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

<ol>
	<li>Del Bigio, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ependymal+cells:+biology+and+pathology.">Ependymal cells: biology and pathology.</a> Acta Neuropathol. 119, 55-73 (2010).</li><li>Johanson, C., 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+distributional+nexus+of+choroid+plexus+to+cerebrospinal+fluid,+ependyma+and+brain:+toxicologic/pathologic+phenomena,+periventricular+destabilization,+and+lesion+spread.">The distributional nexus of choroid plexus to cerebrospinal fluid, ependyma and brain: toxicologic/pathologic phenomena, periventricular destabilization, and lesion spread.</a> Toxicol Pathol. 39, 186-212 (2011).</li><li>Roales-Bujan, R., 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=Astrocytes+acquire+morphological+and+functional+characteristics+of+ependymal+cells+following+disruption+of+ependyma+in+hydrocephalus.">Astrocytes acquire morphological and functional characteristics of ependymal cells following disruption of ependyma in hydrocephalus.</a> Acta Neuropathologica. 124, 531-546 (2012).</li><li>Cserr, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiology+of+the+choroid+plexus.">Physiology of the choroid plexus.</a> Physiol Rev. 51, 273-311 (1971).</li><li>Iliff, J. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+paravascular+pathway+facilitates+CSF+flow+through+the+brain+parenchyma+and+the+clearance+of+interstitial+solutes,+including+amyloid+beta.">A paravascular pathway facilitates CSF flow through the brain parenchyma and the clearance of interstitial solutes, including amyloid beta.</a> Science Translational Medicine. 4, 147ra111 (2012).</li><li>Shook, B. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ventriculomegaly+associated+with+ependymal+gliosis+and+declines+in+barrier+integrity+in+the+aging+human+and+mouse+brain.">Ventriculomegaly associated with ependymal gliosis and declines in barrier integrity in the aging human and mouse brain.</a> Aging Cell. , (2013).</li><li>Xie, 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=Sleep+drives+metabolite+clearance+from+the+adult+brain.">Sleep drives metabolite clearance from the adult brain.</a> Science. 342, 373-377 (2013).</li><li>Fazekas, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathologic+correlates+of+incidental+MRI+white+matter+signal+hyperintensities.">Pathologic correlates of incidental MRI white matter signal hyperintensities.</a> Neurology. 43, 1683-1689 (1993).</li><li>Meier-Ruge, W., Ulrich, J., Bruhlmann, M., Meier, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+white+matter+atrophy+in+the+human+brain.">Age-related white matter atrophy in the human brain.</a> Ann N Y Acad Sci. 673, 260-269 (1992).</li><li>Resnick, S. M., Pham, D. L., Kraut, M. A., Zonderman, A. B., Davatzikos, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+magnetic+resonance+imaging+studies+of+older+adults:+a+shrinking+brain.">Longitudinal magnetic resonance imaging studies of older adults: a shrinking brain.</a> The Journal Of Neuroscience : The Official Journal Of The Society For Neuroscience. 23, 3295-3301 (2003).</li><li>Sener, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Callosal+changes+in+obstructive+hydrocephalus:+observations+with+FLAIR+imaging,+and+diffusion+MRI.">Callosal changes in obstructive hydrocephalus: observations with FLAIR imaging, and diffusion MRI.</a> Comput Med Imaging Graph. 26, 333-337 (2002).</li><li>Sze, G., 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=Foci+of+MRI+signal+(pseudo+lesions)+anterior+to+the+frontal+horns:+histologic+correlations+of+a+normal+finding.">Foci of MRI signal (pseudo lesions) anterior to the frontal horns: histologic correlations of a normal finding.</a> AJR Am J Roentgenol. 147, 331-337 (1986).</li><li>Tisell, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shunt+surgery+in+patients+with+hydrocephalus+and+white+matter+changes.">Shunt surgery in patients with hydrocephalus and white matter changes.</a> Journal of Neurosurgery. 114, 1432-1438 (2011).</li><li>Valdes Hernandez Mdel, C., 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=Automatic+segmentation+of+brain+white+matter+and+white+matter+lesions+in+normal+aging:+comparison+of+five+multispectral+techniques.">Automatic segmentation of brain white matter and white matter lesions in normal aging: comparison of five multispectral techniques.</a> Magn Reson Imaging. 30, 222-229 (2012).