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. 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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. 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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.

El uso de modelos animales para estudiar enfermedades humanas sólo es relevante si existe una superposición entre la respuesta celular y molecular entre humanos y roedores, y que las pruebas de comportamiento realizadas tienen relación directa con los síntomas humanos. Uno de los principales problemas con los estudios de enfermedades pediátricas es que muchos investigadores utilizan roedores adultos para crear el modelo, así como la evaluación del comportamiento de roedores adultos, sin tener en cuenta las diferencias de desarrollo que pueden ser importantes para el proceso de la enfermedad. Debido a estos problemas, es importante que la investigación sobre el uso de enfermedad pediátrica no sólo el ajustada desarrollo puntos de tiempo apropiado (por ejemplo, el desarrollo de CNS humano a las 28 - 32 semanas es equivalente a un día post-natal 2-7 días roedor) 7, sino también las pruebas de comportamiento que se examinarán de motor adecuada, los comportamientos del desarrollo sensorial o reflexivos. Por lo tanto, a medida que se desarrolla cada nuevo modelo de enfermedad neonatal, debe ser probado rigurosamente para asegurar que el celulary las respuestas de comportamiento proporcionarán los datos traducibles más adecuadas entre roedores y humanos. La parálisis cerebral es un trastorno motor, que persisten en la edad adulta. Un problema con muchos de los modelos disponibles en la actualidad la parálisis cerebral es la falta de pruebas de motor repetible, estandarizada que puede correlacionarse con los déficits observados en los pacientes pediátricos. En este nuevo modelo, que combina la hipoxia, la isquemia y la inflamación en un ratón neonatal, el comportamiento motor se evaluó usando una batería de pruebas específicas para ratones neonatales. Con el fin de disminuir la subjetividad y aumentar la información cuantitativa, varias pruebas han sido modificados para incluir muy específico, pero fácil de evaluar las medidas que pueden ser estandarizados. Además, las evaluaciones delanteros y los miembros posteriores se pueden realizar por separado, y las diferencias de izquierda / derecha se pueden determinar. Esta batería de pruebas es específica para ratones recién nacidos hasta dos semanas de edad. Este modelo demuestra CPdificultad para caminar (ambulación, ángulo del pie de las extremidades posteriores), así como la debilidad de las extremidades específica (suspensión de cuatro extremidades, suspensión posterior), y los déficit en los reflejos del desarrollo (agarrar reflejo). Aunque en este estudio se examinó sólo un punto de tiempo, estos déficits pueden ser rastreados a través del tiempo. Hay otras series de pruebas que pueden utilizarse en el recién nacido, tales como la batería del Fox de pruebas o evaluación de hitos del desarrollo de 15 Heyser. Sin embargo, estas pruebas comparan el recién nacido al adulto, cuyas respuestas no son necesariamente lo mismo porque el recién nacido aún está en desarrollo. La batería de Fox y pruebas Assement de Heyser se basan en información subjetiva de observación con evaluación dicotómica (sí o no), en lugar de los datos objetivos (ángulo de la postura, sobre la base de la fuerza, etc). Debido a la subjetividad de estas pruebas, muchos científicos se han adaptado, añadido o eliminado criterios, con lo que sus resultados incomparables a los demás y límiteing la utilidad de los datos en términos de establecer un déficit línea de base para una enfermedad o trastorno particular. Mediante el establecimiento de un conjunto de pruebas de motor estandarizados que son cualitativa y específicamente diseñado para probar los recién nacidos, los resultados de los grupos de investigación individuales pueden ser precisa y fiable y se comparan.

