Este trabalho foi apoiado pelo National Institutes of Health subvenção, R01 AG033620 (BG) e por Robert Draper Inovação Tecnológica Financiamento (DAW).

1. Construção do Dispositivo HIT <ol><li> Fixe a mola para a placa utilizando dois grampos e quatro parafusos (Figura 1A). Centralize os grampos em relação à largura da placa e Butt-los uns contra os outros com o flush braçadeira exterior com a borda da placa. Antes de fixar os grampos, dobre-os usando um alicate para caber firmemente sobre a primavera. NOTA: Consulte a Tabela 1 para obter descrições dos materiais necessários para a construção do dispositivo de HIT. A extremidade apertada da mola deve ser de 1/8 de polegada (3,2 mm) a partir da borda da placa, ea extremidade livre deve estender-se sobre a placa por 3/4 de polegada (19 mm). Ajustar a mola de modo que ela se encontra em paralelo com o comprimento do bordo. </li><li> Enrole a extremidade livre da mola em torno de uma vez com a fita adesiva de loops de velcro. A borda externa do velcro deve estar nivelada com a extremidade da mola. O Velcro é importante porque é usado para proteger o frasco para a mola por CREAting um encaixe de compressão apertado. O velcro permite também uma fácil ligação e remoção de tubos de ensaio, permitindo que muitos frascos a serem processados ​​num curto período de tempo. </li><li> Coloque a tampa do balde de gelo de cabeça para baixo, centrado, apertado contra a placa de madeira. Orientar a região elevada da tampa balde de gelo de modo a que a borda é paralelo ao longo da largura do tabuleiro. Note-se que a região elevada do balde de gelo é de 1/2 polegada (13 mm) maior do que a placa de madeira, de modo que quando um frasco é ligado à mola a mola não vou mentir plana no tabuleiro. </li><li> Deslize todo o dispositivo contra um objeto fixo, como uma parede, de modo que a tampa balde de gelo está encravado entre o conselho eo objeto e não se move. </li><li> Tape o transferidor de papel para o fundo de um cartão voar bandeja frasco e coloque na borda contra a extensão do tabuleiro de modo que a marca de 90 ° está alinhado com a primavera, quando ele é puxado de volta para uma posição perfeitamente vertical. </li></ol> 2.O funcionamento do dispositivo HIT <ol><li> Lugar entre 1 e 60 CO 2 -anesthetized voa em um frasco vazio e tapar o frasco usando uma bola de algodão apertada. </li><li> Confinar as moscas para a parte inferior de 1 polegada (2,5 cm) do frasco, empurrando a bola de algodão para dentro do frasco até que seja de 1 polegada (2,5 cm) a partir do fundo. É útil para desenhar uma linha no frasco na marca de 1 polegada (2,5 cm). Note-se que as moscas para confinar regiões maiores ou menores do frasco pode afectar a gravidade dos fenótipos. </li><li> Esperar 5 min para as moscas para recuperar a mobilidade do CO 2. Note-se que não se sabe se 5 min é suficiente para eliminar completamente os efeitos de CO 2. </li><li> Inserir a extremidade da mola para dentro do frasco até que a extremidade interior do Velcro é nivelada com o topo do frasco (Figura 1B). Quando a primavera está deitado, de 1 polegada (2,5 cm) do frasco para injectáveis ​​deve sobrepor-se a região elevada da tampa do balde de gelo. Os frascos podem ser reutilizados mqualquer hora. </li><li> Ao sentar-se, mantenha o frasco a região de Velcro usando o polegar eo dedo indicador de sua mão esquerda. Segure a placa firmemente à bancada usando sua mão direita. Como alternativa, use C-braçadeiras para segurar a placa apertada para a bancada. </li><li> Puxe a mola perfeitamente em linha reta de volta para o ângulo desejado. Solte a primavera. Permitir que a primavera chegar a uma paragem completa. </li><li> Retire o recipiente da primavera e permitir que as moscas se recuperar para ≥5 min. Apresentar as moscas para outra greve ou transferir as moscas para um frasco com alimentos mosca. NOTA: Uma variedade de ensaios pode ser utilizada para avaliar os efeitos fenotípicos de ataques do dispositivo HIT. Por exemplo, os efeitos sobre a longevidade pode ser determinada através da análise da percentagem de moscas que sobrevivem às vezes após a lesão, os efeitos na morfologia do cérebro pode ser determinada por análise histológica da cabeça, e os efeitos sobre a expressão do gene pode ser determinada por análise quantitativa dos níveis de ARNm 10. </li><li> Determine efeitos do procedimento que não são devido a greves, tratando de forma idêntica moscas de controle que não são sujeitos a ataques. Usar protecção auricular, porque o impacto do frasco contra a cobertura balde de gelo produz um ruído alto. </li></ol>

