Questo lavoro è stato supportato dal programma di scambio FY 2013 ricerca tra JSP e CNRS, JSP KAKENHI Grant numero 26860858 e la Narishige Neuroscience Research Foundation. Ringraziamo la d. ssa Mariko Harada-Shiba e Kyoko Shioya per utili discussioni. Ringraziamo Mari Furuta, Mutsumi Sakamoto, Ritsuko Maki e Dr. Emi Tanaka per l'eccellente assistenza tecnica.

Tutti gli esperimenti sono stati eseguiti secondo protocolli approvati dal comitato di uso della nazionale cerebrale e centro cardiovascolare, Suita, Giappone e cura degli animali sperimentali.
1. preparare i seguenti animali e materiali per la chirurgia MIUH
<ol><li>Preparare temporizzati ratti Sprague-Dawley incinti al giorno gestazionale 17, vale a dire, embrionale giorno 17 (E17). Medio peso corporei del dams sono ± 307.0 40,7 g (media ± DS, n = 9).</li>
	<li>Preparare i microcoils metallici; diametro interno 0,24 mm, in acciaio rivestito in oro.</li>
	<li>Preparare tutti i materiali secondo la tabella dei materiali.</li>
</ol>2. prepararsi all'intervento MIUH
<ol><li>Scaldare una piastra a 37 ° C per il posizionamento di ratto</li>
	<li>Collocare un pannolino sterile sul tappetino di riscaldamento</li>
	<li>Soluzione salina sterile calda a 37 ° C in un bagno d'acqua</li>
</ol>3. eseguire un intervento chirurgico MIUH
<ol><li>Eseguire le procedure chirurgiche in condizioni sterili. Sterilizzare le bobine, pinze, aghi con etanolo al 70% prima dell'uso. Utilizzare guanti sterili e occasionalmente disinfettare applicando spray di etanolo.</li>
	<li>Posizionare il ratto nella casella di induzione di anestesia. Introdurre 4% isoflurane nella casella (circa 5 min).</li>
	<li>Collocare il topo su un pannolino sterile sul tappetino di riscaldamento in posizione supina dopo che l'animale è anestetizzato accuratamente nella finestra di induzione. Verifica per la profondità dell'anestesia dalla mancanza di una risposta a un pizzico di punta. Mantenere isoflurane 1.5-2.0% con un cono di naso.</li>
	<li>Applicare schiuma depilatoria sul addome dall'ombelico verso la regione dell'arco pelvica. Dopo alcuni minuti, togliere la schiuma depilatoria con un tovagliolo di carta. Un rasoio può essere usato come un sostituto per la schiuma depilatoria.</li>
	<li>Applicare una garza imbevuta di soluzione disinfettante iodio nella regione Depilata della pelle.</li>
	<li>Posizionare un telo chirurgico con un'apertura circolare per coprire l'addome, tranne l'area dell'incisione chirurgica. Diversi pezzi di garza possono essere usati come un sostituto per il drappeggio.</li>
	<li>Usando un bisturi chirurgico, fare un'incisione di circa 2,5 cm lungo il basso addome dal bordo superiore del pube verso l'alto verso l'ombelico. Quindi, fare un'incisione attraverso lo strato muscolare sottostante.</li>
	<li>Posto diversi pezzi di garza sterile intorno all'apertura del drappo e bagnare la garza con soluzione salina riscaldata.</li>
	<li>Spingere delicatamente, entrambi i lati laterali dell'addome in modo che una parte del corno uterino emerge dall'incisione.</li>
	<li>Disinfettare le mani del chirurgo con etanolo al 70%.</li>
	<li>Tirare delicatamente l'intero corno uterino dalla cavità addominale manualmente senza usare il forcipe. Usando il forcipe chirurgico non è consigliabile, come possono facilmente danneggiare utero fragile, feti e vasi sanguigni comprimendo loro fortemente e in modo non uniforme.</li>
	<li>Posto il corno uterino su garza umida.</li>
