Bu çalışma Çin Doğa Bilimleri Vakfı (31171355) ve Guangdong Doğa Bilimleri Vakfı (S2011010003403, 2014A030313440) tarafından desteklendi.

Hayvan bakımı, cerrahi prosedür ve kayıt prosedürleri, Güney Çin Normal Üniversitesi Hayvan Bakımı ve Kullanım Komitesi için yönergelere uygundu. 1. Elektrot Hazırlama (Şekil 1A-C) NOT: Bilgisayar pin loküsü, iletişim aygıtlarındaki sinyal arabiriminin bir parçası olarak sadece bir uçlu kontakttır. Dişi konektöre takılan bir erkek konektörden oluşur. <ol><li> Bir kıskaç yardımıyla erkek ve dişi pimleri bilgisayarın pin lokusundan dikkatlice ayırın ( Şekil 1A ). Erkek ve dişi pimleri bir elektrot oluşturacak şekilde birleştirin ve güçlü bir yapışkan bağ oluşturmak için siyanoakrilat uygulayın ( Şekil 1C ). </li><li> Elektrotları, damıtılmış su ile dolu bir behere koyun ve 10 dakika boyunca ultrasonik temizleyiciye yerleştirin. Onları 45 ° C&#39;de 30 dakika kurutma fırına taşıyın. Elektrodları UV ışığı kullanarak 30 dakika sterilize edin. </li></ol> 2. Cerrahi prosedür (Şekil 1D-F) <ol><li> Sterilize cerrahi aletleri ve stereotaksik aparat hazırlayın. Yenidoğan sıçan yavrusuna isofluran anestezi (% 2.5) ile hava ile anestezi uygulayın. Pedi derin anestezi altına alındığında, izofluran dozunu% 1.0&#39;a ayarlayın. Anestezinin doğru derinliğini sağlamak için cerrahiden önce bir kuyruk veya ayak parmakları tutturun. </li><li> Kulak çubuklarını kulak kanallarına yerleştirerek ve hafif sıkıştırarak yavrunun başını stereotaksik aparatta sabitleyin. NOT: Yenidoğan kafatası çok yumuşak olduğu için kulak çubuklarını aşırı derecede sıkmayın. </li><li> Tüm ekipmana% 70 etanol püskürterek steril cerrahi alanı koruyun. Başınızı bir neşter kullanarak 15 mm&#39;lik bir kesi yapın. Forseps kullanarak, dört köşedeki orta hattan kafa derisini yavaşça çekin. İnsizyonun geniş açık kalmasını sağlamak için salinle ıslanmış bir pamuk cildin altına koyun ( Şekil 1D ). </li><li> Kafatasındaki bregma ve lambda noktalarını bulunVe kalemle işaretleyin. Prefrontal kortekse (PFC) ve hipokampusa iki çapak deliği açmak için bir şırınga iğnesi (26 G) kullanın. NOT: Perküsyon, bregma&#39;nın önüne -2.0 mm, orta çizgiye ± 0.5 mm lateralde ( Şekil 1D ve E ) bulunduğu sırada bregma&#39;nın arkasından +1.8 mm ve orta hattan -0.5 mm lateralde bulunur. Beyin hasarını en aza indirgemek için elektrot derinliği korteks yüzeyinin 2 mm&#39;den daha aşağıda olmamalıdır. </li><li> Elektrotları tutmak için forseps kullanın ve sırasıyla referans ve kayıt elektrodlarını