Gli autori desiderano ringraziare Charles River in Germania per la fornitura di CD-1 nei topi. Ringraziamo anche Robin Schneider e il personale del laboratorio centrale per il supporto biologico nel topo alloggi. Ringraziamo Nicholls Flora per un&#39;eccellente assistenza tecnica e il professor Kurt Burki generosamente per fornire servizi di ricerca e risorse.

<ol>
	<li>Cesarovic, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+and+sevoflurane+provide+equally+effective+anesthesia+in+laboratory+mice.">Isoflurane and sevoflurane provide equally effective anesthesia in laboratory mice.</a> Lab. Anim. 44, 329-336 (2010).</li><li>Gross, V., Luft, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercising+restraint+in+measuring+blood+pressure+in+conscious+mice.">Exercising restraint in measuring blood pressure in conscious mice.</a> Hypertension. 41, 879-881 (2003).</li><li>Kramer, K., Kinter, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+and+applications+of+radiotelemetry+in+small+laboratory+animals.">Evaluation and applications of radiotelemetry in small laboratory animals.</a> Physiol. Genomics. 13, 197-205 (2003).</li><li>Kramer, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+telemetry+to+record+electrocardiogram+and+heart+rate+in+freely+moving+mice.">Use of telemetry to record electrocardiogram and heart rate in freely moving mice.</a> J. Pharmacol. Toxicol. Methods. 30, 209-215 (1993).</li><li>Arras, M., Rettich, A., Cinelli, P., Kasermann, H. P., Burki, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+post-laparotomy+pain+in+laboratory+mice+by+telemetric+recording+of+heart+rate+and+heart+rate+variability.">Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability.</a> BMC. Vet. Res. 3, 16-16 (2007).</li><li>Schuler, B., Rettich, A., Vogel, J., Gassmann, ., Arras, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimized+surgical+techniques+and+postoperative+care+improve+survival+rates+and+permit+accurate+telemetric+recording+in+exercising+mice.">Optimized surgical techniques and postoperative care improve survival rates and permit accurate telemetric recording in exercising mice.</a> BMC. Vet. Res. 5, 28-28 (2009).</li><li>Pritchett-Corning, K. R., Mulder, G. B., Luo, Y., White, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+Rodent+Surgery+for+the+New+Surgeon.">Principles of Rodent Surgery for the New Surgeon.</a> J. Vis. Exp. (47), e2586-e2586 (2011).</li><li>Rettich, A., Kasermann, H. P., Pelczar, P., Burki, K., Arras, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+physiological+and+behavioral+impact+of+sensory+contact+among+unfamiliar+adult+mice+in+the+laboratory.">The physiological and behavioral impact of sensory contact among unfamiliar adult mice in the laboratory.</a> J. Appl. Anim. Welf. Sci. 9, 277-288 (2006).</li><li>Spani, D., Arras, M., Konig, B., Rulicke, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Higher+heart+rate+of+laboratory+mice+housed+individually+vs+in+pairs.">Higher heart rate of laboratory mice housed individually vs in pairs.</a> Lab. Anim. 37, 54-62 (2003).</li><li>Zeller, A., Arras, M., Jurd, R., Rudolph, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+the+contribution+of+beta3-containing+GABAA+receptors+to+volatile+and+intravenous+general+anesthetic+actions.">Mapping the contribution of beta3-containing GABAA receptors to volatile and intravenous general anesthetic actions.</a> BMC. Pharmacol. 7, 2-2 (2007).</li><li>Zeller, A., Arras, M., Jurd, R., Rudolph, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+a+molecular+target+mediating+the+general+anesthetic+actions+of+pentobarbital.">Identification of a molecular target mediating the general anesthetic actions of pentobarbital.</a> Mol. Pharmacol. 71, 852-859 (2007).</li><li>Arras, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+of+pain+therapy+in+laboratory+mice.">Improvement of pain therapy in laboratory mice.</a> Altex. 24, 6-8 (2007).</li></ol>

Nessun conflitto di interessi dichiarati.