	<li>Eseguire la stessa procedura per l'altro corno uterino.</li>
	<li>Contare il numero di feti.</li>
	<li>Applicare la soluzione salina riscaldata frequentemente per mantenere l'utero caldo e umido durante tutto l'intervento chirurgico.<ol>
<li>Cambiare il pannolino quando ha assorbito troppo salino. Per mantenere la stabilità, ripetere guanti di disinfezione con etanolo al 70% e punte dello strumento con iodio. Mantenere le dita non sterili lontano le punte sterili dello strumento è cruciale.</li>
	</ol>
</li>
	<li>Identificare la parte prossimale del portico principale delle arterie ovariche ed uterine sotto un microscopio stereoscopico. Nota: È fondamentale distinguere la galleria principale dai rami per le placente. Si noti che l'intervento chirurgico può essere più facile quando il corno uterino è capovolto. Occasionalmente, l'applicazione di una bobina alla parte molto prossimale del porticato è difficile, perché si trova in profondità all'interno della cavità addominale e le arterie alla placenta sono molto vicino alla parte prossimale del arcade. In questo caso, una bobina può essere applicata ad una parte della galleria dopo il prime forcelle di ramo fuori.</li>
	<li>Dividere l'arteria dalla vena che corre parallelo ad esso con una pinza e lo spazio tra di loro.</li>
	<li>Posizionare un pezzo di spago (sutura seta 4.0, 5 cm di lunghezza) sotto l'arteria.</li>
	<li>Sollevare in entrambe le estremità della stringa con una pinza in modo che l'arteria è sollevato e separato dalla vena fragile.</li>
	<li>Tenendo il bordo del microcoil con una pinzetta in altra mano, posizionare un microcoil accanto all'arteria sollevato (Figura 2.1).</li>
	<li>Ruotare l'arteria sollevato utilizzando la stringa intorno alla microcoil per avvolgere il microcoil intorno all'arteria. È più facile ruotare l'arteria intorno alla bobina rispetto al ruotare la bobina intorno all'arteria (Figura 2.2).</li>
	<li>Dopo aver ruotato l'arteria 3 o 4 giri (bobine hanno 5 giri), modificare il sito a cui la bobina è tenuta per l'altro bordo. Tenere premuto il bordo della bobina che è stato avvolto intorno all'arteria con il forcipe. Modificare il sito in cui la stringa solleva l'arteria verso l'altro lato della bobina. Ruotare l'arteria intorno alla bobina affinché la bobina completamente (5 turni) avvolge l'arteria (Figura 2,3, 2.4).</li>
	<li>Rimuovere la stringa.</li>
	<li>Eseguire la stessa procedura per le altre tre arterie.</li>
	<li>Nota che frequentemente l'applicazione riscaldato soluzione fisiologica nell'utero è fondamentale; in caso contrario, aumenta il tasso di mortalità dei feti e ipotermia potrebbe essere neuroprotective.</li>
	<li>Delicatamente e restituire il corno uterino nella cavità addominale. Sollevamento di un bordo dell'incisione della parete addominale con il forcipe può rendere più facile questa procedura.</li>
	<li>Stich il muscolo addominale e quindi stich la pelle dell'addome con sutura (sutura seta 4.0). Legare la sutura dopo ogni stich, cioè interrotte suture singole.</li>
	<li>Applicare una garza imbevuta di soluzione disinfettante iodio su e attorno all'incisione chirurgica.</li>
	<li>Somministrare per via sottocutanea il meloxicam analgesico a 0,5 mg/kg di peso corporeo.</li>
	<li>Consentire per 30 min di recupero in una gabbia riscaldata. Controllare gli animali se completamente sveglio e in movimento intorno alla gabbia, tornare alla gabbia a casa.</li>
</ol>

