Yazarlar fotoğraf çekmek onların yardım için Marin Manuel ve Olivia Goldman-Szwajkajzer teşekkür ederim. Yazarlar ayrıca yazının redaksiyon için Arjun Masukar ve Tobias Bock teşekkür ederiz. Mali destekler la Recherche (HYPER-MND, ANR-2010-BLAN-1429-1401) dökün Agence Nationale tarafından sağlanan, NIH-NINDS (R01NS077863), Thierry Latrán Vakfı (OHEX Projesi), miyopati Fransız derneği ( hibe sayısı 16026) ve Hedef ALS minnetle kabul vardır. Felix Leroy Ecole Normale Supérieure Cachan&#39;da bir &quot;contrat Doktora&quot; layık görüldü.

Deneyler Avrupa direktiflerine (86/609 / CEE ve 2010-63-UE) ve Fransız mevzuatına uygun olarak yapıldı ve Paris Descartes Üniversitesi etik kurul tarafından onaylandı. 1. Omurilik Dilim Hazırlık <ol><li> Aşağıdaki günlük çözümler ya da bir gün önceden hazırlayın. Bir gecede devam ederse,% 95 O 2 ve% 5 CO 2 ile kabarcık ve sıkıca kapalı şişelerde buzdolabında tutun. <ol><li> Düşük Na yapay serebrospinal sıvı (ACSF) + Hazırlama: 3 mM KCI, 1 mM NaH 2 PO 4, 230 mM sakaroz, 26 mM NaHCO 3, 0.8 mM CaCl2, 8 mM MgCI2, 25 mM glükoz, 0.4 mM askorbik asit, 1 mM Na-kynurenate, 2 mM Na-piruvat. % 95 O2 ve% 5 CO2 (pH 7.4) Kabarcık. Na-kynurenate genellikle çözünmesi zor olduğundan, malzeme tabloda listelenen satın almak için emin olun. </li><li> K-glukonat Solu hazırlanmasıtion KOH ile pH 7.4&#39;e ayarlanmış 130 mM K-glukonat, 15 mM KCI, 0.05 mM EGTA, 20 mM HEPES, 25 mM glükoz, 1 mM Na-kynurenate, 2 mM Na-piruvat,. </li><li> ACSF hazırlayın: 130 mM NaCI, 2.5 mM KCI, 2 mM CaCl2, 1 mM MgCl2, 1 mM NaH 2 PO 4, 26 mM NaHCO 3, 25 mM glükoz, 0.4 mM askorbik asit, 2 mM Na-piruvat. % 95 O2 ve% 5 CO2 (pH 7.4) Kabarcık. </li><li> diseksiyon başlamadan önce, K-glukonat çözeltisi 80 ml% 2 agar çözülür ve 60 ° C &#39;de sıcak tutun. </li></ol></li><li> intrakardiyak perfüzyon <ol><li> yaş P2 P11 kadar, dişi ve erkek farenin bu hazırlık gerçekleştirin. </li><li> 25 mM sodyum pentobarbital (50 mg / kg), 0.1 ml intraperitoneal enjeksiyonu ile fare anestezisi. </li><li> iğneler veya bant kullanılarak, silikon ile dolu büyük bir petri sırtında fare hareketsiz. Diseksiyon mikroskop kullanındiseksiyon geri kalanı için e. </li><li> sternum ucu Holding, göğüs kaldırın ve ince makas kullanarak diyafram kesti. Sonra kalp maruz kaburga ile keserek her iki tarafta kutuyu açın. </li><li> 27G iğne ile sol ventrikül delme önce sağ atrium kesin. </li><li> Buz gibi soğuk düşük Na + ACSF serpmek. 30 saniye sonra, düşük Na + ACSF atrium dışarı akan görülmelidir. sodyum düşük miktarda spiking hücreleri önlemek ve diseksiyon sırasında hücresel ölüm azaltır. </li><li> Kan boşaltılır karaciğer sarı olana kadar mikroskop altında kalbinde iğne basılı tutun. </li></ol></li><li> Omurilik diseksiyonu <ol><li> hayvan başını kesmek ve mide üzerine koydu. </li><li> Hızla arkasına (Şekil 1A1) cilt kaldırın. İki kesim omuzları ve göğüs kafesini (Şekil 1A2) aşağı gidiyor olun. Sonra l olarak kablosunu kesiphayvan alt kısmından kaburga başlangıcı ile gezilebilen omurga izole etmek üzere kaudal bölümünde olduğu gibi mümkün olan akış. Yine hayvan çevirmek ve hala kaburga bağlı iç organ kaldırın. </li><li> Başka vertebral sütun transfer daha küçük silikon dolu petri ve dorsal yüzü yukarı (Şekil 1A) tutmak için 4 böcek işaretçilerine kullanın. </li><li> Sırt tarafında bir laminektomi gerçekleştirerek, ve rostral sonunda (Şekil 1B) den omurilik teşhir ederken Sürekli (yaklaşık 4 ° C&#39;de) carbogen-kabarmış ACSF ile hayvan serpmek. Bunu yapmak için, kemik ve omurilik arasındaki ince makas ucu yerleştirin ve beyaz maddeden uzak kalmak için emin olun, rostral ucundan azar azar kemiği kesti. Her iki tarafta alternatif kemik bant zaten kesilmiş (Şekil 1B1) uzak tutmak için cımbız kullanarak. </li><li> en küçük kullanılabilir yaylı makas ve cımbız kullanarak, dura kaldırın ve kesmemakasla omurilik zarar vermemek amacıyla her iki tarafta duranın gevşek kısmını tutarken. postero-kaudal ekseni boyunca kesin. NOT: Dura yarı saydam sürekli zardır; Bu yaşta pia mater çok kırılgan ve diseksiyon ve dilimleme (Şekil 1B2) sırasında ayrı gelecektir. </li><li> Dura kaldırılan kullanımı kez künt cam veya plastik uç hafifçe yarım kesilmiş omurga oluşturduğu oluğun sol tarafındaki kabloyu itin ve rostral taraftan başlayarak, sağ tarafta ventral ve dorsal kökleri kesmek için onlar (en azından birkaç mm Şekil 1B2) kordon girmek yerden uzak. </li><li> her zaman rostral gelen kaudal gidiyor, sol tarafta işlemi tekrarlayın. solak, sol taraftan başlamak ve daha sonra sağ tarafa hareket ettirin. </li></ol></li><li> Agar gömme <ol><li> Vertebral kolon dışına kablosunu Kayma. aşağı pin küçük bir böcek pin kullanındorsal kordon yukarı yüzey ve ince zarın herhangi bir parçası hala ona bağlı (Şekil 1C1) çıkarın. </li><li> Temizlenmiş sonra, her iki ucu (Şekil 1C2) kırpın. Kablosunu işlemek ve yönünü not kord ön kısmında bir bükülmüş böcek pin yerleştirin (Şekil 1C2 ok). Daha sonra, bir buz-soğuğu K-glukonat çözeltisine omurilik aktarın. Not: Bu çözüm BOS hücre içi kompozisyon taklit ve motor nöronlar 26 kesilecektir kez