El desarrollo de nuevos modelos de lesión o enfermedad pediátrica usando roedores es a menudo difícil debido a la sorprendente capacidad de ratas y ratones a recuperarse rápidamente de la lesión neurológica. Por lo tanto, con el fin de validar cualquier nuevo modelo de enfermedad pediátrica, examinando a fondo los cambios celulares y moleculares deben ir mano a mano con los resultados conductuales. En muchos sentidos, la recuperación funcional de la conducta puede ser más importante que los cambios celulares subyacentes en términos de relevancia terapéutica o de traslación. A medida que los investigadores aprenden más sobre lesiones en el adulto y el recién nacido, es evidente que sus respuestas son muy diferentes y no se pueden extrapolar entre los dos. Por ejemplo, los ratones neonatales muestran diferentes niveles de factor de crecimiento nervioso, factor neurotrófico derivado del cerebro, neurotrofina-3 y el factor neurotrófico derivado de la línea celular glial después de una lesión de la médula espinal 1,2. Además, los recién nacidos tienen significativa permeabilidad de la barrera hematoencefálica después del accidente cerebrovascular 3, demonstrate reordenamiento neurona cortical después de la lesión del nervio periférico 4, y tienen una astrogliosis retrasado o desacelerado después de una lesión de la médula espinal y la hipoxia-isquemia 5,6. Por lo tanto, es importante que la investigación pediátrica de traslación utilizar modelos de desarrollo equivalentes y que esos modelos se evalúan para los cambios moleculares / celulares y pruebas de comportamiento apropiadas para la edad. La parálisis cerebral (PC) es un trastorno motor que afecta a 3: 1000 nacidos vivos al año (NIH). Los niños con CP presentan una serie de síntomas y condiciones comórbidas, dependiendo de la gravedad de la enfermedad. Dificultad con el movimiento y la coordinación son los signos más comunes, junto con los retrasos en alcanzar los hitos del desarrollo motor. Otros signos incluyen tono muscular anormal (ya sea aumentado o disminuido), la reducción de las habilidades motoras finas, dificultad para caminar, tragar y babeo excesivo y retrasos en el habla (NIH). La causa subyacente de CP se cree que esuna falta de oxígeno y / o el flujo de sangre al cerebro durante el período de pre- o periparto, o hasta un año después del parto. Además, la inflamación es ahora cree que es un componente clave en el desarrollo de CP. La mayoría de los casos de CP están asociados con daño de la sustancia blanca alrededor de los ventrículos, conocidas como leucomalacia periventricular (PVL). Este sello neurológica sugiere que la lesión inicial que conduce a CP se produce durante el período de desarrollo del cerebro cuando los oligodendrocitos son más vulnerables a los insultos. El período de crecimiento rápido de los oligodendrocitos en un ser humano, también el período en que los oligodendrocitos son los más susceptibles a las lesiones, es entre 24 - 32 semanas de gestación. En los roedores, el período equivalente es días postnatales 2 - 7 7, y es cuando se induce CP en este modelo. El modelo de ratón neonatal de CP que se utilizó para realizar las pruebas descritos aquí combina la hipoxia e isquemia con la inflamación para crear un injUry que mejor imita la neurodegeneración ven en CP humano. Este modelo aborda algunas de las principales deficiencias observadas en otros modelos animales de CP, que carecen de los déficits distintos de motor que se asemeja a los pacientes con PC humanos, así como distintos daño de la materia blanca. Estudios previos realizados por un colaborador con el mismo modelo han demostrado que la adición de la inflamación aumenta el daño de la sustancia blanca, por lo tanto emular mejor el PVL visto en niños con parálisis cerebral 8. Sobre la base de los datos anteriores, este trabajo presenta una batería de pruebas de motor neonatales con el fin de evaluar los cambios en la conducta motora como las edades de los animales.