</li><li>Shook, B. A., Manz, D. H., Peters, J. J., Kang, S., Conover, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatiotemporal+changes+to+the+subventricular+zone+stem+cell+pool+through+aging.">Spatiotemporal changes to the subventricular zone stem cell pool through aging.</a> The Journal of Neuroscience : The Official Journal Of The Society For Neuroscience. 32, 6947-6956 (2012).</li><li>Mirzadeh, Z., Merkle, F. T., Soriano-Navarro, M., Garcia-Verdugo, J. M., Alvarez-Buylla, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+stem+cells+confer+unique+pinwheel+architecture+to+the+ventricular+surface+in+neurogenic+regions+of+the+adult+brain.">Neural stem cells confer unique pinwheel architecture to the ventricular surface in neurogenic regions of the adult brain.</a> Cell Stem Cell. 3, 265-278 (2008).</li><li>Mirzadeh, Z., Doetsch, F., Sawamoto, K., Wichterle, H., Alvarez-Buylla, 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+subventricular+zone+en-face:+wholemount+staining+and+ependymal+flow.">The subventricular zone en-face: wholemount staining and ependymal flow.</a> J Vis Exp. , (2010).</li><li>Luo, J., Daniels, S. B., Lennington, J. B., Notti, R. Q., Conover, J. 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+aging+neurogenic+subventricular+zone.">The aging neurogenic subventricular zone.</a> Aging Cell. 5, 139-152 (2006).</li><li>Luo, J., Shook, B. A., Daniels, S. B., Conover, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subventricular+zone-mediated+ependyma+repair+in+the+adult+mammalian+brain.">Subventricular zone-mediated ependyma repair in the adult mammalian brain.</a> J Neurosci. 28, 3804-3813 (2008).</li><li>Marcus, D. S., Fotenos, A. F., Csernansky, J. G., Morris, J. C., Buckner, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open+access+series+of+imaging+studies:+longitudinal+MRI+data+in+nondemented+and+demented+older+adults.">Open access series of imaging studies: longitudinal MRI data in nondemented and demented older adults.</a> J Cogn Neurosci. 22, 2677-2684 (2010).</li><li>Giorgio, A., De Stefano, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+use+of+brain+volumetry.">Clinical use of brain volumetry.</a> J Magn Reson Imaging. 37, 1-14 (2013).</li><li>Caspers, 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=Studying+variability+in+human+brain+aging+in+a+population-based+German+cohort-rationale+and+design+of+1000BRAINS.">Studying variability in human brain aging in a population-based German cohort-rationale and design of 1000BRAINS.</a> Front Aging Neurosci. 6, 149 (2014).</li><li>Keuken, M. C., 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=Ultra-high+7T+MRI+of+structural+age-related+changes+of+the+subthalamic+nucleus.">Ultra-high 7T MRI of structural age-related changes of the subthalamic nucleus.</a> The Journal of neuroscience : the official journal of the Society for Neuroscience. 33, 4896-4900 (2013).</li><li>Marti-Bonmati, L., Sopena, R., Bartumeus, P., Sopena, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multimodality+imaging+techniques.">Multimodality imaging techniques.</a> Contrast Media Mol Imaging. 5, 180-189 (2010).</li><li>Bergmann, O., 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+age+of+olfactory+bulb+neurons+in+humans.">The age of olfactory bulb neurons in humans.</a> Neuron. 74, 634-639 (2012).</li><li>Sanai, 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=Corridors+of+migrating+neurons+in+the+human+brain+and+their+decline+during+infancy.">Corridors of migrating neurons in the human brain and their decline during infancy.</a> Nature. 478, 382-386 (2011).</li><li>Wang, C., 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=Identification+and+characterization+of+neuroblasts+in+the+subventricular+zone+and+rostral+migratory+stream+of+the+adult+human+brain.">Identification and characterization of neuroblasts in the subventricular zone and rostral migratory stream of the adult human brain.</a> Cell Res. 21, 1534-1550 (2011).</li><li>Carmen Gomez-Roldan, D. e. l., M, , et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroblast+proliferation+on+the+surface+of+the+adult+rat+striatal+wall+after+focal+ependymal+loss+by+intracerebroventricular+injection+of+neuraminidase.">Neuroblast proliferation on the surface of the adult rat striatal wall after focal ependymal loss by intracerebroventricular injection of neuraminidase.</a> The Journal of Comparative Neurology. 507, 1571-1587 (2008).</li></ol>

The authors have nothing to disclose.

