the final phases of neural circuit formation. This interface is a critically important area for investigation. In Drosophila, these phases of functional development occur during a period of &lt;8 hours (at 25&deg;C) during the last third of embryogenesis. This late developmental period was long considered intractable to investigation owing to the deposition of a tough, impermeable epidermal cuticle. A breakthrough advance was the application of water-polymerizing surgical glue that can be locally applied to the cuticle to enable controlled dissection of late-stage embryos. With a dorsal longitudinal incision, the embryo can be laid flat, exposing the ventral nerve cord and body wall musculature to experimental investigation. Whole-cell patch-clamp techniques can then be employed to record from individually-identifiable neurons and somatic muscles. These recording configurations have been used to track the appearance and maturation of ionic currents and action potential propagation in both neurons and muscles. Genetic mutants affecting these electrical properties have been characterized to reveal the molecular composition of ion channels and associated signaling complexes, and to begin exploration of the molecular mechanisms of functional differentiation. A particular focus has been the assembly of synaptic connections, both in the central nervous system and periphery. The glutamatergic neuromuscular junction (NMJ) is most accessible to a combination of optical imaging and electrophysiological recording. A glass suction electrode is used to stimulate the peripheral nerve, with excitatory junction current (EJC) recordings made in the voltage-clamped muscle. This recording configuration has been used to chart the functional differentiation of the synapse, and track the appearance and maturation of presynaptic glutamate release properties. In addition, postsynaptic properties can be assayed independently via iontophoretic or pressure application of glutamate directly to the muscle surface, to measure the appearance and maturation of the glutamate receptor fields. Thus, both pre- and postsynaptic elements can be monitored separately or in combination during embryonic synaptogenesis. This system has been heavily used to isolate and characterize genetic mutants that impair embryonic synapse formation, and thus reveal the molecular mechanisms governing the specification and differentiation of synapse connections and functional synaptic signaling properties.

Electrophysiological Recording in the Drosophila Embryo

Electrophysiological recordings from Drosophila embryos allow analyses of developing muscle and neuron electrical properties, as well as characterization of functional synaptogenesis at the glutamatergic neuromuscular junction and central cholinergic and GABAergic synapses.

Registrazione elettrofisiologiche in Drosophila Embryo

Drosophila is a premier genetic model for the study of both embryonic development and functional neuroscience. Traditionally, these fields are quite ...

Transverse Aortic Constriction in Mice

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model for pressure overload-induced cardiac hypertrophy and heart failure.1 TAC initially leads to compensated hypertrophy of the heart, which often is associated with a temporary enhancement of cardiac contractility. Over time, however, the response to the chronic hemodynamic overload becomes maladaptive, resulting in cardiac dilatation and heart failure.2 The murine TAC model was first validated by Rockman et al.1, and has since been extensively used as a valuable tool to mimic human cardiovascular diseases and elucidate fundamental signaling processes involved in the cardiac hypertrophic response and heart failure development. When compared to other experimental models of heart failure, such as complete occlusion of the left anterior descending (LAD) coronary artery, TAC provides a more reproducible model of cardiac hypertrophy and a more gradual time course in the development of heart failure. Here, we describe a step-by-step procedure to perform surgical TAC in mice. To determine the level of pressure overload produced by the aortic ligation, a high frequency Doppler probe is used to measure the ratio between blood flow velocities in the right and left carotid arteries.3, 4 With surgical survival rates of 80-90%, transverse aortic banding is an effective technique of inducing left ventricular hypertrophy and heart failure in mice.

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model to study mechanisms underlying cardiac hypertrophy and heart failure development. Here, we describe procedures to constrict the aorta to create a reproducible degree of cardiac hypertrophy in mice.

Aortica costrizione trasversale nei topi

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model for pressure overload-induced cardiac hypertrophy and heart ...

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 canale Cell-allegato patch-clamp registrazione

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

Intravenous Injections in Neonatal Mice

Intravenous injection is a clinically applicable manner to deliver therapeutics. For adult rodents and larger animals, intravenous injections are technically feasible and routine. However, some mouse models can have early onset of disease with a rapid progression that makes administration of potential therapies difficult. The temporal (or facial) vein is just anterior to the ear bud in mice and is clearly visible for the first two days after birth on either side of the head using a dissecting microscope. During this window, the temporal vein can be injected with volumes up to 50 &#956;l. The injection is safe and well tolerated by both the pups and the dams. A typical injection procedure is completed within 1-2 min, after which the pup is returned to the home cage. By the third postnatal day the vein is difficult to visualize and the injection procedure becomes technically unreliable. This technique has been used for delivery of adeno-associated virus (AAV) vectors, which in turn can provide almost body-wide, stable transgene expression for the life of the animal depending on the viral serotype chosen.