<table border="0" cellpadding="0" cellspacing="0" width="1354">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">C57BL/6 mice</td>
			<td style="width:239px;">Charles River Laboratories</td>
			<td style="width:103px;">STRAIN CODE: 027&#160;</td>
			<td style="width:741px;">C57BL/6NCrl is the exact strain we use</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:271px;">Anesthesia Dish, PYREX&#8482; Crystallizing Dish</td>
			<td style="width:239px;">Corning Life Sciences Glass&#160;</td>
			<td style="width:103px;">3140125</td>
			<td style="width:741px;">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 height="20">
			<td height="20" style="height:20px;width:271px;">Covered lead ring</td>
			<td style="width:239px;">Fisher Scientific</td>
			<td style="width:103px;">S90139C</td>
			<td style="width:741px;">Lead ring for stablizing flasks in a water bath. It is used inside the anesthesia dish.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Scalpel Blade #11</td>
			<td style="width:239px;">World Precision Instrucments, Inc.</td>
			<td style="width:103px;">500240</td>
			<td style="width:741px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Small Vessel Cauterizer</td>
			<td style="width:239px;">Fine Science Tools</td>
			<td style="width:103px;">18000-00</td>
			<td style="width:741px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Micro Hook</td>
			<td style="width:239px;">Fine Science Tools</td>
			<td style="width:103px;">10064-14</td>
			<td style="width:741px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Vetbond Suture Glue</td>
			<td style="width:239px;">3M</td>
			<td style="width:103px;">1469SB</td>
			<td style="width:741px;">n-butyl cyanoacrylate adhesive</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Lipopolysaccharide</td>
			<td style="width:239px;">Sigma Life Science</td>
			<td style="width:103px;">L4391</td>
			<td style="width:741px;">Lipopolysaccaride from e.coli 0111:B4, gamma irradiated</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:271px;">12x12 inch opaque box</td>
			<td style="width:239px;">Acrylic Display Manufacturing: A division of Piasa Plastics</td>
			<td style="width:103px;">C4022</td>
			<td>Colored Acrylic 5-Sided Cube, 3/16&#34; Colored Acrylic, 12&#34;W x 12&#34;D x 12&#34;H;&#160; http://www.acrylicdisplaymfg.com/html/cubes_19.html</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:271px;">Camera/camcorder</td>
			<td style="width:239px;">JVC</td>
			<td style="width:103px;">GC-PX100BUS</td>
			<td style="width:741px;">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 height="20">
			<td height="20" style="height:20px;width:271px;">Covidien Tendersorb&#8482; Underpads</td>
			<td style="width:239px;">Kendall Healthcare Products Co</td>
			<td style="width:103px;">7174</td>
			<td style="width:741px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">WypAll L40</td>
			<td style="width:239px;">Kimberly-Clark Professional</td>
			<td style="width:103px;">5600</td>
			<td style="width:741px;">Any surface with moderate grip will do</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Surface at 45 degree incline</td>
			<td style="width:239px;"></td>
			<td style="width:103px;"></td>
			<td style="width:741px;">We use a cardboard box.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Thin wire from a pipe cleaner</td>
			<td style="width:239px;">Creatology</td>
			<td style="width:103px;">M10314420</td>
			<td style="width:741px;">Any pipe cleaner from any craft store will work.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">50mL conical tube</td>
			<td style="width:239px;">Falcon</td>
			<td style="width:103px;">352070</td>
			<td style="width:741px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Fiberglass Screen Wire</td>
			<td style="width:239px;">New York Wire&#160; www.lowes.com</td>
			<td style="width:103px;">14436</td>
			<td style="width:741px;">Any supplier can be used as long as their screen is 16x16 or 18x16</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:271px;">Razor blade</td>
			<td style="width:239px;">Fisherbrand</td>
			<td style="width:103px;">12-640</td>
			<td style="width:741px;">A wooden stick applicator or wooden part of a cotton-tipped swab will also work.</td>
		</tr>
		<tr height="100">
			<td height="100" style="height:100px;width:271px;">OPTIX 24-in x 4-ft x 0.22-in Clear Acrylic Sheet to make Clear Acrylic Walkway</td>
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			<td style="width:741px;">Clear acrylic (1/8&#34; 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&#34;x2&#34;; (3) 2&#34;x 18&#34;; (1) 2&#34;x15.5&#34;; (1) 2&#34;x3&#34;. Using clear tape, tape all sides together, with the 15.5&#34; piece on top. Tape the 3&#34; piece to the end of the 15.5&#34; 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>
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			<td height="20" style="height:20px;width:271px;">Protractor</td>
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</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.