PFC&#39;ye ve hipokampusa yerleştirin. Olası enfeksiyonlardan kaçınmak için elektrotun çevresine eritromisin merhemi uygulayın. Elektrodu siyanoakrilat kullanarak sabitleyin. </li><li> Dental akrilik çimentoyu yapışkan ve yapışkan bir kıvamda olacak şekilde hazırlayın. Elektrotları ve kafatasının geri kalanını örtecek diş çimentosunu uygulayın. NOT: Diş çimentosunu uygulamadan önce kafatasını iyice kurutun. </li&gt; <li> Onları korumak için elektrotların üzerine% 5 pikrik asit uygulayın. NOT: Bütün prosedür, steril koşulları korumak için biyogüvenlikli bir kaput içinde yapılmalıdır. </li><li> Hayvanı stereotaksik çerçeveden çıkarın ve subkutan yoldan 300 μL% 10 glikoz enjekte edin. İyileştirmek için onu ısıtmalı bir battaniye koyun. Hayvanın sıcak (37 ° C) ve ayakta ( diğer bir deyişle tamamen iyileşmiş) olduğundan emin olun. Cerrahi sonrası ağrı için buprenorfin intraperitoneal (0.05 mg / kg) uygulanır. NOT: Bir hayvanı yeterince bilinç kazanana kadar gözetimsiz bırakmayın (normal davranış ve hareket). </li><li> Bilinci geri gelip yavruları barajlarıyla evindeki kafese geri getirin. Hayvan tamamen kurtarılana kadar iki gün bekleyin. </li></ol> 3. EEG Kaydı <ol><li> Tam toparlandıktan sonra yavrunun kafatasına implante edilen elektrotları kendi kafesindeki amplifikatöre bağlayın. Amplifikatörü analogdan dijital c&#39;ye bağlayınDönüştürücü ve dönüştürücü bir bilgisayara takın; Bağlantı hatları karışık olmamaları için dikkatle muamele edilmelidir. </li><li> Kayıt için veri toplama ünitesinde en az 10,000 Hz örnekleme hızı seçin (vericinin bant genişliği 1-100 Hz&#39;dir). Verilerin doğru örneklendiğinden emin olun. </li><li> Temel kayıt alınmasından sonra, pupayı epileptik nöbetleri indüklemek için intraperitoneal olarak kainik asit (KA) (2 mg / kg) ile enjekte edin. KA enjeksiyonundan 15 dakika sonra epileptik deşarjları gözlemleyin ve kaydedin. KA aracılığıyla üretilen nöbet genellikle fiziksel. NOT: İitalt tonik süre yaklaşık 15.2 ± 0.9 s, nöbet süresi yaklaşık 62 ± 5 s&#39;dir. Yenidoğan sıçanında, IP enjeksiyonu kloral hidrat (400 mg / kg) vererek nöbetler önlenebilir. </li><li> Dijitalleştirilmiş verileri kaydedin ve spike2 gibi sinyal işleme yazılımı paketlerini kullanarak analiz edin. Yenidoğan EEG sinyalindeki farklı frekans bileşenlerinin güç düzeyini perf ile ortaya koyunBir güç spektrumu analizinin orming edilmesi. 1 ila 100 Hz (EEG bandı) 9 arasındaki kök-ortalama kare genliğini bulmak için gücü 1 dakikalık bir zaman aralığında hesaplayın. </li></ol>

