Supportato da DK065872 (PEV), una famiglia Smith Award Foundation di eccellenza nella ricerca biomedica (PEV), F31 DA023760.

Riassunto Due mesi di età media di topi C57BL/6J età o equivalente o tre mesi età media ratti Sprague Dawley o equivalenti sono anestetizzati con ketamina (60 mg / kg ip per i ratti; 100 mg / kg ip per topi) e xilazina (10 mg / kg, ip per entrambe le specie). La sedazione è controllata mediante un pizzico dolce toe ritirare riflesso dimostrato in <a href="/index/details.stp?ID=236">Walantus et al.</a> (Giove, 6, 2007) e <a href="/index/details.stp?ID=404">Szot et al.</a> (Giove, 9, 2007). Termoregolazione può essere fornita attraverso una piastra elettrica thermostatregulated (ALA Instruments Inc.) e monitorati attraverso un termometro rettale. La testa è rasata di pelliccia e pulito con iodio prima incisione. Dopo l&#39;incisione della pelle (2 cm di lunghezza per i ratti, 1 cm per topi) e la rimozione di tutti i tessuti molli dalla superficie del cranio, il posizionamento della cannula guida è determinato in relazione al bregma. A 6 mm foro si è esercitato attraverso il cranio con un trapano a batteria destinati per la chirurgia roditori (Strumenti Scienza Fine, Inc.). Si fa attenzione in modo che la punta non penetra attraverso le membrane meningee o vasi sanguigni. Cranio è impiantato bilaterale con 5 millimetri 21 alberi in acciaio inox calibro guida che porta al nucleo accumbens posteriore, striato dorsale o corteccia prefrontale. Le coordinate stereotassiche sono stabiliti secondo Franklin e Paxinos, 1997 (Il cervello del mouse in coordinate Stereotassica, Academic Press) o Paxinos e Watson, 2006 (Il cervello di ratto in coordinate Stereotassica, Academic Press). Gli impianti sono garantiti da cemento dentale. Un bolo di Ringer lattato della soluzione salina 0,9% è dato alla fine della chirurgia (5mls SC nei ratti e 1 ml SC nei topi dopo fluidi sono portata a temperatura corporea normale) per prevenire la disidratazione. Buprenorfina (0.1-0.5mg/kg SC) viene somministrato due volte al giorno e, poi, su una base come-necessaria, se l&#39;animale sembra essere in sofferenza. Trattamento antibiotico locale (unguento bacitracina) e un trattamento antibiotico sistemico (penicillina 100.000 UI / kg IM ogni 12 ore per le prime 48 ore post-op) se vengono somministrati infezioni post-operatorie si verificano. Dopo la chirurgia, gli animali sono alloggiati individualmente con il cibo e ad libitum acqua disponibile. Almeno una settimana è consentito per il recupero prima di microdialisi e l&#39;eutanasia. A seguito di recupero da un intervento chirurgico, gli animali vengono trasferiti in una gabbia microdialisi e sonde microdialisi sono inseriti e cementati nei pozzi guida che sono stati installati durante l&#39;intervento. L&#39;inserimento della sonda non provoca dolore o fastidio, perché la sonda è bypassando tessuto cutaneo, muscolare e meningea attraverso l&#39;albero guida. Pertanto, l&#39;inserimento della sonda viene fatto senza anestesia e qualsiasi anestesia effetti indotti sulla neurochimica o comportamenti sono evitati. Lasciamo che le sonde stabilizzazione per 12 ore e poi iniziare il campionamento ogni 30 minuti per un altro 8-12 ore a seconda dell&#39;esperimento. Monitoriamo il comportamento motorio degli animali attraverso fotocellule o la registrazione manuale del movimento dallo sperimentatore. Microdialysate campioni vengono iniettati in una cromatografia liquida ad alte prestazioni con rilevazione elettrochimica (HPLC-EC) strumento per la rilevazione e l&#39;analisi neurochimiche. Siamo alla ricerca di effetti sulla neurochimica basale e il comportamento motorio. Alla fine dell&#39;esperimento l&#39;animale è eutanasia da un overdose di ketamina sistemica (200 mg / kg ip) e xilazina (20 mg / kg, ip). Allora il cuore è perfuso con soluzione salina 0,9% seguito da 4% paraformaldeide. I cervelli sono stati rimossi, congelati e tagliare lungo il tratto della sonda microdialisi per verificare il posizionamento preciso della sonda. Procedura <ol><li> Impostare lo strumento stereotassico e tutti i materiali necessari. Assicurarsi che l&#39;area e gli strumenti sono puliti e sterilizzati. </li><li> Radersi pelliccia con rasoio elettrico. Vai dalle orecchie a soli in mezzo agli occhi, spostare rasoio in diverse direzioni per pulire efficacemente un&#39;area di pelliccia. Applicare povidine / iodio a zona rasata, ma proteggere gli occhi da esso. </li><li> Montare l&#39;animale su l&#39;apparato stereotassico inserendo le barrette orecchio nel condotto uditivo e di serraggio in posizione. Prima cosa si monti una barra orecchio nel canale auricolare, e poi tenerlo in posizione e scivolare nel bar altro orecchio. Sai che sono nella posizione corretta quando la testa non