Los ratones se pusieron a prueba de P7 (24 hr después de la cirugía) para P13 (1 semana después de la cirugía), utilizando ratones diferentes para cada punto de tiempo de manera que el aprendizaje de un paradigma de pruebas no era una variable de confusión. P8 fue seleccionado como resultados representativos, como los ratones mostraron los mayores déficits en este punto temporal. La transición de gatear a caminar es retardado en ratones recién nacidos CP los pacientes con PC humanos tienen alteraciones de la marcha, que van desde caminar de puntillas a una marcha en tijera. Como este modelo CP pantallas de la marcha y los déficits similares a los humanos, se evaluó la deambulación. Los ratones se obtuvieron de la uniformidad de la marcha y el movimiento de las extremidades-pata recta durante un paseo. A las 48 horas después de la cirugía (PND 8), los ratones tenían menos CP movimiento de las extremidades simétricas y un &quot;rastreo&quot; de la marcha, en comparación con sus homólogos sham (puntuación media deambulación: CP 1.083 ±0,6337, n = 12 vs simulacro 1,639 ± 0,4859, n = 9; p &lt;0,05, Figura 10). Por una semana, tanto CP y ratones sham han hecho la transición a caminar (datos no mostrados). <img alt="Figura 10" src="/files/ftp_upload/53569/53569fig10.jpg" /> Figura 10. CP de ratón Los cachorros no Ambulate, así como Shams. Ratones Sham (barra de color negro) tienen una puntuación media de 1.639 ± 0.4859 (n = 9), es decir, su desarrollo se sitúa entre ambulatoria movimiento de las extremidades asimétricas y rastreo lento. ratones CP (barra gris) reciben una puntuación media de 1.083 ± 0.6337 (n = 12), es decir, su deambulación está menos desarrollada y tienden a tener movimiento de las extremidades asimétrica. Los datos se expresan como media ± SEM; * Es p &lt;0,05. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53569/53569fig10large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> Hindlimb Pie de ángulo se incrementa en CP Además de la deambulación, se evaluó extremidad posterior del pie de ángulo. Ocho días de edad, los cachorros ratón del impostor caminar con sus patas traseras hacia adelante, en comparación con los ratones HIL, que han extendidos patas traseras al caminar en una línea recta (Figura 2; ángulo medio: CP 77,48 ± 9,848, n = 9, vs simulacro 54.54 ± 8,043, n = 11; p &lt;0,0001, Figura 11). Este aumento del ángulo se correlaciona con la inestabilidad de la marcha, en la que los cachorros necesitan para aumentar el ángulo de sus patas traseras con el fin de estabilizar su marcha y ayudar con el equilibrio y la coordinación. <img alt="Figura 11" src="/files/ftp_upload/53569/53569fig11.jpg" /> Figura 11. CP ratón Los cachorros Splay sus patas traseras al caminar. Ratones CP (barras negras) tienen un ángulo medio entre sus miembros traseros de 77,48 ± 8,043 (n = 11), mientras que los ratones sham (b gris<html> ARS) tienen un ángulo promedio de 54,54 ± 9,848 (n = 9). Los datos se expresan como media ± SEM; **** Es p &lt;0,0001. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53569/53569fig11large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> CP Los ratones no presentan déficit cuando superficial de enderezamiento Se incluyó la prueba de enderezamiento superficie, ya que algunos pacientes con PC tienen problemas de control del tronco (Heyrman et al., 2013). Además, el sistema vestibular es necesario detectar la necesidad de corregir y hay déficit vestibular en algunos pacientes CP 23. ratones CP no muestran déficits significativos cuando enderezamiento en