<ol>
	<li>Pereda, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+synapses+and+their+functional+interactions+with+chemical+synapses.">Electrical synapses and their functional interactions with chemical synapses.</a> Nat Rev Neurosci. 15 (4), 250-263 (2014).</li><li>Chorev, E., Epsztein, J., Houweling, A. R., Lee, A. K., Brecht, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+recordings+from+behaving+animals--going+beyond+spikes.">Electrophysiological recordings from behaving animals--going beyond spikes.</a> Curr Opin Neurobiol. 19 (5), 513-519 (2009).</li><li>Freeborn, D. L., McDaniel, K. L., Moser, V. C., Herr, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+electroencephalography+(EEG)+to+assess+CNS+changes+produced+by+pesticides+with+different+modes+of+action:+effects+of+permethrin,+deltamethrin,+fipronil,+imidacloprid,+carbaryl,+and+triadimefon.">Use of electroencephalography (EEG) to assess CNS changes produced by pesticides with different modes of action: effects of permethrin, deltamethrin, fipronil, imidacloprid, carbaryl, and triadimefon.</a> Toxicol Appl Pharmacol. 282 (2), 184-194 (2015).</li><li>Werhahn, K. J., Hartl, E., Hamann, K., Breimhorst, M., Noachtar, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Latency+of+interictal+epileptiform+discharges+in+long-term+EEG+recordings+in+epilepsy+patients.">Latency of interictal epileptiform discharges in long-term EEG recordings in epilepsy patients.</a> Seizure. 29, 20-25 (2015).</li><li>Staba, R. J., Stead, M., Worrell, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+biomarkers+of+epilepsy.">Electrophysiological biomarkers of epilepsy.</a> Neurotherapeutics. 11 (2), 334-346 (2014).</li><li>Nimmrich, V., Draguhn, A., Axmacher, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+Network+Oscillations+in+Neurodegenerative+Diseases.">Neuronal Network Oscillations in Neurodegenerative Diseases.</a> Neuromolecular Med. 17 (3), 270-284 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+Continuous+EEG+Monitoring+in+Small+Rodent+Models+of+Human+Disease+Using+the+Epoch+Wireless+Transmitter+System.">Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System.</a> J Vis Exp. (101), e52554 (2015).</li><li>Zayachkivsky, A., Lehmkuhle, M. J., Fisher, J. H., Ekstrand, J. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+EEG+in+immature+rats+with+a+novel+miniature+telemetry+system.">Recording EEG in immature rats with a novel miniature telemetry system.</a> J Neurophysiol. 109 (3), 900-911 (2013).</li><li>Dzhala, V. I., 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=NKCC1+transporter+facilitates+seizures+in+the+developing+brain.">NKCC1 transporter facilitates seizures in the developing brain.</a> Nat Med. 11 (11), 1205-1213 (2005).</li><li>Tucker, A. M., Aquilina, K., Chakkarapani, E., Hobbs, C. E., Thoresen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+amplitude-integrated+electroencephalography+and+interburst+interval+in+the+rat.">Development of amplitude-integrated electroencephalography and interburst interval in the rat.</a> Pediatr Res. 65 (1), 62-66 (2009).</li><li>Savard, 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=Involvement+of+neuronal+IL-1beta+in+acquired+brain+lesions+in+a+rat+model+of+neonatal+encephalopathy.">Involvement of neuronal IL-1beta in acquired brain lesions in a rat model of neonatal encephalopathy.</a> J Neuroinflammation. 10, 110 (2013).</li><li>Cuaycong, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+approach+to+the+study+of+hypoxia-ischemia-induced+clinical+and+subclinical+seizures+in+the+neonatal+rat.">A novel approach to the study of hypoxia-ischemia-induced clinical and subclinical seizures in the neonatal rat.</a> Dev Neurosci. 33 (3-4), 241-250 (2011).</li></ol>