ozmotik şok ölen hücrelerini engeller. </li><li> Omurilik hücre içi çözelti içinde olduğunda, kuru banyo üzerinden agar ile beher almak ve bir buz ve su karışımı üzerine soğuma. </li><li> sıcaklık ölçüm sırasında karıştırmaya devam edin. Sıcaklık 38 ° ulaştığında böcek pimi tutarak, omurilik batırmak ve aşağı rostral tarafı yerleştirin C. Omurilik st olarak olduğundan emin olunraight mümkün olduğunca uzak kaudal kısmı, duvarlar hafifçe yukarı doğru (Şekil 1D1). </li><li> Agar olabildiğince hızlı bir şekilde katılaşmasına izin buz ve su karışımı içinde beher bırakın. Emin kordon mümkün (Şekil 1D1) gibi düz olduğu yerde kalır ve emin olun. </li></ol></li><li> Dilimleme <ol><li> Sertleştikten sonra, (Şekil 1D2 ok) bloğun tabanı kablosunun bel kısmı ile 35 ° &#39;lik açıyla olduğu bir şekilde omuriliği ihtiva eden agar kesti. Dorsal yüzey tabanı (Şekil 1D2) uzağa dönük olmalıdır. Not: Bu çıktıkları ventral köklerine motornöron havuzları sürekliliğini korumak için prosedür kritik bir adımdır. </li><li> siyanoakrilat yapıştırıcı kullanarak vibratome odasına blok Tutkal. K-glukonat solüsyonu içinde daldırın ve korumak için slushed dondurulmuş K-glukonat çözüm ekleyinbanyo (2 ° C&#39;nin altında) soğutulmuş. </li><li> (Onun eğrilik ve büyük çaplı tarafından tanımlanabilir) bel bölgesinde 400 mikron kalınlığında dilimler - 350 kesin. kendi doğru yönde (1.4.1 bakınız.) &#39;de, ventral yüzeyi dorsalden kesip dilimler sürekli daha rostral yapmak için bıçak kullanın. Tipik olarak, 2 mm veya daha fazla uzanan ventral kökleri ile 4-5 uygun dilimler vardır. 10 ° açı, 70 Hz titreşim frekansı ve dilimleme 10 mm / dk hızı: Aşağıdaki parametreleri kullanın. </li></ol></li><li> kuluçka <ol><li> 34 ° C&#39;de ACSF dilimleri aktarın. Yaklaşık 30 dakika sonra, oda sıcaklığına kadar soğumaya dilimleri ve kayıt oturumunu başlatmak. </li></ol></li></ol><img alt="Şekil 1" src="/files/ftp_upload/54525/54525fig1.jpg" /> Şekil 1. Diseksiyon A1:. Dorsal kolon maruz arka derinin çıkarılması A2:. Omuzlar ve kaburga kesme dorsal kolon B1 serbest: silikon dolu petri üzerine tutturulmuş Omurga sütunu (yukarı sırt tarafı, sol kaudal tarafı) B2:. Maruz kalan ve disseke omurilik ile aynı C1:. Omurilik izole . (rostral tarafı sol) C2: yerleştirilmeye hazır Omurilik (yukarı ventral tarafta, sol rostral tarafı). Rostral tarafında küçük böcek pin Not D1:.. Agar beher Omurilik (rostral tarafı dow, dip bakan ventral tarafı) D2: gömülü omurilik ile Kes agar bloğu. 35 ° açısı bloğunun taban ve lomber genişleme (Ok) ile omurilik formları unutmayın. Ölçek çubukları 1 cm. <a href="https://www.jove.com/files/ftp_upload/54525/54525fig1large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> 2. Odası&#39;na Slice Yerleştirme NOT: Ventral kökleri, değişken bir alana yayılır. <ol><li> 40 peşin 170 um arasında değişen uç çapları çeşitli pipetler bir kutu hazırlayın. Emme pipetler hazırlamak uzun konik birçok pipetler hazırlamak. Bir elmas bıçak kullanarak, farklı pozisyonlarda bir kesim yapmak. Sonra bir mikroskop altında, cımbız ile ucu vurarak kırmak. </li><li> Kayıt mikroskop odasına çıkarın ve bir mikroskop altında yerleştirin. </li><li> bir emme stimülasyon elektrodu monte edilmesi için yeterli uzunlukta (2 mm veya daha fazla) bir ventral kök içeren bir dilim seçin. Ventral kökleri ile dilim yukarı (Şekil 2A) doğru yönünü seçin ve dilim (Şekil 2B) etrafında agar geri kalanını terk ederken ince ventral kökleri etrafında agar kesti. NOT: Agar dilim daha sıkı olduğundan, bu dilimin çapa konuları ağar ziyade böylece dilim ve dinlenmek için izin verecektirnchor dokuya zarar vermez. Dilimin çapa ipler (Şekil 2C kırmızı ile gösterilen) motonöron havuzu üzerinde olduğundan emin olun. </li><li> mikroskop üzerine geri bölmesini monte sürekli 1 bir oranda ACSF kayıt odasına serpmek - oda sıcaklığında, 2 ml / dakika. Cam pipet ACSF ile doldurulur ve bir şırıngaya bağlanmış kullanarak, ventral kök (Şekil 2C ok) bir emmeye. ventral kök iyi hareketini elde etmek için, pipet ucu ventral kök etrafında sıkı olması gerekmektedir. Bir kutup uyarıcı elektrot ve (yama-sıkma elektrot referans bağlı veya) banyosunda diğeri olmalıdır. </li><li> Istenen hücre tipi yama-kelepçe kayıt ulaşmak ve previsouly 10 açıklandığı gibi ventral kök stimülasyon etkisi kaydedin. <ol><li> Burada, veri toplama için bir amplifikatör kullanın. 3 kHz tam hücre kayıtları Filtre. 10 kHz Digitize. curre köprü direnci telafint-kelepçe modu. </li></ol></li></ol><img alt="şekil 2" src="/files/ftp_upload/54525/54525fig2.jpg" /> Şekil 2. Ventral Kök Hazırlık A: ventral kök yukarı bakacak şekilde agar gömülü Lomber spinal kord dilim B:.. Agar C kurtulmuş ventral kökleri ile Lomber spinal kord dilim: sıkıca (ok) ventral kökleri etrafında yerleştirilmiş bir uyarıcı elektrot ile Lomber spinal kord dilim . Fare muhabiri R26 Tom 17. Ölçek çubukları 1 mm çapraz chrna2-Cre fare 28 kırmızı floresan ifade Renshaw hücreleri konumunu not alın. <a href="https://www.jove.com/files/ftp_upload/54525/54525fig2large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a>