comparación con controles simulados (datos no mostrados). Los ratones CP realizar la misma como Sham en negativo Prueba geotaxis t &quot;fo: keep-together.within-page =&quot; 1 &quot;&gt; geotaxis negativo se utiliza para probar la coordinación motora en ratones jóvenes cachorros son desafiados por ser el lugar que va cuesta abajo sobre una superficie inclinada retraso o falta de orientar hacia arriba podría indicar déficits.. en coordinación, el equilibrio, o la entrada vestibular. ratones CP no muestran déficits cuando fueron estimulados con geotaxis negativos en comparación con los ratones sham (datos no mostrados). Además, los ratones CP no mostraron una preferencia a girar hacia un lado frente a otro cuando se re-orientación . Frente-Limb suspensión de ensayo es apropiado para ratones más viejos de 10 Días los pacientes con PC han disminución del tono muscular y los déficits en las habilidades motoras finas, tales como agarrar. Para probar la debilidad en este modelo de ratón, se utilizó una prueba de suspensión delantera de las extremidades. Por otra parte, este modelo utiliza la lesión isquémica unilateral y bilateral-dad se pudo determinar el uso de esta prueba de suspensión. Esta pruebaes mejor para los ratones de más de 10 días 15. A los 8 días de edad, dos días después de la lesión, no había diferencias significativas entre CP y ratones sham (datos no mostrados). La fuerza de las extremidades posteriores se reduce en ratones CP los pacientes con PC humanos a menudo necesitan aparatos ortopédicos o dispositivos de soporte para caminar debido a la falta de control del motor y la fuerza. Con el fin de comparar el modelo CP de roedores a los seres humanos, la fuerza del miembro posterior se evaluó mediante la prueba de suspensión posterior. Cuando suspendido desde el lado de un tubo cónico, los ratones CP mostraron debilidad de las extremidades posteriores, como se demuestra por una disminución en colgando puntuación (miembro posterior puntuación colgando: CP 3,468 ± 0,5561, n = 13, vs sham 3,891 ± 0,1329, n = 13; p &lt; 0,05, Figura 12). No se observó ninguna diferencia en el tiempo de suspensión de las extremidades posteriores (datos no mostrados). Por lo tanto, similar a los pacientes CP humanos, ratones CP demuestran hindlIMB (pierna) debilidad. <img alt="Figura 12" src="/files/ftp_upload/53569/53569fig12.jpg" /> Figura 12. Los ratones Sham son ligeramente, pero significativamente más fuerte en sus patas traseras que los ratones CP. En una nota media que cuelga de 3,891 ± 0,1329 (n = 13), los ratones sham (barra de color negro) muestra una separación más extremidad posterior, y por lo tanto una postura más fuerte de las extremidades posteriores, al colgar en el borde de un tubo que los ratones CP (barra gris) con una puntuación media que cuelga de 3.468 ± 0.5561 (n = 13). Los datos se expresan como media ± SEM; * Es p &lt;0,05. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53569/53569fig12large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> La fuerza de agarre se reduce después de una lesión CP Agarrando con las cuatro patas es importante para un roedor entérminos de escalada y correr sobre superficies irregulares. Grip requiere fuerza sostenida significativa, en lugar de destreza o fuerza lineal, principalmente en los dígitos y las patas 24. Los ratones fueron requeridos para mantener su peso corporal en una pantalla de malla de alambre invertida. Ratones CP no fueron capaces de mantener su agarre y estos ratones cayeron en ángulos significativamente más bajos (cuatro extremidades