Radiotelemetry è una valida alternativa ai metodi convenzionali di misura di parametri fisiologici nella ricerca biomedica. Alta qualità, sistemi di telemetria composta da impiantabili trasmettitori, ricevitori e acquisizione dati e analisi di hardware e software sono ora disponibili in commercio, anche per gli animali piccoli come topi. Telemetria rappresenta l&#39;unica tecnica attualmente disponibile per la raccolta dei dati da sfrenato, liberi di muoversi topi. Utilizzando questo metodo, è ora possibile raccogliere dati in modo continuo e / o per periodi più lunghi di tempo da animali che risiedono nel proprio ambiente familiare, riducendo così al minimo lo stress per gli animali e conseguenti artefatti sperimentali. La forma e la posizione dei cavi è stata ottimizzata al fine di ottenere segnali anche durante i movimenti veloci (ad esempio, lotta, corsa, lotta) o in una postura eretta 9. Così, misurazioni accurate possono essere ottenute durante gli esperimenti, ad esempio durante l&#39;anestesia, lo stressproduzione, durante l&#39;esecuzione su un tapis roulant, durante gli esperimenti comportamentali, durante gli esperimenti l&#39;infezione, e molte altre situazioni sperimentali. Tuttavia, al fine di ottenere dati affidabili, ripetibili e prive di artefatti, è fondamentale per escludere influenze ambientali, e attirare l&#39;attenzione in particolare l&#39;importanza di condizioni standardizzate. Si raccomanda che la stanza è isolata dal rumore elettronico e acustico, compresi gli ultrasuoni, a cui i topi sono particolarmente sensibili. Inoltre, non disturbi, come visitatori o non correlati procedure sperimentali, dovrebbe essere consentito lo svolgimento delle misurazioni. Per evitare interferenze influenze (in particolare in caso di misurazioni gabbia di casa), tutte le procedure necessarie allevamento dovrebbero essere completate in sala prima dell&#39;inizio di ogni misurazione. Inoltre, l&#39;alloggiamento di topi, in particolare se i maschi sono utilizzati in gruppi o singolarmente possono avere un impatto sulle misure e devono essere considerati quando si plaesperimenti nning 9. Inoltre, i topi devono essere sani e privi di agenti patogeni murini, dal momento che le infezioni latenti o manifesti, così come le malattie o menomazioni salute, possono avere una notevole influenza sui parametri fisiologici e del comportamento di attività. Di conseguenza, i topi dovrebbe recuperare completamente dopo l&#39;impianto e avere tempo sufficiente per adeguarsi a portare il trasmettitore prima di iniziare qualsiasi sperimentazione. La raccolta dei dati da radiotelemetry nei topi richiede preliminare impianto chirurgico dei trasmettitori telemetrici. Questo dovrebbe essere eseguita solo da personale qualificato con competenze chirurgiche al fine di minimizzare il trauma dei tessuti e il dolore conseguente e angoscia. Per sperimentatori tenendo competenze chirurgiche di base o anche avanzati (micro-), si raccomanda di eseguire le prime prove in cadaveri del mouse fresca utilizzando gli impianti di formazione (ad esempio, manichini, fornito dal produttore) per stabilire le procedure e prendere confidenza con le specifiche di questo tipodella chirurgia. Dopo tale formazione, più sperimentatori sarebbe in grado di impiantare questo tipo di trasmettitori con successo e avrebbe raggiunto una competenza utile dopo un paio di implantologia. Condizioni di asepsi deve essere mantenuta durante l&#39;intervento chirurgico per mantenere il carico microbiologico e il rischio di infezioni basso. Tuttavia, la sterilità completa non possono essere fornite a causa di alcuni particolari, le condizioni di sterilità conflittuali nei topi (ad esempio, effetto di raffreddamento del taglio dei capelli ampia e disinfezione, impraticabilità di bende per proteggere le ferite). Così, profilassi antinfettiva è somministrato durante l&#39;impianto. Bene su misura trattamento analgesico e un piano di monitoraggio ben definito così come un&#39;adeguata assistenza post-operatoria svolgono un ruolo cruciale nel buon esito dell&#39;esperimento. Nel complesso, l&#39;impianto chirurgico di un trasmettitore telemetrico nei topi sarà stressante per l&#39;animale. In particolare, se la modificazione genetica nelle specifichele linee del mouse IFIC influenza il fenotipo e danneggia condizione corporea degli animali &#39;, complicazioni nel peri-operatorio i tempi ei tassi di mortalità aumentati dopo l&#39;impianto potrebbe essere un rischio. Per evitare inutili sofferenze, gli individui espositrici recupero insoddisfacente o convalescenza prolungata devono essere rilasciate da l&#39;esperimento e si sono sacrificati prima di raggiungere un palcoscenico moribondo. A tal fine, una scheda dei dati (Tabella 1: stato di salute generale e la vigilanza foglio dati) facilitare il monitoraggio sistematico dei sintomi critici e fornendo consigli su endpoint umana è stata stabilita. Così, il recupero è documentato nello stile di una cartella clinica o di una rivista di laboratorio, che rende la conduzione di questa metodologia (cioè procedura di impianto e di recupero post-operatorio) trasparente alle autorità competenti e gli enti responsabili del benessere degli animali per la sperimentazione animale (ad esempio, IACUC).

