Bu çalışma, Ulusal Sağlık Enstitüleri (NIH) hibesi R01 EY030998 (J.F.M., A.B. ve S.C.) tarafından desteklenmiştir. Bu yöntem, Coop ve ark. (inceleniyor, 2022; https://www.biorxiv.org/content/10.1101/2022.10.11.511827v2.abstract). Marmoset bakımı ve kullanımı konusunda yardımları için Dina Graf'a ve Mitchell laboratuvarı üyelerine teşekkür ederiz.

Marmosetlerde Hassas Nöral Kayıt için X-Y Elektrot Aşamasının Geliştirilmesi

<ol>
	<li>Mansfield, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marmoset+models+commonly+used+in+biomedical+research.">Marmoset models commonly used in biomedical research.</a> Comparative Medicine. 53 (4), 383-392 (2003).</li><li>Solomon, S. G., Rosa, M. G. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simpler+primate+brain:+the+visual+system+of+the+marmoset+monkey.">A simpler primate brain: the visual system of the marmoset monkey.</a> Frontiers in Neural Circuits. 8, 96 (2014).</li><li>Remington, E. D., Osmanski, M. S., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+operant+conditioning+method+for+studying+auditory+behaviors+in+marmoset+monkeys.">An operant conditioning method for studying auditory behaviors in marmoset monkeys.</a> PLoS One. 7 (10), e47895 (2012).</li><li>Walker, J. 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=Chronic+wireless+neural+population+recordings+with+common+marmosets.">Chronic wireless neural population recordings with common marmosets.</a> Cell Reports. 36 (2), 109379 (2021).</li><li>Jendritza, P., Klein, F. J., Fries, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-area+recordings+and+optogenetics+in+the+awake,+behaving+marmoset.">Multi-area recordings and optogenetics in the awake, behaving marmoset.</a> Nature Communications. 14 (1), 577 (2023).</li><li>Pinotsis, D. 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=Linking+canonical+microcircuits+and+neuronal+activity:+Dynamic+causal+modelling+of+laminar+recordings.">Linking canonical microcircuits and neuronal activity: Dynamic causal modelling of laminar recordings.</a> Neuroimage. 146, 355-366 (2017).</li><li>Ludwig, K. 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=Poly+(3,+4-ethylenedioxythiophene)(PEDOT)+polymer+coatings+facilitate+smaller+neural+recording+electrodes.">Poly (3, 4-ethylenedioxythiophene)(PEDOT) polymer coatings facilitate smaller neural recording electrodes.</a> Journal of Neural Engineering. 8 (1), 014001 (2011).</li><li>Ludwig, K. A., Uram, J. D., Yang, J., Martin, D. C., Kipke, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+neural+recordings+using+silicon+microelectrode+arrays+electrochemically+deposited+with+a+poly+(3,+4-ethylenedioxythiophene)(PEDOT)+film.">Chronic neural recordings using silicon microelectrode arrays electrochemically deposited with a poly (3, 4-ethylenedioxythiophene)(PEDOT) film.</a> Journal of Neural Engineering. 3 (1), 59 (2006).</li><li>Lu, T., Liang, L., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+representations+of+temporally+asymmetric+stimuli+in+the+auditory+cortex+of+awake+primates.">Neural representations of temporally asymmetric stimuli in the auditory cortex of awake primates.</a> Journal of Neurophysiology. 85 (6), 2364-2380 (2001).</li><li>Osmanski, M. S., Song, X., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+harmonic+resolvability+in+pitch+perception+in+a+vocal+nonhuman+primate,+the+common+marmoset+(Callithrix+jacchus).">The role of harmonic resolvability in pitch perception in a vocal nonhuman primate, the common marmoset (Callithrix jacchus).</a> Journal of Neuroscience. 33 (21), 9161-9168 (2013).</li><li>Nummela, S. U., 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=Psychophysical+measurement+of+marmoset+acuity+and+myopia.">Psychophysical measurement of marmoset acuity and myopia.</a> Developmental Neurobiology. 77 (3), 300-313 (2017).</li><li>Paxinos, G., Watson, C., Petrides, M., Rosa, M., Tokuno, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Marmoset Brain in Stereotaxic Coordinates. , (2012).</li><li>Mitchell, J. F., Reynolds, J. H., Miller, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Active+vision+in+marmosets:+A+model+system+for+visual+neuroscience.">Active vision in marmosets: A model system for visual neuroscience.</a> Journal of Neuroscience. 34 (4), 1183-1194 (2014).</li><li>Spitler, K. M., Gothard, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+removable+silicone+elastomer+seal+reduces+granulation+tissue+growth+and+maintains+the+sterility+of+recording+chambers+for+primate+neurophysiology.">A removable silicone elastomer seal reduces granulation tissue growth and maintains the sterility of recording chambers for primate neurophysiology.</a> Journal of Neuroscience Methods. 169 (1), 23-26 (2008).</li><li>Jun, J. 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=Fully+integrated+silicon+probes+for+high-density+recording+of+neural+activity.">Fully integrated silicon probes for high-density recording of neural activity.</a> Nature. 551 (7679), 232-236 (2017).</li><li>Mitzdorf, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+source-density+method+and+application+in+cat+cerebral+cortex:+investigation+of+evoked+potentials+and+EEG+phenomena.">Current source-density method and application in cat cerebral cortex: investigation of evoked potentials and EEG phenomena.</a> Physiological Reviews. 65 (1), 37-100 (1985).</li><li>Coop, S. H., Yates, J. L., Mitchell, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-saccadic+neural+enhancements+in+marmoset+area+MT.">Pre-saccadic neural enhancements in marmoset area MT.</a> bioRxiv. , (2022).</li><li>Okun, M., Lak, A., Carandini, M., Harris, K. D. <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+recordings+with+immobile+silicon+probes+in+the+mouse+cortex.">Long term recordings with immobile silicon probes in the mouse cortex.</a> PloS One. 11 (3), e0151180 (2016).</li></ol>

