Bu çalışma Sağlık hibe, R01 AG033620 (BG) Ulusal Sağlık Enstitüleri tarafından ve Robert Draper Teknoloji İnovasyon Finansman (DAW) tarafından desteklenmiştir.

HIT Cihazın 1. İnşaat <ol><li> İki kelepçeler ve dört vidayı (Şekil 1A) kullanarak kuruluna yayı takın. Yönetim kurulu genişliğine göre kelepçeleri ortalamak ve yönetim kurulu kenarı ile dış kelepçe floş ile birbirlerine karşı onları popo. Önce kelepçeler takılarak için, yay üzerinde sıkıca uyacak şekilde pense ile bükün. NOT: HIT cihazı oluşturmak için gerekli malzemelerin açıklamaları için Tablo 1&#39;e bakınız. Baharın kenetlenmiş uç kurulu kenarından 1/8 inç (3.2 mm) olmalıdır ve serbest ucu 3/4 inç (19 mm) ile tahta üzerinde uzanmalıdır. Bu yönetim kurulu uzunluğu ile paralel yatıyor böylece yayı ayarlayın. </li><li> Velcro döngüler yapışkan bant kez etrafında bahar serbest ucunu sarın. Velcro dış kenarı bahar sonu ile aynı hizada olmalıdır. O CRE tarafından bahar flakon güvenliğini sağlamak için kullanılan, çünkü Velcro önemlidirSıkı bir sıkıştırma uydurma ating. Velcro, aynı zamanda pek çok küçük şişeler kısa bir süre içinde işlenecek sağlayan, kolay bağlantı ve şişeler çıkarılmasını mümkün kılar. </li><li> Sıkı tahta karşı merkezli baş aşağı buz kovası kapağı yerleştirin. Uzun kenarı kurulu genişliğine paralel olacak şekilde buz kovası kapağının kaldırılmış bölgesini yönlendirmek. Bir şişe yayı bağlıyken bahar tahtada düz yalan olmayacak şekilde buz kovası yükseltilmiş bölge, tahta daha 1/2 inç (13 mm) yüksek olduğunu unutmayın. </li><li> Buz kovası kapağı kurulu ve nesne arasında sıkışmış ve hareket etmiyor, böylece, örneğin bir duvar gibi sabit bir nesneye karşı tüm cihaz kaydırın. </li><li> Bant, bir karton altına kağıt iletki flakon tepsiyi sinek ve mükemmel dikey konumuna geri çekildiğinde 90 ° işareti yay ile aynı hizada olacak şekilde kurulu uzunluğuna karşı kenarında stand it. </li></ol> 2.HIT Cihazının Operasyonu <ol><li> Boş bir şişe içinde uçar ve bir dar pamuk kullanarak flakon kapağı kapatılır -anesthetized 1 ile 60 CO 2 arasındaki yeri. </li><li> Bu alttan 1 inç (2.5 cm) kadar şişe içine pamuk iterek şişenin taban 1 inç (2.5 cm) sinekler öngörebilir. Bu 1 inç (2.5 cm) işaretine flakon üzerinde bir çizgi çizmek için yararlıdır. Şişenin büyük veya daha küçük bölgelere sinekler hapsetmesi fenotipleri şiddetini etkileyebilir unutmayın. </li><li> Sinekler CO 2 hareketlilik kurtarmak için 5 dakika bekleyin. O 5 dk tamamen CO 2 etkilerini ortadan kaldırmak için yeterli olup olmadığı bilinmemektedir unutmayın. </li><li> Velcro iç kenarı flakon (Şekil 1B) üst ile aynı hizaya gelene kadar şişenin içine yayın ucunu. Bahar düz yalan olduğunda, flakonun 1 inç (2.5 cm) buz kovası kapağının kaldırılmış bölgeyi örtüşmelidir. Şişeler m yeniden kullanılabilirher zaman. </li><li> Otururken, sol elin baş ve işaret parmağı kullanılarak Velcro bölgede flakon tutun. Sağ elinizi kullanarak Benchtop yönetim kurulu sıkı tutun. Alternatif olarak, masa üstü sıkı yönetim kurulu tutmak için C-kelepçeler kullanın. </li><li> Mükemmel düz geri istenen açıya yayı çekin. Bahar bırakın. Bahar tamamen durmasını bekleyin. </li><li> Bahar şişe çıkarın ve sinekler ≥5 dk kurtarmak için izin verir. Başka bir greve sinekler Konu veya sinek gıda ile flakon sinekler aktarın. Not: birçok analiz HIT cihazından vuruş fenotipik etkilerini değerlendirmek için kullanılabilir. Örneğin, uzun ömürlü etkileri yaralanma sonrası zaman hayatta sinekler yüzde analizi ile tespit edilebilir, beyin morfolojisi üzerinde etkisi kafasının histolojik analiz ile tespit edilebilir, ve gen ifadesi üzerindeki etkisi, mRNA seviyelerinin niceliksel analizi ile belirlenebilir 10. </li><li> Determinaynı grev tabi değildir kontrol sinekler tedavi ederek grev nedeniyle değil prosedürün e etkileri. Buz kovası kapağı karşı flakon etkisi bir gürültü üretir, çünkü işitme koruması giyin. </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>

