An NINDS Grant NS05033 (JCC) supported this work. The University of Connecticut RAC, SURF and OUR programs provided additional support.

<ol>
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The authors have nothing to disclose.

Bu araçlar ve farelerde ve insanlarda beyin ventriküler sisteminin bütünlüğünü değerlendirmek için kullanılabilecek protokoller sunuyoruz. Bu araçlar, ancak, aynı zamanda ya da 14,21,22 yaşlanma sürecinde yaralanma, hastalık nedeniyle değişiklik geçiren diğer beyin yapıları veya organ sistemlerine uygulanabilir. stratejiler kesitsel ve uzunlamasına MRG dizilerinin uyum belirli bölgelerde veya ilgi yapıların 3 boyutlu hacim temsillerini oluşturmasına olanak sağlar yazılım almak a...

Bir ependim hücre tekli-tabakası hatları beyin omurilik sıvısı (CSF) ve interstisyel sıvının (ISF) 1-3 arasında iki yönlü engelleyici ve taşıma fonksiyonları sağlayan beyin ventriküler sistemi. Bu işlevler, beyin toxicant-özgür ve fizyolojik dengede 2,3 tutmak için yardımcı olur. Yaralanma ya da hastalık ile bu kaplama bölümlerinin insan kayıpta diğer epitelyal astarları bulunduğu gibi rejeneratif değiştirilmesine neden görünmemektedir; oldukça ependim hücre kapsama kaybı ventrikül yüzeyinde ependimal hücrelerin arındırılır bölgeleri kapsayan astrositlerde bir ağda ile periventriküler astrogliyozun neden görünmektedir. Önemli BOS / ISF değişimi ve temizlik mekanizmalarında ciddi yansımaları bu epitel tabakasının 1,2,4-7 kaybına yol açması tahmin olacaktır. İnsan yaşlanmanın ortak bir özelliği yanal ventriküller (ventrikülomegali) ve Gözlem olarak ilişkili periventriküler ödem büyütülürMRG ve sıvı-zayıflatılmış inversiyon kurtarma MR (MRG / FLAIR) 8-14 tarafından ed. Ventrikülomegalide ve ventrikül astar hücresel organizasyon arasındaki ilişkiyi araştırmak için, postmortem insan MRG sekansları lateral ventrikül periventriküler dokusunun histolojik preparatları ile eşleştirildi. Ventrikülomegalisi durumlarda, gliozis önemli alanlar lateral ventrikül duvar boyunca ependim hücre kapsama yerini almıştı. Ventrikül genişlemesi MRI tabanlı ses analizi ile tespit edilmedi zaman ependimal hücre astar sağlam ve gliozis ventrikül astar 6 boyunca saptanmadı. Bu kombinasyon yaklaşım bölümlerinin wholemount hazırlıklarını veya tüm lateral ventrikül duvarı ve ventrikül hacimleri 6 3D modelleme kullanılarak lateral ventrikül astar hücresel bütünlük içinde ilk kapsamlı dokümantasyon detaylandırma değişiklikleri temsil eder. Çeşitli hastalıklar (Alzheimer hastalığı, şizofreni), yaralanmalar (travmatik beyin yaralanması)erken nöropatolojik bir özellik olarak ventrikülomegali göstermektedir. Böylece ependim hücre zarının alanlarının denüdasyon, normal ependim hücre fonksiyonu ile müdahale ve BOS / ISF sıvısı ve çözünen değişimi arasındaki homeostatik dengeyi tehlikeye tahmin olacaktır. Böylece, altta yatan ya da komşu beyin yapıları ventriküler sistemi, hücresel kompozisyonu ve sonuç değişiklikler daha kapsamlı bir inceleme sonunda ventrikül genişlemesi ile ilişkili nöropatolojisi hakkında daha fazla ortaya çıkarmak için başlayacak. Birlikte histolojik doku örnekleri karşılık gelen sınırlı erişime sahip multimodal görüntüleme veri eksikliği ve özellikle boyuna veri dizileri, insan beyni patolojilerin analizi zorlaştırır. İnsan yaşlanması veya hastalığa bulunan Modelleme fenotipleri genellikle fare modelleri ile elde edilebilir ve hayvan modelleri, insan hastalık başlangıcında ve ilerlemesinde ilgili soruları keşfetmek için bizim en iyi araçlardan biri haline gelmiştir. Çeşitli çalışmalarSağlıklı genç fareler lateral ventrikül duvarlarının hücre mimarisini ve temel kök hücre niş 4,7-15 tarif var. Bu çalışmalar 6.15 yaşlanma yoluyla ventrikül duvarlarının 3D modelleme ve hücresel analiz içerecek şekilde genişletilmiştir. Farenin nispeten sağlam bir subventicular bölgesi (SVZ) 6,15 astar dokunulmamış ependim hücreden hücreye niş bitişiktir kök göstermek yerine periventriküler gliozis de ventrikülomegali ne, yaşlı farelerde görülür. Böylece, çarpıcı türe özgü farklılıklar 6.15 yaşlanma sürecinde genel bakım ve lateral ventrikül astar bütünlüğü hem de mevcuttur. Bu nedenle, insanlarda bulunan koşullarını sorgulamak için en iyi kullanım farelere, iki türün arasındaki farklar karakterize ve uygun herhangi bir modelleme paradigmasında dikkate alınması gerekir. Burada, her iki insan ve m lateral ventriküllerin uzunlamasına değişiklikler ve ilgili periventriküler doku değerlendirmek için prosedürler sunuyoruzouse. Bizim prosedürler hücresel organizasyon yapısı ve hem de karakterize etmek 3D render ve fare ve insan ventriküllerin hem volumetriye ve periventriküler dokusunun bütün montaj hazırlıkları immünohistokimyasal analizi kullanımını içerir. Birlikte bu işlemler ventriküler sistemindeki değişiklikleri ve ilgili periventriküler doku karakterize bir yol sağlar.

