We would like to thank everyone at Shriners Hospital Pediatric Research Center, in particular Dr. Mickey Seltzer, of whom without his support, this work would not have been funded. In addition, we would like to thank Isha Srivastava, who contributed to early data collection and Amy He, who helped with the figures. This study was funded by Shriners Hospitals for Children. No funding source played a role in experimental design or decision to submit the paper for publication.

<ol>
	<li>Nakamura, M., Bregman, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+neurotrophic+factor+gene+expression+profiles+between+neonate+and+adult+rat+spinal+cord+after+injury.">Differences in neurotrophic factor gene expression profiles between neonate and adult rat spinal cord after injury.</a> Exp Neurol. 169 (2), 407-415 (2001).</li><li>Widenfalk, J., Lundstr&#246;mer, K., Jubran, M., Brene, S., Olson, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurotrophic+factors+and+receptors+in+the+immature+and+adult+spinal+cord+after+mechanical+injury+or+kainic+acid.">Neurotrophic factors and receptors in the immature and adult spinal cord after mechanical injury or kainic acid.</a> J Neurosci. 21 (10), 3457-3475 (2001).</li><li>Fern&#225;ndez-L&#242;pez, D., Faustino, J., 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=Blood-brain+barrier+permeability+is+increased+after+acute+adult+stroke+but+not+neonatal+stroke+in+the+rat.">Blood-brain barrier permeability is increased after acute adult stroke but not neonatal stroke in the rat.</a> J Neurosci. 32 (28), 9588-9600 (2012).</li><li>Cusick, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extensive+cortical+reorganization+following+sciatic+nerve+injury+in+adult+rats+versus+restricted+reorganization+after+neonatal+injury:+implications+for+spatial+and+temporal+limits+on+somatosensory+plasticity.">Extensive cortical reorganization following sciatic nerve injury in adult rats versus restricted reorganization after neonatal injury: implications for spatial and temporal limits on somatosensory plasticity.</a> Prog Brain Res. 108, 379-390 (1996).</li><li>Barrett, C. P., Donati, E. J., Guth, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+between+adult+and+neonatal+rats+in+their+astroglial+response+to+spinal+injury.">Differences between adult and neonatal rats in their astroglial response to spinal injury.</a> Exp Neurol. 84 (2), 374-385 (1984).</li><li>Villapol, S., Gelot, A., Renolleau, S., Charriaut-Marlangue, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Astrocyte+Responses+after+Neonatal+Ischemia:+The+Yin+and+the+Yang.">Astrocyte Responses after Neonatal Ischemia: The Yin and the Yang.</a> Neuroscientist. 14 (4), 339-344 (2008).</li><li>Craig, A., Ling Luo, N., 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=Quantitative+analysis+of+perinatal+rodent+oligodendrocyte+lineage+progression+and+its+correlation+with+human.">Quantitative analysis of perinatal rodent oligodendrocyte lineage progression and its correlation with human.</a> Exp Neurol. 181 (2), 231-240 (2003).</li><li>Shen, Y., Liu, X. B., Pleasure, D. E., Deng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axon-glia+synapses+are+highly+vulnerable+to+white+matter+injury+in+the+developing+brain.">Axon-glia synapses are highly vulnerable to white matter injury in the developing brain.</a> J Neurosci Res. 90 (1), 105-121 (2012).</li><li>Shen, Y., Plane, J. M., Deng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+periventricular+leukomalacia.">Mouse models of periventricular leukomalacia.</a> J Vis Exp. (39), (2010).</li><li>Fox, W. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reflex-ontogeny+and+behavioural+development+of+the+mouse.">Reflex-ontogeny and behavioural development of the mouse.</a> Anim Behav. 13 (2), 234-241 (1965).</li><li>Tremml, P., Lipp, H. P., M&#252;ller, U., Ricceri, L., Wolfer, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobehavioral+development,+adult+openfield+exploration+and+swimming+navigation+learning+in+mice+with+a+modified+beta-amyloid+precursor+protein+gene.">Neurobehavioral development, adult openfield exploration and swimming navigation learning in mice with a modified beta-amyloid precursor protein gene.</a> Behav Brain Res. 95 (1), 65-76 (1998).</li><li>Wahlsten, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+developmental+time+scale+for+postnatal+changes+in+brain+and+behavior+of+B6D2F2+mice.">A developmental time scale for postnatal changes in brain and behavior of B6D2F2 mice.</a> Brain Res. 72 (2), 251-264 (1974).</li><li>Balasubramaniam, J., Xue, M., Del Bigio, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+motor+deficit+following+periventricular+hemorrhage+in+neonatal+rats:+A+potential+model+for+human+cerebral+palsy.">Long-term motor deficit following periventricular hemorrhage in neonatal rats: A potential model for human cerebral palsy.</a> J Cerebr Blood F Met. , (2005).</li><li>Williams, E., Scott, 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=The+Development+of+Social+Behavior+Patterns+in+the+Mouse,+in+Relation+to+Natural+Periods.">The Development of Social Behavior Patterns in the Mouse, in Relation to Natural Periods.</a> Behaviour. 6 (1), 35-65 (1954).</li><li>Heyser, C. J., Crawley, J. Q., 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=Assessment+of+developmental+milestones+in+rodents.">Assessment of developmental milestones in rodents.</a> Current protocols in neuroscience. Chapter 8, (2004).</li><li>Corti, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Grip strength TREAT-NMD: Experimental protocols for SMA animal models. , (2014).</li><li>Corti, S., Nizzardo, 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=Neural+stem+cell+transplantation+can+ameliorate+the+phenotype+of+a+mouse+model+of+spinal+muscular+atrophy.">Neural stem cell transplantation can ameliorate the phenotype of a mouse model of spinal muscular atrophy.</a> J Clin Invest. 118 (10), 3316-3330 (2008).</li><li>Grondard, C., Biondi, O., 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=Regular+exercise+prolongs+survival+in+a+type+2+spinal+muscular+atrophy+model+mouse.">Regular exercise prolongs survival in a type 2 spinal muscular atrophy model mouse.</a> J Neurosci. 25 (33), 7615-7622 (2005).</li><li>El-Khodor, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+Phenotyping+for+Neonates.">Behavioral Phenotyping for Neonates.</a> Experimental Protocols for SMA animal models. , (2011).</li><li>El-Khodor, B. F., Edgar, N., 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=Identification+of+a+battery+of+tests+for+drug+candidate+evaluation+in+the+SMNDelta7+neonate+model+of+spinal+muscular+atrophy.">Identification of a battery of tests for drug candidate evaluation in the SMNDelta7 neonate model of spinal muscular atrophy.</a> Exp Neurol. 212 (1), 29-43 (2008).</li><li>Venerosi, A., Ricceri, L., Scattoni, M. L., Calamandrei, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prenatal+chlorpyrifos+exposure+alters+motor+behavior+and+ultrasonic+vocalization+in+CD-1+mouse+pups.">Prenatal chlorpyrifos exposure alters motor behavior and ultrasonic vocalization in CD-1 mouse pups.</a> Environ Health. 8 (12), (2009).</li><li>Hill, J. M., Lim, M. A., Stone, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+milestones+in+the+newborn+mouse.">Developmental milestones in the newborn mouse.</a> Neuromethods 39: Neuropeptide Techniques. , (2008).</li><li>Yu, Y., Keshner, A. E., Tucker, C. A., Thompson, E. D., Lauer, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+dependence+influences+postural+responses+to+continuous+visual+perturbation+in+adults+with+spastic+cerebral+palsy.">Visual dependence influences postural responses to continuous visual perturbation in adults with spastic cerebral palsy.</a> , (2016).</li><li>Carlson, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+four+limb+hanging+tests+to+monitor+muscle+strength+and+condition+over+time.">The use of four limb hanging tests to monitor muscle strength and condition over time.</a> Experimental Protocols for SMA animal models. , (2011).</li><li>Gordon, A. M., Duff, S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relation+between+clinical+measures+and+fine+manipulative+control+in+children+with+hemiplegic+cerebral+palsy.">Relation between clinical measures and fine manipulative control in children with hemiplegic cerebral palsy.</a> Dev Med Child Neurol. 41 (9), 586-591 (1999).</li><li>Futagi, Y., Toribe, Y., Suzuki, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+grasp+reflex+and+moro+reflex+in+infants:+hierarchy+of+primitive+reflex+responses.">The grasp reflex and moro reflex in infants: hierarchy of primitive reflex responses.</a> Int J Pediat. , (2012).</li></ol>