<ol>
	<li>Harrison-Felix, C. L., Whiteneck, G. G., Jha, A., DeVivo, M. J., Hammond, F. M., Hart, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mortality+over+four+decades+after+traumatic+brain+injury+rehabilitation:+A+retrospective+cohort+study.">Mortality over four decades after traumatic brain injury rehabilitation: A retrospective cohort study.</a> Arch Phys Med Rehabil. 90, 1506-1513 (2009).</li><li>Coronado, V. G., 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=Surveillance+for+traumatic+brain+injury-related+deaths+&#8211;+United+States.">Surveillance for traumatic brain injury-related deaths &#8211; United States.</a> MMWR Surveill Summ. 60, 1-32 (1997).</li><li>Masel, B., DeWitt, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traumatic+brain+injury:+A+disease+process,+not+an+event.">Traumatic brain injury: A disease process, not an event.</a> J. Neurotrauma. 27, 1529-1540 (2010).</li><li>Blennow, K., Hardy, J., Zetterberg, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neuropathology+and+neurobiology+of+traumatic+brain+injury.">The neuropathology and neurobiology of traumatic brain injury.</a> Neuron. 76, 886-899 (2012).</li><li>Prins, M., Greco, T., Alexander, D., Giza, C. 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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unique+challenges+in+clinical+trails+in+traumatic+brain+injury.">Unique challenges in clinical trails in traumatic brain injury.</a> Crit Care Med. 37, S129-S135 (2009).</li><li>Xiong, Y., Mahmood, A., Chopp, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+traumatic+brain+injury.">Animal models of traumatic brain injury.</a> Nature Rev Neurosci. 14, 128-142 (2013).</li><li>Balsiger, Z., Leudkte, J., Mawer, S., Willey, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> HIT device high speed analysis. , (2014).</li><li>Davceva, N., Janevska, V., Illevski, B., Petrushevska, G., Popeska, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+occurrence+of+acute+subdural+haematoma+and+diffuse+axonal+injury+as+two+typical+acceleration+injuries.">The occurrence of acute subdural haematoma and diffuse axonal injury as two typical acceleration injuries.</a> J Forensic Leg Med. 19, 480-484 (2012).</li><li>Katzenberger, R. J., Loewen, C. A., Wassarman, D. R., Petersen, A. J., Ganetzky, B., Wassarman , D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila.+model+of+closed+head+traumatic+brain+injury.">A Drosophila. model of closed head traumatic brain injury.</a> Proc Natl Acad Sci USA. 110, E4152-E4159 (2013).</li><li>Fang, Y., Bonini, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axon+degeneration+and+regeneration:+insights+from+Drosophila+.models+of+nerve+injury.">Axon degeneration and regeneration: insights from Drosophila .models of nerve injury.</a> Annu Rev Cell Biol. 28, 575-597 (2012).</li><li>Babcock, D. 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Não temos conflitos de interesse para divulgar.

O método dispositivo HIT se distingue de outros métodos que infligem lesões traumáticas em moscas pelo fato de que ele faz com que a cabeça fechado, em vez de penetrar TBI 11. Além disso, o método de dispositivo HIT leva menos tempo, esforço e habilidade para infligir TBI em muitas moscas, por isso, o método é mais favorável do que outros métodos para telas genéticos de grande escala. Por fim, o facto de as lesões primárias infligidos pelo dispositivo HIT não estão limitados ao cérebro é ao...

