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Hayvanlar, Güney Danimarka Üniversitesi hayvan bakım tesisi kurumsal yönergelere göre yer. Tüm hayvan deneyleri Ulusal Sağlık Enstitüleri Kılavuzu bakım ve laboratuvar hayvanlarının kullanımı için uygun olarak gerçekleştirilmiştir. Bu akut, Sigara survival cerrahi bir işlemdir.
1. hazırlanması boru, elektrot ve cerrahi işlem için aygıtlar
<ol><li>Mikrocerrahi cihazlar kullanın: Cerrahi makas, açılı mikro makas, doku forseps, Dumont #7 çifti ve #5 eğri mikro forseps, mikro iğne bağı ve traktör. Not: Bu akut bir yordam olarak, aletleri sterilize gerek yoktur. Kullanın sonra temiz ve ipuçları % 70 etanol ile silin.</li>
	<li>Tüp % 70 etanol içinde ıslatın ve sonra steril %0,9 ile flush NaCl ile 100 U/mL heparin kullanmadan önce. Hava kabarcıkları içine belgili tanımlık sistem tanıtımı önlemek için dolu boru bırakın.</li>
	<li>Polietilen (PE)-50 boru bir kateter ICP ölçüm için yapmak için bir 20-30 cm uzunluk parça kesti. Tüp basıncı nemlendirme azaltmak olabildiğince kısa olduğundan emin olun.</li>
	<li>İğne ortada tatili kadar bir steril 24 G iğne yan-yan viraj. Parça eğimi PE boru penis crus içine eklenmek distal ucuna bağlayın. Diğer yarısı bağlantı için basınç detektörler hub'ına ekleyin. Sistem heparinized serum (100 U/mL) doldurun.</li>
	<li>İki kutuplu Teflon kaplı elektrot yapmak için eşit uzunlukta kesilmiş iki 125 µm gümüş teller. Masanın kenarına teller takın ve onları birlikte iş parçacığı için bir parça bant kullanın. Daha sonra elektrot bir siyah plaka ekleyin.</li>
	<li>Teflon içine küçük bir kesi yapmak ve Teflon kaplama elektrot uçları kapalı 4-5 mm uzunluğunda şerit için #5 mikro forseps kullanın. İpuçları bile uzunluğu ulaşmak ve uçları künt kenarında bir neşter bıçak bükme tarafından kanca oluşturmak için bir neşter ile kesti.</li>
	<li>Biraz yukarı doğru işaret kanca ile siyah plaka kenarına uzanan ucuyla elektrot teyp. 10 s ve şal bir tutkal kabarcık elektrot kanca 1-2 mm çapında için plastik tabakta silikon yapıştırıcı karışımı.</li>
	<li>Yaklaşık 5 dakika kullanım (şekil 1) önce kurumasını bekleyin. Şerit Stimülatörü bağlantı için izin vermek için Teflon diğer ucunda, daha uzun bir bölümünden. Not: Distal ucunda kanca yeniden yapıyor ile elektrot birçok kez yeniden kullanılabilir.</li>
</ol>2. hayvan hazırlanması
<ol><li>Hayvan anesthetizing sonra karın, boyun ve perine alt yarısının tıraş. Hayvan povidone-iyot tarafından üç kez takip % 70 alkol ile fırçalayın. Fareyi bir sırtüstü pozisyonda bir ısıtmalı cerrahi pedi yerleştirin. Veteriner göz merhem uygulamak ve % 2.5 isoflurane ve 0, 8 L/dak oksijen taşıyıcı olarak ile burun konisi için anestezi geçmek. Not: İsoflurane ve anestezi kabul edilebilir bir düzeyde sağlamak için oksijen düzeyini ayarlayın.</li>
</ol>3. presurgical hazırlık
<ol><li>Çalışma bir mikroskop altında tüm cerrahi işlem gerçekleştirmek: 3,15 X 20 X arasında değişen bir büyütme yeterlidir. Kullanım eldiven ve ameliyat boyunca temiz bir ortam sağlamak. Fareyi bir örtüsü yerleştirin.</li>
</ol>4. Ischiocavernosus kas diseksiyon ICP ölçümü için
<ol><li>Bir neşter, düz makas kullanın ve Dumont #7 kavisli 1 cm dikey yapmak için mikro forseps cilt kesi 5 mm lateral penis tabanı düzeyinde başlayan ve aşağıya doğru (şekil 2A) uzanan orta hat için. Bir pamuklu çubukla kullanın ve fasya lateral skrotum (şekil 2B) için dikkatlice ayırın. Fasya anatomi sonra makaralara eklemek ve bir pamuk-uçlu-kural yumrunun (şekil 2C) bulmak için takas ile muayene et.</li>