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D., Zytnicki, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+intrinsic+hyperexcitability+does+not+contribute+to+motoneuron+degeneration+in+amyotrophic+lateral+sclerosis.">Early intrinsic hyperexcitability does not contribute to motoneuron degeneration in amyotrophic lateral sclerosis.</a> Elife. 3, (2014).</li><li>Leroy, F., Lamotte d'Incamps, B., Zytnicki, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potassium+currents+dynamically+set+the+recruitment+and+firing+properties+of+F-type+motoneurons+in+neonatal+mice.">Potassium currents dynamically set the recruitment and firing properties of F-type motoneurons in neonatal mice.</a> J Neurophysiol. 114 (3), 1963-1973 (2015).</li><li>Lamotte d'Incamps, B., Ascher, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subunit+composition+and+kinetics+of+the+Renshaw+cell+heteromeric+nicotinic+receptors.">Subunit composition and kinetics of the Renshaw cell heteromeric nicotinic receptors.</a> Biochem Pharmacol. 86 (8), 1114-1121 (2013).</li><li>Lamotte d'Incamps, B., Krejci, E., Ascher, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+shaping+the+slow+nicotinic+synaptic+current+at+the+motoneuron-renshaw+cell+synapse.">Mechanisms shaping the slow nicotinic synaptic current at the motoneuron-renshaw cell synapse.</a> J Neurosci. 32 (24), 8413-8423 (2012).</li><li>Dugue, G. P., Dumoulin, A., Triller, A., Dieudonne, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Target-dependent+use+of+co-released+inhibitory+transmitters+at+central+synapses.">Target-dependent use of co-released inhibitory transmitters at central synapses.</a> J Neurosci. 25 (28), 6490-6498 (2005).</li><li>Mentis, G. Z., Siembab, V. C., Zerda, R., O'Donovan, M. J., Alvarez, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primary+afferent+synapses+on+developing+and+adult+Renshaw+cells.">Primary afferent synapses on developing and adult Renshaw cells.</a> J Neurosci. 26 (51), 13297-13310 (2006).</li><li>Perry, 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=Firing+properties+of+Renshaw+cells+defined+by+Chrna2+are+modulated+by+hyperpolarizing+and+small+conductance+ion+currents+Ih+and+ISK.">Firing properties of Renshaw cells defined by Chrna2 are modulated by hyperpolarizing and small conductance ion currents Ih and ISK.</a> Eur J Neurosci. 41 (7), 889-900 (2015).</li><li>Thurbon, D., Luscher, H. R., Hofstetter, T., Redman, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Passive+electrical+properties+of+ventral+horn+neurons+in+rat+spinal+cord+slices.">Passive electrical properties of ventral horn neurons in rat spinal cord slices.</a> J Neurophysiol. 79 (5), 2485-2502 (1998).</li><li>Zengel, J. E., Reid, S. A., Sypert, G. W., Munson, 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=Membrane+electrical+properties+and+prediction+of+motor-unit+type+of+medial+gastrocnemius+motoneurons+in+the+cat.">Membrane electrical properties and prediction of motor-unit type of medial gastrocnemius motoneurons in the cat.</a> J Neurophysiol. 53 (5), 1323-1344 (1985).</li><li>Cooper, S., Sherington, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gower's+tract+and+spinal+border+cells.">Gower's tract and spinal border cells.</a> Brain. 63, 123-124 (1940).</li><li>Morin, F., Schwartz, H. G., O'Leary, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+study+of+the+spinothalamic+and+related+tracts.">Experimental study of the spinothalamic and related tracts.</a> Acta Psychiatr Neurol Scand. 26 (3-4), 371-396 (1951).</li><li>Sengul, G., Fu, Y., Yu, Y., Paxinos, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+projections+to+the+cerebellum+in+the+mouse.">Spinal cord projections to the cerebellum in the mouse.</a> Brain Struct Funct. 220 (5), 2997-3009 (2015).</li><li>Russier, M., Carlier, E., Ankri, N., Fronzaroli, L., Debanne, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A-,+T-,+and+H-type+currents+shape+intrinsic+firing+of+developing+rat+abducens+motoneurons.">A-, T-, and H-type currents shape intrinsic firing of developing rat abducens motoneurons.</a> J Physiol. 549 (Pt 1), 21-36 (2003).</li><li>Dourado, M., Sargent, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+of+nicotinic+receptors+underlying+Renshaw+cell+excitation+by+alpha-motor+neurons+in+neonatal+rat+spinal+cord).">Properties of nicotinic receptors underlying Renshaw cell excitation by alpha-motor neurons in neonatal rat spinal cord).</a> J Neurophysiol. 87 (6), 3117-3125 (2002).</li><li>Mitra, P., Brownstone, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vitro+spinal+cord+slice+preparation+for+recording+from+lumbar+motoneurons+of+the+adult+mouse.">An in vitro spinal cord slice preparation for recording from lumbar motoneurons of the adult mouse.</a> J Neurophysiol. 107 (2), 728-741 (2012).</li><li>Rothman, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neurotoxicity+of+excitatory+amino+acids+is+produced+by+passive+chloride+influx.">The neurotoxicity of excitatory amino acids is produced by passive chloride influx.</a> J Neurosci. 5 (6), 1483-1489 (1985).</li><li>Olney, J. W., Price, M. T., Samson, L., Labruyere, J. <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+specific+ions+in+glutamate+neurotoxicity.">The role of specific ions in glutamate neurotoxicity.</a> Neurosci Lett. 65 (1), 65-71 (1986).</li></ol>