Biz ifşa herhangi bir çıkar çatışmaları var.

HIT cihaz yöntemi TBI 11 delici yerine kapalı kafa neden gerçeği ile sinekler travmatik yaralanma indirebilmek diğer yöntemlerden ayrılır. Ayrıca, HIT cihaz yöntemi birçok sinekler TBI indirebilmek için daha az zaman, çaba ve beceri alır, bu yüzden yöntem büyük ölçekli genetik ekranlar diğer yöntemlere göre daha müsait. Son olarak, HIT cihazı tarafından uygulanan birincil yaralanmalar, beyin sınırlı değildir aslında bir sınırlama ve bir avantaj niteliğindedir. Ek çalışmala...

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

Travmatik beyin hasarı (TBI), harici bir mekanik kuvvet ile ilgili beyin yaralanması olarak tanımlanmaktadır. En yaygın tür künt kuvvetler ve atalet hızlanma ve beyin kafatasının içini grev neden yavaşlama güçleri olarak kapalı kafa kuvvetlerinden TBI sonuçlar. Amerika Birleşik Devletleri&#39;nde, 50.000 kişi TBI her yıl ölmektedir ve 2,5-6.500.000 bireyler, TBI sonuçları ile yaşayan fiziksel, bilişsel zayıflatıcı dahil ve 1,2 davranış sorunları olduğu tahmin edilmektedir. TBI sonuçları beyne primer mekanik yaralanma için değil, aynı zamanda zaman 3-5 içinde meydana beyin sekonder hücresel ve moleküler yaralanmalar gibi diğer dokularda değil sadece bulunmaktadır. teşhis ve TBI tedavi yaklaşımların geliştirilmesi TBI karmaşık bir hastalık süreci olduğu için zor olduğu kanıtlanmıştır. Birincil yaralanmalar, insan fizyolojisi ve çevresel faktörlerin değişken doğası heterojen ikincil i sonuçlanırnjuries ve patolojileri. Altta yatan değişken faktörler bireyin temel yaralanmanın şiddetini, tekrarlayan primer yaralanma arasındaki süreyi ve yaş ve genotipi içermektedir. Her bir değişken faktör TBI sonuçları nasıl katkıda anlama teşhis ve TBI 6,7 tedavisinde yaklaşımlar geliştirilmesine yardımcı olmak için muhtemeldir. Burada TBI sonuçları değişken faktör katkı tanımlamak için kullanılabileceğini Drosophila melanogaster (meyve sineği) &#39;de kapalı baş TBI kasten için bir yöntem tarif eder. yöntem yoğun bir elin avuç içi karşı bir sinek kültür flakon yan isabet vahşi tip neden bir ilk gözlem dayanmaktadır geçici TBI olası bir sonucu sakat hale uçar. Böylece, el vurma eylem hızlanma ve yavaşlama kuvvetleri özetlemek için Yüksek Etki Travma (HIT) aygıtı inşa. Yüksek hızlı film gösterir tek bir grevCihaz neden HIT onların baş ve gövdesinin 8 flakon duvarı birkaç kez temas uçar. Bir dereceye kadar, tüm kişiler sekerek ve düşme ve araba kazası 9 insanlara ne benzer kafa kapsül, karşı deforme olmasına sinek beyin neden muhtemeldir. Buna göre, beyinde 10 ataksi ardından geçici ehliyetini kaybetmesi, hareketlilik kademeli bir toparlanma, baş gen ifade değişiklikleri ve ilerleyici sinir dejenerasyonuna dahil beyin hasarı, tutarlı HIT cihaz ekran fenotipleri ile tedavi sinekler. Böylece, HIT cihazı mümkün deneysel araçları ve sinekler için geliştirilen tekniklerin büyük cephanelik kullanarak TBI çalışma yapar.