Yazarların açıklayacak hiçbir şeyi yok.

İnsan olmayan primatlarda nörofizyoloji deneyleri yapmak için şu anda çeşitli yöntemler (örneğin, kronik, yarı kronik, akut) mevcuttur. Yaygın marmoset, küçük boyutu ve anatomik işaretler olarak gyri eksikliği nedeniyle nörofizyoloji deneyleri için benzersiz zorluklar ortaya koymaktadır. Bu, araştırmacıların kayıt hedeflerini belirlemek için ilgi alanlarının retinotopisi ve ayar özellikleri gibi nörofizyolojik işaretleri kullanmasını gerektirir. Bu nedenle, başlangıçta bir kortikal alan...

Yaygın marmoset (Callithrix jacchus), son yıllarda beyin fonksiyonlarını incelemek için bir model olarak hızla popülerlik kazanmıştır. Bu artan popülerlik, marmosetin pürüzsüz korteksinin erişilebilirliğinden, insanlarla ve diğer primatlarla sinirsel fonksiyonel organizasyondaki benzerliklerden ve küçük boyut ve hızlı üreme oranındankaynaklanmaktadır 1. Bu model organizmanın popülaritesi arttıkça, marmoset beyninde kullanıma uygun nörofizyolojik tekniklerde hızlı bir gelişme olmuştur. Elektrofizyoloji yöntemleri, sinirbilimde hem kemirgenlerin hem de primatların korteksindeki tek nöronların aktivitesini incelemek için yaygın olarak kullanılmaktadır ve bu da benzersiz zamansal çözünürlük ve konum erişimi ile sonuçlanır. Marmoset maymununun görsel sinirbilimin bir modeli olarak göreceli yeniliği nedeniyle, uyanık davranan elektrofizyoloji tekniklerinin optimizasyonu hala gelişmektedir. Önceki çalışmalar, anestezi altındaki preparatlarda elektrofizyoloji için sağlam protokollerin oluşturulduğunugöstermiştir 2 ve erken uyanık davranan nörofizyoloji çalışmaları, tek kanallı tungsten elektodlarınıngüvenilirliğini göstermiştir 3. Son yıllarda, araştırmacılar uyanık davranan nörofizyoloji için silikon bazlı mikroelektrot dizilerinin kullanımını belirlediler4. Bununla birlikte, marmoset, küçük beyin boyutu ve anatomik işaretlerin olmaması nedeniyle benzersiz hedefleme zorlukları ortaya çıkarmaktadır. Bu protokol, minimum doku hasarı üretirken silikon lineer dizilerle büyük nöron popülasyonlarının kaydedilmesine izin veren marmoset için uygun bir mikro sürücü kayıt sisteminin nasıl oluşturulacağını ve kullanılacağını özetlemektedir.
Marmoset ile çalışmak, görsel korteksteki retinotopik haritaların daha büyük primatlara kıyasla daha küçük ölçekli olması nedeniyle bir zorluk teşkil etmektedir. Elektrotların sadece 1 mm'lik hafif bir kayması, haritalarda önemli değişikliklere neden olabilir. Ayrıca, araştırmacıların görsel kortekste daha geniş bir retinotopik pozisyon aralığı elde etmek için kayıt seansları arasında elektrotların yerleşimini değiştirmeleri gerekir. Mevcut yarı kronik preparatlar, elektrot konumlandırmasının günlük olarak veya milimetre altı ölçeklerde belirli yerleri hedeflemek için yeterli hassasiyetle ayarlanmasına izin vermemektedir5. Bunu akılda tutarak, önerilen mikro sürücü sistemi, hafif bir mikro sürücüyü bir kayıt odasına monte eden ve kortikal bölgelerin milimetre altı hedeflemesine izin veren bir XY elektrot aşaması kullanır. Hareketli XY aşaması bileşenleri, ilgilenilen alanları (retinotopi ve akort özellikleri aracılığıyla ) belirlemek için gerekli olan kortikal alanları sistematik olarak geçmek için doğrusal dizinin dikey ve yatay hareketine izin verir. Kayıt oturumları boyunca araştırmacılar, alan içindeki hedeflenen siteleri kaydırmak için X-Y aşamasını manuel olarak da ayarlayabilirler. Bu, kolay elektrot hedefleme mekanizmalarına sahip olmayan yarı kronik kayıt preparatlarını kullanan alternatif tekniklere göre önemli bir avantajdır.
Mikro sürücü, kortekse indirmek için monte edilecek çeşitli silikon dizilerinin bağlanmasını sağlayan çok yönlü bir araçtır. Bu protokolde, kortikal derinliği kapsayan laminer devrelerin araştırılması için 200 μm aralıklı iki 32 kanallı doğrusal diziye sahip özel bir prob kullanıldı. Nöral devreyi araştırmak için kullanılan yöntemlerin çoğu, tipik olarak, serebral korteksin tüm katmanları boyunca ortalaması alınan elektrik potansiyellerini veya tek birimleri örneklemektedir. Bununla birlikte, son araştırmalar kortikal laminer mikro devreler hakkında ilgi çekici bulgular ortaya çıkarmıştır6. Araştırmacılar, mikro sürücüyü kullanarak laminer probları kullanabilir ve tüm katmanlarda kapsamlı örnekleme sağlamak için kayıt derinliğinde ince ayarlamalar yapabilir.
Bu sistem ticari olarak temin edilebilen bileşenlerle oluşturulabilir ve farklı deneysel teknikler veya problar için kolayca değiştirilebilir. Bu hazırlığın en önemli avantajları, XY kayıt pozisyonunu milimetrenin altında hassasiyetle değiştirme ve korteks içindeki kayıt derinliğini kontrol etme yeteneğidir. Bu protokol, X-Y aşaması mikro sürücü ve nörofizyoloji kayıt tekniklerini oluşturmak için adım adım talimatlar sunar.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1/4 Hp burr drill bit</td>
			<td>McMaster &#38; Carr</td>
			<td>Cat# 43035A32</td>
			<td>Carbide Bur with 1/4&#34; Shank Diameter, Rounded Cylinder Head, trade Number SC-1, single Cut(https://www.mcmaster.com/products/bur-bits/burs-7/?s=1%2F4%22+bur+bits)</td>
		</tr>
		<tr>
			<td>1x1mm Crist Grid</td>
			<td>Crist Instruments</td>
			<td>1 mm x 1 mm Grid</td>
			<td>https://www.cristinstrument.com/products/implant-intro/grids</td>
		</tr>
		<tr>
			<td>91% isopropyl alcohol</td>
			<td>Medline</td>
			<td>N/A</td>
			<td>https://www.medline.com/product/Medline-Isopropyl-Rubbing-Alcohol/Bulk-Alcohol/Z05-PF03807?question=91%25%20isopropyl%20alcohol</td>
		</tr>
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			<td>Acquisition Board</td>
			<td>Open-Ephys</td>
			<td>N/A</td>
			<td>https://open-ephys.org/acquisition-system/eux9baf6a5s8tid06hk1mw5aafjdz1</td>
		</tr>
		<tr>
			<td>Bacitracin Ointment</td>
			<td>Medline: Cosette Pharmaceuticals Inc</td>
			<td>N/A</td>
			<td>https://www.medline.com/product/Bacitracin-Ointment/Antibiotics/Z05-PF86957?question=bacitr</td>
		</tr>
		<tr>
			<td>Blunt straight Forceps</td>
			<td>Medline</td>
			<td>N/A</td>
			<td>https://www.medline.com/category/Central-Sterile/Surgical-Instruments/Forceps/Z05-CA16_02_20/products</td>
		</tr>
		<tr>
			<td>Bone wax</td>
			<td>Medline</td>
			<td>ETHW31G</td>
			<td>https://www.medline.com/product/Ethicon-Bone-Wax/Bone-Wax/Z05-PF61528?question=bonewax</td>
		</tr>
		<tr>
			<td>C&#38;B Metabond Quick Adhesive Cement System</td>
			<td>Parkell, Inc.</td>
			<td>SKU: S380</td>
			<td>https://www.parkell.com/C-B-Metabond-Quick-Adhesive-Cement-System</td>
		</tr>
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			<td>Clavamox</td>
			<td>MWI Animal Health</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Contact lens solution</td>
			<td>Bausch and lomb</td>
			<td></td>
			<td>Various sources available</td>
		</tr>
		<tr>
			<td>Custom Printed 3D printed parts</td>
			<td>ProtoLab</td>
			<td></td>
			<td>https://marmolab.bcs.rochester.edu/resources.html</td>
		</tr>
		<tr>
			<td>DB25-G2 25 Pin Male Plug Port Signal Connector</td>
			<td>Various Sources</td>
			<td>DB25-G2 25</td>
			<td>DB25-G2 25 Pin Male Plug Port Signal 2 Row Terminal Breakout Board Screw Nut Connector</td>
		</tr>
		<tr>
			<td>diamond saw attachement for dremmel</td>
			<td>Dremmel</td>
			<td>545 Diamond Wheel</td>
			<td>https://www.dremel.com/us/en/p/545-26150545ab</td>
		</tr>
		<tr>
			<td>Digitizing Head-stages</td>
			<td>Intan</td>
			<td>RHD 32channel (Part #C3314)</td>
			<td>https://intantech.com/RHD_headstages.html?tabSelect=RHD32ch&#38;yPos=120.80 
			000305175781</td>
		</tr>
		<tr>
			<td>EDOT</td>
			<td>Sigma Aldrich</td>
			<td>Product # 483028</td>
			<td>https://www.sigmaaldrich.com/US/en/product/aldrich/483028</td>
		</tr>
		<tr>
			<td>Helping Hands</td>
			<td>Harbor Freight</td>
			<td>N/A</td>
			<td>https://www.harborfreight.com/helping-hands-60501.html</td>
		</tr>
		<tr>
			<td>Hook Electrical Clips</td>
			<td>Various Sources</td>
			<td>N/A</td>
			<td>Hook test Cable wires</td>
		</tr>
		<tr>
			<td>Interface Cables (RHD 3-ft (0.9 m) ultra thin SPI cable)</td>
			<td>Intan</td>
			<td>&#160;Part #C3213</td>
			<td>https://intantech.com/RHD_SPI_cables.html</td>
		</tr>
		<tr>
			<td>Lab jack</td>
			<td>Various Sources</td>
			<td>N/A</td>
			<td>https://www.amazon.com/Stainless-Steel-Scissor-Stand-Platform/dp/B07T8FM85H/ref=asc_df_B07T8FM85H/?tag=&#38;linkCode=df0&#38;hvadid=366343 
			827267&#38;hvpos=&#38;hvnetw=g&#38;hvrand 
			=2036619536500717246&#38;hvpone 
			=&#38;hvptwo=&#38;hvqmt=&#38;hvdev=c&#38;hv 
			dvcmdl=&#38;hvlocint=&#38;hvlocphy=900 
			5674&#38;hvtargid=pla-795933567991&#38; 
			ref=&#38;adgrpid=71496544770&#38;th=1</td>
		</tr>
		<tr>
			<td>Meloxicam</td>
			<td>MWI Animal Health</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-drive</td>
			<td>Crist Instrument</td>
			<td>3-NRMD</td>
			<td>https://www.cristinstrument.com/products/microdrives/miniature-microdrive-3-nrmd</td>
		</tr>
		<tr>
			<td>Multi-channel linear silicon arrays with 64 channel connector</td>
			<td>NeuroNexus</td>
			<td>A1x32-5mm-25-177</td>
			<td>https://www.neuronexus.com/products/electrode-arrays/up-to-10-mm-depth/</td>
		</tr>
		<tr>
			<td>NanoZ Omentics Adapter- 32 Channel</td>
			<td>NeuraLynx</td>
			<td>ADPT-NZ-N2T-32</td>
			<td>https://neuralynx.com/hardware/adpt-nz-n2t-32</td>
		</tr>
		<tr>
			<td>NanoZ System</td>
			<td>Plexon</td>
			<td>NanoZ Impedence Tester</td>
			<td>https://plexon.com/products/nanoz-impedance-tester/</td>
		</tr>
		<tr>
			<td>Narishige Micromanipulator</td>
			<td>Narishige</td>
			<td>Stereotaxic Micromanipulator</td>
			<td>https://usa.narishige-group.com/</td>
		</tr>
		<tr>
			<td>Open-Ephys GUI</td>
			<td>Open-Ephys</td>
			<td></td>
			<td>https://open-ephys.org/</td>
		</tr>
		<tr>
			<td>Polyimide Tubing (OD(in): 0.021 / ID(in) 0.018 )</td>
			<td>Various Sources (Chamfr)</td>
			<td>Chamfr Cat#HPC01895</td>
			<td>https://chamfr.com/sellers/teleflex-medical-oem-llc/</td>
		</tr>
		<tr>
			<td>Primate Chair</td>
			<td>Custom made by University of Rochester Machine Shop</td>
			<td>Designs online</td>
			<td>https://marmolab.bcs.rochester.edu/resources.html</td>
		</tr>
		<tr>
			<td>Poly(sodium 4-styrenesulfonate) (PSS)</td>
			<td>Sigma Aldrich</td>
			<td>Product # 243051</td>
			<td>https://www.sigmaaldrich.com/US/en/product/aldrich/243051</td>
		</tr>
		<tr>
			<td>RHD USB Interface board</td>
			<td>Intan</td>
			<td>RHD2000 Evaluation Board Version 1.0</td>
			<td>https://intantech.com/RHD_USB_interface_board.html</td>
		</tr>
		<tr>
			<td>Silastic gel</td>
			<td>World Precision Instuments</td>
			<td># KWIK-SIL</td>
			<td>Low Toxicity Silicone Adhesive ((https://www.wpiinc.com/kwik-sil-low-toxicity-silicone-adhesive)</td>
		</tr>
		<tr>
			<td>Slow release buprenorphine</td>
			<td>Compounding Pharmacy</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel wire 36 gauge</td>
			<td>McMaster &#38; Carr</td>
			<td>Cat# 6517K11</td>
			<td>Round Bend-and-Stay Multipurpose 304 Stainless Steel Wire, Matte Finish, 1-Foot Long, 0.008&#34; Diameter</td>
		</tr>
		<tr>
			<td>Stanley 6-Piece Precision Screwdriver Set</td>
			<td>Stanley</td>
			<td>1.4mm flathead screwdriver</td>
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</table>

electrophysiology of laminar cortical activity in the common marmoset

Marmoset maymunu, doğrusal dizilerle kayıtları kolaylaştıran pürüzsüz kortikal yüzeyi nedeniyle laminer kortikal devreleri incelemek için ideal bir model sağlar. Marmoset, diğer primatlara benzer sinirsel fonksiyonel organizasyonu ve kayıt ve görüntüleme için teknik avantajları nedeniyle son zamanlarda popülaritesini artırmıştır. Bununla birlikte, bu modeldeki nörofizyoloji, anatomik işaretler olarak küçük boyut ve gyri eksikliği nedeniyle bazı benzersiz zorluklar ortaya çıkarmaktadır. Araştırmacılar, özel olarak oluşturulmuş mikro sürücüleri kullanarak, doğrusal dizi yerleşimini milimetrenin altında hassasiyetle değiştirebilir ve kayıt günleri boyunca retinotopik olarak hedeflenen aynı konumda güvenilir bir şekilde kayıt yapabilir. Bu protokol, mikro sürücü konumlandırma sisteminin adım adım yapısını ve silikon lineer elektrot dizileri ile nörofizyolojik kayıt tekniğini açıklar. Kayıt seansları boyunca elektrot yerleşiminin hassas kontrolü ile araştırmacılar, retinotopik organizasyonlarına ve kaydedilen nöronların ayar özelliklerine dayalı olarak ilgi alanlarını belirlemek için korteksi kolayca geçebilirler. Ayrıca, bu laminer dizi elektrot sistemini kullanarak, tek tek nöronların kayıt derinliğini belirlemek için bir akım kaynağı yoğunluk analizi (CSD) uygulamak mümkündür. Bu protokol ayrıca, diziler üzerinde birden çok kanalı kapsayan Kilosort'ta izole edilen spike dalga biçimleri de dahil olmak üzere laminer kayıt örneklerini gösterir.

Bu protokol, sahaların milimetre altı hedeflenmesine izin veren ve ayrı kayıt oturumlarında güvenilir konumlandırmayı koruyan bir XY elektrot aşamasının (Şekil 1) nasıl oluşturulacağını açıklar. XY konumlandırmasının güvenilirliği Şekil 6'da gösterilmektedir, bu da bir hafta arayla gerçekleştirilen iki kayıt oturumunun ortalama RF konumlarında %70.8'lik bir örtüşme gösterdiğini göstermektedir (Şekil 6A</s...

Watch this Scientific Journal Video about Electrophysiology of Laminar Cortical Activity in the Common Marmoset at JoVE.com

Yazar Spotlight: Marmoset'te Hassas Elektrot Konumlandırma için Mikro Sürücü Sistemlerini Optimize Etme

Özel olarak üretilen mikro sürücüler, doğrusal silikon dizileri ile kortikal kayıt alanlarının milimetre altı hedeflenmesini sağlar.

Ortak Marmoset'te Laminer Kortikal Aktivitenin Elektrofizyolojisi

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with ...

Yaygın marmoset, küçük boyutu ve anatomik işaretler olarak girdap olmaması nedeniyle nörofizyoloji için benzersiz zorluklar ortaya çıkarmaktadır. Elektrotun sadece bir milimetre kadar hafifçe kayması, retinotopi haritasında önemli değişikliklere neden olabilir. Önerilen mikro tahrik sistemi, milimetre altı ölçekte dikey ve yatay harekete izin veren bir XY elektrot aşaması kullanır.Marmoset maymununun görsel nöron işareti için bir model olarak göreceli yeniliği nedeniyle, uyanık davranan elektrofizyoloji teknikleri hala gelişmektedir. Mevcut preparatlar genellikle konumlandırma mekanizmalarına erişime izin vermeyen yarı kronik problar kullanır. Bu protokol, oda içindeki retinotopiyi haritalamak için seanslar arasında esnek konumlandırmaya izin veren kullanışlı doğrusal dizi kayıtları için hafif bir mikro sürücü gösterir.Güvenli tahrik, dokunun yavaş penetrasyonu ve büyük kan damarlarından kaçınma ile doğru kullanıldığında preparatta uzun süreli kortikal hasar önlenebilir. Ek olarak, bu teknikte enfeksiyonları önlemek için kraniyotomilerde silastik kullanımı optimize edilmiştir.

electrophysiology-of-laminar-cortical-activity-in-the-common-marmoset

Research

JoVE Journal

Neuroscience

Electrophysiology of Laminar Cortical Activity in the Common Marmoset

701 Görüntüleme.  University of Rochester. Yaygın marmoset, küçük boyutu ve anatomik işaretler olarak girdap olmaması nedeniyle nörofizyoloji için benzersiz zorluklar ortaya çıkarmaktadır. Elektrotun sadece bir milimetre kadar hafifçe kayması, retinotopi haritasında önemli değişikliklere neden olabilir. Önerilen mikro tahrik sistemi, milimetre altı ölçekte dikey ve yatay harekete izin veren bir XY elektrot aşaması kullanır.Marmoset maymununun görsel nöron işareti için bir model olarak göreceli yenil...

Video: Yazar Spotlight: Marmoset'te Hassas Elektrot Konumlandırma için Mikro Sürücü Sistemlerini Optimize Etme

Simultaneous fMRI and Electrophysiology in the Rodent Brain

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in which functional MRI data and in vivo intracortical recording can be performed simultaneously. The combination of MRI and electrical recording is technically challenging because the electrodes used for recording distort the MRI images and the MRI acquisition induces noise in the electrical recording. To minimize the mutual interference of the two modalities, glass microelectrodes were used rather than metal and a noise removal algorithm was implemented for the electrophysiology data. In our studies, two microelectrodes were separately implanted in bilateral primary somatosensory cortices (SI) of the rat and fixed in place. One coronal slice covering the electrode tips was selected for functional MRI. Electrode shafts and fixation positions were not included in the image slice to avoid imaging artifacts. The removed scalp was replaced with toothpaste to reduce susceptibility mismatch and prevent Gibbs ringing artifacts in the images. The artifact structure induced in the electrical recordings by the rapidly-switching magnetic fields during image acquisition was characterized by averaging all cycles of scans for each run. The noise structure during imaging was then subtracted from original recordings. The denoised time courses were then used for further analysis in combination with the fMRI data. As an example, the simultaneous acquisition was used to determine the relationship between spontaneous fMRI BOLD signals and band-limited intracortical electrical activity. Simultaneous fMRI and electrophysiological recording in the rodent will provide a platform for many exciting applications in neuroscience in addition to elucidating the relationship between the fMRI BOLD signal and neuronal activity.

We have developed a method for simultaneous functional magnetic resonance imaging and electrophysiological recording in the rodent brain, providing a platform for the investigation of the relationship between neural activity and the blood oxygenation level dependent (BOLD) MRI signal.

Kemirgen Beyin aynı anda fMRI ve Elektrofizyoloji

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in ...

Nörobilim

Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate a method to fabricate suction electrodes that are used to examine CPG activity, or fictive locomotion, in dissected rodent spinal cords. The rodent spinal cords are placed in artificial cerebrospinal fluid and the ventral roots are drawn into the suction electrode. The electrode is constructed by modifying a commercially available suction electrode. A heavier silver wire is used instead of the standard wire given by the commercially available electrode. The glass tip on the commercial electrode is replaced with a plastic tip for increased durability. We prepare hand drawn electrodes and electrodes made from specific sizes of tubing, allowing consistency and reproducibility. Data is collected using an amplifier and neurogram acquisition software. Recordings are performed on an air table within a Faraday cage to prevent mechanical and electrical interference, respectively.

A demonstration of the fabrication and use of an extracellular suction electrode used to measure electrophysiological recordings of neonatal rodent spinal cords in vitro

Omurilik Elektrofizyoloji II: Ekstrasellüler Emiş Elektrot İmalatı

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate ...

Examining Local Network Processing using Multi-contact Laminar Electrode Recording

Cortical layers are ubiquitous structures throughout neocortex1-4 that consist of highly recurrent local networks. In recent years, significant progress has been made in our understanding of differences in response properties of neurons in different cortical layers5-8, yet there is still a great deal left to learn about whether and how neuronal populations encode information in a laminar-specific manner.
Existing multi-electrode array techniques, although informative for measuring responses across many millimeters of cortical space along the cortical surface, are unsuitable to approach the issue of laminar cortical circuits. Here, we present our method for setting up and recording individual neurons and local field potentials (LFPs) across cortical layers of primary visual cortex (V1) utilizing multi-contact laminar electrodes (Figure 1; Plextrode U-Probe, Plexon Inc).
The methods included are recording device construction, identification of cortical layers, and identification of receptive fields of individual neurons. To identify cortical layers, we measure the evoked response potentials (ERPs) of the LFP time-series using full-field flashed stimuli. We then perform current-source density (CSD) analysis to identify the polarity inversion accompanied by the sink-source configuration at the base of layer 4 (the sink is inside layer 4, subsequently referred to as granular layer9-12). Current-source density is useful because it provides an index of the location, direction, and density of transmembrane current flow, allowing us to accurately position electrodes to record from all layers in a single penetration6, 11, 12.

A fundamental issue in our understanding of cortical circuitry is how networks in different cortical layers encode sensory information. Here we describe electrophysiological techniques utilizing multi-contact laminar electrodes to record single-units and local field potentials and present analyses to identify cortical layers.

Multi-contact kullanarak Yerel Ağ İşleme incelenmesi Laminer Elektrot Kayıt

Cortical layers are ubiquitous structures throughout neocortex1-4 that consist of highly recurrent local networks. In recent years, significant progress ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

Locust Sinir Aktivitesi Karakterize Multi-ünite Kayıt Yöntemleri (<em&gt; Schistocerca Americana</em&gt;) Olfaktör Devreler

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety of brain disorders1,2. To better understand the mechanisms by which brain circuits react to an animal's experience requires the ability to monitor the experience-dependent molecular changes in a given set of neurons, over a prolonged period of time, in the live animal. While experience and associated neural activity is known to trigger gene expression changes in neurons1,2, most of the methods to detect such changes do not allow repeated observation of the same neurons over multiple days or do not have sufficient resolution to observe individual neurons3,4. Here, we describe a method that combines in vivo two-photon microscopy with a genetically encoded fluorescent reporter to track experience-dependent gene expression changes in individual cortical neurons over the course of day-to-day experience. 
One of the well-established experience-dependent genes is Activity-regulated cytoskeletal associated protein (Arc)5,6. The transcription of Arc is rapidly and highly induced by intensified neuronal activity3, and its protein product regulates the endocytosis of glutamate receptors and long-term synaptic plasticity7. The expression of Arc has been widely used as a molecular marker to map neuronal circuits involved in specific behaviors3. In most of those studies, Arc expression was detected by in situ hybridization or immunohistochemistry in fixed brain sections. Although those methods revealed that the expression of Arc was localized to a subset of excitatory neurons after behavioral experience, how the cellular patterns of Arc expression might change with multiple episodes of repeated or distinctive experiences over days was not investigated. 
In vivo two-photon microscopy offers a powerful way to examine experience-dependent cellular changes in the living brain8,9. To enable the examination of Arc expression in live neurons by two-photon microscopy, we previously generated a knock-in mouse line in which a GFP reporter is placed under the control of the endogenous Arc promoter10. This protocol describes the surgical preparations and imaging procedures for tracking experience-dependent Arc-GFP expression patterns in neuronal ensembles in the live animal. In this method, chronic cranial windows were first implanted in Arc-GFP mice over the cortical regions of interest. Those animals were then repeatedly imaged by two-photon microscopy after desired behavioral paradigms over the course of several days. This method may be generally applicable to animals carrying other fluorescent reporters of experience-dependent molecular changes4.

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

Experience-dependent molecular changes in neurons are essential for the brain's ability to adapt in response to behavioral challenges. An in vivo two-photon imaging method is described here that allows the tracking of such molecular changes in individual cortical neurons through genetically encoded reporters.

Kortikal Nöronlar In Vivo İki foton Of Görüntüleme Deneyimi bağımlı Moleküler Değişiklikler

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety ...

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich structures suggest alterations in synaptic integrity and connectivity. Here we provide a detailed protocol for culturing primary rat cortical neurons, Phalloidin staining for F-actin puncta, and subsequent quantification techniques. First, the frontal cortex of E18 rat embryos are dissociated into low-density cell culture, then the neurons grown in vitro for at least 12-14 days. Following experimental treatment, the cortical neurons are stained with AlexaFluor 488 Phalloidin (to label the dendritic F-actin puncta) and microtubule-associated protein 2 (MAP2; to validate the neuronal cells and dendritic integrity). Finally, specialized software is used to analyze and quantify randomly selected neuronal dendrites. F-actin rich structures are identified on second order dendritic branches (length range 25-75 &#181;m) with continuous MAP2 immunofluorescence. The protocol presented here will be a useful method for investigating changes in dendritic synapse structures subsequent to experimental treatments.

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

Filamentous actin (F-actin) plays an important role in spinogenesis, synaptic plasticity, and synaptic stability. Quantification of F-actin puncta is therefore a useful tool to study the integrity of synaptic structures. This protocol describes the procedures of quantifying F-actin puncta labeled with Phalloidin in low-density primary cortical neuronal cultures.

Rat Kortikal Nöronlar Filamentli Aktin (F-aktin) puncta Niceleme

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich ...

Prolonged Incubation of Acute Neuronal Tissue for Electrophysiology and Calcium-imaging

Acute neuronal tissue preparations, brain slices and retinal wholemount, can usually only be maintained for 6 - 8 h following dissection. This limits the experimental time, and increases the number of animals that are utilized per study. This limitation specifically impacts protocols such as calcium imaging that require prolonged pre-incubation with bath-applied dyes. Exponential bacterial growth within 3 - 4 h after slicing is tightly correlated with a decrease in tissue health. This study describes a method for&#160;limiting the proliferation of bacteria in acute preparations to maintain viable neuronal tissue for prolonged periods of time (&#62;24 h) without the need for antibiotics, sterile procedures, or tissue culture media containing growth factors. By cycling the extracellular fluid through UV irradiation and keeping the tissue in a custom holding chamber at 15 - 16 &#176;C, the tissue shows no difference in electrophysiological properties, or calcium signaling through intracellular calcium dyes at &#62;24 h postdissection. These methods will not only extend experimental time for those using acute neuronal tissue, but will reduce the number of animals required to complete experimental goals, and will set a gold standard for acute neuronal tissue incubation.

Once removed from the body, neuronal tissue is greatly affected by environmental conditions, leading to eventual degradation of the tissue after 6 - 8 h. Using a unique incubation method, which closely monitors and regulates the extracellular environment of the tissue, tissue viability can be significantly extended for &#62;24 h.

Elektrofizyoloji ve Kalsiyum-görüntüleme için akut nöronal Doku Uzamış İnkübasyon

Acute neuronal tissue preparations, brain slices and retinal wholemount, can usually only be maintained for 6 - 8 h following dissection. This limits the ...

In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice

Two-photon imaging is a powerful tool for the in vivo analysis of neuronal circuits in the mammalian brain. However, a limited number of in vivo imaging methods exist for examining the brain tissue of live newborn mammals. Herein we summarize a protocol for imaging individual cortical neurons in living neonatal mice. This protocol includes the following two methodologies: (1) the Supernova system for sparse and bright labeling of cortical neurons in the developing brain, and (2) a surgical procedure for the fragile neonatal skull. This protocol allows the observation of temporal changes of individual cortical neurites during neonatal stages with a high signal-to-noise ratio. Labeled cell-specific gene silencing and knockout can also be achieved by combining the Supernova with RNA interference and CRISPR/Cas9 gene editing systems. This protocol can, thus, be used for analyzing the developmental dynamics of cortical neurons, molecular mechanisms that control the neuronal dynamics, and changes in neuronal dynamics in disease models.

In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice

We present an in vivo two-photon imaging protocol for imaging the cerebral cortex of neonatal mice. This method is suitable for analyzing the developmental dynamics of cortical neurons, the molecular mechanisms that control the neuronal dynamics, and the changes in neuronal dynamics in disease models.

Vivo Yenidoğan farelerde kortikal nöronların iki fotonlu görüntüleme

Two-photon imaging is a powerful tool for the in vivo analysis of neuronal circuits in the mammalian brain. However, a limited number of in vivo imaging ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

Kronik yaygın Marmoset bütün kortikal Electrocorticographic dizide implantasyonu

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Stimulus-specific Cortical Visual Evoked Potential Morphological Patterns

This paper presents a methodology for the recording and analysis of cortical visual evoked potentials (CVEPs) in response to various visual stimuli using 128-channel high-density electroencephalography (EEG). The specific aim of the described stimuli and analyses is to examine whether it is feasible to replicate previously reported CVEP morphological patterns elicited by an apparent motion stimulus, designed to simultaneously stimulate both ventral and dorsal central visual networks, using object and motion stimuli designed to separately stimulate ventral and dorsal visual cortical networks.&#160; Four visual paradigms are presented: 1. Randomized visual objects with consistent temporal presentation. 2. Randomized visual objects with inconsistent temporal presentation (or jitter).&#160; 3. Visual motion via a radial field of coherent central dot motion without jitter.&#160; 4. Visual motion via a radial field of coherent central dot motion with jitter.&#160; These four paradigms are presented in a pseudo-randomized order for each participant.&#160; Jitter is introduced in order to view how possible anticipatory-related effects may affect the morphology of the object-onset and motion-onset CVEP response.&#160; EEG data analyses are described in detail, including steps of data exportation from and importation to signal processing platforms, bad channel identification&#160;and removal, artifact rejection, averaging, and categorization of average CVEP morphological pattern type based upon latency ranges of component peaks. Representative data show that the methodological approach is indeed sensitive in eliciting differential object-onset and motion-onset CVEP morphological patterns and may, therefore, be useful in addressing the larger research aim. Given the high temporal resolution of EEG and the possible application of high-density EEG in source localization analyses, this protocol is ideal for the investigation of distinct CVEP morphological patterns and the underlying neural mechanisms which generate these differential responses.&#160; &#160; &#160;&#160;

In this paper, we present a protocol to investigate differential cortical visual evoked potential morphological patterns through stimulation of ventral and dorsal networks using high-density EEG. Visual object and motion stimulus paradigms, with and without temporal jitter, are described. Visual evoked potential morphological analyses are also outlined.&#160;&#160;

Stimulus-spesifik kortikal görsel uyarılmış potansiyel morfolojik desenler

This paper presents a methodology for the recording and analysis of cortical visual evoked potentials (CVEPs) in response to various visual stimuli using ...