우리는 도구와 쥐 및 인간에서 뇌의 심실 시스템의 무결성을 평가할 수있다 프로토콜을 제시한다. 이러한 도구는, 그러나, 또한, 14,21,22 또는 노화 과정에서 손상, 질환으로 인한 변화를 겪는 다른 뇌 구조 또는 기관 시스템에 적용될 수있다. 전략의 단면과 길이 MRI 시퀀스들의 정렬이 특정 영역 또는 관심있는 구조의 3 차원 볼륨 표현을 생성 할 수있는 소프트웨어의 인출 이점을 ?...

뇌실막 세포 단층 선 뇌척수액 (CSF) 및 격자 간 유체 (ISF) 1-3 간의 양방향 장벽 및 전송 기능을 제공하는 뇌의 심실 시스템. 이러한 기능은 뇌가 독성이없는 생리적 균형 2,3에서 유지하는 데 도움이됩니다. 부상 또는 질병을 통해이 안감의 일부의 인간 손실에서 다른 상피 라이닝에서 발견 회생 교체에 결과에 표시되지 않습니다; 오히려 뇌실막 셀 커버리지의 손실은 심실 표면에서 뇌실막 세포의 무 결함 영역을 커버하는 성상 세포의 메쉬 작업과 뇌실 주위 백질 as​​trogliosis 될 것으로 보인다. 중요한 CSF / ISF 교환 및 통관 메커니즘에 심각한 영향이 상피 층 1,2,4-7의 손실이 발생할 것으로 예상된다. 인간의 노화의 일반적인 기능은 측면 심실 (뇌실) 및 observ 등 관련 뇌실 주위 부종을 확대MRI 및 유체 감쇠 반전 회복 자기 공명 영상 (MRI / FLAIR) 8-14에 의해 에드. 뇌실과 심실 라이닝의 세포 조직 사이의 관계를 조사하기 위해, 사후 인간의 자기 공명 영상 시퀀스는 측면 뇌실 뇌실 주위 조직의 조직 학적 제제와 일치했다. 뇌실 가지 경우에서, 신경교 증의 실질적인 영역은 측면 뇌실 벽을 따라 뇌실막 세포 커버리지를 교체했다. 뇌실 확장이 자기 공명 영상 기반 볼륨 분석에 의해 감지되지 않은 경우, 뇌실막 세포 안감은 그대로이고 신경교 증은 심실 라이닝 (6)에 따라 검출되지 않았다. 이 조합 방식은 부분의 wholemount 준비 또는 전체 측면 뇌실 벽과 심실 볼륨 (6)의 3D 모델링을 사용하여 측면 뇌실 내벽의 세포 무결성의 첫 번째 포괄적 인 문서 자세히 변경을 나타냅니다. 여러 질환 (알츠하이머 병, 정신 분열증)과 부상 (외상성 뇌 손상)초기 신경 병리학 적 기능으로 뇌실을 보여줍니다. 이에 뇌실막 세포 라이닝의 지역의 삭박 정상 뇌실막 세포 기능을 방해하고 CSF / ISF 유체 및 용질 교환 사이의 항상성 균형을 손상 할 것으로 예상된다. 따라서, 기본 또는 이웃 뇌 구조에 심실 시스템, 그것의 세포 구성하고, 결과에 대한 변경 사항에 대해 더욱 철저한 검사는 궁극적으로 뇌실의 확대와 관련된 신경 병리학에 대한 자세한 내용을 공개하기 시작합니다. 함께 학적 조직 샘플에 대응하는 액세스가 제한된 조영 데이터의 부족, 특히 길이 데이터 시퀀스에서, 인간의 뇌 병리 분석 어렵게 만든다. 인간의 노화 또는 질병에있는 모델링 표현형은 종종 마우스 모델을 달성 할 수있는 동물 모델은 인간의 질병 개시 및 진행에 대한 질문을 탐구하기 위해 최선의 방법 중 하나가. 의 여러 연구건강한 젊은 마우스는 좌우 심실 벽의 cytoarchitecture 및 기본 줄기 세포 틈새 4,7-15을 설명했다. 이들 연구는 -6,15- 노화 통해 좌심실 벽의 3D 모델링 및 세포 분석을 포함하도록 확장되어왔다. 마우스는 비교적 강력한 subventicular 영역 (SVZ) -6,15- 라이닝 그대로 뇌실막 세포에 세포 틈새 바로 아래 줄기 표시 오히려 뇌실 주위 백질 신경교 증이나 뇌실 어느 쪽도, 세 생쥐에서 관찰된다. 따라서, 타격 종 특이 차이 -6,15- 노화 과정에서 일반적인 관리 및 뇌실 라이닝의 무결성에 모두 존재한다. 따라서, 인간에서 발견 조건을 심문하는 것이 가장 사용 마우스로 두 종 사이의 차이가 특징으로 적절 어떤 모델링 패러다임에서 고려 될 필요가있다. 여기, 우리는 모두 인간과 M의 측면 심실에 길이 변화와 관련된 뇌실 주위 조직을 평가하는 절차를 제시여러개. 우리의 절차는 세포의 조직과 구조 모두의 특성을 3D 렌더링 및 마우스와 인간의 심실 모두의 용적 측정, 및 뇌실 주위 조직의 전체 마운트 준비의 면역 조직 화학 분석의 사용을 포함한다. 이러한 절차는 함께 심실 시스템의 변화와 연관된 뇌실 조직을 특성화하는 수단을 제공한다.