The authors declare that they have no competing financial interests.

Utilizzando modelli animali per studiare le malattie umane è rilevante solo se vi è sovrapposizione tra la risposta cellulare e molecolare tra uomo e roditori e che i test comportamentali eseguiti hanno rilevanza diretta per i sintomi umani. Uno dei principali problemi con gli studi sulle malattie pediatriche è che molti ricercatori utilizzano roditori adulti per creare il modello, così come la valutazione del comportamento degli adulti roditore, senza considerare le differenze di sviluppo che possono essere importanti per il processo della malattia. A causa di questi problemi, è importante che la ricerca sull&#39;uso malattia pediatrica non solo la rettificato di sviluppo time-punti appropriata (ad esempio, lo sviluppo del sistema nervoso centrale umano a 28 - 32 settimane è equivalente ad un giorno post-natale 2-7 giorni roditore) 7, ma anche test comportamentali che esamineranno motore del caso, i comportamenti di sviluppo sensoriali o riflessivi. Così, come si sviluppa ogni nuovo modello di malattia neonatale, deve essere rigorosamente testati per assicurare che il cellularee risposte comportamentali forniranno i dati traducibili più adeguati tra roditori e umana. La paralisi cerebrale è un disturbo del motore, che persistono in età adulta. Un problema con molti dei modelli paralisi cerebrale disponibili oggi è la mancanza di ripetibile, test motore normalizzato in grado di correlare con i deficit osservati in pazienti pediatrici. In questo nuovo modello, che unisce ipossia, ischemia e infiammazione in un mouse neonatale, comportamento motorio è stata valutata utilizzando una serie di test specifici per topi neonati. Per ridurre la soggettività e aumentare la segnalazione quantitativa, diverse prove sono state modificate per includere molto specifica, ma facile valutare misure che possono essere standardizzati. Inoltre, le valutazioni anteriori e arti posteriori possono essere eseguiti separatamente, e le differenze sinistra / destra può essere determinato. Questa batteria di test è specifico per topi neonati fino a due settimane di età. Questo modello CP dimostradifficoltà nel camminare (deambulazione, angolo del piede degli arti posteriori), nonché la debolezza specifici degli arti (sospensione di quattro arti, la sospensione degli arti posteriori), e deficit in riflessi di sviluppo (afferrare reflex). Anche se in questo studio solo un timepoint stato esaminato, questi deficit possono essere monitorati nel corso del tempo. Ci sono altre batterie di test che possono essere utilizzati sul neonato, come ad esempio la batteria del Fox di test o di valutazione di 15 tappe dello sviluppo di Heyser. Tuttavia, questi test confrontare il neonato all&#39;adulto, il cui risposte non può essere lo stesso, perché il neonato è ancora in via di sviluppo. La batteria di Fox e test assement di Heyser si basano su informazioni soggettive osservazionale con dicotomico valutazione (sì o no), piuttosto che dati oggettivi (angolo, postura basati sulla forza, ecc). A causa della soggettività di questi test, molti scienziati si sono adattati, aggiunto o rimosso criteri, rendendo così i loro risultati incomparabili agli altri e limiteing l&#39;utilità dei dati in termini di stabilire un deficit di base per una particolare malattia o disturbo. Attraverso la creazione di una serie di test standardizzati a motore che sono qualitative e specificamente progettato per testare i neonati, i risultati dei singoli gruppi di ricerca possono essere preciso e affidabile segnalati e confrontati.