	<li>İschiocavernosus kas görüntülenir kadar bu noktada yağ dokusu medial teşrih (şekil 3A). Dumont #7 eğri mikro forseps bir çift kullanın ve boyuna kas ayrı. Tunica albuginea parlak beyaz yapı (şekil 3B) olarak görünür. Mikro forseps ve mikro makas kullanarak, yeterince onun ders (şekil 3 c) görmek için tunica albuginea ortaya çıkarmak.</li>
	<li>Sistem ayarlarını kalibrasyon sonra tüp penis crus (şekil 4) paralel gidiyor emin perine üzerinde deri yoluyla yerleştirin. Çizginin yerinde bırakmak ve belgili tanımlık kesme serum fizyolojik ile nemli tutmak.</li>
</ol>5. CN diseksiyon uyarılması için
<ol><li>2 cm daha düşük, karın Ensizyon, ilk olarak, bir neşter ve bir çift düz makas ve mikro forseps kullanarak deri yoluyla olun. Linea boyunca fasya ile eşleşen bir giriş oluşturmak alba ve temel kas doku mesane ve prostat duyurmak.</li>
	<li>Çok iyi bir pozlama elde etmek için kullanın. Prostat açık görsel olarak büyük pelvik sinir düğümü (MPG) elde etmek için yağ dokusundan ayırmak için pamuk-uç temizleme bezi kullanın ve CN, prostat (şekil 5) dorsolateral yönü üzerinde çalışan.</li>
	<li>Sonra görselleştirme MPG ve CN, dikkatli bir şekilde sinir 2-5 mm için MPG distal açılı mikro makasla (şekil 6a) örten fasya deşmek. #5 mikro forseps kullanımı ile her iki tarafta bir sinir ve bir 4-mm uzun bölümü (şekil 6b) ücretsiz için altında doku Yayılı ve sinir (şekil 6 c) altında 9-0 dikiş kaydırın.</li>
	<li>Sinir sinir (şekil 7b) çevresinde İki kutuplu elektrot kanca yerleşimini kolaylaştırmak için dikiş (şekil 7a) yardımıyla hafifçe yükseltmek. Sinir için 5 s. kuru iğne ve Tutkal kanca ve sinir (Şekil 7 c, d) çevresi için geçerli bir asistan karışım iki bileşenli silikon yapıştırıcı bir insülin ucu ile izin. Biraz kuru tutkal izin vermek yaklaşık 1 dakika için elektrot çekerek sinir yukarıda tut.</li>
	<li>Dışında herhangi bir çekmek veya elektrot büküm önlemek için sağ tarafta Traktör Traktör kaldırın. Maruz kalan organların serum içinde belgili tanımlık kesme üzerinde batırılmış tuzlu ve yatıyordu gazlı bez ile ıslak.</li>
</ol>6. kavernöz vücut Cannulation ICP ölçümü için
<ol><li>Ekartör kullanarak tunica albuginea görselleştirme geri. Crus tahrif edecek gibi traktör ischiocavernosus kas değil eklemek emin olun.</li>
	<li>İğne takın ve tunica albuginea tanıtımı önce boru heparinized serum fizyolojik ile yıkayın. Gergin, bir elinde Dumont #7 eğri mikro forseps kullanarak tunica albuginea (non-baskın), tunica albuginea ve üstteki kas geri kalanı ekleme noktasına distal basılı tutmak. İğneyi diğer baskın elinde düz mikro Forseps ile sık ve onu tanıtmak kavernöz vücut (şekil 8) ders ile paralel.</li>
	<li>5-8 mm iğne kavernöz vücudun içine itin. Tüp Temizleme ve crus üzerinde yolun (Şekil 9) test etmek için tuşuna basın. Hiçbir sızıntı olduğundan emin olun. Hatta yanlışlıkla çekerek önlemek için bant ile tabloya boru vidaları sıkıştırın. Ekartör kaldırın.</li>
</ol>7. CN uyarılması
<ol><li>Kayıt programı (örneğin, Spike 2) ile çalışan, sürekli olarak her iki intracavernous ve ortalama arter basıncı kaydetmek.</li>
	<li>Bir uyarıcı (örneğin, SD9 çim aletleri, bkz: Malzemeler tablo) CN uyarılması için aşağıdaki parametreleri ayarlayın: 1,5 mA, frekans 16 Hz, 3 V gerilim ve darbe genişliği 1 dk stimülasyon en az 5 Bayan uygulamak 50 s, geçerli Elektrodlar arasında bekletin. Not: İlk elektrodlar genellikle azalmış yanıtın (şekil 10) yol.</li>
</ol>