Corrections in the Protocol and Discussion sections have been made to: Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice
Step 1.2 in the Protocol has been updated from:
1.2 Hair clipping at one day prior to surgery
The day prior to implantation, in order to shave the animals for surgery, mice are anesthetized briefly in a small (8x8x8cm) Perspex chamber using sevoflurane (8%) or isoflurane (5%) in pure oxygen (600 mL/min). After loss of the righting reflex, the mouse is taken out of the chamber and the anterior neck and abdominal hair is clipped with the animal lying in dorsal recumbence; anesthesia is maintained for approximately 5 minutes with a nose mask with sevoflurane 3-4% or isoflurane 1.5-3% in pure oxygen at a flow rate of 600 mL/min. After clipping the hair, the animals are allowed to awaken and are then brought back to their home cage.
to:
After the last health check or directly prior surgery, in order to shave the animals for surgery, mice are anesthetized briefly in a small (8x8x8cm) Perspex chamber using sevoflurane (8%) or isoflurane (5%) in pure oxygen (600 mL/min). Shaving the animals one day before surgery prevents hair stubbles in the operating field. After loss of the righting reflex, the mouse is taken out of the chamber and the anterior neck and abdominal hair is clipped with the animal lying in dorsal recumbence; anesthesia is maintained for approximately 5 minutes with a nose mask with sevoflurane 3-4% or isoflurane 1.5-3% in pure oxygen at a flow rate of 600 mL/min. After clipping the hair, the animals are allowed to awaken and are then brought back to their home cage.
Step 2.3 in the Protocol has been updated from:
2.3 Surgery
The skin of the anterior neck and abdominal region is disinfected with 70% ethanol. A 1- to 1.5-cm-long incision in the skin is made from the lower thorax along the midline to the abdomen. The negative (white/colourless) lead is tunnelled subcutaneously from the thorax to the neck, where a small incision (&#8804;0.5 cm) is made in the longitudinal direction. The skin and underlying tissues are prepared to make space for the fixation of the wire loop of the electrode. The wire loop is fixed between the muscles located to the right of the trachea, using two thin silk sutures (PERMA-Handseide, 6-0, Ethicon, Norderstedt, Germany). The wound in the neck is then closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany) in layers. The abdominal wall is then opened at the linea alba and the body of the telemetric transmitter is placed into the abdominal cavity of the mouse. The wire loop of the positive (red) electrode is sutured to the xiphoid process with silk sutures in such a way that it lies between the liver and the diaphragm in the left upper abdominal region (Figure 2). Then, the muscle layers of the abdominal region are closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany). Before finally closing the abdominal wall, a mixture of Sulfadoxin and Trimethoprim [(30 mg/kg and 6 mg/kg, respectively; dissolved in 1 mL of saline (0.9%) and at approximately body temperature (38-39&#176;C)] is injected into the abdominal cavity for the purposes of anti-infective prophylaxis and to support fluid homeostasis. Finally, the skin of the abdominal region is restored with staples (Precise, 3 M Health Care, St. Paul, MN, USA).
to:
2.3 Surgery
The skin of the anterior neck and abdominal region is disinfected for 5 minutes with 70% ethanol, chlorhexidine or iodine using a soaked cotton swab. A 1- to 1.5-cm-long incision in the skin is made from the lower thorax along the midline to the abdomen. The negative (white/colourless) lead is tunnelled subcutaneously from the thorax to the neck, where a small incision (&#8804;0.5 cm) is made in the longitudinal direction. The skin and underlying tissues are prepared to make space for the fixation of the wire loop of the electrode. The wire loop is fixed between the muscles located to the right of the trachea, using two thin silk sutures (PERMA-Handseide, 6-0, Ethicon, Norderstedt, Germany). The wound in the neck is then closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany) in layers. The abdominal wall is then opened at the linea alba and the body of the telemetric transmitter is placed into the abdominal cavity of the mouse. The wire loop of the positive (red) electrode is sutured to the xiphoid process with silk sutures in such a way that it lies between the liver and the diaphragm in the left upper abdominal region (Figure 2). Then, the muscle layers of the abdominal region are closed with absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany). Before finally closing the abdominal wall, a mixture of Sulfadoxin and Trimethoprim [(30 mg/kg and 6 mg/kg, respectively; dissolved in 1 mL of saline (0.9%) and at approximately body temperature (38-39&#176;C)] is injected into the abdominal cavity for the purposes of anti-infective prophylaxis and to support fluid homeostasis. Finally, the skin of the abdominal region is restored with staples (Precise, 3 M Health Care, St. Paul, MN, USA) or intracutaneous, running, absorbable sutures (VICRYL 6-0, Ethicon, Norderstedt, Germany).