Lo sviluppo di nuovi modelli di infortunio o di malattia pediatrica utilizzando roditori è spesso difficile a causa della sorprendente capacità di entrambi i ratti e topi di recuperare rapidamente da lesioni neurologiche. Pertanto, al fine di convalidare qualsiasi nuovo modello di malattia pediatrica, accuratamente esaminando i cambiamenti cellulari e molecolari devono andare di pari passo con esiti comportamentali. In molti modi, il recupero comportamentale funzionale può essere più importante di cambiamenti cellulari sottostanti in termini di rilevanza terapeutica e traslazionale. Come i ricercatori saperne di più sulla ferita nell&#39;adulto e nel neonato, è chiaro che le loro risposte sono molto diversi e non possono essere estrapolati tra i due. Ad esempio, i topi neonati mostrano diversi livelli di fattore di crescita nervoso, fattore neurotrofico derivato dal cervello, neurotrofina-3 e gliali fattore neurotrofico delle cellule linea di derivazione dopo un trauma del midollo spinale 1,2. Inoltre, i neonati hanno una significativa perdita di barriera emato-encefalica dopo l&#39;ictus 3, demonstrate corticale riassetto neurone dopo periferici lesione del nervo 4, e hanno un astrogliosis ritardato o rallentato in seguito a lesione del midollo spinale e ipossia-ischemia 5,6. Pertanto, è importante che traslazionale ricerca pediatrica utilizzare modelli evolutivamente equivalenti e che tali modelli vengono valutati per entrambi i cambiamenti cellulari / molecolari e test comportamentali adatti alla loro età. Paralisi cerebrale (CP) è un disturbo motorio che colpisce 3: 1000 nati vivi ogni anno (NIH). I bambini con CP presentano una gamma di sintomi e patologie associate, a seconda della gravità della malattia. Difficoltà con il movimento e il coordinamento sono i segni più comuni, insieme a ritardi nel raggiungimento del motore tappe dello sviluppo. Altri segni includono tono muscolare anormale (sia aumentato o diminuito), ridotta capacità motorie, difficoltà a camminare, sbavando eccessiva e la deglutizione, e ritardi del linguaggio (NIH). La causa di fondo di CP si crede di essereuna mancanza di ossigeno e / o il flusso di sangue al cervello durante il periodo di pre o peri-parto, o fino a un anno post-partum. Inoltre, l&#39;infiammazione è ora ritenuta un componente chiave nello sviluppo di CP. La maggioranza dei casi di CP sono associati con il bianco danno la materia intorno ai ventricoli, noto come leucomalacia periventricolare (PVL). Questa caratteristica neurologica suggerisce che l&#39;insulto iniziale che porta al CP si verifica durante il periodo di sviluppo del cervello quando i oligodendrociti sono più vulnerabili agli insulti. Il periodo di crescita rapida degli oligodendrociti in un essere umano, anche il periodo in cui gli oligodendrociti sono i più sensibili al danno, è tra i 24 - 32 settimane di gestazione. Nel roditore, il periodo equivalente è giorni post-natale 2 - 7 7, ed è quando CP è indotta in questo modello. Il modello di topo neonatale di CP che è stato utilizzato per effettuare i test descritti qui combina ipossia e ischemia con l&#39;infiammazione di creare un injury che imita meglio la neurodegenerazione visti in CP umana. Questo modello affronta alcune delle principali carenze riscontrate in altri modelli animali di CP, che mancano di deficit motori distinti che assomiglia pazienti in CP umani, così come distinta danni sostanza bianca. Precedenti studi da un collaboratore utilizzando lo stesso modello hanno dimostrato che l&#39;aggiunta di infiammazione aumenta il danno sostanza bianca, così meglio emulare il PVL osservati nei bambini con CP 8. Sulla base dei dati precedenti, questo documento presenta una serie completa di test del motore neonatali al fine di valutare i cambiamenti nel comportamento del motore, come l&#39;età degli animali.

<table><tbody><tr><td></td><td></td><td></td><td></td></tr><tr><td>C57BL/6 mice</td><td>Charles River Laboratories</td><td>STRAIN CODE: 027&amp;nbsp;</td><td>C57BL/6NCrl is the exact strain we use</td></tr><tr><td>Anesthesia Dish, PYREX&amp;trade; Crystallizing Dish</td><td>Corning Life Sciences Glass&amp;nbsp;</td><td>3140125</td><td>Capacity: 25.03 oz. (740ml); Dia. x H: 4.92 x 2.55 in. (125 x 65mm). However, any small round glass container will work. A 2 cup capacity pyrex food storage bowl with flat bottom will also work and is much cheaper (Pyrex model number: 6017399).</td></tr><tr><td>Covered lead ring</td><td>Fisher Scientific</td><td>S90139C</td><td>Lead ring for stablizing flasks in a water bath. It is used inside the anesthesia dish.</td></tr><tr><td>Scalpel Blade #11</td><td>World Precision Instrucments, Inc.</td><td>500240</td><td></td></tr><tr><td>Small Vessel Cauterizer</td><td>Fine Science Tools</td><td>18000-00</td><td></td></tr><tr><td>Micro Hook</td><td>Fine Science Tools</td><td>10064-14</td><td></td></tr><tr><td>Vetbond Suture Glue</td><td>3M</td><td>1469SB</td><td>n-butyl cyanoacrylate adhesive</td></tr><tr><td>Lipopolysaccharide</td><td>Sigma Life Science</td><td>L4391</td><td>Lipopolysaccaride from E.coli 0111:B4, gamma irradiated</td></tr><tr><td>12 x 12 inch opaque box</td><td>Acrylic Display Manufacturing: A division of Piasa Plastics</td><td>C4022</td><td>Colored Acrylic 5-Sided Cube, 3/16&quot; Colored Acrylic, 12&quot;W x 12&quot;D x 12&quot;H;&amp;nbsp; http://www.acrylicdisplaymfg.com/html/cubes_19.html</td></tr><tr><td>Camera/camcorder</td><td>JVC</td><td>GC-PX100BUS</td><td>Any camcorder that works well in low light and can be imported and edited. We use the JVC GC-PX100 Full HD Everio Camcorder.</td></tr><tr><td>Covidien Tendersorb&amp;trade; Underpads</td><td>Kendall Healthcare Products Co</td><td>7174</td><td></td></tr><tr><td>WypAll L40</td><td>Kimberly-Clark Professional</td><td>5600</td><td>Any surface with moderate grip will do</td></tr><tr><td>Surface at 45 degree incline</td><td></td><td></td><td>We use a cardboard box.</td></tr><tr><td>Thin wire from a pipe cleaner</td><td>Creatology</td><td>M10314420</td><td>Any pipe cleaner from any craft store will work.</td></tr><tr><td>50mL conical tube</td><td>Falcon</td><td>352070</td><td></td></tr><tr><td>Fiberglass Screen Wire</td><td>New York Wire&amp;nbsp; www.lowes.com</td><td>14436</td><td>Any supplier can be used as long as their screen is 16 x 16 or 18 x 16</td></tr><tr><td>Razor blade</td><td>Fisherbrand</td><td>12-640</td><td>A wooden stick applicator or wooden part of a cotton-tipped swab will also work.</td></tr><tr><td>OPTIX 24-in x 4-ft x 0.22-in Clear Acrylic Sheet to make Clear Acrylic Walkway</td><td>PLASKOLITE INC</td><td>1AG2196A</td><td>Clear acrylic (1/8&quot; thick) with sides and a top to limit exploration. We bought a sheet of acrylic from a local hardware store and had them cut it to size. (2) 2&quot; x 2&quot;; (3) 2&quot; x 18&quot;; (1) 2&quot; x 15.5&quot;; (1) 2&quot; x 3&quot;. Using clear tape, tape all sides together, with the 15.5&quot; piece on top. Tape the 3&quot; piece to the end of the 15.5&quot; piece to create a flap/entryway for the mice. Alternatively, part or all of the walkway can be glued together, and only taping on the top pieces. This design will allow for the walkway to be opened for easy cleaning.</td></tr><tr><td>Protractor</td><td>Westscott</td><td>ACM14371</td><td></td></tr></tbody></table>