Yazarlar çıkar çatışması olmadığını beyan eder.

Burada serbestçe hareket eden yenidoğan sıçan yavrularını kablolu yöntemle EEG elde etmek için cerrahi ve kayıt prosedürlerini bildiriyoruz ( Şekil 3 ). P7-P12 sıçan yavrusunun tam dönem insan yenidoğanına karşılık gelen gelişimsel yaşta olduğu öne sürülmüştür 10,11. Bu yaş grubundaki sıçan yavrularıyla çalışırken teknik olarak yüksek kaliteli EEG kayıt verileri elde etmek zordur. Buna ek olarak, özel kullanım ve çok dikkatli olmayı gerektirir ...

Normal beyin fonksiyonunu elde etmek için nöronlar arasındaki sürekli iletişimin gerekli olduğu iyi bilinmektedir. İnterneonal iletişim ilk olarak bir sinir hücresinden gelen bilgilerin ikinci bir sinir hücresine iletildiği sinapslarda gerçekleşir. Bu sinaptik iletim iki tür yapısal düzenlemeyle sağlanır: elektriksel veya kimyasal sinapsler 1 . Elektrofizyoloji, tüm vücut işlevlerini ve davranışını kontrol eden, internöronal iletişim sırasında üretilen elektrik potansiyelini yakalayan alanlardır. EEG birçok elektrofizyolojik teknik arasında en çok kullanılan yöntemdir. EEG, iç veya dış uyaranlar tarafından üretilen elektrik sinyallerindeki değişiklikleri saptamak için kullanılan bir tekniktir. Dahası, epilepsi, Parkinson ve Alzheimer hastalığı gibi çeşitli nörolojik durumların klinik tanı ve sonuç tahmininde önemli bir testtirE, farmakolojik ve toksikolojik ajanların etkileri 3 . Genellikle epileptik bir hastada beyinde aşırı uyarılma ve işlevsel bağlantının bozulması görülür; Bunlar interiktal epileptiform deşarjlar (IED&#39;ler) olarak özetlenmekte ve keskin, geçici sivri uçlar şeklinde EEG ile kaydedilebilmektedir; Keskin dalgalar; Dikenli dalga kompleksleri; Veya polipropikler 4 . Epileptik beynin asıl özelliği epileptik nöbetlerin kendiliğinden ortaya çıkmasıdır; nöbetlerden sorumlu beyin alanını bulmak için kafa derisinden veya beyin parankiminden kaydedilebilir 5 . Ayrıca, EEG&#39;nin Alzheimer hastalığı (AD) gibi nörodejeneratif bozukluklarda da önemli etkileri bulunmaktadır. Araştırmalar AD hastalarında değiştirilmiş EEG kayıtlarının ve bozulmuş salınım ağlarının yaygın olduğunu ileri sürüyor. Bununla birlikte, nörodejeneratif hastalıklarda ağ salınımlarının patofizyolojisi hakkında bilgi iŞaşırtıcı derecede eksik ve daha fazla araştırılması gerekiyor 6 . Bu protokolde hem normal hem de patolojik beyindeki elektriksel iletişimi anlamak için EEG&#39;yi kaydedebilen basit bir elektrot tasarladık. Bu yöntemdeki cerrahi implantasyon diğer kullanılabilir prosedürlerden daha ucuzdur 7 . Dahası, bu yöntem, daha uzun zaman aralıkları için yüksek kaliteli ve kararlı EEG sinyallerinin kaydedilmesi için kullanılabilir ( yani, 1 hafta boyunca her gün 2-4 saat). Buna ek olarak, hayvanların daha doğal davranmalarını sağlayan hafif elektrotlar (yaklaşık 26 mg ağırlığında) kullandık. Bu yöntem, elektrofizyoloji laboratuvarında yaygın olarak kullanılan amplifikatör ve sayısallaştırıcıyı gerektiren yenidoğan sıçan yavrularında EEG çalışması için yaygın bir şekilde uygulanabilir ve herhangi bir ek cihaz gerektirmez.

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			<td>Computer pin</td>
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			<td>Pincer</td>
			<td>DELI Group Co., Ltd.</td>
			<td></td>
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			<td>502 super glue</td>
			<td>DELI Group Co., Ltd.</td>
			<td>7144</td>
			<td></td>
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			<td>Drying oven</td>
			<td>Boxun</td>
			<td>GZX-9140MBE</td>
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			<td>Isofluorane</td>
			<td>RWD Life Science</td>
			<td>902-0000-522</td>
			<td></td>
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		<tr>
			<td>Stereotaxic apparatus</td>
			<td>RWD Life Science</td>
			<td>900-0068-507</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia apparatus</td>
			<td>RWD Life Science</td>
			<td>902-0000-510</td>
			<td></td>
		</tr>
		<tr>
			<td>Homeothermic Heating Device</td>
			<td>Harvard Apparatus</td>
			<td>K 024509</td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier Model 3000&#160;</td>
			<td>A-M Systems</td>
			<td>61558</td>
			<td></td>
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			<td>Micro1401 Analog Digital converter</td>
			<td>Cambridge Electronic Design Ltd.</td>
			<td>4383</td>
			<td>Data acquisition unit</td>
		</tr>
		<tr>
			<td>Spike2</td>
			<td>Cambridge Electronic Design Ltd.</td>
			<td></td>
			<td></td>
		</tr>
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recording eeg in freely moving neonatal rats using a novel method