<ol>
	<li>Ohshima, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mild+intrauterine+hypoperfusion+reproduces+neurodevelopmental+disorders+observed+in+prematurity.">Mild intrauterine hypoperfusion reproduces neurodevelopmental disorders observed in prematurity.</a> Sci Rep. 6, 39377 (2016).</li><li>Anderson, P., Doyle, L. W., Victorian Infant Collaborative Study, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobehavioral+outcomes+of+school-age+children+born+extremely+low+birth+weight+or+very+preterm+in+the+1990s.">Neurobehavioral outcomes of school-age children born extremely low birth weight or very preterm in the 1990s.</a> JAMA. 289 (24), 3264-3272 (2003).</li><li>Levine, T. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+childhood+neurodevelopment+after+intrauterine+growth+restriction:+a+systematic+review.">Early childhood neurodevelopment after intrauterine growth restriction: a systematic review.</a> Pediatrics. 135 (1), 126-141 (2015).</li><li>Sucksdorff, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preterm+Birth+and+Poor+Fetal+Growth+as+Risk+Factors+of+Attention-Deficit/+Hyperactivity+Disorder.">Preterm Birth and Poor Fetal Growth as Risk Factors of Attention-Deficit/ Hyperactivity Disorder.</a> Pediatrics. 136 (3), e599-e608 (2015).</li><li>Volpe, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+injury+in+premature+infants:+a+complex+amalgam+of+destructive+and+developmental+disturbances.">Brain injury in premature infants: a complex amalgam of destructive and developmental disturbances.</a> Lancet Neurol. 8 (1), 110-124 (2009).</li><li>Nardozza, L. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+growth+restriction:+current+knowledge.">Fetal growth restriction: current knowledge.</a> Arch Gynecol Obstet. 295 (5), 1061-1077 (2017).</li><li>Chang, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preterm+birth+and+the+role+of+neuroprotection.">Preterm birth and the role of neuroprotection.</a> Bmj. 350, g6661 (2015).</li><li>Coq, J. O., Delcour, M., Massicotte, V. S., Baud, O., Barbe, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prenatal+ischemia+deteriorates+white+matter,+brain+organization,+and+function:+implications+for+prematurity+and+cerebral+palsy.">Prenatal ischemia deteriorates white matter, brain organization, and function: implications for prematurity and cerebral palsy.</a> Dev Med Child Neurol. 58, 7-11 (2016).</li><li>Jantzie, L. L., Corbett, C. J., Firl, D. J., Robinson, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postnatal+Erythropoietin+Mitigates+Impaired+Cerebral+Cortical+Development+Following+Subplate+Loss+from+Prenatal+Hypoxia-Ischemia.">Postnatal Erythropoietin Mitigates Impaired Cerebral Cortical Development Following Subplate Loss from Prenatal Hypoxia-Ischemia.</a> Cereb Cortex. 25 (9), 2683-2695 (2015).</li><li>Kubo, K. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+of+impaired+neuronal+migration+with+cognitive+deficits+in+extremely+preterm+infants.">Association of impaired neuronal migration with cognitive deficits in extremely preterm infants.</a> JCI Insight. 2 (10), (2017).</li><li>Delcour, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mild+musculoskeletal+and+locomotor+alterations+in+adult+rats+with+white+matter+injury+following+prenatal+ischemia.">Mild musculoskeletal and locomotor alterations in adult rats with white matter injury following prenatal ischemia.</a> Int J Dev Neurosci. 29 (6), 593-607 (2011).</li><li>Gilbert, J. S., Babcock, S. A., Granger, 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=Hypertension+produced+by+reduced+uterine+perfusion+in+pregnant+rats+is+associated+with+increased+soluble+fms-like+tyrosine+kinase-1+expression.">Hypertension produced by reduced uterine perfusion in pregnant rats is associated with increased soluble fms-like tyrosine kinase-1 expression.</a> Hypertension. 50 (6), 1142-1147 (2007).</li><li>Granger, J. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+uterine+perfusion+pressure+(RUPP)+model+for+studying+cardiovascular-renal+dysfunction+in+response+to+placental+ischemia.">Reduced uterine perfusion pressure (RUPP) model for studying cardiovascular-renal dysfunction in response to placental ischemia.</a> Methods Mol Med. 122, 383-392 (2006).</li><li>Mazur, M., Miller, R. H., Robinson, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postnatal+erythropoietin+treatment+mitigates+neural+cell+loss+after+systemic+prenatal+hypoxic-ischemic+injury.">Postnatal erythropoietin treatment mitigates neural cell loss after systemic prenatal hypoxic-ischemic injury.</a> J Neurosurg Pediatr. 6 (3), 206-221 (2010).</li><li>Olivier, P., Baud, O., Evrard, P., Gressens, P., Verney, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prenatal+ischemia+and+white+matter+damage+in+rats.">Prenatal ischemia and white matter damage in rats.</a> J Neuropathol Exp Neurol. 64 (11), 998-1006 (2005).</li><li>Robinson, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+changes+induced+by+graded+prenatal+systemic+hypoxic-ischemic+insults+in+rats.">Developmental changes induced by graded prenatal systemic hypoxic-ischemic insults in rats.</a> Neurobiol Dis. 18 (3), 568-581 (2005).</li><li>Rice, D., Barone, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+periods+of+vulnerability+for+the+developing+nervous+system:+evidence+from+humans+and+animal+models.">Critical periods of vulnerability for the developing nervous system: evidence from humans and animal models.</a> Environ Health Perspect. 108, 511-533 (2000).</li><li>Basilious, A., Yager, J., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurological+outcomes+of+animal+models+of+uterine+artery+ligation+and+relevance+to+human+intrauterine+growth+restriction:+a+systematic+review.">Neurological outcomes of animal models of uterine artery ligation and relevance to human intrauterine growth restriction: a systematic review.</a> Dev Med Child Neurol. 57 (5), 420-430 (2015).</li><li>Delcour, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroanatomical,+sensorimotor+and+cognitive+deficits+in+adult+rats+with+white+matter+injury+following+prenatal+ischemia.">Neuroanatomical, sensorimotor and cognitive deficits in adult rats with white matter injury following prenatal ischemia.</a> Brain Pathol. 22 (1), 1-16 (2012).</li></ol>

