Aşağıdaki finansman kaynaklarını kabul ediyoruz: NIH NINDS – K08NS101122 (JB), R01NS040337 (JK), R01NS044370 (JK), Virginia Üniversitesi Tıp Fakültesi (JB).

Tüm hayvan çalışmaları Virginia Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Komitesi (IACUC) tarafından onaylandı.
1. Elektrot bina / kablo binası
<ol><li>Dişi bir soket konnektörüne bağlı bir elektrot yapmak için tek kutuplu yalıtımlı paslanmaz çelik tel (0,005" çıplak çap, 0,008" kaplamalı) kullanın (dişi kap konnektörü 0,079).</li>
	<li>Hayvanları amplifikatöre bağlamak için özel yapım bir kablo kullanın.<ol>
<li>4 kanallı bir birliğe erkek 4 pinli konnektör (Erkek konnektör 0,079") takın, operasyonel amplifikatörle (op-amp) eşleşen 4 kanallı birlik kazanç empedansı. 9 V bataryaya bağlanan kablolara 10K direnç takın. Op-amp'ye bağlı olmayan bir topraklama teli pilin orta noktası olarak işlev görür.</li>
		<li>Kablonun bir ucunun (AWG, 0,012" OD) op-amp'ye ve kablonun diğer ucunun amplifikatöre bağlanması.</li>
	</ol>
</li>
</ol>2. Elektrot implantasyon ameliyatı
<ol><li>Aşağı doğru bir akış başlığında% 4-5 izofluran ile yavruyu (doğum sonrası gün 9) uyuşturun. İşlemin başlamasından önce, yavrulara bupivacaine enjekte edin (0.02-0.05 mL, % 0.25 deri altı lokal infiltrasyon).</li>
	<li>Hayvan hareketsiz hale geldikten sonra, burun konisi ile stereotaktik bir aşamaya aktarın. Başı sabit tutmak yumuşak olduğu için kulak çubuğunun ters tarafını kullanın. Bu yaşta, yavrular kulak çubuğunun sivri ucını kullanmak için tam olarak gelişmiş bir kulağa sahip değildir.</li>
	<li>Izofluran akışını kısın ve% 2.5-3'te koruyun. Ameliyat prosedürü boyunca yavrunun sabit nefes alışını takip edin. Ağrı yanıtını kontrol etmek için kuyruğu kıstırın ve sonra kesi devam edin.</li>
	<li>Kafatasındaki kesi alanını betadin ve alkolle sterilize edin (3 döngü alternatif iyot ve% 70 etanol). Kesi bölgesinin görülebilecek şekilde çevredeki vücut kısmını örtün.</li>
	<li>Kafa derisini gözlerin biraz üstünden ön-arka açın ve yaklaşık 0,5 cm deriyi geri çek. Fare kafasını stereotaksik sahnede yeniden konumlandırın, böylece cilt açıkta kalan kafatasını dışarı doğru çeker.</li>
	<li>Pamuklu çubuk kullanarak kafatasına hidrojen peroksit uygulayın ve kafatasını neşter bıçağı kullanarak kazıyın. Kafatası çok yumuşaktır; kazıma yaparken dikkatli olun.</li>
	<li>Bir damla (yaklaşık 50 μL) yapıştırıcı uygulayın ve aplikatörünü kullanarak açıkta kalan kafatası bölgesine yayın. Yapıştırıcıyı ayarlamak için 40 sn UV ışığına maruz bırak.</li>