An author's name was corrected in the publication of <a href="http://www.jove.com/video/52905">A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila</a>. The second to last author's was spelled incorrectly. It has been updated from:
Barry Ganetky
to:
Barry Ganetzky

An author's name was corrected in the publication of <a href="http://www.jove.com/video/52905">A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila</a>.

Erratum: A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Traumatismo crânio-encefálico (TCE) é definida como lesão no cérebro de uma força mecânica externa. Mais vulgarmente, a partir de resultados de TCE forças de cabeça fechada, como forças rombas e aceleração inércia e forças de desaceleração que causam o cérebro para atingir o interior do crânio. Nos Estados Unidos, estima-se que 50.000 pessoas morrem a cada ano de TBI e 2,5-6.500.000 indivíduos estão a viver com as consequências do TCE, incluindo debilitante físico, cognitivo e problemas de comportamento 1,2. As conseqüências do TCE não são apenas devido a lesões mecânicas primárias para o cérebro, mas também a lesões celulares e moleculares secundários para o cérebro, bem como outros tecidos que ocorrem ao longo do tempo 3-5. O desenvolvimento de abordagens para diagnosticar e tratar o TCE tem provado ser difícil porque o TCE é um processo de doença complexa. A natureza variável de lesões primárias, fisiologia humana e fatores ambientais resulta em i secundário heterogêneonjuries e patologias. Factores variáveis ​​subjacentes incluem a gravidade da lesão primária, o tempo entre as lesões primárias repetitivas, e da idade e do genótipo do indivíduo. A compreensão de como cada fator variável contribui para as conseqüências de TBI é susceptível de ajudar no desenvolvimento de abordagens para diagnosticar e tratar TBI 6,7. Aqui nós descrevemos um método para infligir TBI cabeça fechada em Drosophila melanogaster (mosca da fruta) que pode ser usado para delinear a contribuição de fatores variáveis ​​para as consequências do TCE. O método baseia-se na observação inicial de que intensamente que bate no lado de um frasco de cultura mosca contra a palma de uma mão causada moscas do tipo selvagem para ficar temporariamente incapacitada, uma consequência provável da TBI. Assim, construímos o dispositivo High-Impact Trauma (HIT) recapitular as forças de aceleração e desaceleração a partir da ação de bater a mão. Um filme de alta velocidade mostra que um único golpe doHIT dispositivo faz com que voa para entrar em contato com a parede do frasco várias vezes com a cabeça eo corpo 8. Em certa medida, todos os contatos são susceptíveis de fazer com que o cérebro da mosca a ricochetear e deformar contra a cápsula cabeça, semelhante ao que acontece com os seres humanos em quedas e acidentes de carro 9. Assim, moscas tratados com os fenótipos visor do dispositivo HIT consistentes com lesão cerebral, incluindo a incapacidade temporária seguido por ataxia, a recuperação gradual da mobilidade, alterações de expressão gênica na cabeça, e neurodegeneração progressiva no cérebro 10. Assim, o dispositivo de HIT torna possível estudar TBI utilizando o enorme arsenal de ferramentas e técnicas desenvolvidas para moscas experimentais.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Zinc plated compression spring</td>
			<td>The Hillman&#160; Group</td>
			<td>540189</td>
			<td>9 7/8 in (length), 15/16 in (outer diameter), 0.12 in (wire size)</td>
		</tr>
		<tr>
			<td>Wooden board</td>
			<td></td>
			<td></td>
			<td>9 in (length), 6.5 in (width), 0.75 in (height)</td>
		</tr>
		<tr>
			<td>Clamps</td>
			<td>Sigma Electrical Manufacturing Corporation</td>
			<td>49822</td>
			<td>3.10 in (length), 0.68 in (width), 1.11 in (height), EMT Two Hole Straps, click on type for 1 inch steel EMT conduit</td>
		</tr>
		<tr>
			<td>Loop half of self-adhesive velcro</td>
			<td></td>
			<td></td>
			<td>3 in (length), (3/4 in width)</td>
		</tr>
		<tr>
			<td>Polyurethane ice bucket cover</td>
			<td>Fisher Scientific</td>
			<td>02-591-45</td>
			<td>9 1/8 in (length), 9 1/8 in (width), 1 1/4 in (height)</td>
		</tr>
		<tr>
			<td>Plastic fly vials</td>
			<td>Applied Scientific</td>
			<td>AS-510</td>
			<td>3 11/16 in (height), 1 1/16 in (inner diameter), 1 1/8 in (outer diameter)</td>
		</tr>
		<tr>
			<td>Large cotton balls</td>
			<td>Fisher Scientific</td>
			<td>22-456-883</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper protractor</td>
			<td></td>
			<td></td>
			<td>10 in (diameter)</td>
		</tr>
	</tbody>
</table>

