Questo lavoro è stato sostenuto dal National Institutes of Health sovvenzione, R01 AG033620 (BG) e di Robert Draper Technology Innovation Funding (DAW).

1. Costruzione del dispositivo di HIT <ol><li> Fissare la molla alla scheda utilizzando due morsetti e quattro viti (Figura 1A). Centrare i morsetti relativi alla larghezza della tavola e allineati l&#39;uno con l&#39;altro con il filo morsetto esterno con il bordo della scheda. Prima di collegare i morsetti, piegarli con una pinza per adattarsi perfettamente durante la primavera. NOTA: Vedi Tabella 1 per le descrizioni dei materiali necessari per la costruzione del dispositivo HIT. L&#39;estremità bloccato della molla deve essere 1/8 di pollice (3,2 mm) dal bordo della tavola, e l&#39;estremità libera sia estesa al bordo da 3/4 di pollice (19 mm). Regolare la molla in modo che risulti parallelo alla lunghezza della tavola. </li><li> Avvolgere l&#39;estremità libera della molla una volta intorno con il nastro adesivo di loop velcro. Il bordo esterno del velcro deve essere a filo con l&#39;estremità della molla. Il Velcro è importante perché è utilizzato per fissare il flacone per la molla creDELL&#39;INDICE DI un raccordo a compressione stretto. Il Velcro consente inoltre una facile connessione e la rimozione di fiale, permettendo molte fiale da trasformare in un breve periodo di tempo. </li><li> Mettere il coperchio del contenitore di ghiaccio a testa in giù, al centro, a contatto con la tavola di legno. Orientare la zona in rilievo del coperchio del contenitore di ghiaccio tale che il lato lungo è parallelo alla larghezza della tavola. Si noti che la zona in rilievo della benna di ghiaccio è di 1/2 di pollice (13 mm) più alto del bordo di legno, in modo che quando una fiala è collegato alla molla la molla non sarà piatte sul bordo. </li><li> Scorrere l&#39;intero dispositivo contro un oggetto fisso, ad esempio un muro, in modo che il coperchio del contenitore di ghiaccio si incunea tra la scheda e l&#39;oggetto e non si muove. </li><li> Nastro il goniometro carta sul fondo del cartone vola vassoio fiala e basamento sul bordo contro la lunghezza della tavola in modo che il marchio di 90 ° sia allineata con la molla quando viene tirato indietro ad una posizione perfettamente verticale. </li></ol> 2.Il funzionamento del dispositivo di HIT <ol><li> Luogo tra 1 e 60 CO 2 -anesthetized vola in una fiala vuota e tappo del flacone con un batuffolo di cotone aderente. </li><li> Confinare linea al fondo 1 pollice (2,5 cm) del flaconcino spingendo il batuffolo di cotone nel flaconcino finché è 1 pollice (2,5 cm) dal fondo. E &#39;utile a tracciare una linea sul flacone al segno 1 pollice (2,5 cm). Si noti che confinare mosche a regioni più grandi o più piccoli della fiala può influenzare la gravità dei fenotipi. </li><li> Attendere 5 minuti per le mosche di recuperare la mobilità della CO 2. Si noti che non è noto se 5 min è sufficiente per eliminare completamente gli effetti di CO 2. </li><li> Inserire l&#39;estremità della molla nel flaconcino finché il bordo interno del Velcro è a filo con la parte superiore della fiala (Figura 1B). Quando la molla è disteso, 1 pollice (2,5 centimetri) del flaconcino deve sovrapporsi regione rialzato del coperchio del secchiello del ghiaccio. Fiale possono essere riutilizzati mogni volta. </li><li> Stando seduti, tenere il flaconcino in corrispondenza della zona in velcro con il pollice e l&#39;indice della mano sinistra. Tenere il bordo stretto al banco con la mano destra. In alternativa, utilizzare C-morsetti per tenere la scheda stretto al banco. </li><li> Tirare la molla perfettamente dritto torna all&#39;angolazione desiderata. Rilasciare la molla. Lasciare che la primavera a venire ad un arresto completo. </li><li> Rimuovere la fiala dal primavera e consentire in linea di recuperare per ≥5 min. Sottoporre linea ad un altro sciopero o trasferire le mosche per una fiala con la mosca cibo. NOTA: Una varietà di saggi possono essere utilizzati per valutare gli effetti fenotipici dei colpi dal dispositivo HIT. Ad esempio, gli effetti sulla longevità può essere determinato analizzando la percentuale di mosche che sopravvivono a volte dopo la lesione, effetti sulla morfologia cerebrale può essere determinata mediante analisi istologica della testa, e gli effetti sulla espressione genica può essere determinata mediante l&#39;analisi quantitativa dei livelli di mRNA 10. </li><li> Determine effetti del procedimento non dovuti ad attacchi trattando identicamente linea di controllo che non sono soggette a colpi. Indossare protezione acustica perché l&#39;impatto della fiala contro il coperchio del contenitore di ghiaccio produce un forte rumore. </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. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathophysiology+of+traumatic+brain+injury+at+a+glance.">The pathophysiology of traumatic brain injury at a glance.</a> Disease Models Mech. 6, 1307-1315 (2013).</li><li>Menon, D. 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. T., Ganetzky, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+method+for+accurate+and+rapid+measurement+of+flight+performance+in+Drosophila.">An improved method for accurate and rapid measurement of flight performance in Drosophila.</a> J Vis Exp. (84), e51223 (2014).</li><li>Tully, T., Preat, T., Boynton, S. C., Vecchio, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+dissection+of+consolidated+memory+in+Drosophila.">Genetic dissection of consolidated memory in Drosophila.</a> Cell. 79, 35-47 (1994).</li><li>Andretic, R., Shaw, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Essentials+of+sleep+recordings+in+Drosophila.:+moving+beyond+sleep+time.">Essentials of sleep recordings in Drosophila.: moving beyond sleep time.</a> Methods Enzymol. 393, 759-772 (2005).</li></ol>