può più essere spostato lateralmente. Fissare la bocca con il supporto anteriore del stereotassica e assicurarsi che la testa sia a livello con un righello. Mettere il righello in posizione verticale rispetto alla piattaforma strumento stereotassico e verificare la presenza di un angolo di 90 ° tra il sovrano e la metà del cuoio capelluto dell&#39;animale). Confermare questo attraverso lo strumento stereotassico se offre funzionalità di questo tipo. </li><li> Fare un&#39;incisione anteriore / posteriore del cuoio capelluto con un bisturi sterile, che si estende dal lambda a soli in mezzo agli occhi dell&#39;animale. Usa hemostats sterilizzati per pizzicare sulla pelle e tenere aperta l&#39;incisione. Utilizzando diversi tamponi di cotone sterile, asciugare la superficie esposta del cranio. <li> Mettere la cannula guida sul suo supporto, trovare bregma sul cranio, e la posizione della cannula guida a destra su questa posizione. Annotare le coordinate anteriore / posteriore e laterale. Da bregma, trovare le coordinate necessarie per il corretto posizionamento del vostro sonda con l&#39;aiuto dell&#39;atlante stereotassico. Posizionare la cannula guida alle coordinate corrette aggiungendo o sottraendo dal bregma. Portate la vostra cannula guida verso il basso fino a toccare il cranio, e poi registrare questa coordinata ventrale. Fare un segno di matita con una matita sterile in questa posizione sul cranio, questo è dove sarete foratura. </li><li> Rimuovere la cannula guida e sterilizzare la tua punta da trapano. Con attenzione praticare un foro al segno a matita fino ad ottenere attraverso la larghezza del cranio. Verificare con la cannula guida per vedere se sarebbe chiaro il foro senza toccare i bordi. Tenere foratura e controllo fino a quando la cannula può cancellare in un percorso rettilineo. Una volta che il buco è fatto, utilizzare un ago sterile a pugni con delicatezza le meningi in modo da permettere l&#39;inserimento della cannula senza ostacoli. </li><li> Successivamente, utilizzando un trapano a mano, rendono sei fori per le viti del cranio: due anteriori al foro della cannula, due laterali al foro della cannula, e posteriori ai lati due. Sterilizzare sei viti e metterli sul cranio, fino a che non sono strettamente ancorate su. </li><li> Pulire la cannula guida con etanolo e saline, montare, e abbassare lentamente alla coordinata corretta ventrale. Assicurarsi che i lati non si toccano e che sta andando in perfetta verticale. </li><li> Posizionare la vite di ancoraggio medialmente e dietro le viti cranio posteriore e tenerlo in posizione con le pinzette. Mescolare un lotto sottile di liquido cemento dentale e coprire la cannula guida, viti, e il resto del cranio con una spatola sterile. Fare un altro lotto, questa volta più spessa, e coprire completamente la zona e la vite cannula e ancora abbastanza per fissarlo. </li><li> Come il cemento diventa più spessa e prima che si solidifica, separare la pelle dalla coppa cemento e stampo la coppa di cemento con la spatola per assicurarsi che il tappo di cemento è tutt&#39;intorno e non irrita la pelle più tardi. </li><li> Lasciare che il cemento dentale si asciughi completamente prima di rimuovere l&#39;animale dall&#39;apparecchio. Rimuovere il hemostats. Applicare tutto il percorso attorno al tappo di cemento bacitracina. </li><li> Una volta che l&#39;animale è spento lo strumento stereotassico, si iniettano con 0,25 ml di penicillina IM (intra-muscolare), seguito da 1 ml di soluzione fisiologica SC (via sottocutanea). </li><li> Posto l&#39;animale nella sua gabbietta e monitorare fino a che non prende coscienza prima di tornare alla sua stanza per recuperare. </li><li> Monitorare gli animali fino a quando non riprendersi dall&#39;anestesia il giorno dell&#39;intervento e giorno post-op, fino alla fine dell&#39;esperimento, eventuali segni di infezione e di valutazione del dolore / disagio. Questo include fine settimana e festivi. Bassa movimento spontaneo, vocalizzazione angoscia sulla gestione, la postura ingobbita, diarrea, gonfiore e scarico nella zona del headmount, e la mancanza di alimentazione / bere sono tutti segni di dolore e di angoscia. Buprenorfina (0.1-0.5mg/kg SC) viene somministrato due volte al giorno, e poi, su una base come-necessaria, se l&#39;animale sembra essere in sofferenza. Trattamento antibiotico locale (unguento bacitracina) e un trattamento antibiotico sistemico (penicillina 100.000 UI / kg IM ogni 12 ore per le prime 48 ore post-op) se vengono somministrati infezioni post-operatorie si verificano. Se qualcuno di questi sintomi persistono dopo la somministrazione di buprenorfina, il liquido supplementare, e il trattamento antibiotico entro le 12 ore di intervento chirurgico, l&#39;animale è eutanasia. </li></ol>