EEG, beyindeki elektriksel aktiviteyi saptamak için yararlı bir yöntemdir. Dahası, epilepsi ve nörodejeneratif bozukluklar gibi çeşitli nörolojik durumlar için yaygın olarak kullanılan bir tanı aracıdır. Bununla birlikte, yenidoğanlarda EEG kayıtları elde etmek teknik olarak zor, özel muamele ve büyük bakım gerektirdiğinden. Burada, neonatal sıçan yavrularında EEG kaydetmek için yeni bir yöntem sunuyoruz (P8-P15). Bilgisayarın pin lokusunu kullanarak basit ve güvenilir bir elektrot tasarladık; Normal ve epileptik beyinde yüksek kaliteli EEG sinyallerinin kaydedilmesi için bir sıçan yavrusunun kafatasına kolayca implante edilebilir. Puplara epileptik nöbetleri indüklemek için nörotoksin kainik asitin (KA) intraperitoneal (ip) enjeksiyonu yapıldı. Bu prosedürde yapılan cerrahi implantasyon yenidoğanlar için diğer EEG prosedürlerinden daha ucuzdur. Bu yöntem, bir kişinin yüksek kaliteli ve istikrarlı EEG sinyallerini 1 haftadan fazla süreyle kaydetmesini sağlar. Dahası, bu prosedür, yetişkin ra&#39;ya da uygulanabilirTs ve farelerden epilepsi veya diğer nörolojik bozuklukları incelemek için kullanılmıştır.

<html> Yukarıdaki cerrahi prosedürler düzgün bir şekilde yürütülürse, bir kanallı sıçan yenidoğan yavrusu EEG kaydı başarıyla gerçekleştirilecektir. KA enjeksiyonundan 10 dakika sonra düzensiz hareketler, kaşınma, titreme ve denge kaybı şeklinde düzenli davranış belirtileri paternleri ortaya çıktı. Şekil 2, temsili ham EEG izlerini ve interiktal, ictal tonik ve ictal-klonik genişletilmiş izleri göstermektedir. KA enjeksiyonundan sonra tekrarlayan interikta...

Watch this Scientific Journal Video about Recording EEG in Freely Moving Neonatal Rats Using a Novel Method at JoVE.com

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

Burada, serbestçe hareket eden neonatal epileptik yavrulara elektroensefalografiyi (EEG) kaydetmek ve prosedürlerini, özelliklerini ve uygulamalarını tanımlamak için tasarlanmış yeni bir teknik sunuyoruz. Bu yöntem, EEG&#39;yi 1 haftadan daha uzun süreyle kayıt ettirmenizi sağlar.

Yeni Bir Yöntemi Kullanarak Serbest Dolaşan Yenidoğan Sıçanlarında EEG Kaydı

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

recording-eeg-in-freely-moving-neonatal-rats-using-a-novel-method

Research

JoVE Journal

Neuroscience

13.1K Görüntüleme.  South China Normal University.  Burada, serbestçe hareket eden neonatal epileptik yavrulara elektroensefalografiyi (EEG) kaydetmek ve prosedürlerini, özelliklerini ve uygulamalarını tanımlamak için tasarlanmış yeni bir teknik sunuyoruz. Bu yöntem, EEG'yi 1 haftadan daha uzun süreyle kayıt ettirmenizi sağlar. 

Video: Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

Novel Apparatus and Method for Drug Reinforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction. Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in animal research to model various components of drug reinforcement, consumption, and addiction in humans. For this study, we used a novel approach to studying drug reinforcement in rats by combining traditional CPP and self-administration methodologies. We assembled an apparatus using two Med Associate operant chambers, sensory stimuli, and a Plexiglas-constructed neutral zone. These modifications allowed our experiments to encompass motivational aspects of drug intake through self-administration and drug-free assessment of drug/cue conditioning strength with the CPP test. In our experiments, rats self-administered cocaine (0.75 mg/kg/inj, i.v.) during either four (e.g., the "short-term") or eight (e.g., the "long-term") alternating-day sessions in an operant environment containing distinctive sensory cues (e.g., olfactory and visual). On the alternate days, in the other (differently-cued) operant environment, saline was available for self-infusion (0.1 ml, i.v.). Twenty-four hours after the last self-administration/cue-pairing session, a CPP test was conducted. Consistent with typical CPP findings, there was a significant preference for the chamber associated with cocaine self-administration. In addition, in animals undergoing the long-term experiment, a significant positive correlation between CPP magnitude and the number of cocaine-reinforced lever responses. In conclusion, this apparatus and approach is time and cost effective, can be used to examine a wide array of topics pertaining to drug abuse, and provides more flexibility in experimental design than CPP or self-administration methods alone.

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

İlaç Takviye için yeni cihaz ve Yöntem

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

Nörobilim

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

Poliimid-temelli Mikroelektrod kullanarak Serbestçe Hareketli Fare Yüksek yoğunluklu EEG Kayıtlar

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

Sıçanlarda Sakkaroz Craving Kuluçka değerlendirilmesi için Genel Bir Yöntem

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

Serbestçe Öğrenmenin üç aşamalı karşısında Güvercinler Hareketli gelen tek nöronların &#39;Aksiyon Potansiyelleri kayıt

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform natural behavior. We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allowed us to record activity from multiple neurons simultaneously while a tethered cockroach turned or altered walking speed. While a major advance, tethered preparations provide access to limited behaviors and often lack feedback processes that occur in freely moving animals. We now present a modified version of that technique that allows us to record from the central complex of freely moving cockroaches as they walk in an arena and deal with barriers by turning, climbing or tunneling. Coupled with high speed video and cluster cutting, we can now relate brain activity to various parameters of the movement of freely behaving insects.

We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allows us to monitor activity in individual units of tethered cockroaches. Here we present a modified version of that technique that allows us to also record brain activity in freely moving insects.

Serbestçe Böcekler Yürüyüş Beyin Ektraselüler Tel Tetrode Kaydı

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

A Novel Davranış Testi (Bumble Bees Serbestçe hareket eden Bireyin Tat Cevapları, Araştırma<em&gt; Bombus terrestris</em&gt;)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

Noninvasive EEG Recordings from Freely Moving Piglets

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel telemetric electroencephalography system in combination with standard self-adhesive hydrogel electrodes. The piglets are calmed down without the use of sedatives. After their release into the pigpen, the piglets behave normally&#8212;they drink and sleep in the same cycle as their siblings. Their sleep phases are used for the EEG recordings.

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

Domuz yavruları serbestçe hareket noninvaziv EEG kayıtları

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

Microdialysis fareler serbestçe hareket EEG kayıtları sırasında eksitatör Amino asit

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

SerbestÇe Hareket Eden Farelerde Çift Alan Yüksek Yoğunluklu Kayıt için Kurtarılabilir Opto-Silikon Prob ve Tetrode ile Hibrid Microdrive Sistemi

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and smaller functional subgroups, it becomes paramount to record from projections and/or genetically-defined subpopulations of neurons. Fiber photometry is an accessible and powerful approach that can overcome these challenges. By combining optical and genetic methodologies, neural activity can be measured in deep brain structures by expressing genetically-encoded calcium indicators, which translate neural activity into an optical signal that can be easily measured. The current protocol details the components of a multi-fiber photometry system, how to access deep brain structures to deliver and collect light, a method to account for motion artifacts, and how to process and analyze fluorescent signals. The protocol details experimental considerations when performing single and dual color imaging, from either single or multiple implanted optic fibers.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

SerbestÇe Hareket Eden Hayvanlarda Nöral AktiviteYi Kaydetmek Için Çoklu Fiber Fotometri

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...