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

면역 염색 50 μm의 관상 섹션과 3D 재구성 (그림 3)에 따라 마우스 측면 심실의 윤곽 추적은 볼륨 데이터가 질병이나 부상에 대한 모델 시스템으로 마우스를 사용하여 다른 실험 패러다임에서 수집 할 수 있습니다. 이 절차에 긴급 뇌실 벽이 서로 접착 영역을 배제한다. 심실의 영역 subsegmenting 각 영역 (도 3C)에 대해 다른 색을 지정함으로써, 인접한 절편을 준수 할 수 있고, ...

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.

인간과 마우스의 뇌실 주위 조직의 측면 심실 및 조직 학적 특성의 3D 모델링

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

14.0K 조회수.  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. 

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

Mouse Models of Periventricular Leukomalacia

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature infants and the most common antecedent of cerebral palsy. PVL is characterized by periventricular white matter damage with prominent oligodendroglial injury. Hypoxia/ischemia with or without systemic infection/inflammation are the primary causes of PVL. We use P6 mice to create models of neonatal brain injury by the induction of hypoxia/ischemia with or without systemic infection/inflammation with unilateral carotid ligation followed by exposure to hypoxia with or without injection of the endotoxin lipopolysaccharide (LPS). Immunohistochemistry of myelin basic protein (MBP) or O1 and electron microscopic examination show prominent myelin loss in cerebral white matter with additional damage to the hippocampus and thalamus. Establishment of mouse models of PVL will greatly facilitate the study of disease pathogenesis using available transgenic mouse strains, conduction of drug trials in a relatively high throughput manner to identify candidate therapeutic agents, and testing of stem cell transplantation using immunodeficiency mouse strains.

We established mouse models of periventricular leukomalacia (PVL), the predominant brain injury in premature infants characterized by periventricular white matter lesions. Hypoxia/ischemia with/without systemic infection are the primary causes of PVL. Unilateral carotid ligation and hypoxia exposure with/without lipopolysaccharide injection creates PVL-like lesions in P6 mice.

Periventricular Leukomalacia의 마우스 모델

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature ...

연구

신경과학

Multiple-mouse Neuroanatomical Magnetic Resonance Imaging

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and evaluating mouse models of human disease. MRI is an excellent modality for investigating genetically altered animals. It is capable of whole brain coverage, can be used in vivo, and provides multiple contrast mechanisms for investigating different aspects of neuranatomy and physiology. The advent of high-field scanners along with the ability to scan multiple mice simultaneously allows for rapid phenotyping of novel mutations.
Effective mouse MRI studies require attention to many aspects of experiment design. In this article, we will describe general methods to acquire quality images for mouse phenotyping using a system that images mice concurrently in shielded transmit/receive radio frequency (RF) coils in a common magnet (Bock et al., 2003). We focus particularly on anatomical phenotyping, an important and accessible application that has shown a high potential for impact in many mouse models at our imaging centre. Before we can provide the detailed steps to acquire such images, there are important practical considerations for both in vivo brain imaging (Dazai et al., 2004) and ex vivo brain imaging (Spring et al., 2007) that should be noted. These are discussed below.

