Questo lavoro è finanziato dall &#39;Organizzazione olandese per la ricerca scientifica (NWO) (no Veni. 916.86.125), assegnato a L. Reneman. Il finanziatore ha avuto alcun ruolo nel disegno dello studio, la raccolta e analisi dei dati, la decisione di pubblicare, o la preparazione del manoscritto. Non ci sono conflitti di interesse.

<ol>
	<li>Abramoff, M. D., Magelhaes, P. J., Ram, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image+Processing+with+ImageJ.">Image Processing with ImageJ.</a> Biophotonics International. 11, 36-42 (2004).</li><li>Anderson, I. M., McKie, S., Elliott, R., Williams, S. R., Deakin, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+human+5-HT+function+in+vivo+with+pharmacoMRI.">Assessing human 5-HT function in vivo with pharmacoMRI.</a> Neuropharmacology. 55, 1029-1037 (2008).</li><li>Choi, J. K., Chen, Y. I., Hamel, E., Jenkins, 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=Brain+hemodynamic+changes+mediated+by+dopamine+receptors:+Role+of+the+cerebral+microvasculature+in+dopamine-mediated+neurovascular+coupling.">Brain hemodynamic changes mediated by dopamine receptors: Role of the cerebral microvasculature in dopamine-mediated neurovascular coupling.</a> Neuroimage. 30, 700-712 (2006).</li><li>Cohen, Z., Bonvento, G., Lacombe, P., Hamel, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serotonin+in+the+regulation+of+brain+microcirculation.">Serotonin in the regulation of brain microcirculation.</a> Prog. Neurobiol. 50, 335-362 (1996).</li><li>Ferrari, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+robust+experimental+protocol+for+pharmacological+fMRI+in+rats+and+mice.">A robust experimental protocol for pharmacological fMRI in rats and mice.</a> J. Neurosci. Methods. 204, 9-18 (2011).</li><li>Gozzi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+effects+of+antipsychotic+and+glutamatergic+agents+on+the+phMRI+response+to+phencyclidine.">Differential effects of antipsychotic and glutamatergic agents on the phMRI response to phencyclidine.</a> Neuropsychopharmacology. 33, 1690-1703 (2008).</li><li>Houston, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+of+brain+activation+in+response+to+pharmacological+agents+using+fMRI+in+the+rat.">Mapping of brain activation in response to pharmacological agents using fMRI in the rat.</a> Magn Reson. Imaging. 19, 905-919 (2001).</li><li>Jenkins, B. G., Sanchez-Pernaute, R., Brownell, A. L., Chen, Y. C., Isacson, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+dopamine+function+in+primates+using+pharmacologic+magnetic+resonance+imaging.">Mapping dopamine function in primates using pharmacologic magnetic resonance imaging.</a> J. Neurosci. 24, 9553-9560 (2004).</li><li>Klomp, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lasting+effects+of+chronic+fluoxetine+treatment+on+the+late+developing+rat+brain:+age-dependent+changes+in+the+serotonergic+neurotransmitter+system+assessed+by+pharmacological+MRI.">Lasting effects of chronic fluoxetine treatment on the late developing rat brain: age-dependent changes in the serotonergic neurotransmitter system assessed by pharmacological MRI.</a> Neuroimage. 59, 218-226 (2012).</li><li>Logothetis, N. K., Pauls, J., Augath, M., Trinath, T., Oeltermann, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurophysiological+investigation+of+the+basis+of+the+fMRI+signal.">Neurophysiological investigation of the basis of the fMRI signal.</a> Nature. 412, 150-157 (2001).</li><li>Martin, C., Sibson, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+MRI+in+animal+models:+A+useful+tool+for+5-HT+research.">Pharmacological MRI in animal models: A useful tool for 5-HT research.</a> Neuropharmacology. 55, 1038-1047 (2008).</li><li>McKie, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+effects+of+acute+citalopram+detected+by+pharmacoMRI.">Neuronal effects of acute citalopram detected by pharmacoMRI.</a> Psychopharmacology (Berl. 180, 680-686 (2005).