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Microcoil stenosi delle arterie sia ovariche che uterine in entrambi i corni uterini produce ipoperfusione intrauterina coerente e riproducibile in tutte le placente e feti. Il livello dell'ipoperfusione può essere modificato utilizzando i microcoils con diversi diametri interni. Cuccioli del ratto, nati da una diga su cui è stata eseguita la stenosi dell'arteria con i microcoils 0,24 mm di diametro interno dimostrano IUGR e parto prematuro (Vedi riferimento 1 per i dettagli). I cuccioli presentano alterazioni neuroana...

Gli infanti con restrizione della crescita intrauterina (IUGR) (noto anche come ritardo di crescita intrauterino) (peso nascita < 10th percentile per l'età gestazionale), nascita pretermine (Nato a < 37 settimane di gestazione), e/o bassa nascita conto peso (< 2500 g) per quasi il 10% di tutti i neonati 2,3. Molti di questi infanti presentano problemi neurologici quali paralisi cerebrale e disturbi dello sviluppo (ad es., attention-deficit/hyperactivity disorder (ADHD) e disturbi dell'apprendimento) 3,4,5. Tali condizioni sono somiglianze e differenze nella loro eziologia ed i risultati. L'eziologia di IUGR è multifattoriale e insufficienza placentare associato ipoperfusione intrauterina è considerato la causa più comune in feti non anomala 7. L'eziologia del pretermine è multifattoriale pure e chorioamnionitis è la più frequente causa di 8.
L'influenza di lieve ipoperfusione intrauterina (MIUH) su sviluppo del cervello è poco chiaro. Attualmente disponibili modelli animali di ipoperfusione/ischemia intrauterina comportano principalmente hypoperfusion severo o grado di pendenza dell'ipoperfusione con o senza riperfusione 9,10,11. In ambito clinico, tuttavia, casi di MIUH sono considerati di essere molto più frequenti rispetto a quelli che coinvolgono tali condizioni gravi. I modelli attualmente disponibili di MIUH sono un modello del roditore che coinvolgono la legatura del suturare dell'uterina o dell'arteria ovarica e un modello del roditore che coinvolge le arterie ovariche bilaterali e l'aorta avendo la pervietà 12,13, di ritaglio 14,15,16,17. Uno degli svantaggi di questi modelli è la notevole variabilità inter-feto, che vanno da feti con profonda ipoperfusione a feti con aspersione quasi intatto, a seconda della posizione del feto all'interno quadriloggiato arteriosa dell'uterina e arterie ovariche. Un altro svantaggio di questi modelli è la loro incapacità di distinguere la posizione di ogni feto dopo la nascita; quindi, i ricercatori non possono distinguere la gravità dell'ipoperfusione intrauterina sperimentato da un pup individuali dopo la sua nascita.
Abbiamo sviluppato un modello del ratto di MIUH che coinvolge più di stenosi dell'arteria 1. Avvolgimento micro-bobine di metallo con un diametro interno di 0,24 mm attorno le arterie ovariche e uterine causa stenosi, ma non ostruzione dei vasi sanguigni (Figura 1). Applicando questi microcoils presso le parti prossimali di tutti delle arterie che assicurano l'utero, vale a dire, le arterie uterine ed ovariche bilaterali, il giorno embrionale 17 (equivalente a embrionali settimane 20-25 in esseri umani 18) induce una diminuzione significativa ma mite flusso di sangue alla placenta e feti. Le diminuzioni nel flusso sanguigno dopo bobine vengono applicate a tutte le quattro arterie che alimentano l'utero sono in gran parte le stesse attraverso ogni placenta ed il feto. Il tasso di mortalità del feto è meno del 20%. I cuccioli sono nati via travaglio spontaneo 1-2 giorni precedenti (embrionale giorno 21-22) rispetto al normale. Quasi tutti i cuccioli sono nati espositrici nascita significativamente bassi pesi 1. Volumi di materia grigia e bianca sono diminuiti senza danni evidente tessuto 1. I cuccioli si presentano con ritardo nella acquisizione della neonato riflessi, debolezza muscolare e alterata attività spontanea 1. Questo modello imita i segni clinici ed i sintomi dei bambini nati prematuramente o con IUGR; espositivo prematuri nati bambini ridotto grigio e volume di materia bianca con o senza materia bianca lesioni 6, presenti le pietre miliari dello sviluppo neurologico in ritardo e può presentare problemi comportamentali come ADHD 3,5; i bambini con IUGR presentano alterazioni neuroanatomiche minime e hanno un rischio aumentato di sviluppo neurologico alterato ad esempio ritardo motorio e cognitivo 4,7. Nascita pretermine e IUGR sono condizioni diverse, ma le due condizioni condividono il meccanismo di base, cioè gli insulti ai cervelli immaturi prima età gestazionale pieno-termine.