Magnetic resonance imaging (MRI) has become an increasingly popular tool for examining the phenotype of genetically altered mice. This article illustrates the methods necessary to achieve high-throughput phenotyping of genetically altered mice using multiple-mouse MRI.

여러 마우스 Neuroanatomical 자기 공명 영상

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and ...

Cerebrovascular Casting of the Adult Mouse for 3D Imaging and Morphological Analysis

Vascular imaging is crucial in the clinical diagnosis and management of cerebrovascular diseases, such as brain arteriovenous malformations (BAVMs). Animal models are necessary for studying the etiopathology and potential therapies of cerebrovascular diseases. Imaging the vasculature in large animals is relatively easy. However, developing vessel imaging methods of murine brain disease models is desirable due to the cost and availability of genetically-modified mouse lines. Imaging the murine cerebral vascular tree is a challenge. In humans and larger animals, the gold standard for assessing the angioarchitecture at the macrovascular (conductance) level is x-ray catheter contrast-based angiography, a method not suited for small rodents.
In this article, we present a method of cerebrovascular casting that produces a durable skeleton of the entire vascular bed, including arteries, veins, and capillaries that may be analyzed using many different modalities. Complete casting of the microvessels of the mouse cerebrovasculature can be difficult; however, these challenges are addressed in this step-by-step protocol. Through intracardial perfusion of the vascular casting material, all vessels of the body are casted. The brain can then be removed and clarified using the organic solvent methyl salicylate. Three dimensional imaging of the brain blood vessels can be visualized simply and inexpensively with any conventional brightfield microscope or dissecting microscope. The casted cerebrovasculature can also be imaged and quantified using micro-computed tomography (micro-CT)1. In addition, after being imaged, the casted brain can be embedded in paraffin for histological analysis.
The benefit of this vascular casting method as compared to other techniques is its broad adaptation to various analytic tools, including brightfield microscopic analysis, CT scanning due to the radiopaque characteristic of the material, as well as histological and immunohistochemical analysis. This efficient use of tissue can save animal usage and reduce costs. We have recently demonstrated application of this method to visualize the irregular blood vessels in a mouse model of adult BAVM at a microscopic level2, and provide additional images of the malformed vessels imaged by micro-CT scan. Although this method has drawbacks and may not be ideal for all types of analyses, it is a simple, practical technique that can be easily learned and widely applied to vascular casting of blood vessels throughout the body.

In this article, we present a simple, practical technique for cerebrovascular casting that is easy to perform and can be utilized to image the vascular tree of the adult mouse brain.

3D 이미징 및 형태학의 분석을위한 성인 마우스 뇌혈관 캐스팅

Vascular imaging is crucial in the clinical diagnosis and management of cerebrovascular diseases, such as brain arteriovenous malformations (BAVMs). ...

Modification of a Colliculo-thalamocortical Mouse Brain Slice, Incorporating 3-D printing of Chamber Components and Multi-scale Optical Imaging

The ability of the brain to process sensory information relies on both ascending and descending sets of projections. Until recently, the only way to study these two systems and how they interact has been with the use of in vivo preparations. Major advances have been made with acute brain slices containing the thalamocortical and cortico-thalamic pathways in the somatosensory, visual, and auditory systems. With key refinements to our recent modification of the auditory thalamocortical slice1, we are able to more reliably capture the projections between most of the major auditory midbrain and forebrain structures: the inferior colliculus (IC), medial geniculate body (MGB), thalamic reticular nucleus (TRN), and the auditory cortex (AC). With portions of all these connections retained, we are able to answer detailed questions that complement the questions that can be answered with in vivo preparations. The use of flavoprotein autofluorescence imaging enables us to rapidly assess connectivity in any given slice and guide the ensuing experiment. Using this slice in conjunction with recording and imaging techniques, we are now better equipped to understand how information processing occurs at each point in the auditory forebrain as information ascends to the cortex, and the impact of descending cortical modulation. 3-D printing to build slice chamber components permits double-sided perfusion and broad access to networks within the slice and maintains the widespread connections key to fully utilizing this preparation.