	<li>Referans olarak maruz kalan bregmayı kullanarak koordinatları ölçün. Hipokampus 'un CA1 bölgesinde [-3,5 mm Dorsal-Ventral (DV), ±2 mm Medial-Lateral (ML), -1,75 mm Derin (D)] ve parietal kortekste iki taraflı implant elektrotları [-1,22 mm DV, ±0,5 mm ML, -1 mm D] ve beyincikte bir referans elektrot17. İşaretli bölgede bir delik oluşturmak için 32 G iğne kullanın.</li>
	<li>Kafatasının yüzeyindeki kanı temizleyin. Kadın soket konektörüne stereotaksik kol yardımıyla takılan daha düşük elektrotlar beyne ve diş akrilik ile yerine sabitlenin. Elektrotları beyne yerleştir. Soket konnektör kulaklığı, diş akrilik ile birbirine yapışmış kafatasının üzerine oturur.</li>
	<li>Elektrot sabitlendikten sonra interskapular bölgeye ketoprofen (5 mg / kg) deri altından enjekte edin. Yavruları annenin arkasına koy. NOT: Çöplerin yarısını teker teker tanıtmak yerine anneye aynı anda kulaklıkla tanıtın. Bu, annenin yavrunun kulaklığını zarar vermesini önleyecektir.</li>
</ol>3. EEG kurulumu ve kaydı (temel/yaralanma öncesi)
<ol><li>Elektrot implantasyonundan sonra 24 saat iyileşmeden sonra, her hayvanı EEG kaydı için ısıtılmış (37 °C) özel yapım pleksiglas bir odaya yerleştirin. Bu oda aynı zamanda bir hipoksi odası olarak da hizmet verecektir.</li>
	<li>Odadaki yavruları esnek bir kablo (özel yapım op-amp kablosu) ile bir video-EEG izleme sistemine bağlayın. NOT: Kulaklık yerindeyken, fareler serbestçe hareket halindedir ve davranışlarda herhangi bir fark sergilemez. Elektrot tellerine bağlandıktan sonra, yavrunun oda boyunca serbestçe hareket edebilmesi için doğru miktarda gevşeklik sağlamak için teller oda bağı içinde ayarlanmalıdır.</li>
	<li>EEG verilerini 1000 Hz'de bir çim amplifikatörü kullanarak 1K kazançla dijitalleştirin. Daha sonra yazılımı (örneğin LabChart Pro) kullanarak EEG sinyalini (3-70 Hz arasında bant geçiş filtresi) gözden geçirin.</li>
	<li>Şahdamar ligasyon prosedürü için hayvanların bağlantısını kesmeden önce 30 dakika boyunca yaralanma öncesi temel EEG kaydedin.</li>
</ol>4. Sol şahdamar ligasyonu
<ol><li>Yavruyu (doğum sonrası gün 10) aşağı doğru bir akış başlığında% 4-5 izofluran ile uyuşturun ve bir su banyosu pedinde özel olarak düzenlenmiş kuruluma yerleştirin. Hayvan supine yerleştirin ve ön ayakları kağıt bantla sabitleyin.<ol>
<li>Izofluran akışını% 2-3'e düşürün. Ağrı tepkisi için kuyruğu kıstırın ve işlem boyunca nefes almayı izleyin.</li>
	</ol>
</li>
	<li>Boynun sol tarafındaki kesi alanını (mandibula ve köprücük kemiği arasında) betadin ve alkol (3 döngü alternatif iyot ve% 70 etanol) ile sterilize edin.</li>