ángulo medio: CP 75.627 ± 24.48, n = 11, vs sham 96,57 ± 10.836, n = 9; p &lt;0,05, Figura 13). Estos datos muestran que hay un déficit significativo en la fuerza de agarre en ratones CP. <img alt="Figura 13" src="/files/ftp_upload/53569/53569fig13.jpg" /> Figura 13. Los ratones tienen CP Grip Más débil que Shams. Ratones Sham (barra de color negro) puede captar un ángulo invertido promedio de 96.57 ± 10.836 (n = 9). ratones CP (barra gris) sólo pueden llegar a un ángulo invertido de 75.627 ± 24,48 (n = 11). Los datos se expresan como media ± SEM; * Es p0; 0.05. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53569/53569fig13large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> Los déficits aferramiento reflejas son evidentes en ratones CP Junto con los déficits de motricidad gruesa, motricidad fina también se vean afectados en pacientes con PC 25,26. El reflejo de agarre en el ser humano está presente al nacer y desaparece alrededor de 5 - 6 En meses. Sin embargo, los cambios en el reflejo de agarre, tales como la velocidad exagerada o la fuerza de agarre, la falta de agarrar, o la reaparición del reflejo de agarre después de 6 meses de edad, todos indican daño en el sistema nervioso. Para comparar captar en el modelo de CP, se determinaron los déficit de agarre reflexivos. A las 48 horas después de la lesión, los ratones CP demuestran una disminución de agarrar refleja (patas medias agarrarona las 48 horas: CP 2,429 ± 0,9376, n = 14, vs simulado 3.214 ± 0,8018, n = 14; p &lt;0,05, Figura 14A). Hubo un aumento ligero, pero no significativo en la preferencia de la pata derecha en las patas delanteras (datos no mostrados). Hubo una preferencia pata derecha significativa en las patas traseras (CP 75.0 ± 42.74, n = 14, vs simulacro 17.86 ± 54.09, n = 14; p &lt;0,005, Figura 14B). Una semana después de la lesión, los ratones CP muestran déficits de agarre (patas medias captados en la semana 1: CP 2,75 ± 1,035, n = 8, vs simulacro 3,80 ± 0,6325, n = 10; p &lt;0,05, Figura 14C), sin preferencia pata notable . <img alt="Figura 14" src="/files/ftp_upload/53569/53569fig14.jpg" /> Figura 14. Los ratones CP tienen déficit agarrar, en la patas traseras, contralateral a la región del cerebro lesionado. (A) 48 horas después de la lesión (PND 8), los ratones CP (barra gris) agarrar un palo con, en promedio, un menor número de patas que anima farsals (barra de color negro). ratones (B) CP (barra gris) muestran una preferencia para agarrar con la pata trasera derecha (contralateral a la lesión) en lugar de utilizar la pata trasera izquierda (ipsilateral a la lesión). ratones Sham (barra de color negro) no muestran esta preferencia pata derecha. Preferencia pata derecha se calcula como ([pata derecha - izquierda pata] / [pata derecha + pata izquierda + ambas patas] * 100) (C) Una semana después de la lesión, los ratones CP (barra gris) muestran todavía agarrando déficit en comparación con. Shams (barra de color negro). Los datos se expresan como media ± SEM; * Es p &lt;0,05, ** es p &lt;0,005. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53569/53569fig14large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> Los ratones CP se alejan del borde del acantilado Durante Aversión La prueba se basa en la aversión al precipicio del miedo inherente de los ratones para convertir awaY desde un acantilado y se dirigen hacia la seguridad. Aunque algunos pacientes con PC tienen dificultades vestibulares, así como problemas de control del motor, los ratones CP no mostraron ningún déficit en esta prueba.