</li><li>Preece, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+that+increased+5-HT+release+evokes+region-specific+effects+on+blood-oxygenation+level-dependent+functional+magnetic+resonance+imaging+responses+in+the+rat+brain.">Evidence that increased 5-HT release evokes region-specific effects on blood-oxygenation level-dependent functional magnetic resonance imaging responses in the rat brain.</a> Neuroscience. 159, 751-759 (2009).</li><li>Schwarz, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+stereotaxic+MRI+template+set+for+the+rat+brain+with+tissue+class+distribution+maps+and+co-registered+anatomical+atlas:+application+to+pharmacological+MRI.">A stereotaxic MRI template set for the rat brain with tissue class distribution maps and co-registered anatomical atlas: application to pharmacological MRI.</a> Neuroimage. 32, 538-550 (2006).</li><li>Schwarz, A. J., Gozzi, A., Reese, T., Bifone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+mapping+of+functional+connectivity+in+neurotransmitter+systems+using+pharmacological+MRI.">In vivo mapping of functional connectivity in neurotransmitter systems using pharmacological MRI.</a> NeuroImage. 34, 1627-1636 (2007).</li><li>Sekar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroadaptive+responses+to+citalopram+in+rats+using+pharmacological+magnetic+resonance+imaging.">Neuroadaptive responses to citalopram in rats using pharmacological magnetic resonance imaging.</a> Psychopharmacology (Berl). 213, 521-531 (2011).</li><li>Smith, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+functional+and+structural+MR+image+analysis+and+implementation+as+FSL.">Advances in functional and structural MR image analysis and implementation as FSL.</a> NeuroImage. 23, S208-S219 (2004).</li><li>Stark, J. A., McKie, S., Davies, K. E., Williams, S. R., Luckman, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=5-HT(2C)+antagonism+blocks+blood+oxygen+level-dependent+pharmacological-challenge+magnetic+resonance+imaging+signal+in+rat+brain+areas+related+to+feeding.">5-HT(2C) antagonism blocks blood oxygen level-dependent pharmacological-challenge magnetic resonance imaging signal in rat brain areas related to feeding.</a> Eur. J. Neurosci. 27, 457-465 (2008).</li><li>Stark, J. A., Davies, K. E., Williams, S. R., Luckman, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+magnetic+resonance+imaging+and+c-Fos+mapping+in+rats+following+an+anorectic+dose+of+m-chlorophenylpiperazine.">Functional magnetic resonance imaging and c-Fos mapping in rats following an anorectic dose of m-chlorophenylpiperazine.</a> NeuroImage. 31, 1228-1237 (2006).</li><li>Steward, C. A., Marsden, C. A., Prior, M. J., Morris, P. G., Shah, Y. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methodological+considerations+in+rat+brain+BOLD+contrast+pharmacological+MRI.">Methodological considerations in rat brain BOLD contrast pharmacological MRI.</a> Psychopharmacology (Berl). 180, 687-704 (2005).</li><li>Strupp, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulate:+A+GUI+based+fMRI+Analysis+Software+Package.">Stimulate: A GUI based fMRI Analysis Software Package.</a> NeuroImage. 3, S607 (1996).</li><li>Tomasi, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methylphenidate+enhances+brain+activation+and+deactivation+responses+to+visual+attention+and+working+memory+tasks+in+healthy+controls.">Methylphenidate enhances brain activation and deactivation responses to visual attention and working memory tasks in healthy controls.</a> Neuroimage. 54, 3101-3110 (2011).</li><li>Woolrich, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bayesian+analysis+of+neuroimaging+data+in+FSL.">Bayesian analysis of neuroimaging data in FSL.</a> NeuroImage. 45, S173-S186 (2009).</li><li>Worsley, K. J., Jezzard, P., Matthews, P. M., Smith, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Statistical+analysis+of+activation+images.">Statistical analysis of activation images.</a> Functional MRI: An Introduction to Methods. , (2001).</li></ol>

Non abbiamo nulla da rivelare.