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			<td>Zinc plated compression spring</td>
			<td>The Hillman&#160; Group</td>
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			<td>9 7/8 in (length), 15/16 in (outer diameter), 0.12 in (wire size)</td>
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			<td>Clamps</td>
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			<td>Loop half of self-adhesive velcro</td>
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a method to inflict closed head traumatic brain injury in drosophila

Travmatik beyin hasarı (TBH), fiziksel, bilişsel, davranışsal ve fonksiyonları ve ölüm bozukluğu neden her yıl milyonlarca insanı etkilemektedir. Drosophila kullanılarak yapılan çalışmalar nörolojik süreçlerin anlaşılmasında önemli atılımlar katkıda bulunmuştur. Böylece, insanlarda TBI patolojilerin hücresel ve moleküler temelini anlamak amacıyla, biz sinek kapalı kafa TBI indirebilmek için High Impact Travma (HIT) cihazı geliştirdi. Sinekler gibi geçici ehliyetini kaybetmesi ve ilerici nörodejenerasyona gibi insan TBI ile uyumlu HIT cihaz ekran fenotipleri tabi. HIT cihaz sinek beyinde mekanik hasara neden bir şişenin duvara sinekleri itmek için bir yay-tabanlı mekanizması kullanır. Cihaz operasyon basit ve hızlı olan, yapımı pahalı olmayan, kolayca ve tekrarlanabilir sonuçlar üretir. Sonuç olarak, HIT cihazı mevcut deneysel araçları ve sinekler temel adrese teknikleri ile kombine edilebilirTBI için teşhis ve tedavilerin gelişmesine yol açabilir TBI ilgili sorular. Özellikle, HIT cihazı TBI patolojilerin genetik temelini anlamak için geniş çaplı genetik ekranlar gerçekleştirmek için kullanılabilir.

Biz sinekler birincil yaralanma kısa bir süre sonra ölür neden anlamak ilgilendi. Ölümü ölçmek için, birincil yaralanma 24 saat içinde ölen sineklerin yüzde 24 saatte Ölüm İndeksi (MI 24), tespit. HIT cihazdan saldırılarından Sinekler uçucu gıda ile bir şişe içinde 25 ° C&#39;de inkübe edilmiştir, ve ölü sinek sayısı, 24 saat sonra sayılmıştır. Biz MI 24 etkileyen faktörleri belirlemek için bu yaklaşımı kullanılır ve MI 24 bir şişeye (10 ila 60...

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

Burada Drosophila kapalı kafa travmatik beyin hasarı (TBI) indirebilmek için bir yöntem açıklanmaktadır. Bu yöntem, sinekler için deneysel araçları ve teknikleri geniş bir dizi kullanarak TBI patolojiler altında yatan hücresel ve moleküler mekanizmaları araştırmak için bir ağ geçidi sağlar.