a battery of motor tests in a neonatal mouse model of cerebral palsy

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, standardized battery of neonatal mouse motor tests. These tests can validate injury or disease models, determine treatment efficacy and/or assess motor behaviors in new transgenic strains. This paper presents a series of neonatal motor tests to evaluate general motor function, including ambulation, hindlimb foot angle, surface righting, negative geotaxis, front- and hindlimb suspension, grasping reflex, four limb grip strength and cliff aversion. Mice between the ages of post-natal day 2 to 14 can be used. In addition, these tests can be used for a wide range of neurological and neuromuscular pathologies, including cerebral palsy, hypoxic-ischemic encephalopathy, traumatic brain injury, spinal cord injury, neurodegenerative diseases, and neuromuscular disorders. These tests can also be used to determine the effects of pharmacological agents, as well as other types of therapeutic interventions. In this paper, motor deficits were evaluated in a novel neonatal mouse model of cerebral palsy that combines hypoxia, ischemia and inflammation. Forty-eight hours after injury, five tests out of the nine showed significant motor deficits: ambulation, hindlimb angle, hindlimb suspension, four limb grip strength, and grasping reflex. These tests revealed weakness in the hindlimbs, as well as fine motor skills such as grasping, which are similar to the motor deficits seen in human cerebral palsy patients.