<ol>
	<li>Bungay, P. M., Newton-Vinson, P., Isele, W., Garris, P. A., Justice, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microdialysis+of+dopamine+interpreted+with+quantitative+model+incorporating+probe+implantation+trauma.">Microdialysis of dopamine interpreted with quantitative model incorporating probe implantation trauma.</a> J. Neurochem. 86, 932-946 (2003).</li><li>Chen, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+tissue+trauma+on+the+characteristics+of+microdialysis+zero-net-flux+method+sampling+neurotransmitters.">Effects of tissue trauma on the characteristics of microdialysis zero-net-flux method sampling neurotransmitters.</a> Journal of Theor. Biology. 238, 863-881 (2006).</li><li>Geiger, B. M., Behr, G. G., Frank, L., Caldera-Siu, A. D., Beinfeld, M. C., Kokkotou, E. G., Pothos, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+defective+mesolimbic+dopamine+exocytosis+in+obesity-prone+rats.">Evidence for defective mesolimbic dopamine exocytosis in obesity-prone rats.</a> FASEB Journal. 8, 2740-2746 (2008).</li><li>Pothos, E. N., Creese, I., Hoebel, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restricted+eating+with+weight+loss+selectively+decreases+extracellular+dopamine+in+the+nucleus+accumbens+and+alters+dopamine+response+to+amphetamine,+morphine+and+food+intake.">Restricted eating with weight loss selectively decreases extracellular dopamine in the nucleus accumbens and alters dopamine response to amphetamine, morphine and food intake.</a> The Journal of Neuroscience. 15, 6640-6650 (1995).</li></ol>

The authors have nothing to disclose.

Microdialisi in vivo è lo strumento di scelta per la misurazione dei neurotrasmettitori multipli e metaboliti in siti diversi del cervello di un animale vivo. Tuttavia, controlla solo i livelli extracellulari di sostanze neurochimiche e non offre la risoluzione temporale di monitorare esocitosi dei neurotrasmettitori in tempo reale. Attraverso una versione della tecnica chiamata &quot;net-flux&quot;, la concentrazione dei neurotrasmettitori effettiva in un determinato sito può essere calcolato, che a loro volta possono dare misurazioni ac...