The authors have nothing to disclose.

Bir, güvenilir, kapsamlı ve belirli bir şekilde tek bir omurga segmentinde motornöron havuzları ve Renshaw hücreleri tek taraflı uyarılması için izin verir çünkü omurilik eğik dilimleme önemlidir. Ayrıca, kaydedilen motor nöronlar, hızlı, zarif ve olmayan belirsiz tanımlanmasına izin verir. Sonra, diğer dilim hazırlama yöntemlerine göre bu tekniğin avantajlarını vurgulamak olacaktır, ve sonra biz bu yordamı gerçekleştirirken önlemek için en yaygın tuzaklar stres olacak. <p class="j...

Tarihsel olarak, keskin elektrodu kullanılarak motonöron kayıtları kedi, sıçan 1 ya da fareler 2 izole edilmiş bir bütün omurilik büyük hayvanlarda in vivo olarak gerçekleştirilmiştir. somas gerekli sızdırmazlık olarak motor nöron doğrudan erişim için çağrıda 1980&#39;li yıllarda patch-kelepçe kayıt tekniğinin ortaya çıkması, görsel rehberliği altında elde edilecek. Böylece, omurilik dilim hazırlık kolayca 1990&#39;ların başından bu yana 3 elde edilmiştir. Ancak, erken dilim hazırlık genellikle ventral köklerinin uyarılması için izin vermedi. Bizim bildiğimiz kadarıyla, sadece iki çalışma enine dilim ventral köklerin başarılı uyarılması bildirilmiştir ve hiçbiri fareler 4,5 elde edilmiştir. motonöron havuzu ventral kök kalkan aksonlar elinde bulundurduğu - (P11 P2) Bu yazıda yenidoğan farelerin canlı omurilik dilim elde etmek için bir teknik sunuyoruz. delikral kök stimülasyon aynı ventral kök çıkan motonöron havuz mis geri antidromik aksiyon potansiyelini tetikler. Aynı zamanda motor nöron teminat hedefleri, diğer motor nöronlar 6-10 ve Renshaw hücreleri 11-13 heyecanlandırıyor. Sadece motor nöronlar ventral kökleri kendi aksonlar aşağı göndermek yana, motor nöronlar 10 belirlemek physiologicaly için basit ve kesin yolu olarak antidromik aksiyon potansiyelleri kayıt kullanın. Motonöron kimliğini doğrulamak için potansiyel olmayan kapsayıcı veya yanıltıcı elektrofizyolojik ve morfolojik kriterler kullanılarak ek olarak, omurilik motor nöronlar üzerinde son çalışmalar da post hoc sıkıcı ve zaman alıcı Renklendirmeler 16 dayanmıştır. Bu tanımlama genellikle kaydedilen bir hücre numunesi üzerinde gerçekleştirilmiştir. Diğer kimlik stratejileri motor nöronlar endojen floresan ifade ettiği fare hatları güveniyor <sup&gt; 17-19. Çalışma zaten transgenik fare hattı kullanarak gerektiriyorsa işaretleyici ifade hala değişken veya Ancak, genetik olarak kodlanmış işaretlerini kullanarak genç yaşta zor olabilir. Seçenek olarak ise, antidromik aksiyon potansiyeli kayıtları hücre kayıt başlangıcından itibaren tüm fareler üzerinde rutin olarak gerçekleştirilebilir. Kedi, sıçan ve fare bozulmamış omurilik hazırlıkları üzerinde çalışan deneyci, güvenilir 1950 1,2,20,21 beri böyle tanımlama teknikleri kullandık. Optimal koşullarda, biz kaydedilen motor nöronlar neredeyse tüm antidromik aksiyon potansiyelleri ortaya çıkarmak için başardık. Ayrıca, ventral kök stimülasyon güvenilir diğer motor nöronlar 22,23 ya da hedef uyarmak için kullanılabilir. Renshaw hücreleri 10,24,25. Burada motonöron mis gelen antidromik aksiyon potansiyeli kayıtlarında şeklinde ventral kök stimülasyon uygulamaları yanı sıra, Renshaw hücre uyarma sunulmuştur.

<table >
	<tbody>
		<tr >
			<td >Na-kynurenate</td>
			<td >ABCAM</td>
			<td >ab120256</td>
			<td >dissolves better then other brands</td>
		</tr>
		<tr >
			<td >KCl</td>
			<td >Sigma</td>
			<td >P3911</td>
			<td></td>
		</tr>
		<tr >
			<td >NaH2PO4</td>
			<td >Sigma</td>
			<td >P5655</td>
			<td></td>
		</tr>
		<tr >
			<td >sucrose&#160;</td>
			<td >Sigma</td>
			<td >S9378</td>
			<td></td>
		</tr>
		<tr >
			<td >NaHCO3&#160;</td>
			<td >Sigma</td>
			<td >S6014</td>
			<td></td>
		</tr>
		<tr >
			<td >CaCl2&#160;</td>
			<td >G Biosciences</td>
			<td >R040</td>
			<td></td>
		</tr>
		<tr >
			<td >MgCl2&#160;</td>
			<td >Quality Biological</td>
			<td >351-033-721</td>
			<td></td>
		</tr>
		<tr >
			<td >glucose&#160;</td>
			<td >Sigma</td>
			<td >G5767</td>
			<td></td>
		</tr>
		<tr >
			<td >ascorbic acid&#160;</td>
			<td >Sigma</td>
			<td >A5960</td>
			<td></td>
		</tr>
		<tr >
			<td >Na-pyruvate&#160;</td>
			<td >Sigma</td>
			<td >P2250</td>
			<td></td>
		</tr>
		<tr >
			<td >K-gluconate&#160;</td>
			<td >Sigma</td>
			<td >P1847</td>
			<td></td>
		</tr>
		<tr >
			<td >EGTA&#160;</td>
			<td >Sigma</td>
			<td >E3889</td>
			<td></td>
		</tr>
		<tr >
			<td >HEPES&#160;</td>
			<td >Sigma</td>
			<td >H4034</td>
			<td></td>
		</tr>
		<tr >
			<td >NaCl</td>
			<td >Sigma</td>
			<td >S9888</td>
			<td></td>
		</tr>
		<tr >
			<td >Agar</td>
			<td>Sigma</td>
			<td>A9799</td>
			<td></td>
		</tr>
		<tr >
			<td >QX-314</td>
			<td >Alomone</td>
			<td>Q150</td>
			<td></td>
		</tr>
		<tr >
			<td >Mg-ATP</td>
			<td>Sigma</td>
			<td >A9187</td>
			<td></td>
		</tr>
		<tr >
			<td >CsOH</td>
			<td>Sigma</td>
			<td >232041</td>
			<td></td>
		</tr>
		<tr >
			<td >Na-GTP</td>
			<td>Sigma</td>
			<td >51120</td>
			<td></td>
		</tr>
		<tr >
			<td >gluconic acid</td>
			<td>Sigma</td>
			<td >G1951</td>
			<td></td>
		</tr>
		<tr >
			<td >Cesium hydroxide solution</td>
			<td>Sigma</td>
			<td >232041</td>
			<td></td>
		</tr>
		<tr >
			<td >KOH</td>
			<td>Sigma</td>
			<td >P5958</td>
			<td></td>
		</tr>
		<tr >
			<td >Vannas Spring Scissors - 2.5mm&#160;</td>
			<td >FST</td>
			<td>15000-08</td>
			<td>only use for cutting the dura, might get damaged if cutting bones</td>
		</tr>
		<tr >
			<td >Stimulator</td>
			<td>A-M Systems</td>
			<td colspan="2">Isolated Pulse Stimulator Model 2100</td>
		</tr>
		<tr >
			<td >Vibratome</td>
			<td >Campden</td>
			<td >Vibrating Microtome 7000 - Model 7000smz-2</td>
			<td></td>
		</tr>
	</tbody>
</table>

the preparation of oblique spinal cord slices for ventral root stimulation

omurilik dilimleri gelen elektrofizyolojik kayıtlar ağ özelliklerine hücresel soruları geniş bir yelpazede araştırmak için değerli bir tekniktir olduğu kanıtlanmıştır. Biz genç farelerde (- P11 P2) spinal kord canlı eğik dilim nasıl hazırlanacağını göstermektedir. Bu hazırlıkta, motor nöronlar aksonları omuriliğin ventral köklerin çıkan korur. Bu akson uyarılması omurilik içinde motornöron Somas ve heyecan verici motor nöron teminatlar istila geri yayılan aksiyon potansiyelleri ortaya çıkarır. antidromik aksiyon potansiyellerinin kaydedilmesi diğer kimlik yöntemlerini aşan motonöron kimliğini, karakterize ivedi, kesin ve zarif bir yoldur. Ayrıca, motor nöron teminatları uyarıcı gibi diğer motor nöronlar veya R omurilik içindeki motor nöronlar teminat hedeflerini, heyecanlandırmak için basit ve güvenilir bir yoldurhücreleri enshaw. Bu protokol, ventral kök uyarılmasından kaynaklanan, motor nöron Somas olarak Renshaw hücre uyarma gelen antidromik kayıtları sunuyoruz.

Antidromic Eylem Potansiyelleri Kullanarak Motonöron Kimlik Onayı Hücre hedefleme Motor nöronlar Karın boynuzuna (Şekil 2C kırmızı görünür) bulunur. ventral kök oluşturan akson demeti başlayın ve paket tamamen dağılır ve bir (20 mikron üzerinde, uzun soma ekseni) büyük hücreler görmeye başlaya...

Watch this Scientific Journal Video about The Preparation of Oblique Spinal Cord Slices for Ventral Root Stimulation at JoVE.com

The Preparation of Oblique Spinal Cord Slices for Ventral Root Stimulation

Biz genç farelerde omurilik eğik dilim nasıl hazırlanacağını göstermektedir. Bu preparat, ventral köklerin uyarılması için izin verir.

Ventral Kök Stimulation için eğik Omurilik Dilimleri hazırlanması

Electrophysiological recordings from spinal cord slices have proven to be a valuable technique to investigate a wide range of questions, from cellular to ...

the-preparation-oblique-spinal-cord-slices-for-ventral-root

Research

JoVE Journal

Neuroscience

9.4K Görüntüleme.  Centre de Neurophysique, Physiologie et Pathologie, Université Paris Descartes.  Biz genç farelerde omurilik eğik dilim nasıl hazırlanacağını göstermektedir. Bu preparat, ventral köklerin uyarılması için izin verir. 

Video: The Preparation of Oblique Spinal Cord Slices for Ventral Root Stimulation

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 ...

Nörobilim

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

To analyze the axonal and dendritic morphology of neurons, it is essential to obtain accurate labeling of neuronal structures. Preparing well labeled samples with little to no tissue damage enables us to analyze cell morphology and to compare individual samples to each other, hence allowing the identification of mutant anomalies.
In the demonstrated dissection method the nervous system remains mostly inside the adult fly. Through a dorsal incision, the abdomen and thorax are opened and most of the internal organs are removed. Only the dorsal side of the ventral nerve cord (VNC) and the cervical connective 
(CvC) containing the big axons of the giant fibers (GFs)1 are exposed, while the brain containing the GF cell body and dendrites remains2 in the 
intact head. In this preparation most nerves of the VNC should remain attached to their muscles. 
Following the dissection, the intracellular filling of the giant fiber (GF) with a fluorescent dye is demonstrated. In the CvC the GF axons are located at the dorsal surface and thus can be easily visualized under a microscope with differential interference contrast (DIC) optics. This allows the injection of the GF axons with dye at this site to label the entire GF including the axons and their terminals in the VNC. This method results in reliable and strong staining of the GFs allowing the neurons to be imaged immediately after filling with an epifluorescent microscope. Alternatively, the fluorescent signal can be enhanced using standard immunohistochemistry procedures3 suitable for high resolution confocal microscopy.

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

An in vivo dissection of the adult Drosophila ventral nerve cord (VNC) is demonstrated. This particular dissection method causes little damage to the VNC allowing the subsequent labeling of the giant fiber neurons with fluorescent dye for high resolution imaging.

Yetişkin Tüm Dağı Hazırlık Drosophila Dev Fiber Boya Enjeksiyon Ventral Sinir Kablosu

To analyze the axonal and dendritic morphology of neurons, it is essential to obtain accurate labeling of neuronal structures. Preparing well labeled ...

Preparation of Parasagittal Slices for the Investigation of Dorsal-ventral Organization of the Rodent Medial Entorhinal Cortex

Computation in the brain relies on neurons responding appropriately to their synaptic inputs. Neurons differ in their complement and distribution of membrane ion channels that determine how they respond to synaptic inputs. However, the relationship between these cellular properties and neuronal function in behaving animals is not well understood. One approach to this problem is to investigate topographically organized neural circuits in which the position of individual neurons maps onto information they encode or computations they carry out1. Experiments using this approach suggest principles for tuning of synaptic responses underlying information encoding in sensory and cognitive circuits2,3.
The topographical organization of spatial representations along the dorsal-ventral axis of the medial entorhinal cortex (MEC) provides an opportunity to establish relationships between cellular mechanisms and computations important for spatial cognition. Neurons in layer II of the rodent MEC encode location using grid-like firing fields4-6. For neurons found at dorsal positions in the MEC the distance between the individual firing fields that form a grid is on the order of 30 cm, whereas for neurons at progressively more ventral positions this distance increases to greater than 1 m. Several studies have revealed cellular properties of neurons in layer II of the MEC that, like the spacing between grid firing fields, also differ according to their dorsal-ventral position, suggesting that these cellular properties are important for spatial computation2,7-10.
Here we describe procedures for preparation and electrophysiological recording from brain slices that maintain the dorsal-ventral extent of the MEC enabling investigation of the topographical organization of biophysical and anatomical properties of MEC neurons. The dorsal-ventral position of identified neurons relative to anatomical landmarks is difficult to establish accurately with protocols that use horizontal slices of MEC7,8,11,12, as it is difficult to establish reference points for the exact dorsal-ventral location of the slice. The procedures we describe enable accurate and consistent measurement of location of recorded cells along the dorsal-ventral axis of the MEC as well as visualization of molecular gradients2,10. The procedures have been developed for use with adult mice (&gt; 28 days) and have been successfully employed with mice up to 1.5 years old. With adjustments they could be used with younger mice or other rodent species. A standardized system of preparation and measurement will aid systematic investigation of the cellular and microcircuit properties of this area.

We describe procedures for preparation and electrophysiological recording from brain slices that maintain the dorsal-ventral axis of the medial entorhinal cortex (MEC). Because neural encoding of location follows a dorsal-ventral organization within the MEC, these procedures facilitate investigation of cellular mechanisms important for navigation and memory.

Kemirgen Medial entorinal Korteksin dorsal-ventral Örgütü Soruşturma parasagital Dilimleri hazırlanması

Computation in the brain relies on neurons responding appropriately to their synaptic inputs. Neurons differ in their complement and distribution of ...

Spinal Cord Transection in the Larval Zebrafish

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability to reinitiate spinal neurogenesis. However, some anamniotes including the zebrafish Danio rerio exhibit both sensory and functional recovery even after complete transection of the spinal cord. The adult zebrafish is an established model organism for studying regeneration following spinal cord injury, with sensory and motor recovery by 6&#160;weeks post-injury. To take advantage of in vivo analysis of the regenerative process available in the transparent larval zebrafish as well as genetic tools not accessible in the adult, we use the larval zebrafish to study regeneration after spinal cord transection. Here we demonstrate a method for reproducibly and verifiably transecting the larval spinal cord. After transection, our data shows sensory recovery beginning at 2&#160;days post-injury (dpi), with the C-bend movement detectable by 3 dpi and resumption of free swimming by 5 dpi. Thus we propose the larval zebrafish as a companion tool to the adult zebrafish for the study of recovery after spinal cord injury.

After spinal transection, adult zebrafish have functional recovery by six weeks post-injury. To take advantage of larval transparency and faster recovery, we present a method for transecting the larval spinal cord. After transection, we observe sensory recovery beginning at 2&#160;days post-injury, and C-bend movement by 3 days post-injury.

Larva Zebra balığı Omurilik Kesisi

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability ...

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

Boyuna için bir Spinal Odası implantasyon için bir Prosedürü<em&gt; İn Vivo</em&gt; Fare Omurilik Görüntüleme

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Preparation of Primary Mixed Glial Cultures from Adult Mouse Spinal Cord Tissue

It has been well-accepted that spinal cord glial responses contribute significantly to the development of neuropathic pain. Tremendous information regarding glial activities at the cellular and molecular levels has been obtained through in vitro cell culture systems. The in vitro systems utilized, mainly include primary glia derived from neonatal brain cortical tissue and immortalized cell lines. However, these systems may not reflect the characteristics of spinal cord glial cells in vivo. In order to further investigate the roles of spinal cord glial cells in the development of peripheral nerve injury-induced neuropathic pain using a culture system that better reflects the in vivo condition, our laboratory has developed a method to establish primary spinal cord mixed glial cultures from adult mice. Briefly, spinal cords are collected from adult mice and processed through papain digestion followed by myelin removal with a density-gradient medium. Single cell suspensions are cultured in complete Dulbecco&#39;s modified Eagle media (cDMEM) supplemented with 2-mercaptoethanol (2-ME) at 35.9 oC. These culture conditions were optimized specifically for the growth of mixed glial cells. Under these conditions, cells are ready to be used for experimentation between 12 - 14 d&#160;(cells are usually in log phase during this time) after the establishment of the culture (D 0) and can be kept in culture conditions up to D 21. This culture system can be used to investigate the responses of spinal cord glial cells upon stimulation with various substances and agents. Besides neuropathic pain, this system can be used to study glial responses in other diseases that involve pathological changes of spinal cord glial cells.

The development of neuropathic pain involves pathological changes of spinal cord glial cells. A reliable glial culture system derived from adult spinal cord tissue and designed for studying these cells in vitro is lacking. Therefore, we show here how&#160;to establish primary mixed glial cultures from adult mouse spinal cord tissue.

Yetişkin Fare Omurilik Doku İlköğretim Karışık glial Kültürler hazırlanması

It has been well-accepted that spinal cord glial responses contribute significantly to the development of neuropathic pain. Tremendous information ...

Hydraulic Extrusion of the Spinal Cord and Isolation of Dorsal Root Ganglia in Rodents

Traditionally, the spinal cord is isolated by laminectomy, i.e. by breaking open the spinal vertebrae one at a time. This is both time consuming and may result in damage to the spinal cord caused by the dissection process. Here, we show how the spinal cord can be extruded using hydraulic pressure. Handling time is significantly reduced to only a few minutes, likely decreasing protein damage. The low risk of damage to the spinal cord tissue improves subsequent immunohistochemical analysis. By performing hydraulic spinal cord extrusion instead of traditional laminectomy, the rodents can further be used for DRG isolation, thereby lowering the number of animals and allowing analysis across tissues from the same rodent. We demonstrate a consistent method to identify and isolate the DRGs according to their localization relative to the costae. It is, however, important to adjust this method to the particular animal used, as the number of spinal cord segments, both thoracic and lumbar, may vary according to animal type and strain. In addition, we illustrate further processing examples of the isolated tissues.

Here, we present a protocol for hydraulic extrusion of the spinal cord as well as identification and isolation of specific dorsal root ganglia (DRGs) in the same rodent. Compared to standard spinal cord isolation methods, this method is significantly faster and reduces the risk of tissue damage.

Kemirgenler dorsal kök gangliyon Omurilik ve İzolasyon hidrolik eldesi

Traditionally, the spinal cord is isolated by laminectomy, i.e. by breaking open the spinal vertebrae one at a time. This is both time consuming and may ...

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

Zebrafish, first introduced as a developmental model, have gained popularity in many other fields. The ease of rearing large numbers of rapidly developing organisms, combined with the embryonic optical clarity, served as initial compelling attributes of this model. Over the past two decades, the success of this model has been further propelled by its amenability to large-scale mutagenesis screens and by the ease of transgenesis. More recently, gene-editing approaches have extended the power of the model.
For neurodevelopmental studies, the zebrafish embryo and larva provide a model to which multiple methods can be applied. Here, we focus on methods that allow the study of an essential property of neurons, electrical excitability. Our preparation for the electrophysiological study of zebrafish spinal neurons involves the use of veterinarian suture glue to secure the preparation to a recording chamber. Alternative methods for recording from zebrafish embryos and larvae involve the attachment of the preparation to the chamber using a fine tungsten pin1,2,3,4,5. A tungsten pin is most often used to mount the preparation in a lateral orientation, although it has been used to mount larvae dorsal-side up4. The suture glue has been used to mount embryos and larvae in both orientations. Using the glue, a minimal dissection can be performed, allowing access to spinal neurons without the use of an enzymatic treatment, thereby avoiding any resultant damage. However, for larvae, it is necessary to apply a brief enzyme treatment to remove the muscle tissue surrounding the spinal cord. The methods described here have been used to study the intrinsic electrical properties of motor neurons, interneurons, and sensory neurons at several developmental stages6,7,8,9.

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

This manuscript describes methods for electrophysiological recordings from spinal neurons of zebrafish embryos and larvae. The preparation maintains neurons in situ and often involves minimum dissection. These methods allow for the electrophysiological study of a variety of spinal neurons, from the initial electrical excitability acquisition through the early larval stages.

Zebra balığı<em&gt; Situ</em&gt; Omurilik Omurilik duyusal ve motor nöronlar gelen Elektrofizyolojik Recordings Hazırlık

Zebrafish, first introduced as a developmental model, have gained popularity in many other fields. The ease of rearing large numbers of rapidly developing ...

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in adult mammalian animals remains elusive. In this study, we describe an experimental procedure for measuring calcium dynamics through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy. This method enables recordings and quantifications of neural activity in bilateral mouse brain regions one at a time in the same experiment for a prolonged period in vivo. Key aspects of this method, which can be completed within an hour, include minimally invasive surgery procedures for creating dual optical windows, and the use of 2P imaging. Although we only demonstrate the technique in the S1 area, the method can be applied to other regions of the living brain facilitating the elucidation of structural and functional complexities of brain neural networks.

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo.

Sinirsel aktivite Thy1kullanarak birincil somatosensor kortekste Imaging-GCaMP6s transgenik fareler

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...

Preparation of Acute Spinal Cord Slices for Whole-cell Patch-clamp Recording in Substantia Gelatinosa Neurons

Recent whole-cell patch-clamp studies from substantia gelatinosa (SG) neurons have provided a large body of information about the spinal mechanisms underlying sensory transmission, nociceptive regulation, and chronic pain or itch development. Implementations of electrophysiological recordings together with morphological studies based on the utility of acute spinal cord slices have further improved our understanding of neuronal properties and the composition of local circuitry in SG. Here, we present a detailed and practical guide for the preparation of spinal cord slices and show representative whole-cell recording and morphological results. This protocol permits ideal neuronal preservation and can mimic in vivo conditions to a certain extent. In summary, the ability to obtain an in vitro preparation of spinal cord slices enables stable current- and voltage-clamp recordings and could thus facilitate detailed investigations into the intrinsic membrane properties, local circuitry and neuronal structure using diverse experimental approaches.

Here, we describe the essential steps for whole-cell patch-clamp recordings made from substantia gelatinosa (SG) neurons in the in vitro spinal cord slice. This method allows the intrinsic membrane properties, synaptic transmission and morphological characterization of SG neurons to be studied.

Akut omurilik dilimleri hazırlanması gözlemliyorum Gelatinosa nöronlar bütün hücreli yama-kelepçe kayıt için

Recent whole-cell patch-clamp studies from substantia gelatinosa (SG) neurons have provided a large body of information about the spinal mechanisms ...

Ventral Kök Stimulation için eğik Omurilik Dilimleri hazırlanması

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