I topi sono stati testati da P7 (24 ore dopo l&#39;intervento chirurgico) a P13 (1 settimana dopo l&#39;intervento chirurgico), usando i mouse diverso per ogni time-point in modo che l&#39;apprendimento di una paradigma di test non era una variabile di confondimento. P8 è stato selezionato come risultati rappresentativi, come topi hanno mostrato i maggiori deficit a questo punto di tempo. Transizione da Crawling a piedi è ritardato di CP topi neonati pazienti in CP umani hanno anomalie dell&#39;andatura, che vanno dalla punta di piedi per un&#39;andatura scissored. Dato che questo modello CP display andatura deficit simili agli esseri umani, la deambulazione è stata valutata. I topi sono stati lanciati sulla simmetria andatura e il movimento degli arti-zampa durante una passeggiata retta. A 48 ore dopo l&#39;intervento chirurgico (PND 8), i topi CP avevano meno movimento simmetrico arto e una &quot;strisciante&quot; andatura rispetto alle loro controparti sham (punteggio medio deambulazione: CP 1.083 ±0,6337, n = 12 vs sham 1.639 ± 0,4859, n = 9; p &lt;0.05, Figura 10). In una settimana, sia CP e topi sham sono passati a camminare (dati non riportati). <img alt="Figura 10" src="/files/ftp_upload/53569/53569fig10.jpg" /> Figura 10. CP mouse Pups non Ambulate così come Shams. Topi Sham (barra nera) hanno un punteggio medio di 1.639 ± 0,4859 (n = 9), vale a dire il loro sviluppo ambulatoriale cade tra il movimento degli arti asimmetrica e lento strisciare. topi CP (barra grigia) ricevono un punteggio medio di 1.083 ± 0,6337 (n = 12), il che significa la loro deambulazione è meno sviluppata e tendono ad avere il movimento degli arti asimmetrica. I dati sono espressi come media ± SEM; * È p &lt;0.05. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig10large.jpg" target="_blank">Cliccate qui per vedere una versione più grande di questa figura.</a> HinAngolo del piede dlimb è aumentata in CP Oltre alla deambulazione, degli arti posteriori piede ad angolo è stata valutata. Otto giorni di età cuccioli farsa del mouse a piedi con i loro hindpaws rivolto in avanti, rispetto ai topi HIL, che hanno divaricate hindpaws quando si cammina in linea retta (figura 2; angolo medio: CP 77.48 ± 9,848, n = 9, vs sham 54.54 ± 8,043, n = 11; p &lt;0,0001, Figura 11). Questo aumento dell&#39;angolo correla con instabilità andatura, dal fatto che i cuccioli bisogno di aumentare l&#39;angolo di loro zampe posteriori per stabilizzare la loro andatura e assistere con equilibrio e coordinazione. <img alt="Figura 11" src="/files/ftp_upload/53569/53569fig11.jpg" /> Figura 11. CP mouse Pups Splay loro hindpaws quando si cammina. Topi CP (barre nere) hanno un angolo medio tra i loro arti posteriori di 77.48 ± 8,043 (n = 11), mentre i topi sham (grigio b<html> ars) hanno un angolo medio di 54,54 ± 9,848 (n = 9). I dati sono espressi come media ± SEM; **** È p &lt;0,0001. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig11large.jpg" target="_blank">Cliccate qui per vedere una versione più grande di questa figura.</a> I topi CP Non mostrare deficit quando Righting Surface Il test di superficie di raddrizzamento è stato incluso come alcuni pazienti CP hanno alterato controllo del tronco (Heyrman et al., 2013). Inoltre, il sistema vestibolare è necessario rilevare la necessità di raddrizzamento e ci sono deficit vestibolari in alcuni pazienti CP 23. topi CP non mostrano deficit significativi quando raddrizzamento rispetto ai controlli sham (dati non riportati). I topi CP Eseguire la stessa come Sham in negativo geotassi Testing t &quot;fo: keep-together.within-page =&quot; 1 &quot;&gt; geotassi negativo è usato per testare la coordinazione motoria nei giovani cuccioli I topi sono in discussione da essere posto in discesa su una superficie inclinata ritardo o mancata salita orientare potrebbe indicare deficit.. in coordinazione, equilibrio, o ingresso vestibolare. topi CP non mostrano deficit quando provocati con geotassi negativi rispetto ai topi sham (dati non riportati). Inoltre, i topi CP non hanno mostrato una preferenza per girare verso un lato rispetto ad un altro, quando ri-orientamento . Front-Limb test sospensione è opportuna per topi anziani di 10 giorni pazienti in CP sono diminuiti del tono muscolare e deficit nelle abilità motorie, come afferrare. Per verificare la debolezza di questo modello di topo, abbiamo usato un test di sospensione anteriore degli arti. Inoltre, questo modello utilizza danno ischemico unilaterale e retro-ness potrebbero essere determinati con questo test sospensione. Questo testè meglio per i topi di età superiore ai 10 giorni 15. A 8 giorni, due giorni dopo un trauma, non vi erano differenze significative tra CP e topi sham (dati non riportati). Forza arti posteriori è diminuito nei topi CP pazienti in CP umani hanno spesso bisogno di parentesi graffe o dispositivi di assistenza a piedi a causa della mancanza di controllo del motore e forza. Al fine di confrontare il modello CP roditore per gli esseri umani, la forza degli arti posteriori è stata valutata utilizzando il test di sospensione degli arti posteriori. Quando sospeso dal lato di un tubo conico, topi CP hanno mostrato la debolezza degli arti posteriori, come dimostrato da una diminuzione della appeso punteggio (arti posteriori punteggio appeso: CP 3.468 ± 0,5561, n = 13, vs sham 3,891 ± 0,1329, n = 13; p &lt; 0,05, Figura 12). Nessuna differenza è stata osservata nel tempo di sospensione hindlimb (dati non mostrati). Così, simile a pazienti in CP umani, topi CP dimostrano hindlIMB (gamba) debolezza. <img alt="Figura 12" src="/files/ftp_upload/53569/53569fig12.jpg" /> Figura 12. I topi Sham sono leggermente ma significativamente più forte nelle loro arti posteriori rispetto ai topi CP. In un punteggio impiccagione di 3,891 ± 0,1329 (n = 13), i topi sham (barra nera) mostrano la separazione più degli arti posteriori, e quindi una più forte presa di posizione degli arti posteriori, quando appeso al bordo di un tubo di topi CP (barra grigia) con un voto medio impiccagione di 3.468 ± 0,5561 (n = 13). I dati sono espressi come media ± SEM; * È p &lt;0.05. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig12large.jpg" target="_blank">Cliccate qui per vedere una versione più grande di questa figura.</a> Grip forza è diminuita Following CP Injury Afferrando con tutte e quattro le zampe è importante per un roditoretermini di arrampicata e correre su superfici irregolari. Grip richiede notevole forza sostenuta, invece di destrezza o forza lineare, soprattutto nelle cifre e zampe 24. I topi sono stati tenuti a tenere il loro peso corporeo su uno schermo di rete metallica rovesciata. Topi CP non sono stati in grado di mantenere la loro presa e questi topi è sceso ad angolo significativamente più bassi (quattro arti angolo medio: CP 75,627 ± 24,48, n = 11, vs sham 96.57 ± 10.836, n = 9; p &lt;0.05, figura 13). Questi dati mostrano che vi è un deficit significativo nella forza di presa in topi CP. <img alt="Figura 13" src="/files/ftp_upload/53569/53569fig13.jpg" /> Figura 13. I topi hanno CP Grip più debole rispetto Shams. Topi Sham (barra nera) può afferrare un angolo invertito media di 96.57 ± 10,836 (n = 9). topi CP (barra grigia) possono raggiungere solo un angolo rovesciata di 75,627 ± 24.48 (n = 11). I dati sono espressi come media ± SEM; * È p0; 0.05. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig13large.jpg" target="_blank">Clicca qui per vedere una versione più grande di questa figura.</a> I deficit Grasping Reflex sono evidenti nei topi CP Insieme con deficit motorie, movimenti motorie sono anche compromesse nei pazienti CP 25,26. Il riflesso cogliere nell&#39;uomo è presente alla nascita e scompare intorno 5-6 mesi. Tuttavia, i cambiamenti riflesso di presa, quali velocità esagerata o la forza di afferramento, mancata afferrare o il riemergere del riflesso afferrare dopo 6 mesi di età, tutti indicano danni al sistema nervoso. Per confrontare afferrare nel modello CP, i deficit di presa riflessivi sono stati determinati. A 48 ore dopo l&#39;infortunio, topi CP dimostrano una diminuzione nel cogliere il riflesso (zampe medie coltein 48 ore: CP 2.429 ± 0,9376, n = 14, vs sham 3.214 ± 0,8018, n = 14; p &lt;0.05, Figura 14A). C&#39;era una leggera, ma non significativo aumento destra preferenza zampa nelle zampe anteriori (dati non riportati). C&#39;è stata una significativa preferenza zampa destra nelle hindpaws (CP 75,0 ± 42,74, n = 14, vs sham 17.86 ± 54.09, n = 14; p &lt;0.005, Figura 14B). Una settimana dopo l&#39;infortunio, topi CP mostrano deficit di presa (zampe medie colte a 1 settimana: CP 2.75 ± 1.035, n = 8, vs sham 3,80 ± 0,6325, n = 10; p &lt;0.05, Figura 14C), senza preferenza zampa notevole . <img alt="Figura 14" src="/files/ftp_upload/53569/53569fig14.jpg" /> Figura 14. I topi CP hanno deficit afferrare, nella hindpaws, controlaterale alla Cervello Regione feriti. (A) 48 ore dopo un trauma (PND 8), i topi CP (barra grigia) afferrare un bastone con, in media, un minor numero di zampe di farsa animaLS (barra nera). topi (B) CP (barra grigia) mostrano una preferenza per afferrare con la hindpaw destra (controlaterale alla lesione) in contrasto con il hindpaw sinistra (omolaterale al danno). topi Sham (barra nera) Non visualizzare questo diritto preferenza zampa. Preferenza zampa destra è calcolato come ([zampa destra - sinistra zampa] / [zampa destra + zampa sinistra + entrambe le zampe] * 100) (C) Una settimana dopo un trauma, i topi CP (barra grigia) mostrano ancora afferrare deficit rispetto al. Shams (barra nera). I dati sono espressi come media ± SEM; * È p &lt;0.05, ** è p &lt;0.005. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53569/53569fig14large.jpg" target="_blank">Fai clic qui per vedere una versione più grande di questa figura.</a> I topi CP si allontanano dal bordo Durante avversione Cliff Il test scogliera avversione si basa sulla paura intrinseca dei topi a girare away da una ripida scogliera e dirigersi verso la sicurezza. Anche se alcuni pazienti in CP hanno difficoltà vestibolari, così come il controllo del motore ridotta, i topi CP non hanno mostrato alcun deficit in questo test.