5-HT phMRI è uno strumento promettente per valutare la funzione dei neurotrasmettitori in animali in vivo. Visualizza la risposta del cervello ad un 5-HT sfida con la RM funzionale. MRI ha il grande vantaggio di avere una elevata risoluzione spaziale e non è necessario che l&#39;iniezione di mezzo di contrasto o di molecole radio-marcate evitando così l&#39;esposizione ripetuta alle radiazioni ionizzanti. Questa tecnica è applicabile sia in soggetti umani e animali, e quindi molto adatto per la ricerca traslazionale dei sistemi neurotrasmettitoriali e disturbi psichiatrici. La sua applicazione non è ovviamente limitato al 5-HT percorso ed è già stata ampiamente utilizzata per valutare gli effetti di farmaci dopaminergici sia negli animali e nell&#39;uomo 5,15 22. Tuttavia, phMRI in piccoli animali rimane difficile, come già sottolineato in articoli di revisione da parte di Martin e Steward Sibson 11 e 20. Una di queste sfide è il mantenimento of stabili i parametri fisiologici durante l&#39;acquisizione dell&#39;immagine. La maggior parte degli anestetici può alterare la funzione cardiovascolare e dato che phMRI è criticamente dipendente sui parametri cardiovascolari / emodinamica è essenziale per garantire che i cambiamenti emodinamici sono esclusivamente attribuibili alla sfida dato farmaco. E &#39;quindi di vitale importanza che pCO 2 livelli rimangono costanti durante l&#39;acquisizione di base. Ventilazione meccanica può per garantire la stabilità fisiologica, ed è spesso usato in questo tipo di esperimenti. Abbiamo però scelto di usare free-respirazione animali lasciare aperta la possibilità di effettuare studi longitudinali in futuro. Invece, ampiamente monitorato (e alterata) frequenza respiratoria e valori dei gas ematici per garantire la stabilità fisiologica all&#39;interno dei normali intervalli prima di iniziare la scansione funzionale e in questo modo di mantenere stabile reattività vascolare e quindi T2 * / T2 segnale. La letteratura sugli effetti degli anestetici generali in materia di emodinamica cerebrale e metabolism è abbondante 20 e oltre la portata di questo manoscritto. Abbiamo scelto di usare l&#39;anestesia gassosa con isoflurano ± 2% in questo specifico protocollo, perché con anestetici inalatori, la profondità dell&#39;anestesia può essere rapidamente e facilmente controllabile. Questo è importante nella nostra configurazione per essere sicuri normali gamma stabili pCO 2 livelli prima dell&#39;inizio della acquisizione delle immagini. Isoflurano è il più comunemente usato inalante anestetico oggi e permette una rapida induzione e recupero, che è importante per gli studi longitudinali. Produce anche minima depressione cardiovascolare e respiratoria e induce buon rilassamento dei muscoli scheletrici. In secondo luogo, la somministrazione endovenosa del farmaco sfida è più complicata in piccoli animali che negli esseri umani. L&#39;intervento che è necessario per l&#39;incannulamento dell&#39;arteria femorale e la vena richiede personale ben addestrato ed esperto. A causa di queste procedure invasive è momentaneamente principalmente utilizzati nelle procedure terminali. Tuttavia, il monitoraggio non invasivo della omeostasi sangue e vena iniezione di coda potrebbero essere utilizzati per studi longitudinali 23. In aggiunta, ci sono alcune limitazioni di carattere più generale alla tecnica, che non sono specifici per animali phMRI. Inoltre, come sottolineato da Martin e Sibson 11, una confondere potenziale di tutti gli studi fMRI è che essa si assume che le variazioni dell&#39;attività cerebrale evocata dalla sfida riflettono i cambiamenti nell&#39;attività neuronale, piuttosto che periferici effetti sistemici. Soprattutto nelle strutture profonde del cervello, una comprensione relativamente povero di accoppiamento neurovascolare (rapporto tra i cambiamenti di attività neuronali e le variazioni