<table border="0" cellpadding="0" cellspacing="0" width="881">
	<tbody>
		<tr height="44">
			<td height="44" style="height:44px;width:227px;">Name of the Materal/Equipment</td>
			<td style="width:363px;">Company</td>
			<td style="width:131px;">Catalog Number</td>
			<td style="width:160px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phosphate buffered saline (PBS)</td>
			<td>Life Technologies</td>
			<td>21600-069</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde (PFA)</td>
			<td>Electron Microscopy Sciences</td>
			<td>19210</td>
			<td>Use at 4% in PBS, 4 &#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Normal Horse Serum</td>
			<td>Life Technologies</td>
			<td>16050</td>
			<td>10% in PBS-TX (v/v)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Normal Goat Serum</td>
			<td>Life Technologies</td>
			<td>16210</td>
			<td>10% in PBS-TX (v/v)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triton X-100 (TX)</td>
			<td>Sigma-Aldrich</td>
			<td>T8787</td>
			<td>0.1% in PBS (v/v)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vibratome</td>
			<td>Leica</td>
			<td>VT1000S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluorescence Microscope</td>
			<td>Zeiss</td>
			<td>Imager.M2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Camera</td>
			<td>Hamamatsu</td>
			<td>ORCA R2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microscope Stage Controller</td>
			<td>Ludl Electronic Products</td>
			<td>MAC 6000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereology software</td>
			<td>MBF Bioscience</td>
			<td colspan="2">Stereo Investigator 11</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereology software</td>
			<td>ImageJ/NIH</td>
			<td>NIH freeware</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3D Reconstruction software</td>
			<td>MBF Bioscience</td>
			<td colspan="2">Neurolucida Explorer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Confocal Microscope</td>
			<td>Leica</td>
			<td>TCS SP2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MRI Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Freesurfer</td>
			<td colspan="2">https://surfer.nmr.mgh.harvard.edu/fswiki/DownloadAndInstall</td>
			<td>Segmentation and Volume</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ITK-Snap</td>
			<td>http://www.itksnap.org/pmwiki/pmwiki.php</td>
			<td></td>
			<td>Segmentation and Volume</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Multi-image Analysis GUI (Mango)</td>
			<td>http://ric.uthscsa.edu/mango/</td>
			<td></td>
			<td>Longitudinal overlay</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Whole Mount Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">22.5&#176; microsurgical straight stab knife</td>
			<td>Fisher Scientific</td>
			<td>NC9854830</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">parafilm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">wax bottom dissecting dish&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pins</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">fine forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">aquapolymount</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissecting Microscope</td>
			<td>Leica</td>
			<td>MZ95</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Whole Mount Antibodies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">mouse anti-b-catenin</td>
			<td>BD Bioschiences, San Jose, CA, USA</td>
			<td></td>
			<td>1:250</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">goat anti-GFAP</td>
			<td>Santa Cruz Biotechnology</td>
			<td></td>
			<td>1:250</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">rabbit anti-AQP4 (aquaporin-4)</td>
			<td>&#160;Sigma-Aldrich</td>
			<td></td>
			<td>1:400</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Coronal Antibodies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anti-S100&#946; antibody</td>
			<td>Sigma-Aldrich</td>
			<td></td>
			<td>1:500</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4&#8217;,6-diamidino-2-phenylindole (DAPI)</td>
			<td>Life Technologies</td>
			<td>D-1306</td>
			<td>10 &#181;g/mL in PBS</td>
		</tr>
	</tbody>
</table>

