这项工作是由NCTR / FDA。

虽然未示出，在视频，第一给定大鼠，用过量的戊巴比妥钠300毫克/公斤，导致在少于3分钟内麻醉，然后断头处死。他们的大脑进行迅速，但仔细从头骨中取出，在冰冷的生理盐水中冷却5分钟。重要的是让大脑冷却这解剖MAV使得MAV是可以分开的皮层的表面（层I的顶部）之前的时间量。大脑，然后放置在底部的玻璃陪替氏培养皿搁在冰满（1厘米深）的冰冷的生理盐水或0.1M磷酸钠缓冲盐水的pH = 7.4。所有的解剖完成与脑的大部分浸入盐水。 1。去除松果体<ol><li>放置在脑背侧向上（相对于陪替氏培养皿的底部）。就到t喙两个半球之间的松果体是位于最尾端腹侧区他小脑（松果体凹槽），并且是正下方的表面或在上丘的脑膜。删除的的松果体第一个使用两个小弯尖镊子。它是粉红色的颜色，如皮质，但经常被包围在从大脑切除残留的血液和血管的残余。 </li></ol> 2。删除的MAV <ol><li>翻转大脑的“倒挂”和椎动脉，小动脉和脑膜，桥前脑脑膜和血管的分离。前脑MAV，然后开始拆除。 重要的是要开始腹部比较大的Willis环的动脉，大脑中动脉（MCA）和前动脉在解剖过程中是最强的，作为“脚手架”，其中较小的表面的软脑膜动脉和微动脉膜然后可以除去从皮层与脑膜的其余部分。审查Scremin 7米矿石的脑表面血管的可视化图形和细节。 </li><li>从任一半球，使用小弯尖镊子切断后交通动脉，颈内动脉关头后的。因此，分离前脑的前部和后部的动脉树。对侧半球重复同样的过程。 </li><li>马华和前动脉的镊子夹住，轻轻拉和周围嗅束前背的方向，使覆盖的腹前部皮层（梨状，嗅结节），MAV解除。这消除了很多的MAV覆盖前皮层（扣和轨道）。 </li><li>转动脑在45〜90°角的陪替氏培养皿。横向的，更靠前的区域周围的MAV外侧皮质区（颗粒孤立的，次要的体感和听觉皮层）可以从皮质中解脱出来，抓住MCA和相关阿尔特指背，轻轻地拉动沿皮层。如果有必要，可以使用的端部的钳子来抬起并从皮质释放的主要动脉剥离时。 </li><li>现在删除的MAV后外侧地区。 （注意，它最好是从一个半球切换的其他在此收获过程，以确保的MAV保持冷和水合。此外，如果从皮质释放的MAV电阻上遇到一个半球开关对侧半球暂时，然后再回到原来的半球）。 </li><li>要删除的最背侧区域周围的初级躯体的MAV和运动皮质转动脑背侧向上（相对于陪替氏培养皿的底部）。为了最终释放，从大脑的MAV，使用的端部的钳子沿矢状窦。 收获MAV的这部分可以被暂时存放在冰冷的生理盐水中，直到测试和分析，或冻结，购买处理。常MAV覆盖最后/尾鳍地区的皮质（嗅，视觉和最尾端的听觉皮层）保持连接的皮质。其去除后视频时，MAV之间的扣带皮质和覆盖丘解剖。 </li></ol> 3。去除脉络丛<ol><li>脑背侧的位置并保持在与较大的钳子的小钳子推通过中线之间的半球，然后使用皮质和胼胝体（≈-3.3毫米来自囟）的端部通过穿刺入的顶部的地方中线的海马状突起。 </li><li>使用镊子远离背海马和隔拉皮质胼胝体，露出最侧脑室的。脉络丛可以定位和识别的红色波浪线划定的主要动脉，运行它的长度。使用的两个端部的钳子撬第三法师的侧壁tricle和扩大它在最尾端（≈-4.3毫米从囟8）。 </li><li>使用小镊子，将这个脉络丛免费。前区≈1.6毫米前囟门）之间的隔膜和尾状核/壳（最喙的程度，然后去和镊子的脑室扩大本节拉脉络丛免费。对侧半球上执行相同的过程，以获得所述第二其它脉络膜丛。 同样，这组织可以被暂时存放在冰冷的生理盐水中，直到测试和分析，或冻结，购买处理。 </li></ol> 4。去除余下的MAV尾鳍地区的Cortex包括扣带，听觉和视觉皮层丘脑和丘以及上覆第三脑室<ol><li>删除整个海马的两个半球暴露在丘脑和丘MAV。的去除这部分的MAV是比除去的MAV皮质不太困难。 </li><li>拉动这部分的MAV免费的把握较大的血管覆丘脑的前部，然后轻轻拉动尾部。此血管连接到上文丘网络和血液供给向背侧海马和背侧丘脑网络。拉了尾部，并逐步supracollicular网络，直到到达最后一个部分的尾动脉环血管。 在这一点上，必须采取更多的照顾，以消除任何剩余的后表面上的颗粒扣带，初级听觉和视觉皮层腹侧MAV。与MAV的其他部分覆盖的前端部分的皮质，这个组织可以暂时留在冰冷的盐水，直到测试和分析或冻结后处理的。