	<li>Mikros makas kullanarak boynun sol tarafında yaklaşık 1 cm uzunluğunda bir kesi yapın. Bir diseksiyon mikroskobu kullanarak, şahdamarını açığa çıkarmak için deri altı dokusunu ve cildi dikkatlice geri çekin. Vagus sinirini (artere yanal çalışan) tanımlamaya ve hassas bir şekilde ayırmaya ve arterden geri çekmeye özen edin.</li>
	<li>Mikroforceps kullanarak arterin altına 5 cm uzunluğunda steril ipek dikiş iplik. Tıkanmış akışa arterin etrafına çift düğümlü bir dikiş bağlayın.</li>
	<li>Fazla dikişi kesin ve deri altı dokusunu ve cildini geri çekerek maruz kalan arteri kapatın. Kesiği kapatmak için veteriner bağını kullanın.</li>
	<li>Hayvanı, ısınan bir şilteye yerleştirilen oda sıcaklığında bir odaya sürekli EEG izlemeye geri yerleştirin. Hazneyi açmamak için yavru çekirdek sıcaklığının nokta kızılötesi sıcaklık kontrollerini alın. Hayvanın hipoksiden önce 1 saat boyunca iyileşmesine izin verin.</li>
</ol>5. EEG ve hipoksi
<ol><li>FiO2'i (ilham alan oksijenin fraksiyonu) bir oksijen monitörü aracılığıyla oda içinde sürekli olarak izleyin.</li>
	<li>Hazneyi % 100 N2 için 60 L / dk N2 ve% 100 O2 için 0.415 L / dk ile yıkayın. Haznedeki oksijen doygunluğu %12'ye ulaştığında, O2 akışını değiştirmeden tutarken N2 akışını 10 L/dk'ya düşürin. Küçük ayarlamalarla FiO2'yi 45 dakika boyunca %8'de koruyun.</li>
	<li>45 dakikalık hipoksi maruziyetinin ardından FiO2'yi %21'e döndürün.</li>
	<li>Yavruların odada iyileşmesini ve hipoksi sonrası 2 saat boyunca EEG'de izlenmesini sağla.</li>
	<li>Kayıt süresi tamamlandıktan sonra, fareleri EEG kaydından çıkarın ve anneye geri dönün.</li>
</ol>6. EEG analizi
<ol><li>EEG dosyasını LabChart Pro'da video ile analiz edin. Kör bir araştırmacının EEG'yi nöbetler ve arka plan desenleri için işaretlemesini söyle17. Nöbetler, net evrim ile yüksek frekanslı ritmik keskin dalga deşarjları (≥3x taban çizgisi) ile 10 saniyeden fazla süren elekrografik bir olay olarak tanımlanır17.</li>
	<li>İkinci bir kör araştırmacı incelemesinin anlaşma için rastgele işaretlenmiş olayları geçirmesini haline geçirin.</li>
	<li>İşaretli her elektrografik olay için ilişkili videoyu gözden geçirin ve yenidoğan kemirgen davranışsal nöbet skoruna göre analiz edin16. Kısaca, bu puan 0-6 arasında değişmektedir (hareketsizlik şiddetli tonik-klonik davranış). Nöbet semiyolojisini daha da karakterize etmek için, yanallık için davranışı analiz edin (multifokal/bilateral hareketler ve odak/tek taraflı vs. karışık).</li>
	<li>Bir güç spektrogramı oluşturun. 1024 veri noktası boyutuna sahip Cosine-Bell veri penceresine sahip hızlı fourier dönüşümü kullanın. %87,5'lik bir pencere çakışması yardımıyla spektrogramda düzgün bir x ekseni oluşturun. Gücü μV218 olarak ifade edin.</li>
</ol>