a method to inflict closed head traumatic brain injury in drosophila

Traumatismo crânio-encefálico (TCE) afeta milhões de pessoas a cada ano, causando comprometimento da saúde física, cognitiva e funções comportamentais e morte. Estudos utilizando Drosophila têm contribuído importantes avanços na compreensão de processos neurológicos. Assim, com o objetivo de compreender a base celular e molecular de patologias TCE em humanos, foi desenvolvido o dispositivo de Trauma de Alto Impacto (HIT) para infligir cabeça fechada TBI em moscas. Moscas submetidos aos fenótipos visor do dispositivo HIT consistentes com TBI humano, tais como incapacidade temporária e neurodegeneração progressiva. O dispositivo HIT usa um mecanismo baseado em mola para impulsionar moscas contra a parede de um tubo de ensaio, provocando danos mecânicos ao cérebro mosca. O dispositivo é barato e fácil de construir, a sua operação é simples e rápida, e que produz resultados reprodutíveis. Por conseguinte, o dispositivo de HIT pode ser combinada com as ferramentas experimentais existentes e técnicas de moscas para tratar fundamentaisdúvidas sobre TCE que podem levar ao desenvolvimento de diagnósticos e tratamentos para o TCE. Em particular, o dispositivo HIT pode ser usado para realizar telas genéticos em grande escala para compreender a base genética de patologias TBI.

Estamos interessados ​​em entender por que as moscas morrem logo após a lesão primária. Para quantificar a morte, determinou-se o Índice de mortalidade às 24 horas (MI 24), que é a percentagem de moscas que morreram dentro de 24 horas da lesão primária. Moscas sujeitos a ataques do dispositivo HIT foram incubadas a 25 ° C num frasco com comida mosca, e o número de moscas mortas foi contado após 24 horas. Utilizou-se esta abordagem para identificar os factores que afectam a MI 24 e de...

Watch this Scientific Journal Video about A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila at JoVE.com

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Aqui nós descrevemos um método para infligir a cabeça fechado traumatismo crânio-encefálico (TCE) em Drosophila. Este método oferece um portal para investigar os mecanismos celulares e moleculares que estão na base patologias TCE, utilizando a vasta gama de ferramentas experimentais e técnicas disponíveis para as moscas.

Um Método para infligir Cabeça Fechada Lesão cerebral traumático em<em&gt; Drosophila</em

Traumatic brain injury (TBI) affects millions of people each year, causing impairment of physical, cognitive, and behavioral functions and death. Studies ...

a-method-to-inflict-closed-head-traumatic-brain-injury-in-drosophila

Research

JoVE Journal

Neuroscience

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

11.4K visualizações.  University of Wisconsin-Madison.  Aqui nós descrevemos um método para infligir a cabeça fechado traumatismo crânio-encefálico (TCE) em Drosophila. Este método oferece um portal para investigar os mecanismos celulares e moleculares que estão na base patologias TCE, utilizando a vasta gama de ferramentas experimentais e técnicas disponíveis para as moscas. 