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Il metodo dispositivo HIT si distingue da altri metodi che infliggono lesione traumatica in linea dal fatto che essa provoca cranico chiuso piuttosto che penetrante TBI 11. Inoltre, il metodo di dispositivo HIT richiede meno tempo, fatica e abilità per infliggere TBI in molte mosche, in modo che il metodo è più suscettibili di altri metodi agli schermi genetici su larga scala. Infine, il fatto che le lesioni primarie inflitte dal dispositivo HIT non si limitano al cervello è un limite e un vantaggio. Si t...

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

Lesione cerebrale traumatica (TBI) è definita come lesioni al cervello da una forza meccanica esterna. Più comunemente, i risultati TBI da forze testa chiusi quali forze smussati e accelerazioni inerziali e decelerazione che causano il cervello per colpire l&#39;interno del cranio. Negli Stati Uniti, si stima che 50.000 persone muoiono ogni anno per TBI e 2,5-6.500.000 individui vivono con le conseguenze del trauma cranico, tra cui debilitante fisico, cognitivo, e problemi comportamentali 1,2. Le conseguenze del trauma cranico non solo a causa di infortuni meccanici primari al cervello, ma anche per le lesioni cellulari e molecolari secondari al cervello e altri tessuti che si verificano nel corso del tempo 3-5. Lo sviluppo di approcci per diagnosticare e trattare TBI ha dimostrato di essere difficile, perché TBI è un processo patologico complesso. La variabilità delle lesioni primarie, la fisiologia umana, e fattori ambientali si traduce in eterogenei i secondarionjuries e patologie. Fattori variabili esistenti includono la gravità della lesione primaria, il tempo tra lesioni primarie ripetitive, e l&#39;età e il genotipo dell&#39;individuo. Capire come ogni fattore variabile contribuisce alle conseguenze di trauma cranico è probabile che per aiutare nello sviluppo di approcci per diagnosticare e trattare TBI 6,7. Qui si descrive un metodo per infliggere testa TBI chiuso in Drosophila melanogaster (moscerino della frutta) che può essere utilizzato per delineare il contributo dei fattori variabili per le conseguenze del trauma cranico. Il metodo si basa su una prima osservazione che intensamente colpire il lato di un flaconcino cultura mosca contro il palmo di una mano causato wild-type vola a diventare temporaneamente inabili, un probabile conseguenza di TBI. Così, abbiamo costruito il Trauma (HIT) dispositivo ad alto impatto di ricapitolare le forze di accelerazione e decelerazione dall&#39;azione incisiva mano. Un film ad alta velocità mostra che un singolo colpo dallaHIT dispositivo fa sì che vola a contattare il muro flacone più volte con la loro testa e il corpo 8. In una certa misura, tutti i contatti sono suscettibili di causare il cervello mosca per rimbalzare e deformare contro la capsula testa, simile a quanto accade per gli esseri umani in cadute e incidenti stradali 9. Di conseguenza, mosche trattato con il display fenotipi dispositivo HIT coerenti con lesioni cerebrali, tra cui incapacità temporanea seguita da atassia, graduale recupero della mobilità, cambiamenti di espressione genica nella testa, e la progressiva neurodegenerazione nel cervello 10. Così, il dispositivo HIT consente di studiare TBI utilizzando l&#39;enorme arsenale di strumenti e tecniche sviluppate per mosche sperimentali.