Using multiple angles to cut the mouse pup brain, we improve upon a previously-described acute brain slice which captures the connections between most of the major auditory midbrain and forebrain structures.

Colliculo-thalamocortical 마우스 뇌 조각의 변경, 3 차원 챔버 구성 요소의 인쇄 및 멀티 스케일 광학 이미징을 통합

The ability of the brain to process sensory information relies on both ascending and descending sets of projections. Until recently, the only way to study ...

Utilizing 3D Printing Technology to Merge MRI with Histology: A Protocol for Brain Sectioning

Magnetic resonance imaging (MRI) allows for the delineation between normal and abnormal tissue on a macroscopic scale, sampling an entire tissue volume three-dimensionally. While MRI is an extremely sensitive tool for detecting tissue abnormalities, association of signal changes with an underlying pathological process is usually not straightforward. In the central nervous system, for example, inflammation, demyelination, axonal damage, gliosis, and neuronal death may all induce similar findings on MRI. As such, interpretation of MRI scans depends on the context, and radiological-histopathological correlation is therefore of the utmost importance. Unfortunately, traditional pathological sectioning of brain tissue is often imprecise and inconsistent, thus complicating the comparison between histology sections and MRI. This article presents novel methodology for accurately sectioning primate brain tissues and thus allowing precise matching between histology and MRI. The detailed protocol described in this article will assist investigators in applying this method, which relies on the creation of 3D printed brain slicers. Slightly modified, it can be easily implemented for brains of other species, including humans.

The overall goal of this protocol is to accurately align magnetic resonance imaging (MRI) image volumes with histology sections via the creation of customized 3D-printed brain holders and slicer boxes.

3D 인쇄 기술을 활용하여 조직학과 MRI를 병합하기 : 두뇌를위한 프로토콜 단면

Magnetic resonance imaging (MRI) allows for the delineation between normal and abnormal tissue on a macroscopic scale, sampling an entire tissue volume ...

Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this challenging. Because the ventricles are negative spaces located deep within the brain, the only way to understand their anatomy is by appreciating their boundaries formed by related structures. Looking at a 2D representation of these spaces, in any of the cardinal planes, will not enable visualisation of all of the structures that form the boundaries of the ventricles. Thus, using 2D sections alone requires students to compute their own mental image of the 3D ventricular spaces. The aim of this study was to develop a reproducible method for dissecting the human brain to create an educational resource to enhance student understanding of the intricate relationships between the ventricles and periventricular structures. To achieve this, we created a video resource that features a step-by-step guide using a fiber dissection method to reveal the lateral and third ventricles together with the closely related limbic system and basal ganglia structures. One of the advantages of this method is that it enables delineation of the white matter tracts that are difficult to distinguish using other dissection techniques. This video is accompanied by a written protocol that provides a systematic description of the process to aid in the reproduction of the brain dissection. This package offers a valuable anatomy teaching resource for educators and students alike. By following these instructions educators can create teaching resources and students can be guided to produce their own brain dissection as a hands-on practical activity. We recommend that this video guide be incorporated into neuroanatomy teaching to enhance student understanding of the morphology and clinical relevance of the ventricles.

This paper demonstrates the effective use of a fiber dissection method to reveal the superficial white matter tracts and periventricular structures of the human brain, in three-dimensional space, to aid student comprehension of ventricular morphology.

깊은 공간-인간 두뇌의 측면 심 공개 뇌 구조 해부학 적 탐구

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this ...

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.

유도 및 마우스 모델에서 대뇌 동굴 같은 기형의 마이크로-CT 영상

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 ...

의학

Large-scale Three-dimensional Imaging of Cellular Organization in the Mouse Neocortex

The mammalian neocortex is composed of many types of excitatory and inhibitory neurons, each with specific electrophysiological and biochemical properties, synaptic connections, and in vivo functions, but their basic functional and anatomical organization from cellular to network scale is poorly understood. Here we describe a method for the three-dimensional imaging of fluorescently-labeled neurons across large areas of the brain for the investigation of the cortical cellular organization. Specific types of neurons are labeled by the injection of fluorescent retrograde neuronal tracers or expression of fluorescent proteins in transgenic mice. Block brain samples, e.g., a hemisphere, are prepared after fixation, made transparent with tissue clearing methods, and subjected to fluorescent immunolabeling of the specific cell types. Large areas are scanned using confocal or two-photon microscopes equipped with large working distance objectives and motorized stages. This method can resolve the periodic organization of the cell type-specific microcolumn functional modules in the mouse neocortex. The procedure can be useful for the study of three-dimensional cellular architecture in the diverse brain areas and other complex tissues.