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 batería de pruebas de motor en un modelo de ratón neonatal de la parálisis cerebral

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

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

Research

JoVE Journal

Medicine

24.0K vistas.  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

Making Sense of Listening: The IMAP Test Battery

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the abilities to extract and use the information encoded in the auditory signal. We refer to these as listening skills (or auditory processing AP). Deficits in these skills are associated with delayed language and literacy development, though the nature of the relevant deficits and their causal connection with these delays is hotly debated.
When a child is referred to a health professional with normal hearing and unexplained difficulties in listening, or associated delays in language or literacy development, they should ideally be assessed with a combination of psychoacoustic (AP) tests, suitable for children and for use in a clinic, together with cognitive tests to measure attention, working memory, IQ, and language skills. Such a detailed examination needs to be relatively short and within the technical capability of any suitably qualified professional. Current tests for the presence of AP deficits tend to be poorly constructed and inadequately validated within the normal population. They have little or no reference to the presenting symptoms of the child, and typically include a linguistic component. Poor performance may thus reflect problems with language rather than with AP. To assist in the assessment of children with listening difficulties, pediatric audiologists need a single, standardized child-appropriate test battery based on the use of language-free stimuli.
We present the IMAP test battery which was developed at the MRC Institute of Hearing Research to supplement tests currently used to investigate cases of suspected AP deficits. IMAP assesses a range of relevant auditory and cognitive skills and takes about one hour to complete. It has been standardized in 1500 normally-hearing children from across the UK, aged 6-11 years. Since its development, it has been successfully used in a number of large scale studies both in the UK and the USA. IMAP provides measures for separating out sensory from cognitive contributions to hearing. It further limits confounds due to procedural effects by presenting tests in a child-friendly game-format. Stimulus-generation, management of test protocols and control of test presentation is mediated by the IHR-STAR software platform. This provides a standardized methodology for a range of applications and ensures replicable procedures across testers. IHR-STAR provides a flexible, user-programmable environment that currently has additional applications for hearing screening, mapping cochlear implant electrodes, and academic research or teaching.

A test battery (IMAP) for performing an in-depth assessment of auditory and cognitive abilities contributing to listening skills is described. It is quick to administer, child-friendly and free from linguistic confounds. Stimulus generation and protocol management are controlled via a software platform (IHR-STAR) to ensure replicable procedures.

Tener sentido de la escucha: la batería de pruebas IMAP

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the ...

Investigación

Medicina

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

Modelo de ratón de la oclusión arteria cerebral media

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Multi-fotón imágenes de invasión de células tumorales en un modelo de ratón ortotópico de carcinoma oral de células escamosas

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

A Swine Model of Neonatal Asphyxia

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, perfusion deficit, hypoxic-ischemic encephalopathy, pulmonary hypertension, vasculopathic enterocolitis, renal failure and thrombo-embolic complications. Animal models are developed to help us understand the patho-physiology and pharmacology of neonatal asphyxia. In comparison to rodents and newborn lambs, the newborn piglet has been proven to be a valuable model. The newborn piglet has several advantages including similar development as that of 36-38 weeks human fetus with comparable body systems, large body size (&tilde;1.5-2 kg at birth) that allows the instrumentation and monitoring of the animal and controls the confounding variables of hypoxia and hemodynamic derangements.
We here describe an experimental protocol to simulate neonatal asphyxia and allow us to examine the systemic and regional hemodynamic changes during the asphyxiating and reoxygenation process as well as the respective effects of interventions. Further, the model has the advantage of studying multi-organ failure or dysfunction simultaneously and the interaction with various body systems. The experimental model is a non-survival procedure that involves the surgical instrumentation of newborn piglets (1-3 day-old and 1.5-2.5 kg weight, mixed breed) to allow the establishment of mechanical ventilation, vascular (arterial and central venous) access and the placement of catheters and flow probes (Transonic Inc.) for the continuously monitoring of intra-vascular pressure and blood flow across different arteries including main pulmonary, common carotid, superior mesenteric and left renal arteries. Using these surgically instrumented piglets, after stabilization for 30-60 minutes as defined by Z&lt;10% variation in hemodynamic parameters and normal blood gases, we commence an experimental protocol of severe hypoxemia which is induced via normocapnic alveolar hypoxia. The piglet is ventilated with 10-15% oxygen by increasing the inhaled concentration of nitrogen gas for 2h, aiming for arterial oxygen saturations of 30-40%. This degree of hypoxemia will produce clinical asphyxia with severe metabolic acidosis, systemic hypotension and cardiogenic shock with hypoperfusion to vital organs. The hypoxia is followed by reoxygenation with 100% oxygen for 0.5h and then 21% oxygen for 3.5h. Pharmacologic interventions can be introduced in due course and their effects investigated in a blinded, block-randomized fashion.