Vídeo: A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic Brain Injury (mTBI). The mechanisms responsible for its complex pathological changes remain largely elusive, despite a recent consensus to define the neuropathological criteria. Here, we describe a novel method to develop a model of CTE in Drosophila melanogaster&#160;(Drosophila&#160;)&#160;in an attempt to identify the key genes and pathways that lead to the characteristic hyperphosphorylated tau accumulation and neuronal death in the brain. Adjustable-strength impacts to inflict mild closed injury are delivered directly to the fly head, subjecting the head to rapid acceleration and deceleration. Our method eliminates the potential problems inherent with other Drosophila mTBI models (e.g.,animal death might be induced by damage to other parts of the body or to internal organs). The less labor- and cost-intensive animal care, short life span, and extensive genetic tools make the fruit fly ideal to study CTE pathogenesis and make it possible to perform large-scale, genome-wide forward genetic and pharmacological screens. We anticipate that the ongoing characterization of the model will generate important mechanistic insights on disease prevention and therapeutic approaches.

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Here, we describe a new approach to inflict closed-head traumatic brain injury in Drosophila melanogaster. Our method has the advantage of directly delivering repetitive impacts with adjustable strength to the head alone. Further exploration of the invertebrate system will help to illuminate the pathogenesis of chronic traumatic encephalopathy.

Um novo método para modelar a encefalopatia traumática crônica em<em&gt; Drosophila</em

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic ...

Neurociência

A Novel In Vitro Model of Blast Traumatic Brain Injury

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the detonation of explosive devices, however, the mechanisms that underlie the brain damage resulting from blast overpressure exposure are not entirely understood and are believed to be unique to this type of brain injury. Preclinical models are crucial tools that contribute to better understand blast-induced brain injury. A novel in vitro blast TBI model was developed using an open-ended shock tube to simulate real-life open-field blast waves modelled by the Friedlander waveform. C57BL/6N mouse organotypic hippocampal slice cultures were exposed to single shock waves and the development of injury was characterized up to 72 h using propidium iodide, a well-established fluorescent marker of cell damage that only penetrates cells with compromised cellular membranes. Propidium iodide fluorescence was significantly higher in the slices exposed to a blast wave when compared with sham slices throughout the duration of the protocol. The brain tissue injury is very reproducible and proportional to the peak overpressure of the shock wave applied.

A Novel In Vitro Model of Blast Traumatic Brain Injury

This paper describes a novel model of primary blast traumatic brain injury. A compressed-air driven shock tube is used to expose in vitro mouse hippocampal slice cultures to a single shock wave. This is a simple and rapid protocol generating a reproducible brain tissue injury with a high throughput.

Um modelo de romance In Vitro de explosão traumatismo crânio-encefálico

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the ...

Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral vascular effects. Impact (i.e., non-blast) traumatic brain injury (TBI) is known to decrease pressure autoregulation in the cerebral vasculature in both humans and experimental animals. The hypothesis that blast-induced traumatic brain injury (bTBI), like impact TBI, results in impaired cerebral vascular reactivity was tested by measuring myogenic dilatory responses to reduced intravascular pressure in rodent middle cerebral arterial (MCA) segments from rats subjected to mild bTBI using an Advanced Blast Simulator (ABS) shock tube. Adult, male Sprague-Dawley rats were anesthetized, intubated, ventilated and prepared for Sham bTBI (identical manipulation and anesthesia except for blast injury) or mild bTBI. Rats were randomly assigned to receive Sham bTBI or mild bTBI followed by sacrifice 30 or 60 min post-injury. Immediately after bTBI, righting reflex (RR) suppression times were assessed, euthanasia at the time points post-injury was completed, the brain was harvested and the individual MCA segments were collected, mounted and pressurized. As the intraluminal pressure perfused through the arterial segments was reduced in 20 mmHg increments from 100 to 20 mmHg, MCA diameters were measured and recorded. With decreasing intraluminal pressure, MCA diameters steadily increased significantly above baseline in the Sham bTBI groups while MCA dilator responses were significantly reduced (p &#60; 0.05) in both bTBI groups as evidenced by the impaired, smaller MCA diameters recorded for the bTBI groups. In addition, RR suppression in the bTBI groups was significantly (p &#60; 0.05) higher than in the Sham bTBI groups. MCA&#39;s collected from the Sham bTBI groups exhibited typical vasodilatory properties to decreases in intraluminal pressure while MCA&#39;s collected following bTBI exhibited significantly impaired myogenic vasodilatory responses to reduced pressure that persisted for at least 60 min after bTBI.