Watch this Scientific Journal Video about A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy at JoVE.com

A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy

Presented is a concise battery of mouse neonatal motor tests. Using these tests, neonatal motor deficits can be demonstrated in a variety of neonatal motor disorders. By having a standardized set of tests, results from different studies can be compared, allowing for better and accurate reporting between groups.

Una serie di test a motore in neonatale topo modello di paralisi cerebrale

As the sheer number of transgenic mice strains grow and rodent models of pediatric disease increase, there is an expanding need for a comprehensive, ...

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Universidad de Cartagena UDEC

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24.0K Views.  Lewiz Katz School of Medicine at Temple University. Presented is a concise battery of mouse neonatal motor tests. Using these tests, neonatal motor deficits can be demonstrated in a variety of neonatal motor disorders. By having a standardized set of tests, results from different studies can be compared, allowing for better and accurate reporting between groups.

Video: A Battery of Motor Tests in a Neonatal Mouse Model of Cerebral Palsy

Assessment of Sensorimotor Function in Mouse Models of Parkinson's Disease

Sensitive and reliable behavioral outcome measures are essential to the evaluation of potential therapeutic treatments in preclinical trials for many neurodegenerative diseases. In Parkinson's disease, sensorimotor tests sensitive to varying degrees of nigrostriatal dysfunction are fundamental for testing the efficacy of potential therapeutics. Reliable and quite elegant sensorimotor measures exist for rats, however many of these tests measure sensorimotor asymmetry within the rat and are not entirely suitable for the newer genetic mouse models of PD. We have put together a battery of sensorimotor tests inspired by the sensitive tests in rats and adapted for mice. The test battery highlighted in this study is chosen for a) its sensitivity in a wide variety of mouse models of PD, b) its ease in implementing into a study, and c) its low expense. These tests have proven useful in characterizing novel genetic mouse models of PD as well as in testing potential disease-modifying therapies.

In Parkinson's disease and movement disorders in general, sensitive and reliable behavioral assays are essential for testing novel potential therapeutics. Here, we describe a manageable battery of sensorimotor tests for mice that are sensitive to varying degrees of injury to the nigrostriatal system and useful for preclinical studies.

Valutazione della Sensorimotor funzione in modelli murini di malattia di Parkinson

Sensitive and  reliable behavioral outcome measures are essential to the evaluation of  potential therapeutic treatments in preclinical trials for many  ...

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Comportamento

Paw-Dragging: a Novel, Sensitive Analysis of the Mouse Cylinder Test

The cylinder test is routinely used to predict focal ischemic damage to the forelimb motor cortex in rodents. When placed in the cylinder, rodents explore by rearing and touching the walls of the cylinder with their forelimb paws for postural support. Following ischemic injury to the forelimb sensorimotor cortex, rats rely more heavily on their unaffected forelimb paw for postural support resulting in fewer touches with their affected paw which is termed forelimb asymmetry. In contrast, focal ischemic damage in the mouse brain fails to result in comparable consistent deficits in forelimb asymmetry. While forelimb asymmetry deficits are infrequently observed, mice do demonstrate a novel behaviour post stroke termed &#8220;paw-dragging&#8221;. Paw-dragging is the tendency for a mouse to drag its affected paw along the cylinder wall rather than directly push off from the wall when dismounting from a rear to a four-legged stance. We have previously demonstrated that paw-dragging behaviour is highly sensitive to small cortical ischemic injuries to the forelimb motor cortex. Here we provide a detailed protocol for paw-dragging analysis. We define what a paw-drag is and demonstrate how to quantify paw-dragging behaviour. The cylinder test is a simple and inexpensive test to administer and does not require pre-training or food deprivation strategies. In using paw-dragging analysis with the cylinder test, it fills a niche for predicting cortical ischemic injuries such as photothrombosis and Endothelin-1 (ET-1)-induced ischemia &#8211; two models that are ever-increasing in popularity and produce smaller focal injuries than middle cerebral artery occlusion. Finally, measuring paw-dragging behaviour in the cylinder test will allow studies of functional recovery after cortical injury using a wide cohort of transgenic mouse strains where previous forelimb asymmetry analysis has failed to detect consistent deficits.