Step 3 in the Protocol has been updated from:
3. Post-operative care
After completion of surgery and anesthesia, 0.1 mg/kg of buprenorphine (Temgesic, Essex Chemie AG, Lucerne, Switzerland) and 5 mg/kg of meloxicam (Metacam, Boehringer Ingelheim, Basel, Switzerland) is administered subcutaneously for pain treatment, and the animals are left on the warm (39&#176;C +/-1) surface of the work bench to recover for approximately 2h. Together with pain relief (twice daily: buprenorphine, 0.1 mg/kg and meloxicam 5 mg/kg), supportive therapy consisting of 300 &#956;L glucose (5%) and 300 &#956;L saline (0.9%) warmed to body temperature, is applied subcutaneously twice daily for 4 days. For further recovery support, it is worthwhile providing the animals with an additional drinking bottle containing 15% glucose solution During the recovery period of 4-10 days, it is recommended that the animals are kept warm. Therefore, in our case, the mice are housed in a warming cabinet (30&#176;C +/- 1). Monitoring of general condition and body weight, as well as food and water consumption, is performed once daily according to the general condition and health monitoring data sheet (Table 1) for 10 days post-operatively. Humane endpoints, i.e. the sacrifice of an animal to avoid unnecessary suffering and pain if progression of recovery is unsatisfactory, are realised under the following conditions:
i. If in poor general condition, i.e. the animal is substantially apathetic (no movement after being touched/pushed) and its body surface feels cold despite warming, the animal should be euthanatized immediately
ii. If, on day 4 after transmitter implantation, the animal shows clear signs of apathy, is extremely aggressive or does not show any food intake, it should be euthanatized immediately.
iii. On day 8 after transmitter implantation, the animal has to display a clear increase in body weight in comparison to the preceding post-operative days. Moreover, it has to consume at least 80% of the pre-operative daily food intake. If one of these conditions is not met, the animal should be euthanatized immediately.
At 10 days after implantation, the animal is transferred back to the animal room under standard housing conditions. Mice should be housed in compatible groups to allow social interaction and to prevent the adverse effects of long-term individual housing, which can have substantial impacts on the read-out of subsequent experiments8, 9. Mice should have a period of at least 4 weeks convalescence after transmitter implantation before the first experiment is conducted and data acquisition begins.
to:
After completion of surgery and anesthesia, 0.1 mg/kg of buprenorphine (Temgesic, Essex Chemie AG, Lucerne, Switzerland) and 5 mg/kg of meloxicam (Metacam, Boehringer Ingelheim, Basel, Switzerland) is administered subcutaneously for pain treatment, and the animals are left on the warm (39&#176;C +/-1) surface of the work bench to recover for approximately 2h. Together with pain relief (twice daily: buprenorphine, 0.1 mg/kg and meloxicam 5 mg/kg), supportive therapy consisting of 300 &#956;L glucose (5%) and 300 &#956;L saline (0.9%) warmed to body temperature, is injected subcutaneously twice daily for 4 days. For further recovery support, it is worthwhile