Here, we present a protocol to describe methods for ex vivo vascular reactivity determination following a primary blast traumatic brain injury (bTBI) using isolated, pressurized, rodent middle cerebral arterial (MCA) segments. bTBI induction is accomplished using a shock tube, also known as an Advanced Blast Simulator (ABS) device.

Efeitos de Neurotrauma induzida por explosão em roedores pressurizado artérias cerebrais de médio

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral ...

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also demonstrate that traumatic brain injuries cause changes to the gut microbiota. However, mechanisms underlying the bidirectional regulation of the brain-gut axis remain unknown. Currently, few models exist for studying the changes in gut microbiota after traumatic brain injury. Therefore, the presented study combines protocols for inducing traumatic brain injury using a lateral fluid percussion device and analysis of caecum samples following injury for investigating alterations in the gut microbiome. Alterations of the gut microbiota composition after traumatic brain injury are determined using 16S-rDNA sequencing. This protocol provides an effective method for studying the relationships between enteric microorganisms and traumatic brain injury.

Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome analysis.

Investigando alterações na microbiota de caecum após traumatismo cranioencefálico em camundongos

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also ...

Low-intensity Blast Wave Model for Preclinical Assessment of Closed-head Mild Traumatic Brain Injury in Rodents

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of medical visits in the United States. There are currently no FDA-approved treatments available for TBI. The increased incidence of military-related, blast-induced TBI further accentuates the urgent need for effective TBI treatments. Therefore, new preclinical TBI animal models that recapitulate aspects of human blast-related TBI will greatly advance the research efforts into the neurobiological and pathophysiological processes underlying mild to moderate TBI as well as the development of novel therapeutic strategies for TBI.
Here we present a reliable, reproducible model for the investigation of the molecular, cellular, and behavioral effects of mild to moderate blast-induced TBI. We describe a step-by-step protocol for closed-head, blast-induced mild TBI in rodents using a bench-top setup consisting of a gas-driven shock tube equipped with piezoelectric pressure sensors to ensure consistent test conditions. The benefits of the setup that we have established are its relative low-cost, ease of installation, ease of use and high-throughput capacity. Further advantages of this non-invasive TBI model include the scalability of the blast peak overpressure and the generation of controlled reproducible outcomes. The reproducibility and relevance of this TBI model has been evaluated in a number of downstream applications, including neurobiological, neuropathological, neurophysiological and behavioral analyses, supporting the use of this model for the characterization of processes underlying the etiology of mild to moderate TBI.

We present here a protocol of a blast wave model for rodents to investigate neurobiological and pathophysiological effects of mild to moderate traumatic brain injury. We established a gas-driven, bench-top setup equipped with pressure sensors allowing for reliable and reproducible generation of blast-induced mild to moderate traumatic brain injury.

Modelo de onda de explosão de baixa intensidade para avaliação pré-clínica de lesão cerebral traumática leve de cabeça fechada em roedores

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of ...

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

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

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

Modelo de Lesão Cerebral Traumática Leve em Camundongos com Controle Eletromagnético

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

Assessing Changes in Synaptic Plasticity Using an Awake Closed-Head Injury Model of Mild Traumatic Brain Injury