Classical forelimb asymmetry analysis of the cylinder test is routinely used to assess behavioural deficits in rats following brain injury or stroke; however, it fails to detect consistent deficits in mice. This study demonstrates that quantifying paw-dragging behaviour is a more sensitive analysis of brain injury in mice.

Paw-trascinamento: un romanzo, analisi sensibile del cilindro di prova del mouse

The cylinder test is routinely used to predict focal ischemic damage to the forelimb motor cortex in rodents. When placed in the cylinder, rodents explore ...

Measuring Motor Coordination in Mice

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such as emotionality or cognition can be assessed, however, it is vital to determine whether the motor capabilities of e.g. a mutant or lesioned mouse allow such an assessment. Performance on a maze task requiring strength and coordination, such as the Morris water maze, might well be impaired in a mouse by motor, rather than cognitive, impairments, so it is essential to selectively dissect the latter from the former. For example, sensorimotor impairments caused by NMDA antagonists have been shown to impair water maze performance2. 
Motor coordination has traditionally been assessed in mice and rats by the rotarod test, in which the animal is placed on a horizontal rod that rotates about its long axis; the animal must walk forwards to remain upright and not fall off. Both set speed and accelerating versions of the rotarod are available.
The other three tests described in this article (horizontal bar, static rods and parallel bars) all measure coordination on static apparatus. The horizontal bar also requires strength for adequate performance, particularly of the forelimbs as the mouse initially grips the bar just with the front paws. 
Adult rats do not perform well on tests such as the static rods and parallel bars (personal observations); they appear less well coordinated than mice. I have only tested male rats, however, and male mice seem generally less well coordinated than females. Mice appear to have a higher strength:weight ratio than rats; the Latin name, Mus musculus, seems entirely appropriate. The rotarod, the variations of the foot fault test12 or the Catwalk (Noldus)15 apparatus are generally used to assess motor coordination in rats.

Protocols are presented for two established motor coordination tasks, the accelerating rotarod and horizontal bar, also two tests developed in Oxford recently, the static rods and parallel bars. These tests can detect motor impairments potentially of interest in their own right, as well as being possible variables in tests of other areas of behavior.

Misurare Coordinamento motore in Mouse

Mice are increasingly being used in behavioral neuroscience, largely replacing rats as the behaviorist's animal of choice. Before aspects of behavior such ...

Neuroscienze

Neurodevelopmental Reflex Testing in Neonatal Rat Pups

Neurodevelopmental reflex testing is commonly used in clinical practice to assess the maturation of the nervous system. Neurodevelopmental reflexes are also referred to as primitive reflexes. They are sensitive and consistent with later outcomes. Abnormal reflexes are described as an absence, persistence, reappearance, or latency of reflexes, which are predictive indices of infants that are at high risk for neurodevelopmental disorders. Animal models of neurodevelopmental disabilities, such as cerebral palsy, often display aberrant developmental reflexes, as would be observed in human infants. The techniques described assess a variety of neurodevelopmental reflexes in neonatal rats. Neurodevelopmental reflex testing offers the investigator a testing method that is not otherwise available in such young animals. The methodology presented here aims to assist investigators in examining developmental milestones in neonatal rats as a method of detecting early-onset brain injury and/or determining the effectiveness of therapeutic interventions. The methodology presented here aims to provide a general guideline for investigators.

Behavioral testing is the gold standard for determining outcomes following brain injury, and can identify the presence of developmental disabilities in infants and children. Neurodevelopmental reflexes are an early indicator of these abnormalities. A host of easily accomplished developmental reflex tests in the neonatal rodent were developed and described here.

Neurologico Reflex Test in neonatali Rat Pups

Neurodevelopmental reflex testing is commonly used in clinical practice to assess the maturation of the nervous system. Neurodevelopmental reflexes are ...

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the animal&#39;s wellbeing and health. There are numerous behavioral tests that may be applied. We selected a test for general locomotion, the open field test (OFT); a test for muscular strength, the mesh test (MT); and a test for coordination, the rotarod test (RR). Testing can be accomplished on a weekly or monthly basis. As a test for general locomotion, the OFT works by objectively monitoring movement parameters while the mouse is in an open field apparatus. The field is generally a 2&#39; x 2&#39; box, and the movements are recorded through laser sensing or through video capture. The mouse is placed in the center of the open field and allowed to move freely for the test. The MT uses the latency for a mouse to fall off an inverted screen as a measure of muscular strength. A mouse is placed on a screen, which is inverted over a clear box, and is timed for their latency to fall. Three trials are performed, with the best of the three trials scored for that day. A score of 60 s is the maximum time a mouse is left inverted. Mice are given a 5-min rest period between mesh test trials. Lastly, an accelerated protocol on the RR assesses motor coordination and endurance. During a trial, a mouse walks on a rotating rod as it increases in speed from 4 rpm to 40 rpm over 5 min. The trial ends when the mouse touches the magnetized pressure sensor upon falling. Each mouse undergoes three trials, and the best trial is scored for that day. This combined behavioral data allows for the global assessment of mobility, coordination, strength, and movement of the test animals. At least two out of the three behavioral testing measures must show improvement for an animal to qualify as having overall improved motor function.

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects of a disease or treatment on an animal's wellbeing. In order to globally assess motor functioning, we selected tests for general locomotion, muscular strength, and coordination: the open field test, the mesh test, and the rotarod test, respectively.