<ol>
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Childhood outcomes after hypothermia for neonatal encephalopathy.</a> New England Journal of Medicine. 366 (22), 2085-2092 (2012).</li><li>Pappas, A., 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=Cognitive+outcomes+after+neonatal+encephalopathy.">Cognitive outcomes after neonatal encephalopathy.</a> Pediatrics. 135 (3), 624-634 (2015).</li><li>van Schie, P. 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Hipoksik-iskemik nöbetler sırasında yenidoğan farelerde sürekli video-EEG izleme için bir model sunduk. EEG ile birlikte video analizi, nöbet semiyolojisinin karakterizasyonuna izin verir. EEG analizi, güç spektrogramlarının çıkarılmasına ve arka plan genlik analizine izin verir.
Elektrotların doğru ve dikkatli yerleştirilmesi bu protokolde çok önemlidir, çünkü elektrot yerleştirme veya yanlış yerleştirme sırasında yaralanma sonuçları önemli ölçüde etkileyebi...

Hipoksik iskemik ensefalopati (HIE) yılda 1000 yenidoğanda 1,5'ini etkileyen ve yenidoğan nöbetlerinin en sık nedeni olan bir durumdur1,2. Hayatta kalan bebekler serebral palsi, zihinsel engellilik ve epilepsi gibi çeşitli nörolojik engelliler için risk altındadır3,4,5.
Hayvan modelleri hipoksi iskemi ve yenidoğan nöbetlerinin patofizyolojisini anlamada ve araştırmada kritik bir rol oynar6,7. Modifiye vannucci modeli, doğum sonrası 10 (p10)7,8'de hipoksi iskemisini (HI) indük etmek için kullanılır. Bu yaştaki fare yavruları nörolojik olarak kabaca tam insan yenidoğanı terimine çevirir9.
Bu yaralanma modeli ile birlikte kullanılan sürekli video elektroensefalografi (EEG) izleme, yenidoğan hipoksik iskemik nöbetlerin daha iyi anlaşılmasına ve karakterizasyonuna olanak tanır. Önceki çalışmalar kemirgenlerdeki yenidoğan nöbetlerini analiz etmek için video kayıtları, sınırlı EEG kayıtları ve telemetri EEG kayıtları dahil olmak üzere çeşitli yöntemler kullanmıştır10,11,12,13,14,15,16. Aşağıdaki yazıda, hipoksi-iskemi sırasında fare yavrularında sürekli video EEG kaydetme sürecini derinlemesine ele alıyoruz. Yenidoğan fare yavrularında sürekli video EEG izleme için bu teknik çeşitli hastalık ve nöbet modellerine uygulanabilir.

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continuous video electroencephalogram during hypoxia-ischemia in neonatal mice

Hipoksi iskemi yenidoğan nöbetlerinin en sık nedenidir. Hayvan modelleri, yenidoğan nöbetleri ve hipoksi iskemisinin altında kalan mekanizmaları ve fizyolojiyi anlamak için çok önemlidir. Bu makalede, hipoksi iskemisi sırasında nöbetleri tespit etmek ve EEG arka planını analiz etmek için yenidoğan farelerinde sürekli video elektroensefalogram (EEG) izleme yöntemi açıklanmaktadır. Video ve EEG'nin birlikte kullanılması, nöbet semiyolojisinin tanımlanmasına ve nöbetlerin onaylanmasına izin verir. Bu yöntem ayrıca deneysel zaman dilimi boyunca güç spektrogramlarının ve EEG arka plan desen eğilimlerinin analizine izin verir. Bu hipoksi iskemi modelinde yöntem, yaralanmadan önce EEG kaydının normatif bir taban çizgisi elde etmesine ve yaralanma ve iyileşme sırasında iyileşmesine izin verir. Toplam izleme süresi, yavruların anneden dört saatten uzun süre ayrılamaması ile sınırlıdır. Bu yazıda hipoksik-iskemik nöbetlerin bir modelini kullanmış olsak da, yenidoğan video EEG izleme için bu yöntem kemirgenlerde çeşitli hastalık ve nöbet modellerine uygulanabilir.

Nöbet semiyolojisi
Yenidoğan hipoksi-iskemi maruziyeti farelerde hem genelleştirilmiş hem de odak nöbetleriyle sonuçlanır (Şekil 1A-C). Video EEG kayıtları, elekrografik bulguların videodaki davranışla ilişkilendirilmesini sağlar. Bu davranışlar daha önce yayınlanan yenidoğan kemirgen davranışsal nöbet skoru (BSS)16 kullanılarak puanlanmıştır. BSS'ye ek olarak, olayları odak/tek taraflı...

Watch this Scientific Journal Video about Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice at JoVE.com

Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice

Bu makalede hipoksi-iskemi geçiren yenidoğan farelerde birden fazla derinlik elektrodu kullanılarak sürekli video EEG kayıtları için bir yöntem açıklanmaktadır.