I materiali sono descritti nel documento protocollo

survivable stereotaxic surgery in rodents 

La capacità di misurare i livelli basali extracellulari di neurotrasmettitori nel cervello di animali svegli permette per la determinazione degli effetti di diverse sfide sistemiche (farmacologici o fisiologico) al sistema nervoso centrale. Per esempio, si può misurare direttamente come proiezioni dopamina dell&#39;animale mesencefalo rispondere a rilascio di dopamina farmaci come gli stimoli d-anfetamina o naturale come il cibo. In questo video vi mostriamo come impianto cannule guida targeting siti specifici nel cervello del ratto, come inserire e impiantare una sonda microdialisi e come utilizzare cromatografia liquida ad alta prestazione accoppiata con rilevazione elettrochimica (HPLC-EC) per misurare i livelli extracellulari di neurotrasmettitori ossidabili e metaboliti. Locali introduzione preciso dei farmaci attraverso la sonda di microdialisi permette di raffinato lavoro sulle specificità del sito in un composto s meccanismo di azione. Questa tecnica è eccellente e la risoluzione anatomica chimiche, ma solo i tempi di risoluzione modesta come campioni microdialisi sono di norma elaborata ogni 20-30 minuti per assicurare i livelli di neurotrasmettitore rilevabili. Strumenti complementari ex vivo (cioè fetta e coltura cellulare elettrofisiologia) può aiutare con il monitoraggio in tempo reale la neurotrasmissione.

Watch this Scientific Journal Video about Survivable Stereotaxic Surgery in Rodents at JoVE.com

Survivable Stereotaxic Surgery in Rodents

Il monitoraggio dei livelli del neurotrasmettitore extracellulare nelle regioni cerebrali distinte di muoversi liberamente animali offre approfondimenti sul legame tra il rilascio dei neurotrasmettitori e comportamento. In microdialisi in vivo accoppiato con rilevazione elettrochimica fornisce anatomica eccellente e la risoluzione chimici e informazioni su come neurotrasmissione basale è alterato da manipolazioni farmacologiche o fisiologiche.

Survivable Chirurgia Stereotassica in roditori

The ability to measure extracellular basal levels of neurotransmitters in the brain of awake animals allows for the determination of effects of different ...

survivable-stereotaxic-surgery-in-rodents

Research

JoVE Journal

Biology

Survivable Stereotaxic Surgery in Rodents 

68.3K Views.  Tufts University. Il monitoraggio dei livelli del neurotrasmettitore extracellulare nelle regioni cerebrali distinte di muoversi liberamente animali offre approfondimenti sul legame tra il rilascio dei neurotrasmettitori e comportamento. In microdialisi in vivo accoppiato con rilevazione elettrochimica fornisce anatomica eccellente e la risoluzione chimici e informazioni su come neurotrasmissione basale è alterato da manipolazioni farmacologiche o fisiologiche.

Video: Survivable Stereotaxic Surgery in Rodents

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 ...

A Craniotomy Surgery Procedure for Chronic Brain Imaging

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an extremely useful method, because it allows the study of the live intact brain. With appropriate preparations, this technique allows the observation of the same cortical area chronically, from minutes to months. In this video, we show a preparation for chronic in vivo imaging of the brain using two-photon microscopy. This technique was initially pioneered by Dr. Karel Svoboda, who is now a Howard Hughes Medical Institute Investigator at Janelia Farm. Preparations like the one shown here can be used for imaging of neocortical structure (e.g., dendritic and axonal dynamics),  to record neuronal activity using calcium-sensitive dyes, to image cortical blood flow dynamics, or for intrinsic optical imaging studies. Deep imaging of the neocortex is possible with optimal cranial window surgeries. Operating under the most sterile conditions possible to avoid infections, together with using extreme care to do not damage the dura mater during the surgery, will result in successful and long-lasting glass-covered cranial windows.


This video and protocol demonstrate how to implant a glass-covered cranial window in rodents. These preparations can be used for chronic in vivo two-photon imaging of the neocortex over time scales of months. It may also be used for other types of imaging, including optical intrinsic signal imaging.

Un Chirurgia Procedura Craniotomia per l&#39;imaging cerebrale cronica

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an ...

Dissecting the Non-human Primate Brain in Stereotaxic Space

The use of non-human primates provides an excellent translational model for our understanding of developmental and aging processes in humans1-6. In addition, the use of non-human primates has recently afforded the opportunity to naturally model complex psychiatric disorders such as alcohol abuse7. Here we describe a technique for blocking the brain in the coronal plane of the vervet monkey (Chlorocebus aethiops sabeus) in the intact skull in stereotaxic space. The method described here provides a standard plane of section between blocks and subjects and minimizes partial sections between blocks. Sectioning a block of tissue in the coronal plane also facilitates the delineation of an area of interest. This method provides manageable sized blocks since a single hemisphere of the vervet monkey yields more than 1200 sections when slicing at 50&mu;m. Furthermore by blocking the brain into 1cm blocks, it facilitates penetration of sucrose for cyroprotection and allows the block to be sliced on a standard cryostat.