<table>
	<tbody>
		<tr>
			<td>Stereomicroscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane anesthesia machine</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia induction box</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Diaper 30x40 cm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Depilatory foam or shaver</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine disinfectant solution</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze 10x20 cm&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical drape 45x45 cm with a round opening 5 cm in diameter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spray bottle with ethanol for disinfection</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swab</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps with large blunt tips</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps with angled fine tips</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scalpel, blade size is 27mm long (no.10, Axel, AS ONE Corporation, Osaka, Japan)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical suture needle</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Metal microcoils; inner diameter 0.24 mm, made from gold-coated steel (SAMINI Co. Ltd., Shizuoka, Japan)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Silk suture 4-0</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile saline (0.9% sodium chloride)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Heating water bath</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic syringes (50ml) and needles (18G)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

a rat model of mild intrauterine hypoperfusion with microcoil stenosis

Ipoperfusione/ischemia intrauterina è una delle principali cause di restrizione della crescita intrauterina/fetale, parto prematuro e basso peso alla nascita. Maggior parte degli studi di questo fenomeno sono stata effettuata in entrambi modelli con ischemia intrauterina severa o con grado di pendenza dell'ipoperfusione intrauterina. Nessuno studio è stato condotto in un modello su uniforme lieve ipoperfusione intrauterina (MIUH). Due modelli sono stati utilizzati per gli studi di MIUH: un modello basato sulla legatura del suturare di entrambi i lati del porticato arterioso formata con le arterie uterine ed ovariche, e un modello transitorio basato sul ritaglio l'arterie ovariche bilaterali e l'aorta con l'evidenza. Quei due modelli del roditore di MIUH hanno alcune limitazioni, ad esempio, non tutti i feti sono sottoposti a MIUH, a seconda della loro posizione nel corno uterino. Nel nostro modello MIUH, tutti i feti sono sottoposti ad un livello paragonabile di ipoperfusione intrauterina. MIUH è stato realizzato da stenosi delicata di tutte le quattro arterie d'alimentazione dell'utero, cioè, arterie uterine ed ovariche bilaterali.
Stenosi arteriosa è stata indotta da metallo microcoils avvolto intorno le arterie d'alimentazione. Produzione di stenosi arteriosa con i microcoils ci ha permessi di controllare, ottimizzare e riprodurre la diminuzione del flusso sanguigno con pochissima variabilità e un tasso di mortalità basso, consentendo in tal modo una valutazione accurata. Quando si usavano i microcoils con un diametro interno di 0,24 mm, il flusso di sangue nella placenta e feto era leggermente diminuita (circa 30% dal livello pre-stenosi nella placenta). La prole del nostro modello MIUH dimostra chiaramente di lunga durata alterazioni nei risultati del test neurologici, neuroanatomical e comportamentali.

Dopo aver applicato i microcoils a tutti delle arterie che alimentano l'utero, vale a dire, le arterie uterine ed ovariche bilaterali, tutti i feti sono sottoposti a livelli comparabili di ipoperfusione. L'applicazione di microcoils con un diametro interno di 0,24 mm provoca lieve stenosi di quelle arterie, causando così una lieve diminuzione nel flusso di sangue alla placenta e feti (Figura 3; circa il 30% dal livello pre-stenosi nella placenta, vedere rife...

Watch this Scientific Journal Video about A Rat Model of Mild Intrauterine Hypoperfusion with Microcoil Stenosis at JoVE.com

A Rat Model of Mild Intrauterine Hypoperfusion with Microcoil Stenosis

Lieve ipoperfusione intrauterino è stato prodotto da stenosi dell'arteria con i microcoils metallici avvolti intorno le arterie uterine ed ovariche in ratti al giorno embrionale 17. Questa procedura ha prodotto prenatale ipoperfusione e restrizione della crescita intrauterina.

Un modello del ratto dell'ipoperfusione lieve intrauterina con stenosi Microcoil

Intrauterine hypoperfusion/ischemia is one of the major causes of intrauterine/fetal growth restriction, preterm birth, and low birth weight. Most studies ...

a-rat-model-of-mild-intrauterine-hypoperfusion-with-microcoil-stenosis

Research

JoVE Journal

Neuroscience

10.2K Views.  National Cerebral and Cardiovascular Center. Lieve ipoperfusione intrauterino è stato prodotto da stenosi dell'arteria con i microcoils metallici avvolti intorno le arterie uterine ed ovariche in ratti al giorno embrionale 17. Questa procedura ha prodotto prenatale ipoperfusione e restrizione della crescita intrauterina.