Yenidoğan Farelerde Hipoksi-İskemi Sırasında Sürekli Video Elektroensefalogram

Hypoxia ischemia is the most common cause of neonatal seizures. Animal models are crucial for understanding the mechanisms and physiology underlying ...

continuous-video-electroencephalogram-during-hypoxia-ischemia

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

3.3K Görüntüleme.  University of Virginia. Bu makalede hipoksi-iskemi geçiren yenidoğan farelerde birden fazla derinlik elektrodu kullanılarak sürekli video EEG kayıtları için bir yöntem açıklanmaktadır.

Video: Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice

Delivery of In Vivo Acute Intermittent Hypoxia in Neonatal Rodents to Prime Subventricular Zone-derived Neural Progenitor Cell Cultures

Extended culture of neural stem/progenitor cells facilitates in vitro analyses to understand their biology while enabling expansion of cell populations to adequate numbers prior to transplantation. Identifying approaches to refine this process, to augment the production of all CNS cell types (i.e., neurons), and to possibly contribute to therapeutic cell therapy protocols is a high research priority. This report describes an easily applied in vivo &#8220;pre-conditioning&#8221; stimulus which can be delivered to awake, non-anesthetized animals. Thus, it is a non-invasive and non-stressful procedure. Specifically described are the procedures for exposing mouse or rat pups (aged postnatal day 1-8) to a brief (40-80 min) period of intermittent hypoxia (AIH). The procedures included in this video protocol include calibration of the whole-body plethysmography chamber in which pups are placed during AIH and the technical details of AIH exposure. The efficacy of this approach to elicit tissue-level changes in the awake animal is demonstrated through the enhancement of subsequent in vitro expansion and neuronal differentiation in cells harvested from the subventricular zone (SVZ). These results support the notion that tissue level changes across multiple systems could be observed following AIH, and support the continued optimization and establishment of AIH as a priming or conditioning modality for therapeutic cell populations.

Delivery of In Vivo Acute Intermittent Hypoxia in Neonatal Rodents to Prime Subventricular Zone-derived Neural Progenitor Cell Cultures

This article describes the methodology for administering short periods of intermittent hypoxia to postnatal day 1-8 mouse or rat pups. This approach effectively elicits a robust tissue level &#8220;priming effect&#8221; on cultured neural progenitor cells that are harvested within 30 min of hypoxia exposure.

Teslimatı<em&gt; İn Vivo</em&gt; Akut Aralıklı hipoksi Yenidoğan Kemirgenler Başbakan subventricular bölgesi kökenli Sinir Progenitör Hücre Kültürleri için

Extended culture of neural stem/progenitor cells facilitates in vitro analyses to understand their biology while enabling expansion of cell populations to ...

Gelişim Biyolojisi

A Swine Model of Neonatal Asphyxia

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

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

Yenidoğan Asfiksi Bir Domuz Modeli

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

Tıp

Simultaneous Video-EEG-ECG Monitoring to Identify Neurocardiac Dysfunction in Mouse Models of Epilepsy

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can potentially increase risk of sudden unexpected death in epilepsy (SUDEP). Electroencephalography (EEG) and electrocardiography (ECG) are widely used clinical diagnostic tools to monitor for abnormal brain and cardiac rhythms in patients. Here, a technique to simultaneously record video, EEG, and ECG in mice to measure behavior, brain, and cardiac activities, respectively, is described. The technique described herein utilizes a tethered (i.e., wired) recording configuration in which the implanted electrode on the head of the mouse is hard-wired to the recording equipment. Compared to wireless telemetry recording systems, the tethered arrangement possesses several technical advantages such as a greater possible number of channels for recording EEG or other biopotentials; lower electrode costs; and greater frequency bandwidth (i.e., sampling rate) of recordings. The basics of this technique can also be easily modified to accommodate recording other biosignals, such as electromyography (EMG) or plethysmography for assessment of muscle and respiratory activity, respectively. In addition to describing how to perform the EEG-ECG recordings, we also detail methods to quantify the resulting data for seizures, EEG spectral power, cardiac function, and heart rate variability, which we demonstrate in an example experiment using a mouse with epilepsy due to Kcna1 gene deletion. Video-EEG-ECG monitoring in mouse models of epilepsy or other neurological disease provides a powerful tool to identify dysfunction at the level of the brain, heart, or brain-heart interactions.