emodinamiche) rimane. Studi di tipo eseguita da Logothetis 10 per determinare accoppiamento neurovascolare nella corteccia non sono ancora stati eseguiti in altre parti del cervello. È quindi noto che un aumento del segnale BOLD in strutture importanti come il corpo striato o amigdala è telling chi siamo l&#39;attività neuronale. Il meglio potremmo dire in questo momento è che la regione del cervello reagisce alla sfida dato e che a seconda del trattamento e / o le condizioni, siamo in grado di monitorare i cambiamenti significativi della reattività del cervello. Questo può essere verificato in gran parte, cercando in entrambi i dati MRI e le risposte fisiologiche. Lo schema generale di attivazione cervello dovrebbe essere regione specifica e con limitato alle superfici, in questo caso, un elevato 5-HT innervazione, e non tanto una risposta generale vascolare. Inoltre, un diverso profilo temporale tra cambiamenti vascolari ed emodinamici è previsto. Considerando che le variazioni della pressione del sangue ritorneranno ai valori basali pochi minuti, l&#39;effetto del farmaco sulla attivazione BOLD è nel caso di visibile fluoxetina fino alla fine della acquisizione dell&#39;immagine e corrispondono alle note proprietà farmacocinetiche di questo farmaco. Infine, le risposte fisiologiche di tutti gli animali dovrebbero essere simili al fine di rendere inter-soggetto confronti. Nonetheless, è noto che un regolamento neurogena del flusso ematico locale da 5-HT esistono 4. Pertanto non si può escludere che variazioni locali di segnale BOLD possono attribuire ai cambiamenti vascolari dovute al rilascio di 5-HT nella vicinanza di navi. Sebbene questi effetti non sono associati a locali attivazione neuronale e possono pertanto essere considerate come falsi risultati positivi, è anche un indice del funzionamento complessivo specifico di 5-HT sistema (vedi anche 3). Passi critici di questa tecnica sono quindi di monitorare risposte fisiologiche ampiamente e assicurarsi che le condizioni fisiologiche dell&#39;animale sono stabili prima e durante l&#39;acquisizione dell&#39;immagine. Anche le condizioni di scanner deve essere il più stabile possibile ed esattamente lo stesso per ogni animale. La stabilità del segnale della sequenza deve essere verificata e confermata prima dell&#39;inizio dell&#39;esperimento. Inoltre, assicurarsi di avere sempre grandi abbastanza potenza statistica, anche con Grou soggetto piccolops. Per una bella recensione sulle considerazioni sperimentali di animali phMRI in generale, vedere Steward 20 e per un ulteriore esempio di un protocollo sperimentale per fMRI farmacologica nel ratto e nel topo, vedere la Ferrari 5. Eventuali modifiche della tecnica qui descritti sono numerosi. Si potrebbe: <ol><li> utilizza un farmaco diverso per 5-HT sfida, come un altro SSRI o recettore 5-HT (ant) agonisti 16,13,18,7 o anche una doppia sfida, al fine di rivelare i meccanismi alla base di azione dei farmaci 6,19; </li><li> usare una diversa configurazione sperimentale, come ad esempio un diverso regime di anestesia, ventilazione meccanica, sangue piscina agenti di contrasto, invece di BOLD 15, studi longitudinali (animale ha bisogno di essere mantenuto in vita, quindi non invasiva della pressione sanguigna / sangue di misura gas e / o ventilazione meccanica sono possibili), o anche combinazioni con altri invasive) i metodi, quali la registrazione dell&#39;attività neuronale utilizzando MR-compatibile electrodes 10 o PET / SPECT studi 4; </li><li> utilizzare diversi metodi di analisi dei dati di risonanza magnetica come metodo &#39;p-block&#39; di McKie 12 o analisi di connettività funzionale 15. </li></ol> Quali scelte si fanno nel setup sperimentale è fortemente dipendente dalle possibilità di e / o l&#39;esperienza all&#39;interno del vostro laboratorio e il tipo di domanda di ricerca che si desidera rispondere.