Here we describe a procedure for tissue clearing, fluorescent labeling, and large-scale imaging of mouse brain tissue which, thereby, enables visualization of the three-dimensional organization of cell types in the neocortex.

마우스 피 질 세포 조직의 대형 3-차원 영상

The mammalian neocortex is composed of many types of excitatory and inhibitory neurons, each with specific electrophysiological and biochemical ...

A Micro-CT-based Method for Characterizing Lesions and Locating Electrodes in Small Animal Brains

Lesion and electrode location verification are traditionally done via histological examination of stained brain slices, a time-consuming procedure that requires manual estimation. Here, we describe a simple, straightforward method for quantifying lesions and locating electrodes in the brain that is less laborious and yields more detailed results. Whole brains are stained with osmium tetroxide, embedded in resin, and imaged with a micro-CT scanner. The scans result in 3D digital volumes of the brains with resolutions and virtual section thicknesses dependent on the sample size (12&#8211;15 and 5&#8211;6 &#181;m&#160;per voxel for rat and zebra finch brains, respectively). Surface and deep lesions can be characterized, and single tetrodes, tetrode arrays, electrolytic lesions, and silicon probes can also be localized. Free and proprietary software allows experimenters to examine the sample volume from any plane and segment the volume manually or automatically. Because this method generates whole brain volume, lesions and electrodes can be quantified to a much higher degree than in current methods, which will help standardize comparisons within and across studies.

This article describes a straightforward method to prepare small animal brains for micro-CT imaging, in which lesions can be quantified and electrodes located with high precision in the context of the whole brain.

병 변 특성화 및 작은 동물 두뇌에 전극을 위치에 대 한 마이크로-CT 기반 방법

Lesion and electrode location verification are traditionally done via histological examination of stained brain slices, a time-consuming procedure that ...

A Needle Model for Vascular Cognitive Impairment Studies in Mice

A Mouse Model for Vascular Cognitive Impairment and Dementia Based on Needle-guided Asymmetric Bilateral Common Carotid Artery Stenosis

Vascular cognitive impairment and dementia (VCID) results from vascular brain injury. Given VCID&#39;s high incidence, which is expected to continue rising as the population ages, it is critical to establish a robust animal model for the disease. This paper presents a novel method of creating a mouse model of VCID that is based on asymmetric bilateral common carotid artery stenosis, which mimics human chronic cerebral hypoperfusion caused by carotid atherosclerosis.
Briefly, common carotid arteries (CCAs) are ligated to different gauge needles (32 G for the right CCA and 34 G for the left CCA) using 7-0 silk sutures followed by immediate needle removal. The remaining suture rings cause persistent blood flow reduction and long-term cognitive impairment associated with white matter injury, microinfarcts, and reactive gliosis, thus closely mimicking the pathogenesis of VCID. Importantly, in this needle model, the clinical representations do not revert with time, providing reliable long-term cognitive impairment. Moreover, the survival rate 24 weeks post surgery was 81.6%, which is higher compared to the other established models of VCID with a similar level of blood flow reduction.
Additional advantages include low material cost and compatibility with MRI to monitor brain injury in live animals since no metal is implanted. The main challenge in employing the needle model of VCID is the requirement for developing advanced surgical skills since mouse CCAs are less than 0.6 mm in diameter and are very fragile. High-quality visual representation of the surgery will thus help researchers to master this technique and advance our understanding of VCID, potentially leading to the development of novel therapeutic modalities to decrease the devastating cognitive decline associated with VCID.

The needle method of asymmetric bilateral common carotid artery stenosis is proposed to create a mouse model for vascular cognitive impairment and dementia. It results in longer-term outcomes compared to previously established models and is compatible with live MRI. Visual representation demonstrating the procedure provides guidance for mastering the surgery.

Vascular cognitive impairment and dementia (VCID) results from vascular brain injury. Given VCID&#39;s high incidence, which is expected to continue ...