Here, we present a protocol to record brain and heart bio signals in mice using simultaneous video, electroencephalography (EEG), and electrocardiography (ECG). We also describe methods to analyze the resulting EEG-ECG recordings for seizures, EEG spectral power, cardiac function, and heart rate variability.

Eş zamanlı Video-EEG-EKG Neurocardiac disfonksiyon epilepsi fare modelleri tanımlamak için izleme

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can ...

Genetik

The Hypoxic Ischemic Encephalopathy Model of Perinatal Ischemia

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe and permanent neuropsychological sequelae, including mental retardation, cerebral palsy, and epilepsy. The outcomes of HIE are devastating and permanent, making it critical to identify and develop therapeutic strategies to reduce brain injury in newborns with HIE. To that end, the neonatal rat model for hypoxic-ischemic brain injury has been developed to model this human condition. The HIE model was first validated by Vannucci et al 1 and has since been extensively used to identify mechanisms of brain injury resulting from perinatal hypoxia-ischemia 2 and to test potential therapeutic interventions 3,4. The HIE model is a two step process and involves the ligation of the left common carotid artery followed by exposure to a hypoxic environment. Cerebral blood flow (CBF) in the hemisphere ipsilateral to the ligated carotid artery does not decrease because of the collateral blood flow via the circle of Willis; however with lower oxygen tension, the CBF in the ipsilateral hemisphere decreases significantly and results in unilateral ischemic injury. The use of 2,3,5-triphenyltetrazolium chloride (TTC) to stain and identify ischemic brain tissue was originally developed for adult models of rodent cerebral ischemia 5, and is used to evaluate the extent of cerebral infarctin at early time points up to 72 hours after the ischemic event 6. In this video, we demonstrate the hypoxic-ischemic injury model in postnatal rat brain and the evaluation of the infarct size using TTC staining.

The postnatal rat model for hypoxic-ischemic brain injury is a well-established model of human neonatal hypoxic ischemic encephalopathy (HIE). In this article, we describe the model of HIE in post-natal rat pups.

Perinatal İskemi Hipoksik İskemik Ensefalopati Modeli

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe ...

Biyoloji

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

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

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

Yenidoğan Hipoksik-İskemik Ensefalopati A Piglet Modeli

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

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

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

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

Yenidoğan hipoksik-iskemik beyin hasarı, fare modeli neurobehavioral değerlendirmesi

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

Nörobilim

Noninvasive Electrocardiography in the Perinatal Mouse

Electrocardiography (ECG) has long been relied upon as an effective and reliable method of assessing cardiovascular (and cardiopulmonary) function in both human and animal models of disease. Individual heart rate, rhythm, and regularity, combined with quantitative parameters collected from ECG, serve to assess the integrity of the cardiac conduction system as well as the integrated physiology of the cardiac cycle. This article provides a comprehensive description of the methods and techniques used to perform a noninvasive ECG on perinatal and neonatal mouse pups as early as the first postnatal day, without requiring the use of anesthetics. This protocol was designed to directly address a need for a standardized and repeatable method for obtaining ECG in newborn mice. From a translational perspective, this protocol proves to be entirely effective for characterization of congenital cardiopulmonary defects generated using transgenic mouse lines, and particularly for analysis of defects causing lethality at or during the first postnatal days. This protocol also aims to directly address a gap in the scientific literature to characterize and provide normative data associated with maturation of the early postnatal cardiac conduction system. This method is not limited to a specific postnatal timepoint, but rather allows for ECG data collection in neonatal mouse pups from birth to postnatal day 10 (P10), a window that is of critical importance for modeling human diseases in vivo, with particular emphasis on congenital heart disease (CHD).

Here, we present a noninvasive electrocardiography (ECG) protocol, optimized for early postnatal mice, that does not require the use of anesthetics.