Video: A Rat Model of Mild Intrauterine Hypoperfusion with Microcoil Stenosis

Investigations on Alterations of Hippocampal Circuit Function Following Mild Traumatic Brain Injury

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological impairments 1. Two pervasive and disabling symptoms experienced by TBI survivors, memory deficits and a reduction in seizure threshold, are thought to be mediated by TBI-induced hippocampal dysfunction 2,3. In order to demonstrate how altered hippocampal circuit function adversely affects behavior after TBI in mice, we employ lateral fluid percussion injury, a commonly used animal model of TBI that recreates many features of human TBI including neuronal cell loss, gliosis, and ionic perturbation 4-6.
	Here we demonstrate a combinatorial method for investigating TBI-induced hippocampal dysfunction. Our approach incorporates multiple ex vivo physiological techniques together with animal behavior and biochemical analysis, in order to analyze post-TBI changes in the hippocampus. We begin with the experimental injury paradigm along with behavioral analysis to assess cognitive disability following TBI. Next, we feature three distinct ex vivo recording techniques: extracellular field potential recording, visualized whole-cell patch-clamping, and voltage sensitive dye recording. Finally, we demonstrate a method for regionally dissecting subregions of the hippocampus that can be useful for detailed analysis of neurochemical and metabolic alterations post-TBI.
	These methods have been used to examine the alterations in hippocampal circuitry following TBI and to probe the opposing changes in network circuit function that occur in the dentate gyrus and CA1 subregions of the hippocampus (see Figure 1). The ability to analyze the post-TBI changes in each subregion is essential to understanding the underlying mechanisms contributing to TBI-induced behavioral and cognitive deficits. 
	The multi-faceted system outlined here allows investigators to push past characterization of phenomenology induced by a disease state (in this case TBI) and determine the mechanisms responsible for the observed pathology associated with TBI.

A multi-faceted approach to investigating functional changes to hippocampal circuitry is explained. Electrophysiological techniques are described along with the injury protocol, behavioral testing and regional dissection method. The combination of these techniques can be applied in similar fashion for other brain regions and scientific questions.

Indagini su Alterazioni della funzione del circuito ippocampale seguito lieve trauma cranico

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological ...

Ricerca

Neuroscienze

A Protocol for Measuring Cue Reactivity in a Rat Model of Cocaine Use Disorder

Cocaine use disorder (CUD) follows a trajectory of repetitive self-administration during which previously neutral stimuli gain incentive value. Cue reactivity, the sensitivity to cues previously linked with the drug-taking experience, plays a prominent role in human craving during abstinence. Cue reactivity can be assessed as the attentional orientation toward drug-associated cues that is measurable as appetitive approach behavior in both preclinical and human studies. Herein describes an assessment of cue reactivity in rats trained to self-administer cocaine. Cocaine self-administration is paired with the presentation of discrete cues that act as conditioned reinforcers (i.e., house light, stimulus light, infusion pump sounds). Following a period of abstinence, lever presses in the cocaine self-administration context accompanied by the discrete cues previously paired with cocaine infusion are measured as cue reactivity. This model is useful to explore neurobiological mechanisms underlying cue reactivity processes as well as to assess pharmacotherapies to suppress cue reactivity and therefore, modify relapse vulnerability. Advantages of the model include its translational relevance, and its face and predictive validities.&#160;The primary limitation of the model is that the cue reactivity task can only be performed infrequently and must only be used in short duration (e.g., 1 hour), otherwise rats will begin to extinguish the pairing of the discrete cues with the cocaine stimulus. The model is extendable to any positively reinforcing stimulus paired with discrete cues; though particularly applicable to drugs of abuse, this model may hold future applications in fields such as obesity, where palatable food rewards can act as positively reinforcing stimuli.

Cue reactivity is conceptualized as sensitivity to cues linked with drug-taking experiences that contribute to craving and relapse in abstinent humans. Cue reactivity is modeled in rats by measuring attentional orientation toward drug-associated cues that results in appetitive approach behavior in a cue reactivity test following self-administration and forced abstinence.

Un protocollo per la misurazione Cue reattività in un modello del ratto di disturbo da uso di cocaina

Cocaine use disorder (CUD) follows a trajectory of repetitive self-administration during which previously neutral stimuli gain incentive value. Cue ...

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed as a water tank with narrow platforms on the bottom. The model rats were put into the tank and stood on the platforms for 14 h (18:00 - 8:00) per day for a consecutive 21 days, with a blank control group set for contrast. At the end of modeling, rats in the model group showed an obvious fatigued appearance. To assess the model, we performed several behavioral tests, including the open field test (OFT), the elevated plus maze (EPM) test, and the exhaustive swimming (ES) test. The results showed that anxiety, spatial cognition impairment, poor muscle performance, and declined voluntary activity presented in model rats confirm the diagnosis of&#160;central fatigue. Changes of the central neurotransmitters also verified the result. In conclusion, the model successfully simulated central fatigue, and future study with the model may help reveal the pathological mechanism of the disease.

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

Un ratto di modello di affaticamento centrale utilizzando una versione modificata più piattaforma metodo

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed ...