Mild traumatic brain injuries (mTBIs) are a prevalent health issue in North America. There is increasing pressure to utilize ecologically valid models of closed-head mTBI in the preclinical setting to increase translatability to the clinical population. The awake closed-headed injury (ACHI) model uses a modified controlled cortical impactor to deliver closed-headed injury, inducing clinically relevant behavioral deficits without the need for a craniotomy or the use of an anesthetic.
This technique does not normally induce fatalities, skull fractures, or brain bleeds, and is more consistent with being a mild injury. Indeed, the mild nature of the ACHI procedure makes it ideal for studies investigating repetitive mTBI (r-mTBI). Growing evidence indicates that r-mTBI can result in a cumulative injury that produces behavioral symptoms, neuropathological changes, and neurodegeneration. r-mTBI is common in youths playing sports, and these injuries occur during a period of robust synaptic reorganization and myelination, making the younger population particularly vulnerable to the long-term influences of r-mTBI.
Further, r-mTBI occurs in cases of intimate partner violence, a condition for which there are few objective screening measures. In these experiments, synaptic function was assessed in the hippocampus in juvenile rats that had experienced r-mTBI using the ACHI model. Following the injuries, a tissue slicer was utilized to make hippocampal slices to evaluate bidirectional synaptic plasticity in the hippocampus at either 1 or 7 days following the r-mTBI. Overall, the ACHI model provides researchers with an ecologically valid model to study changes in synaptic plasticity following mTBI and r-mTBI.

Here, it is demonstrated how an awake closed-head injury model can be used for examining the effects of repeated mild traumatic brain injury (r-mTBI) on synaptic plasticity in the hippocampus. The model replicates important features of r-mTBI in patients and is used in conjunction with in vitro electrophysiology.

Avaliação das Alterações na Plasticidade Sináptica Utilizando um Modelo de Traumatismo Cranioencefálico Leve

Mild traumatic brain injuries (mTBIs) are a prevalent health issue in North America. There is increasing pressure to utilize ecologically valid models of ...

Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope

The goal of this protocol is to demonstrate how to longitudinally visualize the expression and localization of a protein of interest within specific cell types of an animal's brain, upon exposure to exogenous stimuli. Here, the administration of a closed-skull traumatic brain injury (TBI) and simultaneous implantation of a cranial window for subsequent longitudinal intravital imaging in mice is shown. Mice are intracranially injected with an adeno-associated virus (AAV) expressing enhanced green fluorescent protein (EGFP) under a neuronal specific promoter. After 2 to 4 weeks, the mice are subjected to a repetitive TBI using a weight drop device over the AAV injection location. Within the same surgical session, the mice are implanted with a metal headpost and then a glass cranial window over the TBI impacting site. The expression and cellular localization of EGFP is examined using a two-photon microscope in the same brain region exposed to trauma over the course of months.

This study demonstrates delivery of a repetitive traumatic brain injury to mice and simultaneous implantation of a cranial window for subsequent intravital imaging of a neuron-expressed EGFP using two-photon microscopy.

Imagem Intravital da Expressão de Proteínas Fluorescentes em Camundongos com Traumatismo Cranioencefálico de Crânio Fechado e Janela Craniana Usando um Microscópio de Dois Fótons

The goal of this protocol is to demonstrate how to longitudinally visualize the expression and localization of a protein of interest within specific cell ...

Validando a resposta a lesões e avaliações comportamentais em um modelo de rato mTBI

Rat Model of Closed-Head Mild Traumatic Injury and its Validation

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful insights, developing a stable and reproducible animal model is essential. In this study, we report a detailed description of a closed-head mTBI model and a representative validation method using Sprague-Dawley rats to verify the modeling effect. The model involves dropping a 550 g mass weight from a height of 100 cm directly onto the head of a rat on a destructible surface, followed by a 180-degree turn. To assess the injury, rats underwent a series of neurobehavioral assessments 10 min post-injury, including time of loss of consciousness, first seeking-behavior time, escape ability, and beam balance ability test. During the acute and subacute stages following the injury, behavioral tests were conducted to assess motor coordination ability (Beam task), anxiety (Open Field test), and learning and memory abilities (Morris Water Maze test). The closed-head mTBI model produced a consistent injury response with minimal mortality and replicated real-life situations. The validation method effectively verified the model development and ensured the stability and consistency of the model.

Autor em destaque: Avançando na pesquisa de lesões cerebrais traumáticas - um modelo de cabeça fechada para replicação precisa e avaliação rápida 

Here, we present a closed-head mild traumatic brain injury (mTBI) rat model and its validation exhibiting remarkable similarity to human mTBI concerning behavioral manifestations during the acute and subacute stages.

Modelo de Lesão Traumática Leve de Cabeça Fechada em Rato e sua Validação

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful ...