Bu makalenin yazımına ve özenle okunmasına katkılarından ötürü tüm laboratuar üyelerine teşekkür ediyoruz. Tartışmaları teşvik ettiği için Paul Franken&#39;e, cerrahi protokol hakkında faydalı yorumlar için Dr. Gisèle Ferrand&#39;a ve gürültüye maruz kalma için orijinal Labview çalıştırılabilir dosyalarını sunma konusunda Dr. Jean-Yves Chatton&#39;a minnettarız. Finansman İsviçre Ulusal Bilim Vakfı (Hibe 31003A_146244 ve 31003A_166318) ve Etat de Vaud tarafından sağlandı.

<ol>
	<li>Lecci, S., 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=Coordinated+infra-slow+neural+and+cardiac+oscillations+mark+fragility+and+offline+periods+in+mammalian+sleep.">Coordinated infra-slow neural and cardiac oscillations mark fragility and offline periods in mammalian sleep.</a> Sci Adv. 3 (2), 1602026 (2017).</li><li>Rechtschaffen, A., Kales, 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+manual+of+standardized+terminology,+techniques+and+scoring+system+for+sleep+of+human+subjects.">A manual of standardized terminology, techniques and scoring system for sleep of human subjects.</a> U.S. Department of Health, Education, and Welfare. , (1968).</li><li>Blake, H., Gerard, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+potentials+during+sleep.">Brain potentials during sleep.</a> Am J Physiol. 119, 692-703 (1937).</li><li>Rechtschaffen, A., Hauri, P., Zeitlin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auditory+awakening+threshold+in+REM+and+NREM+sleep+stages.">Auditory awakening threshold in REM and NREM sleep stages.</a> Percept Mot Skills. 22 (3), 927-942 (1966).</li><li>Achermann, P., Borb&#233;ly, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-frequency+(&lt;+1+Hz)+oscillations+in+the+human+sleep+electroencephalogram.">Low-frequency (&lt; 1 Hz) oscillations in the human sleep electroencephalogram.</a> Neuroscience. 81 (1), 213-222 (1997).</li><li>Aeschbach, D., Borb&#233;ly, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=All-night+dynamics+of+the+human+sleep+EEG.">All-night dynamics of the human sleep EEG.</a> J. Sleep Res. 2 (2), 70-81 (1993).</li><li>Astori, S., Wimmer, R. D., L&#252;thi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Manipulating+sleep+spindles--expanding+views+on+sleep,+memory,+and+disease.">Manipulating sleep spindles--expanding views on sleep, memory, and disease.</a> Trends Neurosci. 36 (12), 738-748 (2013).</li><li>Brown, R. E., Basheer, R., McKenna, J. T., Strecker, R. E., McCarley, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+sleep+and+wakefulness.">Control of sleep and wakefulness.</a> Physiol Rev. 92 (3), 1087-1187 (2012).</li><li>Buzs&#225;ki, G., Wang, X. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+gamma+oscillations.">Mechanisms of gamma oscillations.</a> Annu Rev Neurosci. 35, 203-225 (2012).</li><li>Rasch, B., Born, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=About+sleep's+role+in+memory.">About sleep's role in memory.</a> Physiol Rev. 93 (2), 681-766 (2013).</li><li>Hal&#225;sz, P., B&#242;dizs, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Dynamic structure of NREM sleep. , (2013).</li><li>Terzano, M. G., Parrino, L., Spaggiari, M. 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+cyclic+alternating+pattern+sequences+in+the+dynamic+organization+of+sleep.">The cyclic alternating pattern sequences in the dynamic organization of sleep.</a> Electroencephalogr Clin Neurophysiol. 69 (5), 437-447 (1988).</li><li>Gottesmann, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+seven+sleep-waking+stages+in+the+rat.">Detection of seven sleep-waking stages in the rat.</a> Neurosci Biobehav Rev. 16 (1), 31-38 (1992).</li><li>Benington, J. H., Kodali, S. K., Heller, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scoring+transitions+to+REM+sleep+in+rats+based+on+the+EEG+phenomena+of+pre-REM+sleep:+an+improved+analysis+of+sleep+structure.">Scoring transitions to REM sleep in rats based on the EEG phenomena of pre-REM sleep: an improved analysis of sleep structure.</a> Sleep. 17 (1), 28-36 (1994).</li><li>Sullivan, D., Mizuseki, K., Sorgi, A., Buzs&#225;ki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+sleep+spindles+and+theta+oscillations+in+the+hippocampus.">Comparison of sleep spindles and theta oscillations in the hippocampus.</a> J Neurosci. 34 (2), 662-674 (2014).</li><li>Vas, S., 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=Differential+adaptation+of+REM+sleep+latency,+intermediate+stage+and+theta+power+effects+of+escitalopram+after+chronic+treatment.">Differential adaptation of REM sleep latency, intermediate stage and theta power effects of escitalopram after chronic treatment.</a> J Neural Transm (Vienna). 120 (1), 169-176 (2013).</li><li>Schiffelholz, T., Aldenhoff, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+object+presentation+affects+sleep-wake+behavior+in+rats.">Novel object presentation affects sleep-wake behavior in rats.</a> Neurosci Lett. 328 (1), 41-44 (2002).</li><li>Wimmer, R. D., 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=Sustaining+sleep+spindles+through+enhanced+SK2-channel+activity+consolidates+sleep+and+elevates+arousal+threshold.">Sustaining sleep spindles through enhanced SK2-channel activity consolidates sleep and elevates arousal threshold.</a> J Neurosci. 32 (40), 13917-13928 (2012).</li><li>Fernandez, L. M. J., 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=Highly+dynamic+spatiotemporal+organization+of+low-frequency+activities+during+behavioral+states+in+the+mouse+cerebral+cortex.">Highly dynamic spatiotemporal organization of low-frequency activities during behavioral states in the mouse cerebral cortex.</a> Cereb Cortex. , (2016).</li><li>Franken, P. <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+vs.+short-term+processes+regulating+REM+sleep.">Long-term vs. short-term processes regulating REM sleep.</a> J Sleep Res. 11 (1), 17-28 (2002).</li><li>Feige, B., 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=The+microstructure+of+sleep+in+primary+insomnia:+an+overview+and+extension.">The microstructure of sleep in primary insomnia: an overview and extension.</a> Int J Psychophysiol. 89 (2), 171-180 (2013).</li><li>Parrino, L., Halasz, P., Tassinari, C. A., Terzano, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CAP,+epilepsy+and+motor+events+during+sleep:+the+unifying+role+of+arousal.">CAP, epilepsy and motor events during sleep: the unifying role of arousal.</a> Sleep Med Rev. 10 (4), 267-285 (2006).</li><li>Akinseye, O. 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=Sleep+as+a+mediator+in+the+pathway+linking+environmental+factors+to+hypertension:+a+review+of+the+literature.">Sleep as a mediator in the pathway linking environmental factors to hypertension: a review of the literature.</a> Int J Hypertens. 2015, 926414 (2015).</li><li>Campbell, K., Muller-Gass, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+extent+of+processing+of+near-hearing+threshold+stimuli+during+natural+sleep.">The extent of processing of near-hearing threshold stimuli during natural sleep.</a> Sleep. 34 (9), 1243-1249 (2011).</li><li>Nir, Y., Vyazovskiy, V. V., Cirelli, C., Banks, M. I., Tononi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auditory+responses+and+stimulus-specific+adaptation+in+rat+auditory+cortex+are+preserved+across+NREM+and+REM+sleep.">Auditory responses and stimulus-specific adaptation in rat auditory cortex are preserved across NREM and REM sleep.</a> Cereb Cortex. 25 (5), 1362-1378 (2015).</li><li>Dang-Vu, T. T., 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=Interplay+between+spontaneous+and+induced+brain+activity+during+human+non-rapid+eye+movement+sleep.">Interplay between spontaneous and induced brain activity during human non-rapid eye movement sleep.</a> Proc Natl Acad Sci USA. 108 (37), 15438-15443 (2011).</li><li>Elton, 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=Event-related+potentials+to+tones+in+the+absence+and+presence+of+sleep+spindles.">Event-related potentials to tones in the absence and presence of sleep spindles.</a> J Sleep Res. 6 (2), 78-83 (1997).</li><li>McKinney, S. M., Dang-Vu, T. T., Buxton, O. M., Solet, J. M., Ellenbogen, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Covert+waking+brain+activity+reveals+instantaneous+sleep+depth.">Covert waking brain activity reveals instantaneous sleep depth.</a> PLoS One. 6 (3), 17351 (2011).</li><li>Mang, G. M., Franken, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+and+EEG+phenotyping+in+mice.">Sleep and EEG phenotyping in mice.</a> Curr Protoc Mouse Biol. 2 (1), 55-74 (2012).</li><li>Borb&#233;ly, A. A., Tobler, I., Hanagasioglu, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+sleep+deprivation+on+sleep+and+EEG+power+spectra+in+the+rat.">Effect of sleep deprivation on sleep and EEG power spectra in the rat.</a> Behav Brain Res. 14 (3), 171-182 (1984).</li><li>Jurysta, F., 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=The+impact+of+chronic+primary+insomnia+on+the+heart+rate--EEG+variability+link.">The impact of chronic primary insomnia on the heart rate--EEG variability link.</a> Clin Neurophysiol. 120 (6), 1054-1060 (2009).</li><li>Silvani, A., Calandra-Buonaura, G., Benarroch, E. E., Dampney, R. A. L., Cortelli, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bidirectional+interactions+between+the+baroreceptor+reflex+and+arousal:+an+update.">Bidirectional interactions between the baroreceptor reflex and arousal: an update.</a> Sleep Med. , (2015).</li></ol>

Yazarlar, rakip mali çıkarlarının olmadığını beyan ettiler.

Burada, EEG, EMG ve EKG değişkenlerini bütünleyen REM dışı uykunun sürekli bir zamansal profilini nasıl oluşturacağımızı göstereceğiz. Bu, REM uykusu olmayan 1 sırasında yüksek ve düşük gürültüye karşı düzenlenen daha önce tanınmayan zaman ölçeğini tanımlamaya yardımcı olabilen, fare uykusunun bütünleyici bir tanımını geliştirmeye yönelik ilk adımdır. Benzer bir geçici yapı, insan olmayan REM uykusunda benzer bir analizle tarif edildi <sup class="xre...

Memeli uykusu, dinlenme davranışsal bir halidir ve çevresel uyarılara karşı dirençlidir. Bu belirgin tekdüzeliğe rağmen, polisomnografik ve otonomik parametreler, uykunun çeşitli temporal ve mekansal ölçeklerde niteliksel ve niceliksel olarak farklı sinirsel ve somatik durumlar arasında hareket ettiğini göstermektedir 1 . On dakika sonra onlarca dakika REM ve REM uyku modu arasında geçiş olur. REM uykusu, EEG&#39;de büyük amplitüd, düşük frekans aktivitesi ve yaklaşık 0.5-4 Hz civarında bir spektral zirve eşliğinde bulunurken, REM uykusu teta bandında (6-10 Hz) düzenli EEG aktivitesini gösterir Kas atoni 2 . REM dışı uykuda, insanlar ışıktan (S2) ve derin yavaş dalga uykudan (SWS) dönerler. Bunların adlandırma anlaşıldığı gibi, bu iki aşama daha düşük gösterir ve daha yüksek uyarılma, sırasıyla 4, 3 eşikleri ve bunlar düşük frequ yoğunluğu yönünden farklılıkYavaş dalga aktivitesi (SWA; 0.75 - 4 Hz) olarak adlandırılan kortikal EEG gücünü saptayın. Düzensizlik, S2 ve SWS&#39;nin bireysel sezişleri boyunca, dakikadan saniyeye kadar zaman ölçeğinde devam eder ve SWA&#39;nın değişken varlığı ile 5 , 6 sezonu boyunca kapsamlı olarak belgelenmiş olduğu gibi, aynı zamanda EEG ve alan potansiyel ritimleri ile de belgelenmiştir Sigma bandında (10-15 Hz) ve gama ritimlerinde (80 - 120 Hz) iş mili dalgaları da dahil olmak üzere daha yüksek frekanslar (bir inceleme için bkz. 7 , 8 , 9 , 10 ). İnsanlardaki uyku kortikal durumunu, ince olmaktan ziyade spektrumun uç noktalarına kaydırır. REM dışı uyku için bunlar SWA&#39;nın baskınlığından yaklaşık uyandırma benzeri aktiviteye kadar geniş bir aralıkta yüksek frekanslı bileşen 11 <suP&gt;, 12 . Kemirgenler ve kedilerde, REM dışındaki uyku aşamasına bölünmese de, ara uyku (IS) olarak adlandırılan kısa bir süre REM uykusu başlangıcından 13 önce ortaya çıkar. Sırasında, iki uyku arasında bir karışımı gösteren, hipokamp teta aktivitesi ve ponto-geniculo-oksipital dalgalar, örneğin mil dalgaları ve SWA olmayan REM uyku imzalar, hala mevcut olsa da, başlatmak olarak REM uykusu özellikleri, 15, 14 belirtmektedir. Yine de, antidepresanlar 16 tarafından modüle edildiğinden ve önceki uyanma esnasında yeni nesne sunumu vasıtasıyla işlevsel olarak ayırt edilebilir 17 ve uyarılma eşiğinin ayarlanmasına katkıda bulunur 18 . Dahası, serbestçe hareket eden sıçanların EEG ve EMG parametrelerinin durum uzayı grafikleri, REM olmayan uyku, REM uykusu ve uyanıklık arasında kesintisiz 14 noktaların bir kümesini göstermektedir. SWA&#39;da, uyanıklık ya da REM uykusuna girmeden, 14 , 19 ve 20 numaralı konsolide olmayan bir REM uykusu sırasında düşük frekans bileşenlerinin ve yüksek frekans bileşenlerinin göreli varlığında önemli dalgalanmalara neden olan sporadik düşüşler de vardır. Son olarak, non-REM uykusu sırasında SWA değişken frekansları ve daha yüksek frekanslı ritimler, yalnızca zamanla değil, aynı zamanda kortikal alanlar arasındaki genlik ve senkronizasyonda bölgesel farklılıkları göstermektedir 19 . Memeli non-REM uykusu üniforma uzaktır. Bununla birlikte, bu tür düzensizliğin işlev ve davranışsal özniteliklerden farklı durumlara yol açıp açmadığı açık değildir. Birkaç uyku bozukluğu türünde, sürekli uyku, kendiliğinden uyanma ve uygunsuz motor davranışı ile bozulur. Dahası, spektral analizler, EEG 21&#39;deki daha yüksek frekansların göreli varlığında değişikliklerVe nefes alma ve kalp atışı oranları gibi otonomik parametrelerde 22 . Dengeli uyku durumlarının düzenli sırası bozulur ve kortikal ve / veya otonomik uyanış unsurları kontrolsüz bir şekilde girer. Bu nedenle, uyku durumlarının sürekliliğini anlamak, hastalık için muhtemel bir önem taşır. Ek olarak, kentsel çevrede uykunun çevresel gürültüye maruz kalması genel sağlık riskleri ile ilişkilidir ve uyku esnasında artan hassasiyet anlarını belirlemek çok önemlidir 23 . Uykuda insanlarda davranışsal uyarılma deneyleri, hafif non-REM uykusu (S2 evresi) ve REM uykusu ile karşılaştırılabilir ve daha düşük uyarılma eşikleri 4 gösterirken, SWA baskınlı olmayan REM uykusundan (S3 evresi) uyanmanın en zor olduğunu göstermektedir. Kısa ses uyarılarının kortikal işlenmesi, REM uykusu, S2 ve S3 24 ,25 , bu duruma özgü kortikal aktivite kalıplarının duyusal işlemenin ilk evrelerini modüle ettiğini gösterir. İnsanlarda REM dışındaki uyku için, gürültüye tepki olarak uyanma eğilimi, EEG 26 , 27 , 28&#39;de işmili dalgalarının ve alfa ritimlerinin varlığına göre değişir. Iğ sırasında Talamokortikal ritmisitesi duyusal işleme 7&#39;nin zayıflama katkıda bulunduğu düşünülen her iki talamik ve kortikal seviyelerde geliştirilmiş sinaptik inhibisyonu ile eşlik eder. Gürültüye karşı dirençli ve hassas dönemler nasıl düzenlenir ve belirleyicileri nelerdir? Hem farelerde hem de insanda nöral ritimlerde kızılötesi yavaş, 0.02 Hz&#39;lik bir salınım tespit ettik. Bu 0,02 Hz&#39;lik salınımın fazına bağlı olarak, fareler harekete geçiren uyarılara değişken uyarı gösterdi, uyanma ya da uyumaGürültü. İlginç olarak, bu salınım otonom sinir sistemi, harici uyaranlara 1 Sleep&#39;in açığına modülasyonu katılır belirten kalp atışı oranı ile doğru orantılıydı. Hafıza ile ilgili hipokampal ritimler de bu ritim içinde organize edildi ve en çarpıcı biçimde, gücünün insanlardaki bellek konsolidasyonunun kalitesi ile korelasyona girdi. 0.02 Hz salınım, çevreye duyarlılığı ve dahili bellek işlemeyi modüle eden, kemirgen ve insan olmayan REM uykusunun düzenleyici bir ilkesi gibi görünmektedir. Bu durum, işlevselliğini tanımak ve potansiyel güvenlik açığı alanlarını belirlemek için uyku durumlarının çok parametrik ve sürekli değerlendirilmesine olan ihtiyacı vurguluyor. Burada, kombinasyon halinde EEG / ECoG ve EMG-EKG ölçümleri için farelerin cerrahi implantasyonu, duyusal uyaranlara maruz kalma,Analiz rutinleri. Bu prosedür, uykuyu farklı temel uyku fonksiyonlarının ard arda yürütülürken sürekli olarak değişen ancak son derece organize edilmiş uyanıklık hali olarak görüntülemek için bir temel oluşturmaktadır. Daha genel olarak, bu prosedür, hem sağlık hem de hastalık hallerinde uyku esnasında davranışsal bir sonucun öncesinde bulunan spektral ve otonom özellikleri elde etmeyi amaçlayan yaklaşımlara uygulanabilir.

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		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:316px;">2-components epoxy glue</td>
			<td style="width:128px;">Henkel</td>
			<td style="width:177px;">Loctite EA 3450</td>
			<td style="width:605px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Absorbable Suturing Fiber (Prolene)</td>
			<td>Ethicon</td>
			<td>5-0 FS-3</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Adson Forceps</td>
			<td>FST</td>
			<td>11006-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Antiseptic swab</td>
			<td>VWR</td>
			<td>149-0332</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Attane Isoflurane</td>
			<td>Piramal</td>
			<td>Isoflurane 250mL</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Connectors 3x2-channels</td>
			<td>ENA AG</td>
			<td>2.316</td>
			<td>Raster 2.00 x 2.00 mm; size 5x8x9 mm; pin size 5mm; http://www.ena.ch/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dragonfly commutator</td>
			<td>Dragonfly</td>
			<td>Model #SL-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">EMBLA amplifier</td>
			<td>EMBLA</td>
			<td>A10 amplifier</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fine scissors</td>
			<td>FST</td>
			<td>14108-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Flat Head Gold-plated steel screw</td>
			<td>J.I. Morris</td>
			<td>FF00CE125</td>
			<td>https://jimorrisco.com/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gold wire</td>
			<td>CMSA</td>
			<td>T.69 5gr</td>
			<td>http://www.cmsa.ch/en/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hemostatic sponge</td>
			<td>Pfizer</td>
			<td>Gelfoam</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">iodine-based disinfectant (Betadine)</td>
			<td>Mundipharma</td>
			<td>standart solution 60mL</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Komet drill steel 1/005PM104</td>
			<td>UNOR AG</td>
			<td>22310</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Matlab Analysis Software</td>
			<td>MathWorks</td>
			<td>R2016b</td>
			<td>https://ch.mathworks.com/products/matlab.html</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microdrill</td>
			<td>Fine Science Tools</td>
			<td>96758</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse Gas Anesthesia Head Holder</td>
			<td>Kopf Instruments</td>
			<td>Model 923-B</td>
			<td>http://kopfinstruments.com/product/model-923-b-mouse-gas-anesthesia-head-holder/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ophtalmic ointment</td>
			<td>Pharmamedica</td>
			<td>VITA-POS</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paladur (liquid)</td>
			<td>UNOR AG</td>
			<td>2260215</td>
			<td>for dental cement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Palavit (powder)</td>
			<td>UNOR AG</td>
			<td>5410929</td>
			<td>for dental cement</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Small Animal Stereotaxic Frame</td>
			<td>Kopf Instruments</td>
			<td>Model 930</td>
			<td>http://kopfinstruments.com/product/model-930-small-animal-stereotaxic-frame-assembly/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Soldering wire</td>
			<td>Stannol</td>
			<td>593072</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Temperature controller - Mini rectal probe</td>
			<td>Phymep</td>
			<td>4090502</td>
			<td>http://www.phymep.com/produit/dc-temperature-controller/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Temperature controller- heating pad</td>
			<td>Phymep</td>
			<td>4090205</td>
			<td>http://www.phymep.com/produit/dc-temperature-controller/</td>
		</tr>
	</tbody>
</table>

quantifying infra-slow dynamics of spectral power and heart rate in sleeping mice

Uyanıklık, hızlı olmayan göz hareketi (non-REM) uykusu ve REM uykusu: Üç ihtiyatlılık durumu memelilerin yaşamına hâkim. Serbest hareket eden hayvanlarda davranışın daha fazla sinirsel korelasyonu tanımlandığından, bu üçlü alt bölüm çok basitleşir. Uyanıklık sırasında, küresel ve lokal kortikal aktivitelerin toplulukları, pupil çapı ve sempatovagal denge gibi çevresel parametrelerle birlikte çeşitli derecelerde uyarılma tanımlar. Uykunun da beyin devletlerinin bir sürekliliğini oluşturduğu belirsizliğini koruyor; duyusal uyaranlara ve uyanmaya eğilimli olma derecesine ve belki de diğer uyku işlevlerine kademeli olarak değişen ve periferal fizyolojik durumların nasıl değiştiği. Uyku esnasında birden çok parametrenin izlenmesine yönelik yöntemlerin geliştirilmesi ve bu işlevsel niteliklerin takımyıldızlarına atıf yapılmasının araştırılması, uykudaki uyumu çok işlevli bir süreç olarak inceltmenin merkezinde olup, bunun sonucunda pek çok faydalı etki önceden bulunmalıdırecuted. Uyku durumlarını karakterize eden yeni parametrelerin belirlenmesi, uyku bozukluklarında yeni teşhis yollarına fırsatlar açacaktır. Standart polisomnografik kayıt teknikleri kullanılarak elektroensefalogram (EEG) / elektrokortikogram (ECoG), elektromyogram (EMG) ve elektrokardiyogram (EKG) sinyallerinin birleşik izleme ve analizi vasıtasıyla fare olmayan REM uyku durumlarının dinamik varyasyonlarını tanımlamak için bir prosedür sunmaktayız. Bu yaklaşımı kullanarak, fare olmayan REM uykusunun, harici uyaranlara yüksek ve düşük kırılganlık aralıklarıyla 25 sn aralıklarla eşzamanlı sinir ve kalp salınımları döngüleri halinde düzenlendiğini bulduk. Bu nedenle, merkezi ve otonom sinir sistemleri, konsolide non-REM uykusu sırasında davranışsal olarak farklı uyku durumlarını oluşturmak üzere koordine edilir. Biz, serbestçe uyku fareindeki bu döngüleri izlemek için polisomnografi ( yani EKG ile birlikte EEG / EMG) izlem için cerrahi manipülasyonlar sunuyoruz, quanti için analizDinamikleri ve uyanma olasılıklarındaki rollerini değerlendirmek için akustik uyarı protokolleri. Yaklaşımımız insan uykusuna zaten genişletilmiş ve memelilerde REM dışı uyku durumlarının ortak düzenleyici ilkelerini ortaya çıkarmayı vaat ediyor.

Şekil 2 A (üst panel), açıklandığı gibi implante edilen polisomnografik elektrotlar aracılığıyla kaydedilen 100 dakikalık spontan uyku-uyanıklık davranışlarını göstermektedir (bkz. Şekil 1 ). REM dışı uyku başlangıcında EEG ve EMG genliklerinde artış ve azalmalar açıkça görülebilir. Aralıklı REM uykusu, EEG amplitüdünde bir azalma ve EMG tonusunda bu sıkıştırılmış zama...

Watch this Scientific Journal Video about Quantifying Infra-slow Dynamics of Spectral Power and Heart Rate in Sleeping Mice at JoVE.com

Quantifying Infra-slow Dynamics of Spectral Power and Heart Rate in Sleeping Mice

Burada, akustik uyarılara karşı uyku tepkisini modüle eden, farelerde non-REM uykusunda sinirsel ve kardiyak değişkenlerin zamansal dinamiklerini tanımlamak için deneysel ve analitik prosedürleri sunuyoruz.

Uyku Faresinde Spektral Gücün ve İnsanın Yavaş yavaş Dinamiklerinin Nicelleştirilmesi

Three vigilance states dominate mammalian life: wakefulness, non-rapid eye movement (non-REM) sleep, and REM sleep. As more neural correlates of behavior ...

quantifying-infra-slow-dynamics-spectral-power-heart-rate-sleeping

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

9.9K Görüntüleme.  University of Lausanne.  Burada, akustik uyarılara karşı uyku tepkisini modüle eden, farelerde non-REM uykusunda sinirsel ve kardiyak değişkenlerin zamansal dinamiklerini tanımlamak için deneysel ve analitik prosedürleri sunuyoruz. 

Video: Quantifying Infra-slow Dynamics of Spectral Power and Heart Rate in Sleeping Mice

Multi-electrode Array Recordings of Neuronal Avalanches in Organotypic Cultures

The cortex is spontaneously active, even in the absence of any particular input or motor output. During development, this activity is important for the migration and differentiation of cortex cell types and the formation of neuronal connections1. In the mature animal, ongoing activity reflects the past and the present state of an animal into which sensory stimuli are seamlessly integrated to compute future actions. Thus, a clear understanding of the organization of ongoing i.e. spontaneous activity is a prerequisite to understand cortex function. 
Numerous recording techniques revealed that ongoing activity in cortex is comprised of many neurons whose individual activities transiently sum to larger events that can be detected in the local field potential (LFP) with extracellular microelectrodes, or in the electroencephalogram (EEG), the magnetoencephalogram (MEG), and the BOLD signal from functional magnetic resonance imaging (fMRI). The LFP is currently the method of choice when studying neuronal population activity with high temporal and spatial resolution at the mesoscopic scale (several thousands of neurons). At the extracellular microelectrode, locally synchronized activities of spatially neighbored neurons result in rapid deflections in the LFP up to several hundreds of microvolts. When using an array of microelectrodes, the organizations of such deflections can be conveniently monitored in space and time. 
Neuronal avalanches describe the scale-invariant spatiotemporal organization of ongoing neuronal activity in the brain2,3. They are specific to the superficial layers of cortex as established in vitro4,5, in vivo in the anesthetized rat 6, and in the awake monkey7. Importantly, both theoretical and empirical studies2,8-10 suggest that neuronal avalanches indicate an exquisitely balanced critical state dynamics of cortex that optimizes information transfer and information processing.
In order to study the mechanisms of neuronal avalanche development, maintenance, and regulation, in vitro preparations are highly beneficial, as they allow for stable recordings of avalanche activity under precisely controlled conditions. The current protocol describes how to study neuronal avalanches in vitro by taking advantage of superficial layer development in organotypic cortex cultures, i.e. slice cultures, grown on planar, integrated microelectrode arrays (MEA; see also 11-14).

A robust way to study neuronal avalanches, i.e. scale-invariant spatio-temporal activity bursts, indicative of critical state dynamics in cortex. Avalanches emerge spontaneously in developing superficial layers of cultured cortex which allows for long-term measurements of the activity with planar integrated multi-electrode arrays (MEA) under precisely controlled conditions.

Organotipik Kültürleri nöronal Çığ Çok elektrot Dizi Kayıtlar

The cortex is spontaneously active, even in the absence of any particular input or motor output.  During development, this activity is important for the ...

Nörobilim

Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration

Ligand-gated ion channels in the central nervous system (CNS) are implicated in numerous conditions with serious medical and social consequences. For instance, addiction to nicotine via tobacco smoking is a leading cause of premature death worldwide (World Health Organization) and is likely caused by an alteration of ion channel distribution in the brain1. Chronic nicotine exposure in both rodents and humans results in increased numbers of nicotinic acetylcholine receptors (nAChRs) in brain tissue1-3. Similarly, alterations in the glutamatergic GluN1 or GluA1 channels have been implicated in triggering sensitization to other addictive drugs such as cocaine, amphetamines and opiates4-6.

Consequently, the ability to map and quantify distribution and expression patterns of specific ion channels is critically important to understanding the mechanisms of addiction. The study of brain region-specific effects of individual drugs was advanced by the advent of techniques such as radioactive ligands. However, the low spatial resolution of radioactive ligand binding prevents the ability to quantify ligand-gated ion channels in specific subtypes of neurons. 

Genetically encoded fluorescent reporters, such as green fluorescent protein (GFP) and its many color variants, have revolutionized the field of biology7.By genetically tagging a fluorescent reporter to an endogenous protein one can visualize proteins in vivo7-10. One advantage of fluorescently tagging proteins with a probe is the elimination of antibody use, which have issues of nonspecificity and accessibility to the target protein. We have used this strategy to fluorescently label nAChRs, which enabled the study of receptor assembly using F&ouml;rster Resonance Energy Transfer (FRET) in transfected cultured cells11.More recently, we have used the knock-in approach to engineer mice with yellow fluorescent protein tagged &alpha;4 nAChR subunits (&alpha;4YFP), enabling precise quantification of the receptor ex vivo at submicrometer resolution in CNS neurons via spectral confocal microscopy12. The targeted fluorescent knock-in mutation is incorporated in the endogenous locus and under control of its native promoter, producing normal levels of expression and regulation of the receptor when compared to untagged receptors in wildtype mice. This knock-in approach can be extended to fluorescently tag other ion channels and offers a powerful approach of visualizing and quantifying receptors in the CNS.

In this paper we describe a methodology to quantify changes in nAChR expression in specific CNS neurons after exposure to chronic nicotine. Our methods include mini-osmotic pump implantation, intracardiac perfusion fixation, imaging and analysis of fluorescently tagged nicotinic receptor subunits from &alpha;4YFP knock-in mice (Fig. 1). We have optimized the fixation technique to minimize autofluorescence from fixed brain tissue.We describe in detail our imaging methodology using a spectral confocal microscope in conjunction with a linear spectral unmixing algorithm to subtract autofluoresent signal in order to accurately obtain &alpha;4YFP fluorescence signal. Finally, we show results of chronic nicotine-induced upregulation of &alpha;4YFP receptors in the medial perforant path of the hippocampus.

We have developed a novel technique of quantifying nicotinic acetylcholine receptor changes within subcellular regions of specific subtypes of CNS neurons to better understand the mechanisms of nicotine addiction by using a combination of approaches including fluorescent protein tagging of the receptor using the knock-in approach and spectral confocal imaging.

Kronik Nikotin İdaresi fareler Knock-in içinde floresan etiketli Nikotinik Reseptör Spektral Konfokal Görüntüleme

Ligand-gated ion channels in the central nervous system (CNS) are implicated in numerous conditions with serious medical and social consequences.  For ...

Behavioral Determination of Stimulus Pair Discrimination of Auditory Acoustic and Electrical Stimuli Using a Classical Conditioning and Heart-rate Approach

Acute animal preparations have been used in research prospectively investigating electrode designs and stimulation techniques for integration into neural auditory prostheses, such as auditory brainstem implants1-3 and auditory midbrain implants4,5. While acute experiments can give initial insight to the effectiveness of the implant, testing the chronically implanted and awake animals provides the advantage of examining the psychophysical properties of the sensations induced using implanted devices6,7.
Several techniques such as reward-based operant conditioning6-8, conditioned avoidance9-11, or classical fear conditioning12 have been used to provide behavioral confirmation of detection of a relevant stimulus attribute. Selection of a technique involves balancing aspects including time efficiency (often poor in reward-based approaches), the ability to test a plurality of stimulus attributes simultaneously (limited in conditioned avoidance), and measure reliability of repeated stimuli (a potential constraint when physiological measures are employed).
Here, a classical fear conditioning behavioral method is presented which may be used to simultaneously test both detection of a stimulus, and discrimination between two stimuli. Heart-rate is used as a measure of fear response, which reduces or eliminates the requirement for time-consuming video coding for freeze behaviour or other such measures (although such measures could be included to provide convergent evidence). Animals were conditioned using these techniques in three 2-hour conditioning sessions, each providing 48 stimulus trials. Subsequent 48-trial testing sessions were then used to test for detection of each stimulus in presented pairs, and test discrimination between the member stimuli of each pair. 
This behavioral method is presented in the context of its utilisation in auditory prosthetic research. The implantation of electrocardiogram telemetry devices is shown. Subsequent implantation of brain electrodes into the Cochlear Nucleus, guided by the monitoring of neural responses to acoustic stimuli, and the fixation of the electrode into place for chronic use is likewise shown.

The application of a classical fear conditioning behavioral paradigm for auditory prosthetic research in rats is described. This paradigm provides a mechanism for identifying both detection of, and discrimination between, distinct acoustic and electrical stimuli using heart-rate as an outcome measure.

Bir Klasik Koşullanma ve Kalp hızı Yaklaşımı ile İşitsel Akustik ve Elektrik uyarıcı Stimulus Çifti Ayrımcılığın Davranış Belirlenmesi

Acute animal preparations have been used in research prospectively investigating electrode designs and stimulation techniques for integration into neural ...

Applications of EEG Neuroimaging Data: Event-related Potentials, Spectral Power, and Multiscale Entropy

When considering human neuroimaging data, an appreciation of signal variability represents a fundamental innovation in the way we think about brain signal. Typically, researchers represent the brain's response as the mean across repeated experimental trials and disregard signal fluctuations over time as "noise". However, it is becoming clear that brain signal variability conveys meaningful functional information about neural network dynamics. This article describes the novel method of multiscale entropy (MSE) for quantifying brain signal variability. MSE may be particularly informative of neural network dynamics because it shows timescale dependence and sensitivity to linear and nonlinear dynamics in the data.

Neuroimaging researchers typically consider the brain's response as the mean activity across repeated experimental trials and disregard signal variability over time as "noise". However, it is becoming clear that there is signal in that noise. This article describes the novel method of multiscale entropy for quantifying brain signal variability in the time domain.

Olay İlişkili Potansiyeller, Spektral Güç ve Çok Ölçekli Entropi: EEG beyin görüntüleme Veri Uygulamaları

When considering human neuroimaging data, an appreciation of signal variability represents a fundamental innovation in the way we think about brain ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

Fareler Soğuk Hassasiyeti ve Uyum belirlenmesi için Basit ve ucuz Yöntem

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Determination of Photoreceptor Cell Spectral Sensitivity in an Insect Model from In Vivo Intracellular Recordings

Intracellular recording is a powerful technique used to determine how a single cell may respond to a given stimulus. In vision research, intracellular recording has historically been a common technique used to study sensitivities of individual photoreceptor cells to different light stimuli that is still being used today. However, there remains a dearth of detailed methodology in the literature for researchers wishing to replicate intracellular recording experiments in the eye. Here we present the insect as a model for examining eye physiology more generally. Insect photoreceptor cells are located near the surface of the eye and are therefore easy to reach, and many of the mechanisms involved in vision are conserved across animal phyla. We describe the basic procedure for in vivo intracellular recording of photoreceptor cells in the eye of a butterfly, with the goal of making this technique more accessible to researchers with little prior experience in electrophysiology. We introduce the basic equipment needed, how to prepare a live butterfly for recording, how to insert a glass microelectrode into a single cell, and finally the recording procedure itself. We also explain the basic analysis of raw response data for determining spectral sensitivity of individual cell types. Although our protocol focuses on determining spectral sensitivity, other stimuli (e.g., polarized light) and variations of the method are applicable to this setup.

Determination of Photoreceptor Cell Spectral Sensitivity in an Insect Model from In Vivo Intracellular Recordings

The electrophysiological technique of intracellular recording is demonstrated and used to determine spectral sensitivities of single photoreceptor cells in the compound eye of a butterfly.

bir Böcek Modeli fotoreseptör hücre spektral Duyarlılık Belirlenmesi<em&gt; İn Vivo</em&gt; Hücre içi Kayıtlar

Intracellular recording is a powerful technique used to determine how a single cell may respond to a given stimulus. In vision research, intracellular ...

Electroencephalographic, Heart Rate, and Galvanic Skin Response Assessment for an Advertising Perception Study: Application to Antismoking Public Service Announcements

The evaluation of advertising, products, and packaging is traditionally performed through methods based on self-reports and focus groups, but these approaches often appear poorly accurate in scientific terms. Neuroscience is increasingly applied to the investigation of the neurophysiological bases of the perception of and reaction to commercial stimuli to support traditional marketing methods. In this context, a particular sector or marketing is represented by public service announcements (PSAs). The objective of this protocol is to apply electroencephalography (EEG) and autonomic signal analysis to study responses to selected antismoking PSAs. Two EEG indices were employed: the frontal alpha band EEG asymmetry (the Approach Withdrawal (AW) index) and the frontal theta (effort index). Furthermore, the autonomic Emotional Index (EI) was calculated, as derived from the Galvanic Skin Response (GSR) and Heart Rate (HR) signals. The present protocol describes a series of operational and computational steps required to properly estimate, through the aforementioned indices, the emotional and cerebral reaction of a group of subjects towards a selected number of antismoking PSAs. In particular, a campaign characterized by a symbolic communication style (classified as "awarded" on the basis of the prizes received by specialized committees) obtained the highest approach values, as estimated by the AW index. A spot and an image belonging to the same PSA campaign based on the "fear arousing appeal" and with a narrative/experiential communication style (classified as "effective" on the basis of the economical/health-related improvements promoted) reported the lowest and highest effort values, respectively. This is probably due to the complexity of the storytelling (spot) and to the immediateness of the image (a lady who underwent a tracheotomy). Finally, the same "effective" campaign showed the highest EI values, possibly because of the empathy induced by the testimonial and the explicitness of the message.

The following protocol describes a series of operational and computational steps required to properly estimate the emotional and cerebral reaction of a group of subjects towards a selected number of Public Service Announcements (PSAs) against smoking, aired in the USA and Europe during the period between 1998 and 2015.

Electroencephalographic, kalp hızı ve galvanik deri tepkisini değerlendirmesi için bir reklam algı çalışma: Sigara karşıtı kamu hizmet duyuruları uygulamaya

The evaluation of advertising, products, and packaging is traditionally performed through methods based on self-reports and focus groups, but these ...

Spectral Reflectometric Microscopy on Myelinated Axons In Situ

In a mammalian nervous system, myelin provides an electrical insulation by enwrapping the axon fibers in a multilayered spiral. Inspired by its highly-organized subcellular architecture, we recently developed a new imaging modality, named spectral reflectometry (SpeRe), which enables unprecedented label-free nanoscale imaging of the live myelinated axons in situ. The underlying principle is to obtain nanostructural information by analyzing the reflectance spectrum of the multilayered subcellular structure. In this article, we describe a detailed step-by-step protocol for performing a basic SpeRe imaging of the nervous tissues using a commercial confocal microscopic system, equipped with a white-light laser and a tunable filter. We cover the procedures of sample preparation, acquisition of spectral data, and image processing for obtaining nanostructural information.

Spectral Reflectometric Microscopy on Myelinated Axons In Situ

Here, we present a step-by-step protocol for imaging myelinated axons in a fixed brain slice using a label-free nanoscale imaging technique based on spectral reflectometry.

Myelinated akson in Situ üzerinde spektral Reflectometric mikroskobu

In a mammalian nervous system, myelin provides an electrical insulation by enwrapping the axon fibers in a multilayered spiral. Inspired by its ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.

Interstimulus Aralık kemirgenler zamansal işleme değerlendirilmesi için güç

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

Quantifying Subcellular Ubiquitin-proteasome Activity in the Rodent Brain

The ubiquitin-proteasome system is a key regulator of protein degradation and a variety of other cellular processes in eukaryotes. In the brain, increases in ubiquitin-proteasome activity are critical for synaptic plasticity and memory formation and aberrant changes in this system are associated with a variety of neurological, neurodegenerative and psychiatric disorders. One of the issues in studying ubiquitin-proteasome functioning in the brain is that it is present in all cellular compartments, in which the protein targets, functional role and mechanisms of regulation can vary widely. As a result, the ability to directly compare brain ubiquitin protein targeting and proteasome catalytic activity in different subcellular compartments within the same animal is critical for fully understanding how the UPS contributes to synaptic plasticity, memory and disease. The method described here allows collection of nuclear, cytoplasmic and crude synaptic fractions from the same rodent (rat) brain, followed by simultaneous quantification of proteasome catalytic activity (indirectly, providing activity of the proteasome core only) and linkage-specific ubiquitin protein tagging. Thus, the method can be used to directly compare subcellular changes in ubiquitin-proteasome activity in different brain regions in the same animal during synaptic plasticity, memory formation and different disease states. This method can also be used to assess the subcellular distribution and function of other proteins within the same animal.

This protocol is designed to efficiently quantify ubiquitin-proteasome system (UPS) activity in different cellular compartments of the rodent brain. Users are able to examine UPS functioning in nuclear, cytoplasmic and synaptic fractions in the same animal, reducing the amount of time and number of animals needed to perform these complex analyses.

Subcellular Ubiquitin ölçümü-kemirgen beyinde proteasom aktivitesi

The ubiquitin-proteasome system is a key regulator of protein degradation and a variety of other cellular processes in eukaryotes. In the brain, increases ...