A Novel and Translational Rat Model of Concussion Combining Force and Rotation with In Vivo Cerebral Microdialysis

Persistent cognitive and motor symptoms are known consequences of concussions/mild traumatic brain injury (mTBIs) that can be partly attributable to altered neurotransmission. Indeed, cerebral microdialysis studies in rodents have demonstrated an excessive extracellular glutamate release in the hippocampus within the first 10 min following trauma. Microdialysis offers the clear advantage of in vivo neurotransmitter continuous sampling while not having to sacrifice the animal. In addition to the aforementioned technique, a closed head injury model that exerts rapid acceleration and deceleration of the head and torso is needed, as such a factor is not available in many other animal models. The Wayne State weight-drop model mimics this essential component of human craniocerebral trauma, allowing the induction of an impact on the head of an unrestrained rodent with a falling weight. Our novel and translational rat model combines cerebral microdialysis with the Wayne State weight-drop model to study, in lightly anesthetized and unrestrained adult rats, the acute changes in extracellular neurotransmitter levels following concussion. In this protocol, the microdialysis probe was inserted inside the hippocampus as region of interest, and was left inserted in the brain at impact. There is a high density of terminals and receptors in the hippocampus, making it a relevant region to document altered neurotransmission following concussion. When applied to adult Sprague-Dawley rats, our combined model induced increases in hippocampal extracellular glutamate concentrations within the first 10 min, consistent with the previously reported post-concussion symptomology. This combined weight-drop model provides a reliable tool for researchers to study early therapeutic responses to concussions in addition to repetitive brain injury, since this protocol induces a closed-head mild trauma.

Neurotransmitter alteration is a mechanism of neural dysfunction that occurs after concussion and contributes to the sometimes-catastrophic long-term consequences. This rat model combines microdialysis, allowing in vivo neurotransmitter quantification, with a weight-drop technique exerting rapid acceleration and deceleration of the head and torso, an important factor of human craniocerebral trauma.

Un nuovo e traslazionale modello di commozione cerebrale che combina forza e rotazione con la microdialisi cerebrale in vivo

Persistent cognitive and motor symptoms are known consequences of concussions/mild traumatic brain injury (mTBIs) that can be partly attributable to ...

Advanced Diffusion Imaging in The Hippocampus of Rats with Mild Traumatic Brain Injury

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous variability and heterogeneity (age, gender, type of trauma, other possible pathologies, etc.), animal models play a key role in unraveling factors that are limitations in clinical research. They provide a standardized and controlled setting to investigate the biological mechanisms of injury and repair following TBI. However, not all animal models mimic the diffuse and subtle nature of mTBI effectively. For example, the commonly used controlled cortical impact (CCI) and lateral fluid percussion injury (LFPI) models make use of a craniotomy to expose the brain and induce widespread focal trauma, which are not commonly seen in mTBI. Therefore, these experimental models are not valid to mimic mTBI. Thus, an appropriate model should be used to investigate mTBI. The Marmarou weight drop model for rats induces similar microstructural alterations and cognitive impairments as seen in patients who sustain mild trauma; therefore, this model was selected for this protocol. Conventional computed tomography and magnetic resonance imaging (MRI) scans commonly show no damage following a mild injury, because mTBI induces often only subtle and diffuse injuries. With diffusion weighted MRI, it is possible to investigate microstructural properties of brain tissue, which can provide more insight into the microscopic alterations following mild trauma. Therefore, the goal of this study is to obtain quantitative information of a selected region-of-interest (i.e., hippocampus) to follow up disease progression after obtaining a mild and diffuse brain injury.

The overall goal of this procedure is to obtain quantitative microstructural information of the hippocampus in a rat with mild traumatic brain injury. This is done using an advanced diffusion-weighted magnetic resonance imaging protocol and region-of-interest based analysis of parametric diffusion maps.

Advanced Diffusion Imaging nell'ippocampo di ratti con lievi lesioni cerebrali traumatiche

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous ...

Applying the RatWalker System for Gait Analysis in a Genetic Rat Model of Parkinson's Disease

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by the loss of dopaminergic (DA) neurons in the substantia nigra pars compacta. Gait abnormalities, including decreased arm swing, slower walking speed, and shorter steps are common in PD patients and appear early in the course of disease. Thus, the quantification of motor patterns in animal models of PD will be important for phenotypic characterization during disease course and upon therapeutic treatment. Most cases of PD are idiopathic; however, the identification of hereditary forms of PD uncovered gene mutations and variants, such as loss-of-function mutations in Pink1 and Parkin, two proteins involved in mitochondrial quality control that could be harnessed to create animal models. While mice are resistant to neurodegeneration upon loss of Pink1 and Parkin (single and combined deletion), in rats, Pink1 but not Parkin deficiency leads to nigral DA neuron loss and motor impairment. Here, we report the utility of FTIR imaging to uncover gait changes in freely walking young (2 months of age) male rats with combined loss of Pink1 and Parkin prior to the development of gross visually apparent motor abnormality as these rats age (observed at 4-6 months), characterized by hindlimb dragging as previously reported in Pink1 knockout (KO) rats.