providing the animals with an additional drinking bottle containing 15% glucose solution During the recovery period of 4-10 days, it is recommended that the animals are kept warm. Therefore, in our case, the mice are housed in a warming cabinet (30&#176;C +/- 1). Monitoring of general condition and body weight, as well as food and water consumption, is performed once daily according to the general condition and health monitoring data sheet (Table 1) for 10 days post-operatively. Humane endpoints, i.e. the sacrifice of an animal to avoid unnecessary suffering and pain if progression of recovery is unsatisfactory, are realised under the following conditions:
i. If in poor general condition, i.e. the animal is substantially apathetic (no movement after being touched/pushed) and its body surface feels cold despite warming, the animal should be euthanatized immediately
ii. If, on day 4 after transmitter implantation, the animal shows clear signs of apathy, is extremely aggressive or does not show any food intake, it should be euthanatized immediately.
iii. On day 8 after transmitter implantation, the animal has to display a clear increase in body weight in comparison to the preceding post-operative days. Moreover, it has to consume at least 80% of the pre-operative daily food intake. If one of these conditions is not met, the animal should be euthanatized immediately.
At 10 days after implantation, the animal is transferred back to the animal room under standard housing conditions. In case staples have been used, these should be removed 7-10 days after surgery; absorbable sutures have not to be removed. Mice should be housed in compatible groups to allow social interaction and to prevent the adverse effects of long-term individual housing, which can have substantial impacts on the read-out of subsequent experiments8, 9. Mice should have a period of at least 4 weeks convalescence after transmitter implantation before the first experiment is conducted and data acquisition begins.
The 4th paragraph in the Discussion has been updated from:
Aseptic conditions should be maintained during surgery to keep the microbiological burden and the risk of infections low. However, complete sterility cannot be provided because of some specific, sterility conflicting conditions in mice (e.g., cooling effect of extensive hair clipping and disinfection, impracticality of bandages to protect the wounds). Thus, anti-infective prophylaxis is administered during the implantation. Well tailored analgesic treatment and a clearly defined monitoring plan as well as adequate post-operative care play a crucial role in the satisfactory outcome of the experiment.
to:
Aseptic conditions should be maintained during surgery to keep the microbiological burden and the risk of infections low. However, if there are doubts that asepsis was breached because of some specific, sterility conflicting conditions in mice (e.g., cooling effect of extensive hair clipping and disinfection, impracticality of bandages to protect the wounds). Anti-infective prophylaxis should be administered during the implantation. Well-tailored analgesic treatment and a clearly defined monitoring plan as well as adequate post-operative care play a crucial role in the satisfactory outcome of the experiment.