Batteria di comportamentali test valutazione generale locomozione, forza muscolare e coordinazione in topi

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects and outcomes that a particular disease or treatment has on the ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

Un neonatale mouse Spinal Cord Injury compressione Modello

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

Medicina

Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury

We performed unilateral carotid artery occlusion on CD-1 mice to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of neonatal HI brain injury by studying neurobehavioral functions in these mice compared to non-operated (i.e., normal) mice. During the study, Rice-Vannucci&#39;s method was used to induce neonatal HI brain damage in postnatal day 7-10 (P7-10) mice. The HI operation was performed on the pups by unilateral carotid artery ligation and exposure to hypoxia (8% O2 and 92% N2 for 90 min). One week after the operation, the damaged brains were evaluated with the naked eye through the semi-transparent skull and were categorized into subgroups based on the absence (&#34;no cortical injury&#34; group) or presence (&#34;cortical injury&#34; group) of cortical injury, such as a lesion in the right hemisphere. On week 6, the following neurobehavioral tests were performed to evaluate the cognitive and motor functions: passive avoidance task (PAT), ladder walking test, and grip strength test. These behavioral tests are helpful in determining the effects of neonatal HI brain injury and are used in other mouse models of neurodegenerative diseases. In this study, neonatal HI brain injury mice showed motor deficits that corresponded to right hemisphere damage. The behavioral test results are relevant to the deficits observed in human neonatal HI patients, such as cerebral palsy or neonatal stroke patients. In this study, a mouse model of neonatal HI brain injury was established and showed different degrees of motor deficits and cognitive impairment compared to non-operated mice. This work provides basic information on the HI mouse model. MRI images demonstrate the different phenotypes, separated according to the severity of brain damage by motor and cognitive tests.

We performed unilateral carotid artery occlusion on postnatal day 7-10 CD-1 mouse pups to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of HI brain injury. We studied neurobehavioral functions in these mice compared to non-operated normal mice.

Valutazioni di neurobehavioral in un modello murino di ferita di cervello Hypoxic-ischemica neonatale

We performed unilateral carotid artery occlusion on CD-1 mice to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of neonatal HI ...

A Controlled Mouse Model for Neonatal Polymicrobial Sepsis

Neonatal sepsis remains a global burden. A preclinical model to screen effective prophylactic or therapeutic interventions is needed. Neonatal mouse polymicrobial sepsis can be induced by injecting cecal slurry intraperitoneally into day of life 7 mice and monitoring them for the following week. Presented here are the detailed steps necessary for the implementation of this neonatal sepsis model. This includes making a homogeneous cecal slurry stock, diluting it to a weight- and litter-adjusted dose, an outline of the monitoring schedule, and a definition of observed health categories used to define humane endpoints. The generation of a homogeneous cecal slurry stock from pooled donors allows for the administration into many litters over time, reducing the variation between donors, and preventing the use of potentially toxic glycerol. The monitoring strategy used allows for the anticipation of survival outcome and the identification of mice that would later progress to death, allowing for an earlier identification of the humane endpoint. Two main behavioral features are used to define the health scores, namely, the ability of the neonatal mice to right themselves when placed on their back and their level of mobility. These criteria could potentially be applied to address humane endpoints in other studies of neonatal disease in mice, as long as a pilot study is performed to confirm accuracy. In conclusion, this approach provides a standardized method to model newborn sepsis in mice, while providing resources to assess animal welfare used to define early humane endpoints for challenged animals.

This protocol provides the necessary steps to establish and evaluate neonatal sepsis in 7-day-old mice.

Un modello di Mouse controllata per sepsi polimicrobica neonatale

Neonatal sepsis remains a global burden. A preclinical model to screen effective prophylactic or therapeutic interventions is needed. Neonatal mouse ...

Immunologia e infezioni

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal neurological deficits and even amnesia. Athletes participating in contact sports are at high risk of exposure to large number of mTBIs. In terms of the level of injury in a sporting athlete, a mTBI is defined as a mild injury that does not cause gross pathological changes, but does cause short-term neurological deficits that are spontaneously resolved. Despite previous attempts to model mTBI in mice and rats, many have reported gross adverse effects including skull fractures, intracerebral bleeding, axonal injury and neuronal cell death. Herein, we describe our highly reproducible animal model of mTBI that reproduces clinically relevant symptoms. This model uses a custom made pneumatic impactor device to deliver a closed-head trauma. This impact is made under precise velocity and deformation parameters, creating a reliable and reproducible model to examine the mechanisms that contribute to effects of single or repetitive concussive mTBI.

Athletes absorb several hundred mild traumatic brain injuries (mTBI)/concussions every year; however, the consequence of these on the brain is poorly understood. Therefore, an animal model of single and repetitive mTBI that consistently replicates clinically relevant symptoms provides the means to advance the study of mTBI and concussion.

Un modello di mouse del singolo e ripetitivo dolore cerebrale traumatico lieve

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal ...

Quantificazione della disfunzione locomotoria nei topi con lesione del midollo spinale utilizzando MouseWalker

Using the MouseWalker to Quantify Locomotor Dysfunction in a Mouse Model of Spinal Cord Injury

The execution of complex and highly coordinated motor programs, such as walking and running, is dependent on the rhythmic activation of spinal and supra-spinal circuits. After a thoracic spinal cord injury, communication with upstream circuits is impaired. This, in turn, leads to a loss of coordination, with limited recovery potential. Hence, to better evaluate the degree of recovery after the administration of drugs or therapies, there is a necessity for new, more detailed, and accurate tools to quantify gait, limb coordination, and other fine aspects of locomotor behavior in animal models of spinal cord injury. Several assays have been developed over the years to quantitatively assess free-walking behavior in rodents; however, they usually lack direct measurements related to stepping gait strategies, footprint patterns, and coordination. To address these shortcomings, an updated version of the MouseWalker, which combines a frustrated total internal reflection (fTIR) walkway with tracking and quantification software, is provided. This open-source system has been adapted to extract several graphical outputs and kinematic parameters, and a set of post-quantification tools can be to analyze the output data provided. This manuscript also demonstrates how this method, allied with already established behavioral tests, quantitatively describes locomotor deficits following spinal cord injury.

Autore in primo piano: Utilizzo del MouseWalker per quantificare la disfunzione locomotoria in un modello murino di lesione del midollo spinale  

An experimental pipeline to quantitatively describe the locomotor pattern of freely walking mice using the MouseWalker (MW) toolbox is provided, ranging from initial video recordings and tracking to post-quantification analysis. A spinal cord contusion injury model in mice is employed to demonstrate the usefulness of the MW system.