3d modeling of the lateral ventricles and histological characterization of periventricular tissue in humans and mouse

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional units of the ventricles are ciliated epithelial cells termed ependymal cells, which line the ventricles and through ciliary action are capable of generating laminar flow of CSF at the ventricle surface. This monolayer of ependymal cells also provides barrier and filtration functions that promote exchange between brain interstitial fluids (ISF) and circulating CSF. Biochemical changes in the brain are thereby reflected in the composition of the CSF and destruction of the ependyma can disrupt the delicate balance of CSF and ISF exchange. In humans there is a strong correlation between lateral ventricle expansion and aging. Age-associated ventriculomegaly can occur even in the absence of dementia or obstruction of CSF flow. The exact cause and progression of ventriculomegaly is often unknown; however, enlarged ventricles can show regional and, often, extensive loss of ependymal cell coverage with ventricle surface astrogliosis and associated periventricular edema replacing the functional ependymal cell monolayer. Using MRI scans together with postmortem human brain tissue, we describe how to prepare, image and compile 3D renderings of lateral ventricle volumes, calculate lateral ventricle volumes, and characterize periventricular tissue through immunohistochemical analysis of en face lateral ventricle wall tissue preparations. Corresponding analyses of mouse brain tissue are also presented supporting the use of mouse models as a means to evaluate changes to the lateral ventricles and periventricular tissue found in human aging and disease. Together, these protocols allow investigations into the cause and effect of ventriculomegaly and highlight techniques to study ventricular system health and its important barrier and filtration functions within the brain.

Immunohistokimyasal 50 mikron koronal kesitler ve 3D rekonstrüksiyon (Şekil 3) dayalı fare lateral ventriküllerin Kontur takibi hacmi verileri hastalık ya da yaralanma için bir model sistem olarak fareyi kullanarak farklı deneysel paradigmalar toplanan sağlar. Bu prosedüre Kritik lateral ventrikül duvarları birbirine yapışır bölgelerin dışlama olduğunu. Ventriküllerin bölgeleri subsegmenting ve her bir bölgede (Şekil 3C) için farklı bir renk tayin, bitişik böl?...

Watch this Scientific Journal Video about 3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse at JoVE.com

3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse

Using MRI scans (human), 3D imaging software, and immunohistological analysis, we document changes to the brain&#8217;s lateral ventricles. Longitudinal 3D mapping of lateral ventricle volume changes and characterization of periventricular cellular changes that occur in the human brain due to aging or disease are then modeled in mice.

İnsanlar ve Mouse Periventriküler Doku Yanal ventrikülleri ve histolojik Karakterizasyonu 3D Modelleme

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional ...

3d-modeling-lateral-ventricles-histological-characterization

Research

JoVE Journal

Neuroscience

13.9K Görüntüleme.  University of Connecticut. Using MRI scans (human), 3D imaging software, and immunohistological analysis, we document changes to the brain’s lateral ventricles. Longitudinal 3D mapping of lateral ventricle volume changes and characterization of periventricular cellular changes that occur in the human brain due to aging or disease are then modeled in mice. 