Corrections in the Protocol and Discussion sections have been made to: Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

Erratum: Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

implantation of radiotelemetry transmitters yielding data on ecg, heart rate, core body temperature and activity in free-moving laboratory mice

Il topo di laboratorio è la specie animale di scelta per la maggior parte della ricerca biomedica, sia nel mondo accademico e l&#39;industria farmaceutica. I topi sono di dimensioni gestibili e relativamente facile da casa. Questi fattori, insieme con la disponibilità di un patrimonio di mutanti spontaneo e indotto sperimentalmente, fanno topi di laboratorio ideale per una vasta gamma di aree di ricerca. Nella ricerca cardiovascolare, farmacologiche e tossicologiche, la misurazione accurata dei parametri relativi al sistema circolatorio degli animali da laboratorio è spesso richiesto. Determinazione della frequenza cardiaca, variabilità della frequenza cardiaca, e la durata del PQ e gli intervalli QT sono basate su elettrocardiogramma (ECG) registrazioni. Tuttavia, ottenere affidabile tracciati ECG, nonché i dati fisiologici come la temperatura corporea nei topi possono essere difficili utilizzando tecniche di misura convenzionali, che richiedono il collegamento di sensori e cavi ad una trattenuta, legati, o anche unaesthetized animali. I dati ottenuti in questo modo devono essere interpretati con cautela, in quanto è ben noto che il contenimento e l&#39;anestesia può avere una grande influenza sui parametri fisiologici artefatta 1, 2. Radiotelemetry permette di dati da raccogliere da animali coscienti e untethered. Le misure possono essere condotte anche in animali di muoversi liberamente, e senza richiedere l&#39;investigatore di essere in prossimità dell&#39;animale. Così, fonti note di artefatti sono evitati, e misure accurate ed affidabili sono assicurate. Questa metodologia riduce anche la variabilità interanimal, riducendo così il numero di animali utilizzati, rendendo questa tecnologia il metodo più umano di monitoraggio dei parametri fisiologici in animali da laboratorio 3, 4. Progressi costanti nella tecnologia di acquisizione dati e la miniaturizzazione degli impianti significa che è ora possibile registrare i parametri fisiologici e attività locomotoria in modo continuo e in tempo reale su più periods come ore, giorni o addirittura settimane 3, 5. Qui, descriviamo una tecnica chirurgica per l&#39;impianto di un trasmettitore di telemetria disponibili in commercio utilizzati per misure in continuo della temperatura corporea, l&#39;attività locomotoria e biopotenziale (cioè onelead ECG), da cui la frequenza cardiaca, variabilità della frequenza cardiaca, e PQ e gli intervalli QT può essere fondata nel freeroaming, topi untethered. Siamo presenti anche pre-operatoria procedure e protocolli per la post-operatorio di terapia intensiva e terapia del dolore che migliorare il recupero, il benessere e la sopravvivenza in topi impiantati 5, 6.

Watch this Scientific Journal Video about Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice at JoVE.com

Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

Una tecnica chirurgica per l&#39;impianto di trasmettitori telemetrici disponibili in commercio utilizzati per la misura continua della biopotenziale (un ECG), frequenza cardiaca, temperatura corporea e l&#39;attività locomotoria nei topi liberamente in movimento è mostrato. Raccomandazioni e protocolli per il post-operatorio e del dolore, migliorare il recupero, il benessere e il tasso di sopravvivenza sono presentati.

L&#39;impianto di trasmettitori Radiotelemetry Cedendo dati su ECG, frequenza cardiaca, temperatura corporea e attività di movimento libero topi di laboratorio

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a ...

implantation-radiotelemetry-transmitters-yielding-data-on-ecg-heart

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JoVE Journal

Medicine

40.3K Views.  University Hospital Zurich. Una tecnica chirurgica per l'impianto di trasmettitori telemetrici disponibili in commercio utilizzati per la misura continua della biopotenziale (un ECG), frequenza cardiaca, temperatura corporea e l'attività locomotoria nei topi liberamente in movimento è mostrato. Raccomandazioni e protocolli per il post-operatorio e del dolore, migliorare il recupero, il benessere e il tasso di sopravvivenza sono presentati.