<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

Lesione cerebrale traumatica (TBI) colpisce milioni di persone ogni anno, causando perdita di valore di fisico, cognitivo, e funzioni comportamentali e la morte. Gli studi che utilizzano Drosophila hanno contribuito importanti passi avanti nella comprensione dei processi neurologici. Così, con l&#39;obiettivo di comprendere le basi cellulari e molecolari di patologie TBI negli esseri umani, abbiamo sviluppato il dispositivo High Impact Trauma (HIT) di infliggere alla testa chiusa TBI in mosche. Le mosche sottoposte alla visualizzazione fenotipi dispositivo HIT coerenti con TBI umana, come incapacità temporanea e progressiva neurodegenerazione. Il dispositivo HIT utilizza un meccanismo a molla per spingere basato mosche contro la parete di una fiala, causando danni meccanici al cervello mosca. Il dispositivo è economico e facile da costruire, il suo funzionamento è semplice e rapida, e produce risultati riproducibili. Di conseguenza, il dispositivo HIT può essere combinato con strumenti sperimentali esistenti e tecniche per mosche per affrontare fondamentaledomande su TBI che possono portare allo sviluppo di strumenti diagnostici e terapie per TBI. In particolare, il dispositivo HIT può essere utilizzato per eseguire schermi genetici su larga scala per comprendere la base genetica di patologie TBI.

Siamo interessati a capire il motivo per cui le mosche muoiono poco dopo la lesione primaria. Per quantificare la morte, abbiamo determinato l&#39;indice di mortalità in 24 ore (MI 24), che è la percentuale di mosche che sono morti entro 24 ore della lesione primaria. Mosche sottoposti a colpi dal dispositivo HIT sono state incubate a 25 ° C in una fiala con fly cibo, e il numero di mosche morte è stato contato dopo 24 ore. Abbiamo usato questo approccio per identificare fattori che influenzano il MI <sub...

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

Qui si descrive un metodo per infliggere cranico chiuso lesione cerebrale traumatica (TBI) in Drosophila. Questo metodo fornisce un gateway per studiare i meccanismi cellulari e molecolari che sono alla base delle patologie TBI utilizzando la vasta gamma di strumenti sperimentali e le tecniche disponibili per le mosche.

Un metodo per infliggere Chiuso Testa trauma cranico in<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

11.3K Views.  University of Wisconsin-Madison.  Qui si descrive un metodo per infliggere cranico chiuso lesione cerebrale traumatica (TBI) in Drosophila. Questo metodo fornisce un gateway per studiare i meccanismi cellulari e molecolari che sono alla base delle patologie TBI utilizzando la vasta gamma di strumenti sperimentali e le tecniche disponibili per le mosche. 

Video: 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.