<table border="1"><tbody><tr><td> Nome del reagente / apparecchiatura </td><td> Azienda </td><td> Numero di catalogo </td><td> Comments </td></tr><tr><td> Isoflurano </td><td> ABBOTT <a href="http://www.abbott.co.uk/" target="_blank">Abbott Laboratories Ltd., Maidenhead, UK</a> </td><td> No B506 </td><td> Mescolare con aria medicale </td></tr><tr><td> Aria medicale </td><td> <a href="http://www.bochealthcare.co.uk/" target="_blank">BOC Healthcare</a> </td><td></td><td></td></tr><tr><td> Riscaldamento pad </td><td> <a href="http://www.harvardapparatus.co.uk/" target="_blank">Harvard Apparatus</a> </td><td> 507223F <a href="http://www.harvardapparatus.co.uk/webapp/wcs/stores/servlet/haisku3_10001_11555_39108_-1_HAUK_ProductDetail_N_37610_37611_37613" target="_blank">Sistema completo Blanket omeotermi con sonda flessibile, Medium, 230 VAC, 50 Hz</a> </td><td></td></tr><tr><td> Silk l<html> igature </td><td> <a href="http://www.harvardapparatus.com/" target="_blank">http://www.harvardapparatus.com/</a> </td><td> <a href="http://www.harvardapparatus.com/webapp/wcs/stores/servlet/product_11051_10001_43051_-1_HAI_ProductDetail" target="_blank">2-0 di seta nera intrecciata non assorbibile</a> Num Cat 51-7631 </td><td></td></tr><tr><td> PE-50 Cannula </td><td> <a href="http://www.scientificlabs.co.uk/" target="_blank">http://www.scientificlabs.co.uk/~~V</a> </td><td> <a href="http://www.scientificlabs.co.uk/product/TUB3662" target="_blank">Portex tubi PE 0.58x0.96mm</a> </td><td> 0,58 ID 0,96 millimetri OD </td></tr><tr><td> Eparina sodica </td><td> <a href="http://www.leo-pharma.com/Home/Products/List-of-LEO-Pharma-products.aspx" target="_blank">Leo Laboratories Ltd, Bucks., UK</a> </td><td> Eparina di sodio 1000IU/ml </td><td> 15 U / ml </td></tr><tr><td> Tessuto adesivo Vetbond </td><td> <a href="http://solutions.3m.co.uk" target="_blank">3M</a> </td><td>ttp :/ / solutions.3m.co.uk/wps/portal/3M/en_GB/HealthCare/Home/ProdInfo/VetProducts / &quot;target =&quot; _blank &quot;&gt; M tessuto adesivo Vetbond </td><td></td></tr><tr><td> Sistema di controllo </td><td> <a href="http://www.i4sa.com/" target="_blank">SA Instruments</a> </td><td> <a href="http://www.i4sa.com" target="_blank">http://www.i4sa.com</a> <a href="http://69.20.42.253/web_app/main/defaultProduct.aspx?ID=16&amp;PT=3" target="_blank">Modello sistema di monitoraggio 1025L</a> </td><td> Monitor respirazione e la temperatura </td></tr><tr><td> Trasduttore di pressione </td><td> <a href="http://www.biopac.com/" target="_blank">BIOPAC Systems Corp</a> </td><td> <a href="http://www.biopac.com/blood-pressure-transducer" target="_blank">BLOOD TRASDUTTORE DI PRESSIONE - TSD104A</a> <a href="http://www.biopac.com/data-acquisition-analysis-system-mp150-system-windows" target="_blank">MP150 SISTEMA DI ACQUISIZIONE DATI - WIN - MP150WSW</a> </td><td> Monitor pressione arteriosa </td></tr><tr><td> RapidLab sangue analizzatore di gas </td><td> <a href="http://www.medical.siemens.com/" target="_blank">Siemens Diagnostics</a> </td><td> <a href="http://www.medical.siemens.com/webapp/wcs/stores/servlet/ProductDisplay~q_catalogId~e_-111~a_catTree~e_100001,1023069,1016115~a_langId~e_-111~a_productId~e_172940~a_storeId~e_10001.htm" target="_blank">RAPIDLab 248/348 Sistemi</a> </td><td></td></tr><tr><td> Animale scanner 4.7T </td><td> Agilent Technologies (precedentemente Magnex) 4.7T frequenza di 199,845 MHz </td><td></td><td></td></tr><tr><td> MR letto stereotassico compatibile </td><td> <a href="http://www.m2mimaging.com/products/preclinical.html" target="_blank">m2m Imaging Corp</a> </td><td> Rat letto: elemento PA Multi AHS 50-72-1003/100 </td><td></td></tr><tr><td> Bobina </td><td> <a href="http://www.m2mimaging.com/products/preclinical.html" target="_blank">m2m Imaging Corp</a> </td><td> Volume TH / Rx RQD1 72/112 200 </td><td></td></tr><tr><td> Fluoxetina cloridrato </td><td> Sigma-Aldrich </td><td> F-132 </td><td> 5mg/kg in soluzione salina </td></tr></tbody></table>

the use of pharmacological-challenge fmri in pre-clinical research: application to the 5-ht system

Farmacologico MRI (phMRI) è un metodo nuovo e promettente per studiare gli effetti delle sostanze sulla funzione del cervello che potrà poi essere usati per svelare sottostanti meccanismi neurobiologici alla base dell&#39;azione dei farmaci e neurotrasmettitori disturbi, quali depressione e ADHD. Come la maggior parte dei metodi di imaging (PET, SPECT, CT) rappresenta un progresso nello studio delle patologie cerebrali e la relativa funzione di percorsi neurotrasmettitori in modo non invasivo nel rispetto della connettività complessiva neuronale. Inoltre fornisce anche lo strumento ideale per la traduzione alle indagini cliniche. MRI, mentre ancora dietro in strategie di imaging molecolare rispetto al PET e SPECT, ha il grande vantaggio di avere una elevata risoluzione spaziale e senza necessità per l&#39;iniezione di un mezzo di contrasto o molecole radio-marcate, evitando l&#39;esposizione a radiazioni ionizzanti ripetitiva . Risonanza magnetica funzionale (fMRI) è ampiamente utilizzato in ambiente di ricerca e clinica, dove è generally combinato con un compito psico-motoria. phMRI è un adattamento del fMRI che consente lo studio di un sistema neurotrasmettitore specifico, come la serotonina (5-HT), in condizioni fisiologiche o patologiche dopo l&#39;attivazione tramite somministrazione di un farmaco specifico impegnativo. L&#39;obiettivo del metodo qui descritto è di valutare cervello 5-HT funzione in free-respirazione animali. Sfidando il 5-HT sistema, mentre allo stesso tempo l&#39;acquisizione di immagini di risonanza magnetica funzionale nel corso del tempo, la risposta del cervello a questa sfida può essere visualizzato. Diversi studi su animali hanno già dimostrato che il farmaco-indotto un aumento dei livelli extracellulari di esempio 5-HT (distaccanti, selettivi del re-uptake bloccanti, ecc) evocano regionali specifici cambiamenti nel livello di ossigenazione del sangue segnali MRI dipendenti (BOLD) (segnale dovuto ad una variazione dei livelli emoglobina ossigenata / deossigenata si verificano durante l&#39;attivazione del cervello attraverso un aumento dell&#39;apporto ematico per fornire l&#39;ossigeno e glucose ai neuroni esigenti) fornendo un indice di funzione neurotrasmettitore. È stato inoltre dimostrato che questi effetti possono essere invertito trattamenti che diminuzione 5-HT disponibilità 16,13,18,7. In ratti adulti, i cambiamenti segnale BOLD acuta dopo somministrazione degli SSRI sono stati descritti in diversi 5-HT regioni del cervello collegate, vale a dire le aree corticali, ippocampo, ipotalamo ed il talamo 9,16,15. La stimolazione del sistema 5-HT e la sua risposta a questo problema può essere quindi usato come una misura della sua funzione sia negli animali e nell&#39;uomo 2,11.

Watch this Scientific Journal Video about The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System at JoVE.com

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

L&#39;obiettivo di questa tecnica è quello di valutare serotonina (5-HT) in funzione di neurotrasmettitore l&#39;animale vivo e respiro libero con farmacologica risonanza magnetica (phMRI) e una sfida per via endovenosa con un inibitore selettivo della ricaptazione della serotonina (SSRI), fluoxetina.

L&#39;uso di Farmacologiche-sfida fMRI in ricerca pre-clinica: Applicazione del sistema 5-HT

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

the-use-pharmacological-challenge-fmri-pre-clinical-research

Research

JoVE Journal

Medicine

15.3K Views.  Academic Medical Center Amsterdam. L'obiettivo di questa tecnica è quello di valutare serotonina (5-HT) in funzione di neurotrasmettitore l'animale vivo e respiro libero con farmacologica risonanza magnetica (phMRI) e una sfida per via endovenosa con un inibitore selettivo della ricaptazione della serotonina (SSRI), fluoxetina.

Video: The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

Uso di un modello animale di sepsi per valutare nuove terapie a base di erbe

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Ricerca

Medicina

Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact. For this strategy to be successful, recent studies have shown that involvement of the host immune system is essential. Therefore, oncolytic virotherapy should be evaluated within the context of an immunocompetent model. Furthermore, the study of antitumor therapies in tolerized animal models may better recapitulate results seen in clinical trials. Cotton rats, commonly used to study respiratory viruses, are an attractive model to study oncolytic virotherapy as syngeneic models of mammary carcinoma and osteosarcoma are well established. However, there is a lack of published information on the proper handling procedure for these highly excitable rodents. The handling and capture approach outlined minimizes animal stress to facilitate experimentation. This technique hinges upon the ability of the researcher to keep calm during handling and perform procedures in a timely fashion. Finally, we describe how to prepare cotton rat mammary tumor cells for consistent subcutaneous tumor formation, and how to perform intratumoral and intraperitoneal injections. These methods can be applied to a wide range of studies furthering the development of the cotton rat as a relevant pre-clinical model to study antitumor therapy.

Cotton rats are extremely excitable and have a strong flight-or-fight response. A handling method optimized to reduce the stress of the animals is described which will make cotton rats more accessible as a preclinical model.

Manipolazione del Cotton Rat in studi per la valutazione pre-clinica di oncolitici virus

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact.  For this strategy to be ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

Pre-clinica ortotopico Modello murino di Human cancro alla prostata

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Use of a Low-flow Digital Anesthesia System for Mice and Rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

L&#39;utilizzo di un basso flusso di sistema per anestesia digitale per topi e ratti

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

Ultrahigh Resolution Mouse Optical Coherence Tomography to Aid Intraocular Injection in Retinal Gene Therapy Research

HR-SD-OCT is utilized to monitor the progression of photoreceptor degeneration in live mouse models, assess the delivery of therapeutic agents into the subretinal space, and to evaluate toxicity and efficacy in vivo. HR-SD-OCT uses near infrared light (800-880 nm) and has optics specifically designed for the unique optics of the mouse eye with sub-2-micron axial resolution. Transgenic mouse models of outer retinal (photoreceptor) degeneration and controls were imaged to assess the disease progression. Pulled glass microneedles were used to deliver sub retinal injections of adeno-associated virus (AAV) or nanoparticles (NP) via a trans-scleral and trans-choroidal approach. Careful positioning of the needle into the subretinal space was required prior to a calibrated pressure injection, which delivers fluid into the sub retinal space. Real time subretinal surgery was conducted on our retinal imaging system (RIS). HR-SD-OCT demonstrated progressive uniform retinal degeneration due to expression of a toxic mutant human mutant rhodopsin (P347S) (RHOP347S) transgene in mice. HR-SD-OCT allows rigorous quantification of all the retinal layers. Outer nuclear layer (ONL) thickness and photoreceptor outer segment length (OSL) measurements correlate with photoreceptor vitality, degeneration, or rescue. The RIS delivery system allows real-time visualization of subretinal injections in neonatal (~P10-14) or adult mice, and HR-SD-OCT immediately determines success of delivery and maps areal extent. HR-SD-OCT is a powerful tool that can evaluate the success of subretinal surgery in mice, in addition to measuring vitality of photoreceptors in vivo. HR-SD-OCT can also be used to identify uniform animal cohorts to evaluate the extent of retinal degeneration, toxicity, and therapeutic rescue in preclinical gene therapy research studies.