The non-human primate is an important translational species for our understanding of the normal processing of the brain. The anatomical organization of the primate brain can provide important insights into normal and pathological conditions in humans.

Dissezione di non-umano cervello dei primati nello spazio Stereotassica

The use of non-human primates provides an excellent translational model for our understanding of developmental and aging processes in humans1-6. In ...

Establishing Primary Adult Fibroblast Cultures From Rodents

The importance of using primary cells, rather than cancer cell lines, for biological studies is becoming widely recognized. Primary cells are preferred in studies of cell cycle control, apoptosis, and DNA repair, as cancer cells carry mutations in genes involved in these processes. Primary cells cannot be cultured indefinitely due to the onset of replicative senescence or aneuploidization. Hence, new cultures need to be established regularly. The procedure for isolation of rodent embryonic fibroblasts is well established, but isolating adult fibroblast cultures often presents a challenge. Adult rodent fibroblasts isolated from mouse models of human disease may be a preferred control when comparing them to fibroblasts from human patients. Furthermore, adult fibroblasts are the only available material when working with wild rodents where pregnant females cannot be easily obtained. Here we provide a protocol for isolation and culture of adult fibroblasts from rodent skin and lungs. We used this procedure successfully to isolate fibroblasts from over twenty rodent species from laboratory mice and rats to wild rodents such as beaver, porcupine, and squirrel.

This article describes a protocol for isolation and maintenance of primary fibroblast cultures from skin and lung tissue of wild rodents.

Stabilire colture primarie di fibroblasti adulti dai roditori

The importance of using primary cells, rather than cancer cell lines, for biological studies is becoming widely recognized.  Primary cells are preferred ...

Principles of Rodent Surgery for the New Surgeon

For both scientific and animal welfare reasons, training in basic surgical concepts and techniques should be undertaken before ever seeking to perform surgery on a rodent. Students, post-doctoral scholars, and others interested in performing surgery on rodents as part of a research protocol may not have had formal surgical training as part of their required coursework. Surgery itself is a technical skill, and one that will improve with practice. The principles of aseptic technique, however, often remain unexplained or untaught. For most new surgeons, this vital information is presented in piecemeal fashion or learned on the job, neither of which is ideal. It may also make learning how to perform a particular surgery difficult, as the new surgeon is learning both a surgical technique and the principles of asepsis at the same time. This article summarizes and makes recommendations for basic surgical skills and techniques necessary for successful rodent surgery. This article is designed to supplement hands-on training by the user's institution.

Before attempting surgery, a new surgeon should have training in basic surgical techniques and concepts. This article will present basic surgical considerations with an emphasis on rodents.

Principi di Chirurgia roditori per il chirurgo nuovo

For both scientific and animal welfare reasons, training in basic surgical concepts and techniques should be undertaken before ever seeking to perform ...

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. ...

Inhibition of Wound Epidermis Formation via Full Skin Flap Surgery During Axolotl Limb Regeneration

Classical experiments in salamander regenerative biology over the last century have long established that the wound epidermis is a crucial signaling structure that forms rapidly post-amputation and is required for limb regeneration. However, methods to study its precise function at the molecular level over the last decades have been limited due to a paucity of precise functional techniques and genomic information available in salamander model systems. Excitingly, the recent plethora of sequencing technologies coupled with the release of various salamander genomes and the advent of functional genetic testing methods, including CRISPR, makes it possible to re-visit these foundational experiments at unprecedented molecular resolution. Here, I describe how to perform the classically developed full skin flap (FSF) surgery in adult axolotls in order to inhibit wound epidermis formation immediately following amputation. The wound epidermis normally forms via distal migration of epithelial cells in the skin proximal to the amputation plane to seal off the wound from the outside environment. The surgery entails immediately suturing full thickness skin (which includes both epidermal and dermal layers) over the amputation plane to hinder epithelial cell migration and contact with the underlying damaged mesenchymal tissues. Successful surgeries result in the inhibition of blastema formation and limb regeneration. By combining this surgery method with contemporary downstream molecular and functional analyses, researchers can begin to uncover the molecular underpinnings of wound epidermis function and biology during limb regeneration.