Perinatal Farede Noninvaziv Elektrokardiyografi

Electrocardiography (ECG) has long been relied upon as an effective and reliable method of assessing cardiovascular (and cardiopulmonary) function in both ...

Echocardiographic Characterization of Left Ventricular Structure, Function, and Coronary Flow in Neonate Mice

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular disease and is used to assess the impact of potential treatments in preclinical studies. Echocardiographic studies are usually conducted in young adult mice (i.e., 4-6 weeks of age). The evaluation of early neonatal cardiovascular function is not usually performed because of the small size of the mouse pups and the associated technical difficulties. One of the most important challenges is that the short length of the pups&#39; limbs prevents them from reaching the electrodes in the echocardiography platform. Body temperature is the other challenge, as pups are very susceptible to changes in temperature. Therefore, it is important to establish a practical guide for performing echocardiographic studies in small mouse pups to help researchers detect early pathological changes and study the progression of cardiovascular disease over time. The current work describes a protocol for performing echocardiography in mouse pups at the early age of 7 days old. The echocardiographic characterization of cardiac morphology, function, and coronary flow in neonatal mice is also described.

The present protocol describes the echocardiographic assessment of left ventricular morphology, function, and coronary blood flow in 7-day old neonate mice.

Yenidoğan Farelerde Sol Ventrikül Yapısının, Fonksiyonunun ve Koroner Akımın Ekokardiyografik Karakterizasyonu

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular ...

Early Pathological and Magnetic Resonance Detection of Cerebral Injury Using a Rat Model of Neonatal Hypoxic Ischemic Encephalopathy

Perinatal hypoxic-ischemic encephalopathy (HIE) is an acute disease that may afflict newborns, resulting in variable long- and short-term neurodevelopmental outcomes. Early diagnosis is critical to identifying infants who may benefit from intervention; however, early diagnosis relies heavily on clinical criteria. No molecular or radiological tests have shown promise in detecting early cerebral injury. Studies have shown that magnetic resonance imaging (MRI) can show changes in both blood flow/ischemia and metabolic disruption. However, they have all been used to evaluate the secondary phase of the disease (&gt;12 h) after the onset of the injury. Early diagnosis is critical to rapidly starting therapeutic hypothermia in eligible infants, which is currently recommended to be initiated within 6 h of birth. The rat model of hypoxic-ischemic injury was developed in 1981 and has been validated and used extensively to study changes in brain perfusion, cerebral injury markers, and morphology. However, it has primarily been used as a "late model", evaluating injury several days after the initial ischemic insult. The model has been known to have poor sensitivity in evaluating reliable and reproducible early cerebral changes. The objective of this study was to develop a reliable model to study early gross morphological and radiological markers of HIE using pathological staining and cerebral magnetic resonance imaging/magnetic resonance spectroscopy.

The present protocol describes a rodent model of newborn hypoxic-ischemic injury for identifying early changes in cerebral tissue by gross morphology and magnetic resonance imaging. This has benefits over existing models, which can be used to study late injury but do not allow the evaluation of reproducible early changes.

Perinatal hypoxic-ischemic encephalopathy (HIE) is an acute disease that may afflict newborns, resulting in variable long- and short-term ...

Establishing a Mouse Model of Neonatal Hypoxic-Ischemic Brain Injury

Source: Kim, M., et al. <a href="https://appjove.unicartagenaproxy.elogim.com/v/55838">Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury.</a> J. Vis. Exp. (2017).
The video demonstrates the procedure for creating a mouse model to study neonatal hypoxic-ischemic brain injury. It outlines the steps for surgically inducing ischemia and subjecting the animal to hypoxia to exacerbate brain damage. Finally, it shows how to assess the resulting brain lesions, ensuring the mouse model is prepared for further scientific investigation.

Yenidoğan Farelerde Hipoksi-İskemi Sırasında Sürekli Video Elektroensefalogram

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