Animal models of pediatric disease can experience early onset and aggressive disease progression. Clinically relevant therapy delivery to young mouse models can be technically difficult. This protocol describes a non-invasive intravenous injection method for newborn mice within the first two postnatal days of life.

Le iniezioni per via endovenosa nel topo neonatale

Intravenous injection is a clinically applicable manner to deliver therapeutics. For adult rodents and larger animals, intravenous injections are ...

Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents

Erectile dysfunction (ED) is defined as the inability to attain or keep an erection of the penis, and this has become a prevalent male sexual disorder. Rodents are employed by many studies to research the physiology/pathology of erectile function. Erectile function in rodents can be evaluated by measuring the intracavernosal pressure (ICP). In practice, ICP can be monitored following electrical stimulation of the cavernous nerves (CNs). The arterial pressure of the carotid artery (the mean arterial pressure) is used as the reference for ICP. Using ICP recording protocols, many key parameters of erectile function can be measured from the ICP response curve. The ICP measurement provides more information than the apomorphine-induced penile erection test, and is cheaper than telemetric monitoring of the corpus spongiosum penis, making this method the most popular one to evaluate erectile function. However, compared to the easily-performed APO-induced erectile function test, successful ICP recordings require attention to detail, practice, and adherence to the operation method. In this work, an introduction to ICP recording in rats is provided to complement the procedure efficiently.

Intracavernosal pressure recording (ICP) is an important method to evaluate the erectile function of experimental animals. Here, a detailed protocol is demonstrated for the recording procedure of ICP by catheterizing the crura penis and then electrically stimulating the cavernous nerves in rats.

Registrazione di pressione intracavernosa per valutare la funzione erettile in roditori

Erectile dysfunction (ED) is defined as the inability to attain or keep an erection of the penis, and this has become a prevalent male sexual disorder. ...

Inducing Apical Periodontitis in Mice

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical periodontitis (AP). Apical periodontitis is a common pathology of an inflammatory nature in the periodontal tissues surrounding the tooth root. In order to better understand the nature and mechanism of this pathology it is advantageous to perform the procedure in mice. The induction of this odontogenic inflammation is achieved by drilling into the mouse tooth until the dental pulp is exposed. Next, the tooth pulp remains exposed to be contaminated by the natural oral flora over time, causing apical periodontitis. After this time period, the animal is sacrificed, and the tooth and the jaw bone can be analyzed in various ways. Typical analyses include micro-CT imaging (to evaluate bone resorption), histological staining, immunohistochemistry, and RNA expression. This protocol is useful for research in the field of oral biology to better understand this inflammatory process in an in vivo experimental setting with uniform conditions. The procedure requires a careful handling of the mice and the isolated jaw, and a visual demonstration of the technique is useful. All technical aspects of the procedures leading to induced apical periodontitis and its characterization in a mouse model are demonstrated.

Here, we present a protocol to locally induce apical periodontitis in mice. We show how to drill a hole in the mouse&#39;s tooth and expose its pulp, in order to cause local inflammation. Analysis methods to investigate the nature of this inflammation, such as micro-CT and histology, are also demonstrated.

Indurre la parodontite apicale nei topi

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical ...

Establishing a Diaphyseal Femur Fracture Model in Mice

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age and overall health status of the patient, failures can occur, leading to delayed union or nonunion. Due to the increasing number of fractures resulting from high-energy trauma and aging, the development of innovative therapeutic strategies to improve bone repair based on the combination of skeletal/mesenchymal stem/stromal cells and biomimetic biomaterials is urgently needed. To this end, the use of reliable animal models is fundamental to better understanding the key cellular and molecular mechanisms that determine the healing outcomes. Of all the models, the mouse is the preferred research model because it offers a wide variety of transgenic strains and reagents for experimental analysis. However, the establishment of fractures in mice may be technically challenging due to their small size. Therefore, this article aims to demonstrate the procedures for the surgical establishment of a diaphyseal femur fracture in mice, which is stabilized with an intramedullary wire and resembles the most common bone repair process, through cartilaginous callus formation.

This protocol describes a surgical procedure for the establishment of a diaphyseal fracture in the femur of mice, which is stabilized with an intramedullary wire, for fracture healing studies.

Stabilire un modello di frattura del femore diafisario nei topi

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age ...