Un nuovo metodo per modellare l&#39;encefalopatia traumatica cronica in<em&gt; Drosophila</em

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

Ricerca

Neuroscienze

Stereotactic Atlas-Guided Laser Capture Microdissection of Brain Regions Affected by Traumatic Injury

The ability to isolate specific brain regions of interest can be impeded in tissue disassociation techniques that do not preserve their spatial distribution. Such techniques also potentially skew gene expression analysis because the process itself can alter expression patterns in individual cells. Here we describe a laser capture microdissection (LCM) method to selectively collect specific brain regions affected by traumatic brain injury (TBI) by using a modified Nissl (cresyl violet) staining protocol and the guidance of a rat brain atlas. LCM provides access to brain regions in their native positions and the ability to use anatomical landmarks for identification of each specific region. To this end, LCM has been used previously to examine brain region specific gene expression in TBI. This protocol allows examination of TBI-induced alterations in gene and microRNA expression in distinct brain areas within the same animal. The principles of this protocol can be amended and applied to a wide range of studies examining genomic expression in other disease and/or animal models.

We describe the use of laser capture microdissection to obtain samples of distinct cell populations from different brain regions for gene and microRNA analysis. This technique allows the study of differential effects of traumatic brain injury in specific regions of the rat brain.

Atlas-guida stereotassica del Laser Capture Microdissection di regioni del cervello colpite da lesione traumatica

The ability to isolate specific brain regions of interest can be impeded in tissue disassociation techniques that do not preserve their spatial ...

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.

Un modello di romanzo In Vitro di ferita di cervello traumatica di scoppio

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.

Effetti del Neurotrauma indotta da Blast il roditore pressurizzato Middle arterie cerebrali

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.

Indagare sulle alterazioni nel caecum Microbiota dopo una lesione cerebrale traumatica nei topi

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.

Modello di onda d'esplosione a bassa intensità per la valutazione preclinica della lesione cerebrale traumatica lieve a testa chiusa nei roditori

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 ...

Systems Analysis of the Neuroinflammatory and Hemodynamic Response to Traumatic Brain Injury

Mild traumatic brain injuries (mTBIs) are a significant public health problem. Repeated exposure to mTBI can lead to cumulative, long-lasting functional deficits. Numerous studies by our group and others have shown that mTBI stimulates cytokine expression and activates microglia, decreases cerebral blood flow and metabolism, and impairs cerebrovascular reactivity. Moreover, several works have reported an association between derangements in these neuroinflammatory and hemodynamic markers and cognitive impairments. Herein we detail methods to characterize the neuroinflammatory and hemodynamic tissue response to mTBI in mice. Specifically, we describe how to perform a weight-drop model of mTBI, how to longitudinally measure cerebral blood flow using a non-invasive optical technique called diffuse correlation spectroscopy, and how to perform a Luminex multiplexed immunoassay on brain tissue samples to quantify cytokines and immunomodulatory phospho-proteins (e.g., within the MAPK and NF&#954;B pathways) that respond to and regulate activity of microglia and other neural immune cells. Finally, we detail how to integrate these data using a multivariate systems analysis approach to understand the relationships between all of these variables. Understanding the relationships between these physiologic and molecular variables will ultimately enable us to identify mechanisms responsible for mTBI.

This protocol presents methods to characterize the neuroinflammatory and hemodynamic response to mild traumatic brain injury and to integrate these data as part of a multivariate systems analysis using partial least squares regression.

Analisi dei sistemi della risposta neuroinfiammatoria ed emodinamica alla lesione cerebrale traumatica

Mild traumatic brain injuries (mTBIs) are a significant public health problem. Repeated exposure to mTBI can lead to cumulative, long-lasting functional ...

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

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

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

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

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

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.

Valutazione dei cambiamenti nella plasticità sinaptica utilizzando un modello di trauma cranico chiuso sveglio di lieve lesione cerebrale traumatica

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.

Imaging intravitale dell'espressione di proteine fluorescenti nei topi con una lesione cerebrale traumatica a cranio chiuso e una finestra cranica utilizzando un microscopio a due fotoni

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