<ol>
	<li>Eckhard, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Untersuchungen &#252;ber die Erektion des Hundes In: Beitr&#228;ge zur Anatomie und Physiologie. , 123-166 (1863).</li><li>Quinlan, D. M., Nelson, R. J., Partin, A. W., Mostwin, J. L., Walsh, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+rat+as+a+model+for+the+study+of+penile+erection.">The rat as a model for the study of penile erection.</a> J Urol. 141 (3), 656-661 (1989).</li><li>Heaton, J. P., Varrin, S. J., Morales, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+characterization+of+a+bio-assay+of+erectile+function+in+a+rat+model.">The characterization of a bio-assay of erectile function in a rat model.</a> J Urol. 145 (5), 1099-1102 (1991).</li><li>Martinez-Pineiro, L., 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=Rat+model+for+the+study+of+penile+erection:+pharmacologic+and+electrical-stimulation+parameters.">Rat model for the study of penile erection: pharmacologic and electrical-stimulation parameters.</a> Eur Urol. 25 (1), 62-70 (1994).</li><li>Hayashi, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+FK1706+on+erectile+function+following+bilateral+cavernous+nerve+crush+injury+in+a+rat+model.">The effect of FK1706 on erectile function following bilateral cavernous nerve crush injury in a rat model.</a> J Urol. 176 (2), 824-829 (2006).</li><li>Burnett, A. L., Becker, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunophilin+ligands+promote+penile+neurogenesis+and+erection+recovery+after+cavernous+nerve+injury.">Immunophilin ligands promote penile neurogenesis and erection recovery after cavernous nerve injury.</a> J Urol. 171 (1), 495-500 (2004).</li><li>Yamashita, 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=Nerve+injury-related+erectile+dysfunction+following+nerve-sparing+radical+prostatectomy:+a+novel+experimental+dissection+model.">Nerve injury-related erectile dysfunction following nerve-sparing radical prostatectomy: a novel experimental dissection model.</a> Int J Urol. 16 (11), 905-911 (2009).</li><li>Burnett, A. L., 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=GGF2+is+neuroprotective+in+a+rat+model+of+cavernous+nerve+injury-induced+erectile+dysfunction.">GGF2 is neuroprotective in a rat model of cavernous nerve injury-induced erectile dysfunction.</a> J Sex Med. 12 (4), 897-905 (2015).</li><li>Lin, H., 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=Nanoparticle+Improved+Stem+Cell+Therapy+for+Erectile+Dysfunction+in+a+Rat+Model+of+Cavernous+Nerve+Injury.">Nanoparticle Improved Stem Cell Therapy for Erectile Dysfunction in a Rat Model of Cavernous Nerve Injury.</a> J Urol. 195 (3), 788-795 (2016).</li><li>Kapoor, M. S., Khan, S. A., Gupta, S. K., Choudhary, R., Bodakhe, S. 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Yazarlar ifşa gerek yok.

Penis crus cannulation ICP kayıt için basitleştirilmiş bir cerrahi teknik ve yalıtım CN elektrostimülasyon için açıklamak için bu çalışmanın asıl hedefi oldu. Biz ameliyat basitleştirmek ve CN stimülasyon ile tekrarlanabilir kayıtları ICP artış sağlamak kavernöz vücut diseksiyon değişiklikler tanıttı. 1 cm dikey cilt kesi ile aşikar kural yumrunun kılavuz kullanarak penis tabanına yanal ischiocavernosus kas ve tunica albuginea iyi bir pozlama elde. Bu yordamı daha hızlıdır (küçük 15...

Vivo çalışmada deneysel hayvanlarda erektil fonksiyonunun 1863 Eckhard1öncü deneysel çalışmaları ile başladı. Elektrostimülasyon pelvik sinir köpeklerde artan ICP ikna etmek için kullanıldı. 20 yy benzer deneysel protokoller kediler ve tavşan, köpek, maymun gibi büyük hayvanlar kullanılmaya başlanmıştır. Bir sıçan erektil işlev değerlendirmek ilk Quinlan ve ark. 19892tarafından geliştirilmiştir. Yöntem beri modifiye edilmiş ve birkaç diğer gruplar34tarafından güncelleştirildi. Bugün, en yaygın kullanılan hayvan modeli patoloji erektil disfonksiyon eğitim ve gelişmekte olan tedavi seçenekleri değerlendirmek için faredir. Sistemik kan basıncı ICP bir artış ikna etmek için Şah damarı, ölçmek ICP penis crus cannulation ve CN uyarılması bir satırını kullanarak kayıt yordamı ana adımları içerir. Çeşitli araştırmacılar model iyileştirdiyseniz rağmen onun tekrarlanabilirlik bir sorun olmaya devam etmektedir ve değişken sonuçları farklı Laboratuvarları tarafından bildirilmiştir. Çeşitli tuzaklar, devam.
Önceki Makaleler5,6,7,8,9,10 penis kavernöz vücut cannulation için degloving ile tam penis pozlama kullanımını açıklar. Bu manipülasyon olarak en iyi bir yaklaşım değildir ve yıkıcı diseksiyon erektil işlev için gerekli olan yapılar için yaralanma neden olur. CN diseksiyon de açıklanan10,11oldu, ama sinir stimülasyonu deneysel sonuçları etkileyebilecek Multipl faktörler nedeniyle en uygun değil. CN stimülasyon tekniği çevreleyen doku sinir sinir konumlandırılmış, bipolar kanca elektrodu, çekerek ve her stimülasyon önce sinir kurutma tarafından kaldırma içerir. Bu sinir hasarı ve elektrik geçerli sızıntı, azalmış yanıt veya ICP alternatif yolları örneğin, Pelvik Taban kasları, mesane uyarılması yoluyla yanlış artış elde edilen ve gastrointestinal değişik derecelerde açabilir yolu12. Tüm bu faktörlerin tekrarlanabilirlik sınırlayın.
Bizim çalışma sırasında derinlik ve anestezi türünü ICP üzerinde büyük bir etkiye sahip görülmektedir. Anestezikler kullanılan sodyum Fentobarbital, ketamin/xylazine veya ketamin/midazolam enjeksiyon veya isoflurane/oksijen zehirlenmesi olabilir.
Burada biz basitleştirilmiş bir cerrahi yöntem tanımlamak ve veri deneysel protokol standardizasyon destek sağlamak.

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		<tr height="21">
			<td height="21" style="height:21px;width:257px;">NaCl 0,9 % 100 mL</td>
			<td style="width:199px;">Local pharmacy</td>
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			<td height="21" style="height:21px;width:257px;">Heparin</td>
			<td style="width:199px;">Local pharmacy</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">25 mL Syringe</td>
			<td style="width:199px;">Odense University Hospital</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Vet eye ointment - Viscotears</td>
			<td style="width:199px;">Local pharmacy</td>
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			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">silver wires&#160;</td>
			<td colspan="2">Science Products GmbH, Heidelberg, Germany</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Silicon Glue&#160;</td>
			<td colspan="3">Kwik-Sil, World Precision Instruments, Sarasota, FL, USA</td>
		</tr>
	</tbody>
</table>

cavernous nerve stimulation and recording of intracavernous pressure in a rat

Kavernöz sinir (CN) uyarılması ve intracavernous basınç (ICP) ölçümü kullanılan kapsamlı bir şekilde test etmek ve değerlendirmek için erektil disfonksiyon terapiler. Ancak, kullanılan yöntemleri laboratuvarlar arasında değişir ve tuzaklar hareketsiz var olmak. Bu çalışmanın amacı bir güvenilir ve tekrarlanabilir model sağlayacak bir cerrahi teknik açıklamak için oldu. İschiocavernosus kas, kural yumrunun tarihinde ekleme noktasında teşhir ederek penis crus en az diseksiyon ve yapıları erektil işlev ilgili yaralanma ile cannulated. Kaldırma ve kurutma, gerek kalmadan CN tekrarlanan uyarılması elektrot-sinir kompleksi yalıtmak için bir 125 µm manik depresif gümüş elektrot ve Biyouyumlu silikon yapıştırıcı kullanarak sağlanır. Bu yöntem germe ve sinir kurutma azaltarak neuropraxia engeller ve elektrik kaçağı inkâr ve alternatif yollar uyarılması önleme sinirin tam izolasyon sağlar.

Kullanılması önerilen stimülasyon ayarları bu protokolle inhalasyon anestezi isoflurane %2.0 oksijen ile altında 0, 8 L/dak, üretmek sonuçları Şekil 11 ve şekil12', gösterildiği gibi olmadığı bir yerde birden fazla arka arkaya Elektrodlar arasında 75-80 mm Hg. şekil 13 gösterir aynı kararlı yanıt 20 dk stimülasyon yanıtı ile üzerinde istikrarlı 73-77 mm Hg. Test hattı ...

Watch this Scientific Journal Video about Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat at JoVE.com

Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

Bu çalışmada bir basitleştirilmiş cerrahi müdahale ve kavernöz sinir stimülasyonu sinir-elektrot yalıtım ile gerçekleştirmek için teknik açıklar karmaşık silikon yapıştırıcı ve intracavernous basınç ölçüm kullanarak.

Kavernöz sinir stimülasyonu ve kayıt bir sıçan Intracavernous basınç

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for ...

cavernous-nerve-stimulation-recording-intracavernous-pressure

Research

JoVE Journal

Medicine

12.4K Görüntüleme.  University of Southern Denmark. Bu çalışmada bir basitleştirilmiş cerrahi müdahale ve kavernöz sinir stimülasyonu sinir-elektrot yalıtım ile gerçekleştirmek için teknik açıklar karmaşık silikon yapıştırıcı ve intracavernous basınç ölçüm kullanarak.

Video: Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and cardiac arrest. This elevation may result in further neurological injury, in the form of transtentorial herniation1,2,3,4, midbrain compression, neurological deficit or increased cerebral infarct2,4. Current therapies are often inadequate to control elevated ICP in the clinical setting5,6,7 . Thus there is a need for accurate methods of ICP measurement in animal models to further our understanding of the basic mechanisms and to develop new treatments for elevated ICP.
In both the clinical and experimental setting ICP cannot be estimated without direct measurement. Several methods of ICP catheter insertion currently exist. Of these the intraventricular catheter has become the clinical 'gold standard' of ICP measurement in humans8. This method involves the partial removal of skull and the instrumentation of the catheter through brain tissue. Consequently, intraventricular catheters have an infection rate of 6-11%9. For this reason, subdural and epidural cannulations have become the preferred methods in animal models of ischemic injury. 
Various ICP measurement techniques have been adapted for animal models, and of these, fluid-filled telemetry catheters10 and solid state catheters are the most frequently used11,12,13,14,15. The fluid-filled systems are prone to developing air bubbles in the line, resulting in false ICP readings. Solid state probes avoid this problem (Figure 1). An additional problem is fitting catheters under the skull or into the ventricles without causing any brain injury that might alter the experimental outcomes. Therefore, we have developed a method that places an ICP catheter contiguous with the epidural space, but avoids the need to insert it between skull and brain. 
An optic fibre pressure catheter (420LP, SAMBA Sensors, Sweden) was used to measure ICP at the epidural location because the location of the pressure sensor (at the very tip of the catheter) was found to produce a high fidelity ICP signal in this model. There are other manufacturers of similar optic fibre technologies13 that may be used with our methodology. Alternative solid state catheters, which have the pressure sensor located at the side of the catheter tip, would not be appropriate for this model as the signal would be dampened by the presence of the monitoring screw. 
Here, we present a relatively simple and accurate method to measure ICP. This method can be used across a wide range of ICP related animal models.

A novel technique to record the pressures within the skull is described. The minimally invasive method uses a fibre-optic pressure sensing system to accurately measure intracranial pressure (ICP) in anaesthetized rats without causing significant brain trauma. The technique may be used in a wide range of experimental models.

Bir Fiber optik Basıç kullanarak Sıçanlarda Epidural İntrakraniyal Basınç Ölçümü

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

Tıp

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As such, primary or &#39;neuronal-like&#39; cell culture systems are commonly utilized. While&#160;these systems are relatively easy to work with, and are useful model systems in which various functional outcomes (such as cell death) can be readily quantified, the examined outcomes and pathways in cultured immature neurons (such as excitotoxicity-mediated cell death pathways) are not necessarily the same as those observed in mature brain, or in intact tissue. Therefore, there is the need to develop models in which cellular mechanisms in mature neural tissue can be examined. We have developed an in vitro technique that can be used to investigate a variety of molecular pathways in intact nervous tissue. The technique described herein utilizes rat cortical tissue, but this technique can be adapted to use tissue from a variety of species (such as mouse, rabbit, guinea pig, and chicken) or brain regions (for example, hippocampus, striatum, etc.). Additionally, a variety of stimulations/treatments can be used (for example, excitotoxic, administration of inhibitors, etc.). In conclusion, the brain slice model described herein can be used to examine a variety of molecular mechanisms involved in excitotoxicity-mediated brain injury.

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

We have developed a brain slice model which can be used to examine molecular mechanisms involved in excitotoxicity-mediated brain injury.&#160;This technique generates viable mature brain tissue and reduces animal numbers required for experimentation, whilst keeping the neuronal circuitry, cellular interactions, and postsynaptic compartments partly intact.

Bir olarak Beyin Microslices arasında excitotoxic uyarılması<em&gt; In vitro</em&gt; İnme Model

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

Rahim içi Telemetri Fare Kontraktil Basıncını Ölçmek<em&gt; İn Vivo</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

A Multimodal Imaging- and Stimulation-based Method of Evaluating Connectivity-related Brain Excitability in Patients with Epilepsy

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations over time in the blood-oxygenation level dependent signal. rs-fcMRI has been applied extensively to identify abnormalities in brain connectivity in different neurologic and psychiatric diseases. However, the relationship among rs-fcMRI connectivity abnormalities, brain electrophysiology and disease state is unknown, in part because the causal significance of alterations in functional connectivity in disease pathophysiology has not been established. Transcranial Magnetic Stimulation (TMS) is a technique that uses electromagnetic induction to noninvasively produce focal changes in cortical activity. When combined with electroencephalography (EEG), TMS can be used to assess the brain's response to external perturbations. Here we provide a protocol for combining rs-fcMRI, TMS and EEG to assess the physiologic significance of alterations in functional connectivity in patients with neuropsychiatric disease. We provide representative results from a previously published study in which rs-fcMRI was used to identify regions with abnormal connectivity in patients with epilepsy due to a malformation of cortical development, periventricular nodular heterotopia (PNH). Stimulation in patients with epilepsy resulted in abnormal TMS-evoked EEG activity relative to stimulation of the same sites in matched healthy control patients, with an abnormal increase in the late component of the TMS-evoked potential, consistent with cortical hyperexcitability. This abnormality was specific to regions with abnormal resting-state functional connectivity. Electrical source analysis in a subject with previously recorded seizures demonstrated that the origin of the abnormal TMS-evoked activity co-localized with the seizure-onset zone, suggesting the presence of an epileptogenic circuit. These results demonstrate how rs-fcMRI, TMS and EEG can be utilized together to identify and understand the physiological significance of abnormal brain connectivity in human diseases.

Resting-state functional-connectivity MRI has identified abnormalities in patients with a wide range of neuropsychiatric disorders, including epilepsy due to malformations of cortical development. Transcranial Magnetic Stimulation in combination with EEG can demonstrate that patients with epilepsy have cortical hyperexcitability in regions with abnormal connectivity.

Epilepsi Hastalarında Beyin uyarılabilmenin Bağlantı ile ilgili değerlendirmek için bir Multimodal imaging- ve Uyarım tabanlı Yöntemi

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations ...

Novel Approach for Simultaneous Recording of Renal Sympathetic Nerve Activity and Blood Pressure with Intravenous Infusion in Conscious, Unrestrained Mice.

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity (RSNA) is of great interest in many areas of research such as chronic kidney disease, hypertension, heart failure, diabetes and obesity. Unequivocal assessment of the role of the sympathetic nervous system is thus imperative for proper interpretation of experimental results and understanding of disease processes. RSNA has been traditionally measured in anesthetized rodents, including mice. However, mice usually exhibit very low systemic blood pressure and hemodynamic instability for several hours during anesthesia and surgery. Meaningful interpretation of RSNA is confounded by this non-physiological state, given the intimate relationship between sympathetic nervous tone and cardiovascular status. To address this limitation of traditional approaches, we developed a new method for measuring RSNA in conscious, freely-moving mice. Mice were chronically instrumented with radio-telemeters for continuous monitoring of blood pressure as well as a jugular venous infusion catheter and custom-designed bipolar electrode for direct recording of RSNA. Following a 48-72 hour recovery period, survival rate was 100% and all mice behaved normally. At this time-point, RSNA was successfully recorded in 80% of mice, with viable signals acquired up to 4 and 5 days post-surgery in 70% and 50% of mice, respectively. Physiological blood pressures were recorded in all mice (116&plusmn;2 mmHg; n=10). Recorded RSNA increased with eating and grooming, as well-established in the literature. Furthermore, RSNA was validated by ganglionic blockade and modulation of blood pressure with pharmacological agents. Herein, an effective and manageable method for clear recording of RSNA in conscious, freely-moving mice is described.

Anesthetized mice exhibit non-physiological systemic blood pressure, which precludes meaningful assessment of autonomic tone given the intimate relationship between blood pressure and the autonomic nervous system. Thus, a novel method to simultaneously record renal sympathetic nerve activity and blood pressure with intravenous infusion in conscious mice is outlined.

Böbrek sempatik sinir etkinliği ve kan basıncı bilinçli, intravenöz infüzyon ile aynı anda kayıt için yeni yaklaşımın kontrolsüz fareler.

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity ...

Subcutaneous Trigeminal Nerve Field Stimulation for Refractory Facial Pain

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. Neuromodulation techniques for chronic facial pain include deep brain stimulation and motor cortex stimulation, which are complex to perform. Subcutaneous nerve field stimulation is certified for chronic back pain and is the least invasive form of neuromodulation. We applied this technique to treat chronic and neuropathic trigeminal pain as an individual therapy concept. First, trial stimulation is performed. Subcutaneous leads are placed in the painful trigeminal dermatome under local anesthesia. The leads are connected to an external neurostimulator that applies constant stimulation. Patients undergo a 12 day outpatient trial to assess the effect of the stimulation. Electrodes are removed after the trial. If the patient reports pain reduction of at least 50% in intensity and/or attack frequency, a reduction in medication or increase in quality of life, permanent implantation is scheduled. New electrodes are implanted under general anesthesia and are subcutaneously tunneled to an infraclavicular internal pulse generator. Patients are able to turn stimulation on and off and to increase or decrease the stimulation amplitude as needed.
This technique represents a minimal invasive alternative to other more invasive means of neuromodulation for trigeminal pain such as motor cortex stimulation or deep brain stimulation.

Treating chronic or neuropathic facial pain can be challenging when medical or standard treatment fails. Subcutaneous nerve field stimulation is the least invasive form of neuromodulation and is used for chronic back pain. We applied this technology to treat chronic and neuropathic trigeminal facial pain.

Refrakter Yüz Ağrısı İçin Subkutan Trigeminal Sinir Sahası Stimülasyonu

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. ...

Induction and Micro-CT Imaging of Cerebral Cavernous Malformations in Mouse Model

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have been established by tamoxifen induced deletion of Ccm genes in postnatal animals. These mouse models provide invaluable tools to investigate molecular mechanism and therapeutic approaches for CCM disease. An accurate and quantitative method to assess lesion burden and progression is essential to harness the full value of these animal models. Here, we demonstrate the induction of CCM disease in a mouse model and the use of the contrast enhanced X-ray micro computed tomography (micro-CT) method to measure CCM lesion burden in mouse brains. At postnatal day 1 (P1), we used 4-hydroxytamoxifen (4HT) to activate Cre recombinase activity from the Cdh5-CreErt2 transgene to cleave the floxed allele of Ccm2. CCM lesions in mouse brains were analyzed at P8. For micro-CT, iodine based Lugol&#39;s solution was used to enhance contrast in brain tissue. We have optimized the scan parameters and utilized a voxel dimension of 9.5 &#181;m, which lead to a minimum feature size of approximately 25 &#181;m. This resolution is sufficient to measure CCM lesion volume and number globally and accurately, and provide high-quality 3-D mapping of CCM lesions in mouse brains. This method enhances the value of the established mouse models to study the molecular basis and potential therapies for CCM and other cerebrovascular diseases.

This protocol demonstrates the induction of cerebral cavernous malformation disease in a mouse model and uses contrast enhanced micro computed tomography to measure lesion burden. This method enhances the value of established mouse models to study the molecular basis and potential therapies for cerebral cavernous malformation and other cerebrovascular diseases.

İndüksiyon ve fare modeli serebral derin malformasyon mikro-CT görüntüleme

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have ...

Vagus Nerve Stimulation As an Adjunctive Neurostimulation Tool in Treatment-resistant Depression

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression (TRD). VNS exhibits antidepressive and stabilizing effects. This method is particularly useful as a long-term treatment, in which up to two-thirds of patients respond. The vagus nerve stimulator is positioned on the left vagus nerve during a surgical procedure and is activated telemetrically by a wand connected to a handheld computerized device. The treating physician can perform various adjustments of the vagus nerve stimulator during in-office visits (e.g., by modifying stimulation intensity or stimulation frequency) to achieve maximum therapeutic effects with low side effects. Set-up of the device usually takes several months. Typical side effects include wound infection, temporary salivation, coughing, paralysis of the vocal cords, bradycardia, or even asystole. The patient can stop the VNS by placing a magnet over the generator. The current protocol describes delivery of the specific stimulation tool and methods for adjusting the tuning parameters to achieve the best remission rates in patients with TRD.

Vagus nerve stimulation (VNS) has been shown to be effective as an adjunct treatment for treatment-resistant depression (TRD). VNS leads to antidepressive and antisuicidal effects and quality of life improvements. This protocol offers a step-by-step guide to managing and adjusting a vagus nerve stimulator for efficient treatment of TRD.

Vagus sinir stimülasyonu ek bir Neurostimulation aracı olarak tedaviye dirençli depresyon

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression ...

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. Research focusing on cardiovascular disease models, potential cardiovascular therapeutic targets or pharmaceutical agents requires assessment of systemic arterial pressure and heart rhythm changes. In situations where radio telemetry systems are not available or affordable, the technique of femoral artery cannulation is an alternative way to obtain intra-arterial pressure waveform recordings and systemic BP measurements. This technique is economical and can be performed with standard equipment in animal facilities. However, invasive arterial pressure recording requires cannulation of small arteries, which can be a challenging surgical skill. Here, we present step-by-step protocols for femoral artery cannulation procedures. Key procedures include the calibration of the data acquisition system, tissue dissection and femoral artery cannulation, and setup of the arterial cannulation system for pressure recording. Surface electrocardiography recording procedures are also included. We also present examples of BP recordings from normotensive and hypertensive rats. This protocol allows reliable direct recordings of systemic BP with simultaneous electrocardiography.

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

Eşzamanlı elektrokardiyografi kayıt ve invaziv kan basınç ölçüm sıçanlarda

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. ...

Updated Technique for Reliable, Easy, and Tolerated Transcranial Electrical Stimulation Including Transcranial Direct Current Stimulation

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of brain stimulation presents several potential advantages compared to other techniques, as it is noninvasive, cost-effective, broadly deployable, and well-tolerated provided proper equipment and protocols are administered. Even though tDCS is apparently simple to perform, correct administration of the tDCS session, especially the electrode positioning and preparation, is vital for ensuring reproducibility and tolerability. The electrode positioning and preparation steps are traditionally also the most time consuming and error-prone. To address these challenges, modern tDCS techniques, using fixed-position headgear and pre-assembled sponge electrodes, reduce complexity and setup time while also ensuring that the electrodes are consistently placed as intended. These modern tDCS methods present advantages for research, clinic, and remote-supervised (at home) settings. This article provides a comprehensive step-by-step guide for administering a tDCS session using fixed-position headgear and pre-assembled sponge electrodes. This guide demonstrates tDCS using commonly applied montages intended for motor cortex and dorsolateral prefrontal cortex (DLPFC) stimulation. As described, selection of the head size and montage-specific headgear automates electrode positioning. Fully assembled pre-saturated snap-electrodes are simply affixed to the set position snap-connectors on the headgear. The modern tDCS method is shown to reduce setup time and reduce errors for both novice and expert operators. The methods outlined in this article can be adapted to different applications of tDCS as well as other forms of transcranial electrical stimulation (tES) such as transcranial alternating current stimulation (tACS) and transcranial random noise stimulation (tRNS). However, since tES is application specific, as appropriate, any methods recipe is customized to accommodate subject, indication, environment, and outcome specific features.

When administering transcranial direct current stimulation (tDCS), reproducible electrode preparation and placement are vital for a tolerated and effective session. The purpose of this article is to demonstrate updated modern setup procedures for the administration of tDCS and related transcranial electrical stimulation techniques, such as transcranial alternating current stimulation (tACS).

Transkraniyal Doğru Akım Stimülasyonu Dahil Olmak Üzere Güvenilir, Kolay ve Tolere Edilen Transkraniyal Elektriksel Stimülasyon için Güncellenmiş Teknik

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of ...