Here we describe the RatWalker system, built by redesigning the MouseWalker apparatus to accommodate for the increased size and weight of rats. This system uses frustrated total internal reflection (FTIR), high-speed video capture, and open-access analysis software to track and quantify gait parameters.

Applicazione del sistema RatWalker per l'analisi dell'andatura in un modello genetico di ratto della malattia di Parkinson

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by the loss of dopaminergic (DA) neurons in the substantia nigra pars ...

Establishment of a Rat Model of Superior Sagittal-Sinus Occlusion via a Thread-Embolism Method

The mechanisms contributing to the natural onset of cerebral venous sinus thrombosis (CVST) are mostly unknown, and a variety of uncontrollable factors are involved in the course of the disease, resulting in great limitations in clinical research. Therefore, the establishment of stable CVST animal models that can standardize a variety of uncontrollable confounding factors have helped to circumvent shortcomings in clinical research. In recent decades, a variety of CVST animal models have been constructed, but the results based on these models have been inconsistent and incomplete. Hence, in order to further explore the pathophysiological mechanisms of CVST, it is necessary to establish a novel and highly compatible animal model, which has important practical value and scientific significance for the diagnosis and treatment of CVST. In the present study, a novel Sprague-Dawley (SD) rat model of superior sagittal sinus (SSS) thrombosis was established via a thread-embolization method, and the stability and reliability of the model were verified. Additionally, we evaluated changes in cerebral venous blood flow in rats after the formation of CVST. Collectively, the SD-rat SSS-thrombosis model represents a novel CVST animal model that is easily established, minimizes trauma, yields good stability, and allows for accurately controlling ischemic timing and location.

Here, we establish a novel Sprague-Dawley (SD) rat model of superior sagittal sinus (SSS) thrombosis via a thread-embolization method, and the stability and reliability of the model were verified.

Istituzione di un modello di ratto di occlusione sagittale-seno superiore tramite un metodo thread-embolismo

The mechanisms contributing to the natural onset of cerebral venous sinus thrombosis (CVST) are mostly unknown, and a variety of uncontrollable factors ...

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

Un modello di ratto di infezione cerebrale EcoHIV

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements known as L-DOPA-induced dyskinesias (LIDs) develop with prolonged use of this dopamine precursor.&#160;It is estimated that the incidence of LIDs escalates to approximately 90% of individuals with PD within 10&#8211;15 years of treatment. Understanding the mechanisms of this malady and developing both novel and effective anti-dyskinesia treatments requires consistent and accurate modeling for pre-clinical testing of therapeutic interventions. A detailed method for reliable induction and comprehensive rating of LIDs following 6-OHDA-induced nigral lesioning in a rat model of PD is presented here. Dependable LID assessment in rats provides a powerful tool that can be readily utilized across laboratories to test emerging therapies focused on reducing or eliminating this common treatment-induced burden for individuals with PD.

This article describes methods to induce and evaluate levodopa-induced dyskinesias in a rat model of Parkinson&#39;s disease. The protocol offers detailed information regarding the intensity and frequency of a range of dyskinetic behaviors, both dystonic and hyperkinetic, providing a reliable tool to test treatments targeting this unmet medical need.

Induzione e valutazione delle discinesie indotte da levodopa in un modello di ratto della malattia di Parkinson

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements ...

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and understanding the mechanisms of how a TBI alters brain function. The availability of multiple animal models of TBI is necessary to model the different aspects and severities of TBI seen in people. This manuscript describes the use of a midline closed head injury (CHI) to develop a mouse model of mild TBI. The model is considered mild because it does not produce structural brain lesions based on neuroimaging or gross neuronal loss. However, a single impact creates enough pathology that cognitive impairment is measurable at least 1 month after injury. A step-by-step protocol to induce a CHI in mice using a stereotaxically guided electromagnetic impactor is defined in the paper. The benefits of the mild midline CHI model include the reproducibility of the injury-induced changes with low mortality. The model has been temporally characterized up to 1 year after the injury for neuroimaging, neurochemical, neuropathological, and behavioral changes. The model is complementary to open skull models of controlled cortical impact using the same impactor device. Thus, labs can model both mild diffuse TBI and focal moderate-to-severe TBI with the same impactor.

The protocol describes mild traumatic brain injury in a mouse model. In particular, a step-by-step protocol to induce a mild midline closed head injury and the characterization of the animal model is fully explained.

Modello elettromagnetico controllato a testa chiusa di lieve lesione cerebrale traumatica nei topi

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...