This article describes how to perform a surgical method to inhibit wound epidermis formation during axolotl limb regeneration by immediately suturing full thickness skin over the amputation plane. This method allows researchers to investigate the functional roles of the wound epidermis during the early stages of limb regeneration.

Inibizione della formazione dell'epidermide della ferita tramite chirurgia del lembo della pelle completa durante la rigenerazione degli arti Axolotl

Classical experiments in salamander regenerative biology over the last century have long established that the wound epidermis is a crucial signaling ...

Application of Passive Head Motion to Generate Defined Accelerations at the Heads of Rodents

Exercise is widely recognized as effective for various diseases and physical disorders, including those related to brain dysfunction. However, molecular mechanisms behind the beneficial effects of exercise are poorly understood. Many physical workouts, particularly those classified as aerobic exercises such as jogging and walking, produce impulsive forces at the time of foot contact with the ground. Therefore, it was speculated that mechanical impact might be implicated in how exercise contributes to organismal homeostasis. For testing this hypothesis on the brain, a custom-designed ''passive head motion'' (hereafter referred to as PHM) system was developed that can generate vertical accelerations with controlled and defined magnitudes and modes and reproduce mechanical stimulation that might be applied to the heads of rodents during treadmill running at moderate velocities, a typical intervention to test the effects of exercise in animals. By using this system, it was demonstrated that PHM recapitulates the serotonin (5-hydroxytryptamine, hereafter referred to as 5-HT) receptor subtype 2A (5-HT2A) signaling in the prefrontal cortex (PFC) neurons of mice. This work provides detailed protocols for applying PHM and measuring its resultant mechanical accelerations at rodents' heads.

The present protocol describes a custom-designed ''passive head motion'' system, which reproduces mechanical accelerations at rodents' heads generated during their treadmill running at moderate velocities. It allows dissecting mechanical factors/elements from the beneficial effects of physical exercise.

Applicazione del movimento passivo della testa per generare accelerazioni definite alle teste dei roditori

Exercise is widely recognized as effective for various diseases and physical disorders, including those related to brain dysfunction. However, molecular ...

Performing In Vivo and Ex Vivo Electrical Impedance Myography in Rodents

Electrical impedance myography (EIM) is a convenient technique that can be used in preclinical and clinical studies to assess muscle tissue health and disease. EIM is obtained by applying a low-intensity, directionally focused, electrical current to a muscle of interest across a range of frequencies (i.e., from 1 kHz to 10 MHz) and recording the resulting voltages. From these, several standard impedance components, including the reactance, resistance, and phase, are obtained. When performing ex vivo measurements on excised muscle, the inherent passive electrical properties of the tissue, namely the conductivity and relative permittivity, can also be calculated. EIM has been used extensively in animals and humans to diagnose and track muscle alterations in a variety of diseases, in relation to simple disuse atrophy, or as a measure of therapeutic intervention. Clinically, EIM offers the potential to track disease progression over time and to assess the impact of therapeutic interventions, thus offering the opportunity to shorten the clinical trial duration and reduce sample size requirements. Because it can be performed noninvasively or minimally invasively in living animal models as well as humans, EIM offers the potential to serve as a novel translational tool enabling both preclinical and clinical development. This article provides step-by-step instructions on how to perform in vivo and ex vivo EIM measurements in mice and rats, including approaches to adapt the techniques to specific conditions, such as for use in pups or obese animals.

Performing In Vivo and Ex Vivo Electrical Impedance Myography in Rodents

This article details how to perform in vivo (using surface and needle electrode arrays) and ex vivo (using a dielectric cell) electrical impedance myography on the rodent gastrocnemius muscle. It will demonstrate the technique in both mice and rats and detail the modifications available, (i.e., obese animals, pups).

Esecuzione della miografia ad impedenza elettrica in vivo ed ex vivo nei roditori

Electrical impedance myography (EIM) is a convenient technique that can be used in preclinical and clinical studies to assess muscle tissue health and ...