Video: Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

Impianto di un bracciale carotidea per innescare Shear-stress aterosclerosi indotta in topi

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

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Medicina

Implantation of the Syncardia Total Artificial Heart

With advances in technology, the use of mechanical circulatory support devices for end stage heart failure has rapidly increased. The vast majority of such patients are generally well served by left ventricular assist devices (LVADs). However, a subset of patients with late stage biventricular failure or other significant anatomic lesions are not adequately treated by isolated left ventricular mechanical support. Examples of concomitant cardiac pathology that may be better treated by resection and TAH replacement includes: post infarction ventricular septal defect,&#160;aortic root aneurysm / dissection, cardiac allograft failure, massive ventricular thrombus, refractory malignant arrhythmias (independent of filling pressures), hypertrophic / restrictive cardiomyopathy, and complex congenital heart disease. Patients often present with cardiogenic shock and multi system organ dysfunction. Excision of both ventricles and orthotopic replacement with a total artificial heart (TAH) is an effective, albeit extreme, therapy for rapid restoration of blood flow and resuscitation. Perioperative management is focused on end organ resuscitation and physical rehabilitation. In addition to the usual concerns of infection, bleeding, and thromboembolism common to all mechanically supported patients, TAH patients face unique risks with regard to renal failure and anemia. Supplementation of the abrupt decrease in brain natriuretic peptide following ventriculectomy appears to have protective renal effects. Anemia following TAH implantation can be profound and persistent. Nonetheless, the anemia is generally well tolerated and transfusion are limited to avoid HLA sensitization. Until recently, TAH patients were confined as inpatients tethered to a 500 lb pneumatic console driver. Recent introduction of a backpack sized portable driver (currently under clinical trial) has enabled patients to be discharged home and even return to work. Despite the profound presentation of these sick patients, there is a 79-87% success in bridge to transplantation.

The purpose of this manuscript is to briefly review indications, management, and outcomes for the total artificial heart. Video operative techniques for device implantation are presented.

L&#39;impianto di Cuore Artificiale Totale SynCardia

With advances in technology, the use of mechanical circulatory support devices for end stage heart failure has rapidly increased. The vast majority of ...

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

La valutazione ecocardiografica del cuore destro in topi

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease and injury regional estimates, World Health Organization, 2004). Surgeries that often compromise the quality of life are required to correct heart defects, reminding us of the importance of finding the causes of CHD. Mutant mouse models and live imaging technology have become essential tools to study the etiology of this disease. Although advanced methods allow live imaging of abnormal hearts in embryos, the physiological and hemodynamic states of the latter are often compromised due to surgical and/or lengthy procedures. Noninvasive ultrasound imaging, however, can be used without surgically exposing the embryos, thereby maintaining their physiology. Herein, we use simple M-mode ultrasound to assess heart rates of embryos at E18.5 in utero. The detection of abnormal heart rates is indeed a good indicator of dysfunction of the heart and thus constitutes a first step in the identification of developmental defects that may lead to heart failure.

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Ultra-high frequency ultrasound is a powerful live imaging tool to examine cardiac abnormalities in small animals. Its noninvasiveness allows maintaining the physiologic state of embryos. Herein, we show the use of M-mode ultrasound to measure the heart rates of embryos at E18.5 in utero.

In utero Misurazione della frequenza cardiaca in mouse da non invasiva ecocardiografia M-mode

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease ...

Implantation of Total Artificial Heart in Congenital Heart Disease

In patients with end-stage heart failure (HF), a total artificial heart (TAH) may be implanted as a bridge to cardiac transplant. However, in congenital heart disease (CHD), the malformed heart presents a challenge to TAH implantation. 
In the case presented here, a 17 year-old patient with congenital transposition of the great arteries (CCTGA) experienced progressively worsening HF due to his congenital condition. He was hospitalized multiple times and received an implantable cardioverter defibrillator (ICD). However, his condition soon deteriorated to end-stage HF with multisystem organ failure. 
Due to the patient's grave clinical condition and the presence of complex cardiac lesions, the decision was made to proceed with a TAH. The abnormal arrangement of the patient's ventricles and great arteries required modifications to the TAH during implantation.
With the TAH in place, the patient was able to return home and regain strength and physical well-being while awaiting a donor heart. He was successfully bridged to heart transplantation 5 months after receiving the device. This report highlights the TAH is feasible even in patients with structurally abnormal hearts, with technical modification.