Here we demonstrate a novel approach to using high resolution spectral-domain optical coherence tomography (HR-SD-OCT) to assist delivery of gene therapy agents into the subretinal space, assess its areal coverage, and characterize photoreceptor vitality.

Tomografia ottica di coerenza di risoluzione ultraelevata Mouse per aiutare l'iniezione intraoculare nella ricerca retinica terapia genica

HR-SD-OCT is utilized to monitor the progression of photoreceptor degeneration in live mouse models, assess the delivery of therapeutic agents into the ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

Un modello pre-clinico di trapianto di cuore ortotopico

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Pre-clinical Model of Cardiac Donation after Circulatory Death

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after circulatory death (DCD) is a solution to address this limited availability, but due to a period of prolonged warm ischemia and the risk of tissue injury, its routine use in cardiac transplantation is seldom seen. In this manuscript we provide a detailed protocol closely mimicking current clinical practices in the context of DCD with continuous monitoring of heart function, allowing for the evaluation of novel cardioprotective strategies and interventions to decrease ischemia-reperfusion injury.
In this model, the DCD protocol is initiated in anesthetized Lewis rats by stopping ventilation to induce circulatory death. When systolic blood pressure drops below 30 mmHg, the warm ischemic time is initiated. After a pre-set warm ischemic period, hearts are flushed with a normothermic cardioplegic solution, procured, and mounted onto a Langendorff ex vivo heart perfusion system. Following 10 min of initial reperfusion and stabilization, cardiac reconditioning is continuously evaluated for 60 min using intraventricular pressure monitoring. A heart injury is assessed by measuring cardiac troponin T and the infarct size is quantified by histological staining. The warm ischemic time can be modulated and tailored to develop the desired amount of structural and functional damage. This simple protocol allows for the evaluation of different cardioprotective conditioning strategies introduced at the moment of cardioplegia, initial reperfusion and/or during ex vivo perfusion. Findings obtained from this protocol can be reproduced in large models, facilitating clinical translation.

This protocol shows a simple and flexible approach for the evaluation of new conditioning agents or strategies to increase the feasibility of cardiac donation after circulatory death.

Modello preclinico di donazione cardiaca dopo la morte circolatoria

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

Valutazione emodinamica invasiva per il sistema ventricolare destro e l'ipertensione arteriosa polmonare indotta dall'ipossia nei topi

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

Un modello di ratto preclinico per lo studio delle lesioni ischemia-reperfusione in microchirurgia ricostruttiva

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare recommendations, animals are warmed and monitored during procedures requiring anesthesia. Common anesthesia and physiological monitoring equipment include gas tanks, anesthetic vaporizers and stands, warming controllers and pads, mechanical ventilators, and pulse oximeters. A computer is also necessary for data collection and to run equipment software. In smaller spaces or when performing field work, it can be challenging to configure all this equipment in limited space.
The goal of this protocol is to demonstrate best practices for use of a low-flow digital vaporizer using both compressed oxygen and room air, along with an integrated mechanical ventilator, pulse oximeter, and far infrared warming as an all-inclusive anesthesia and physiological monitoring suite ideal for rodents.

Here, we present a protocol to safely and effectively administer anesthetic gas to mice using a digital, low flow anesthesia system with integrated ventilator and physiological monitoring modules.

Utilizzo di un vaporizzatore anestetico integrato a basso flusso, ventilatore e sistema di monitoraggio fisiologico per roditori

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare ...

L'uso di Farmacologiche-sfida fMRI in ricerca pre-clinica: Applicazione del sistema 5-HT

Riepilogo

Esplora Altri Video

Capitoli in questo video