Kapalı Başkanı Travmatik Beyin yaralayabilecek Bir Yöntem<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 Görüntüleme.  University of Wisconsin-Madison.  Burada Drosophila kapalı kafa travmatik beyin hasarı (TBI) indirebilmek için bir yöntem açıklanmaktadır. Bu yöntem, sinekler için deneysel araçları ve teknikleri geniş bir dizi kullanarak TBI patolojiler altında yatan hücresel ve moleküler mekanizmaları araştırmak için bir ağ geçidi sağlar. 

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

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and mortality. Based on the nature of primary injury following TBI, complex and heterogeneous secondary consequences result, which are followed by regenerative processes 1,2. Primary injury can be induced by a direct contusion to the brain from skull fracture or from shearing and stretching of tissue causing displacement of brain due to movement 3,4. The resulting hematomas and lacerations cause a vascular response 3,5, and the morphological and functional damage of the white matter leads to diffuse axonal injury 6-8. Additional secondary changes commonly seen in the brain are edema and increased intracranial pressure 9. Following TBI there are microscopic alterations in biochemical and physiological pathways involving the release of excitotoxic neurotransmitters, immune mediators and oxygen radicals 10-12, which ultimately result in long-term neurological disabilities 13,14. Thus choosing appropriate animal models of TBI that present similar cellular and molecular events in human and rodent TBI is critical for studying the mechanisms underlying injury and repair.
Various experimental models of TBI have been developed to reproduce aspects of TBI observed in humans, among them three specific models are widely adapted for rodents: fluid percussion, cortical impact and weight drop/impact acceleration 1. The fluid percussion device produces an injury through a craniectomy by applying a brief fluid pressure pulse on to the intact dura. The pulse is created by a pendulum striking the piston of a reservoir of fluid. The percussion produces brief displacement and deformation of neural tissue 1,15. Conversely, cortical impact injury delivers mechanical energy to the intact dura via a rigid impactor under pneumatic pressure 16,17. The weight drop/impact model is characterized by the fall of a rod with a specific mass on the closed skull 18. Among the TBI models, LFP is the most established and commonly used model to evaluate mixed focal and diffuse brain injury 19. It is reproducible and is standardized to allow for the manipulation of injury parameters. LFP recapitulates injuries observed in humans, thus rendering it clinically relevant, and allows for exploration of novel therapeutics for clinical translation 20.
We describe the detailed protocol to perform LFP procedure in mice. The injury inflicted is mild to moderate, with brain regions such as cortex, hippocampus and corpus callosum being most vulnerable. Hippocampal and motor learning tasks are explored following LFP.

Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is described.

Yanal Akışkan Perküsyon: Travmatik Beyin Hasarı Farelerde Modeli

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

Nörobilim

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

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

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

Hafif Travmatik Beyin Hasarı takiben Hipokampal Devre İşlev değişimler üzerine araştırmalar

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

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.

Kronik Travmatik Ensefalopatiyi Modelleyen Yeni Bir Yöntem<em&gt; Drosophila</em

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

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.

Stereotaksik Atlas güdümlü lazer yakalama mikrodiseksiyon travma yaralanma tarafından etkilenen beyin bölgeleri

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.

Patlama travmatik beyin hasarı roman Vitro modeli

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.

Serebral arterler patlama kaynaklı Neurotrauma etkilerini basınçlı kemirgen orta

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.

Farelerde Travmatik Beyin Hasarı Sonrası Caecum Mikrobiyotasında Ki Değişikliklerin Araştırılması

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.

Kemirgenlerde Kapalı Başlı Hafif Travmatik Beyin Hasarının Preklinik Değerlendirmesi için Düşük Yoğunluklu Patlama Dalgası Modeli

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.

Travmatik Beyin Hasarına Nöroinflamatuar ve Hemodinamik Yanıtın Sistem Analizi

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.

Farelerde Hafif Travmatik Beyin Hasarının Elektromanyetik Kontrollü Kapalı Kafa Modeli

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

Kapalı Başkanı Travmatik Beyin yaralayabilecek Bir Yöntem

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