Large animal models have good translational values in the examination of physiology and pharmacology of neonatal asphyxia. Using newborn piglets, we develop an experimental protocol to simulate neonatal asphyxia which has advantages of studying the systemic and regional hemodynamics, oxygen transport with biochemical and pathologic pathways and correlations.

Un modelo porcino de la asfixia neonatal

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, ...

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

No invasiva de medición óptica del metabolismo cerebral en lactantes y Hemodinámica

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

Un modelo del cochinillo de Neonatología encefalopatía hipóxico-isquémica

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

Assessment of Perigenital Sensitivity and Prostatic Mast Cell Activation in a Mouse Model of Neonatal Maternal Separation

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under the age of 50, yet it is the least understood and studied chronic pelvic pain disorder. A significant subset of patients with chronic pelvic pain report having experienced early life stress or abuse, which can markedly affect the functioning and regulation of the hypothalamic-pituitary-adrenal (HPA) axis. Mast cell activation, which has been shown to be increased in both urine and expressed prostatic secretions of CP/CPPS patients, is partially regulated by downstream activation of the HPA axis. Neonatal maternal separation (NMS) has been used for over two decades to study the outcomes of early life stress in rodent models, including changes in the HPA axis and visceral sensitivity. Here we provide a detailed protocol for using NMS as a preclinical model of CP/CPPS in male C57BL/6 mice. We describe the methodology for performing NMS, assessing perigenital mechanical allodynia, and histological evidence of mast cell activation. We also provide evidence that early psychological stress can have long-lasting effects on the male urogenital system in mice.

We are measuring perigenital mechanical sensitivity and mast cell activation in the prostate of male C57BL/6 mice that underwent an early life stress paradigm &#8211; neonatal maternal separation, in order to induce a preclinical model of chronic prostatitis/chronic pelvic pain syndrome.

Evaluación de perigenital Sensibilidad y Prostática mástil activación celular en un modelo de ratón de la separación materna neonatal

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a lifetime prevalence of 14% and is the most common urological diagnosis for men under 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 ratón neonatal lesión de médula espinal de compresión Modelo

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

Myocardial Infarction in Neonatal Mice, A Model of Cardiac Regeneration

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and regeneration, and to define new targets for therapeutics. For decades, models of complete heart regeneration existed in amphibians and fish, but a mammalian counterpart was not available. The recent discovery of a postnatal window during which mice possess regenerative capabilities has led to the establishment of a mammalian model of cardiac regeneration. A surgical model of mammalian cardiac regeneration in the neonatal mouse is presented herein. Briefly, postnatal day 1 (P1) mice are anesthetized by isoflurane and placed on an ice pad to induce hypothermia. After the chest is opened, and the left anterior descending coronary artery (LAD) is visualized, a suture is placed around the LAD to inflict myocardial ischemia in the left ventricle. The surgical procedure takes 10-15 min. Visualizing the coronary artery is crucial for accurate suture placement and reproducibility. Myocardial infarction and cardiac dysfunction are confirmed by triphenyl-tetrazolium chloride (TTC) staining and echocardiography, respectively. Complete regeneration 21 days post myocardial infarction is verified by histology. This protocol can be used to as a tool to elucidate mechanisms of mammalian cardiac regeneration after myocardial infarction.

This protocol describes a highly reproducible model of cardiac regeneration by surgical induction of myocardial infarction in the left ventricle of postnatal day 1 mice. The method involves induction of hypothermic anesthesia and ligation of the left anterior descending coronary artery.

El infarto de miocardio en ratones recién nacidos, un modelo de regeneración cardiaca

Myocardial infarction induced by coronary artery ligation has been used in many animal models as a tool to study the mechanisms of cardiac repair and ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

Transitoria de la arteria cerebral media oclusión Modelo de accidente cerebrovascular neonatal en ratas P10

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...