This is a case report of a patient with congenitally corrected transposition of the great arteries (CCTGA) who received a total artificial heart (TAH) as a bridge to heart transplant. The TAH was successfully implanted with modifications to accommodate the patient's congenitally malformed heart.

L&#39;impianto di Cuore Artificiale Totale nelle cardiopatie congenite

In patients with end-stage heart failure (HF), a total artificial heart (TAH) may be implanted as a bridge to cardiac transplant. However, in congenital ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Cloruro ferrico-indotta Trombosi modello murino sulla carotide e Mesentere Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

Eterotopico cervicale trapianto cardiaco nei topi

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a wide variety of disorders including metabolic diseases, premature aging syndromes, and neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD). Maintenance of mitochondrial health depends on mitochondrial biogenesis and the efficient clearance of dysfunctional mitochondria through mitophagy. Experimental methods to accurately detect autophagy/mitophagy, especially in animal models, have been challenging to develop. Recent progress towards the understanding of the molecular mechanisms of mitophagy has enabled the development of novel mitophagy detection techniques. Here, we introduce several versatile techniques to monitor mitophagy in human cells, Caenorhabditis elegans (e.g., Rosella and DCT-1/ LGG-1 strains), and mice (mt-Keima). A combination of these mitophagy detection techniques, including cross-species evaluation, will improve the accuracy of mitophagy measurements and lead to a better understanding of the role of mitophagy in health and disease.

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

Mitophagy, the process of clearing damaged mitochondria, is necessary for mitochondrial homeostasis and health maintenance. This article presents some of the latest mitophagy detection methods in human cells, Caenorhabditis elegans, and mice.

In Vitro e In Vivo rilevamento di Mitophagy in cellule umane, C. Eleganse topi

Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a ...

Methodology for Sputum Induction and Laboratory Processing

The technique of sputum induction and processing is a recognized non-invasive method allowing the collection and analysis of cells from the airways, which is interesting in various respiratory diseases like asthma, chronic obstructive pulmonary disease (COPD), chronic cough, or idiopathic pulmonary fibrosis. This technique is well tolerated, safe and non-invasive, but is currently limited to research services and specialized centers in clinical practice because it is technically demanding, time-consuming, and requires trained staff. The success rate of sputum induction and analysis is about 80%.
Here, we describe the induction and laboratory processing of sputum samples. Sputum is induced by inhalation of hypertonic or isotonic saline with salbutamol. For the processing, we use the whole sputum technique. Dithiothreitol (DTT) is used to allow mucolysis of sputum samples. The primary aim of sputum processing is to obtain a differential cell count to study the cell types present in the airway lumen. Additional analyses may also be performed on sputum supernatant and sputum cells, which may allow further investigation into inflammatory processes and immune mechanisms. Examples include studying mediators in sputum supernatant and performing a large spectrum of analysis on sputum cells such as flow cytometry, genomics, or proteomics.
Finally, representative results of sputum analysis in healthy controls, asthmatics, and COPD patients are presented.

Here we describe the technique of sputum induction. This protocol also explains the sputum processing to perform a differential cell count and to collect sputum supernatant and cells for further analysis.

Metodologia per l'induzione dell'espettorato e laboratorio di lavorazione

The technique of sputum induction and processing is a recognized non-invasive method allowing the collection and analysis of cells from the airways, which ...

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.

Il modello di pilocarpina di epilessia del lobo temporale e monitoraggio EEG utilizzando il sistema di radiotelemetria in topi

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