任何剩余的后腹MAV收获第二个MAV部分的，应该被删除D添加到第一解剖MAV覆盖皮层部分，如果它们是单独进行分析。 </li></ol> 5。代表性的成果当清扫正确执行，导致的MAV组织应该在两个完整的实体，重约35至45毫克总。初步解剖MAV周围的皮质，重约25毫克和的MAV覆盖的丘脑，丘和枕叶皮质重约15毫克。它应该是稍微带粉红色的颜色从残留的血液在血管。脉络膜丛双边收获应该是在两个完整的组织样本与每个称量1〜2毫克。虽然多余的水可以被删除前的MAV储存或处理，如用纸巾在清洁镜头使用，以避免组织损失，这是不是一个好主意，以消除多余的水分从解剖脉络丛。生成图1比较的表达访问过的<html>在三个区域（纹状体，顶叶皮层和MAV）在控制条件下，使用安捷伦ES-014879全大鼠基因组4x44K 60mer寡核苷酸阵列（G4131F，安捷伦科技公司，帕洛阿尔托，加利福尼亚州NCBI GEO加入GPL7294; GSE23093和GSE29733存在NCBI的GEO库）。图中的两条曲线图示显示1）中的表达相比，顶叶皮质纹状体和2）相比的MAV顶叶皮质。很显然，纹状体和顶叶皮质的关系更加紧密地相互比MAV。脉络丛的MAV比较，数据将被呈现在未来的出版物。但是，它很可能在MAV和脉络丛的表达谱的关系更为密切，相互比他们的纹状体和顶叶皮质。 图2显示了基因的差异表达热疗（EIH）与AMPH相比，MAV控制和对方。 内容“&gt;超过11,000个基因与官方NCBI基因符号对照组动物在MAV相比，那些记录在纹状体和顶叶皮质的基因表达。以上这些基因中有2000年的2.5倍或更多不同的表达在MAV相比，两个神经元丰富的组织，和550的基因有一个大10倍的表达差异。略多于40％（253），这些基因的表达减少10倍或更MAV有关神经功能。相比，顶叶皮层和纹状体表达的MAV超过10倍或以上的343个基因，这种基因列表为出发点，以确定基因在MAV具有非常高的富集。 1列出了大于15倍的表达MAV基因相比，顶叶皮层和纹状体和分类 ​​方面的功能。其中许多基因都与血管系统和/或免疫系统。这些TYPES应富集的基因约5 - 至10 - 倍，因为增加的血管本身和血液本内。然而，即使是与已知的一般血管功能的基因，同比增长15倍以上，表明富集在MAV。也有倍的变化是非常大的多的基因; 1）胞外基质蛋白（不是特别相关，脉管系统），2）溶质转运和3）脂质和视黄酸代谢。由于好，一些生长和分化的基因或转录因子被发现。 <img alt="图1" src="/files/ftp_upload/4285/4285fig1.jpg" /> 图1。 MAV与顶叶皮层和纹状体基因表达的比较。火山情节比较基因表达水平的顶叶皮层和纹状体（上图）和顶叶皮层和MAV（下图）。红色的符号表示的基因，是显着不同的，即可连续通连接P &lt;0.05和地区的1.5倍或更大的区域差异表达。黑色符号代表不显着的区域之间不同（P&gt; 0.05）的基因，并小于1.5倍不同的表达式中。粉红色的符号表示的基因不到一个1.5倍的表达差异，但在统计学上显着性差异，P &lt;0.05。黄色符号识别基因表达的1.5倍或更高，但这种变化在统计上并不显着，P &lt;0.05。 缩略语 AMPH安非他明 EIH环境引起的高热 MAV的脑膜和相关脑血管 规范常温控制<img alt="图2" src="/files/ftp_upload/4285/4285fig2.jpg" /> 图2。热疗和安非他明MAV基因表达的影响。火山情节比较基因表达水平的MAV后，在3小时的时间点的生理盐水，EIH或AMPH。红色符号表示的基因被认为是显着不同，而黑点/圆圈代表不显着不同地区之间的基因。其他统计分析，以验证其表达控制和治疗之间的差异其实是统计上显著。 <table border="1"><tbody><tr><td fo:width="1in"> MAV表达脑表达 </td><td fo:width="4.5in"> NCBI基因符号 </td><td fo:width="1.5in"> 组织特异性或报告功能（S） </td></tr><tr><td> &gt; 50倍</td><td> ANXA2 一个 ，COL8A1，德，Esm1，Glycam1，KL 一个 ，Lect1，LEPR，LUM，MYH11，OGN，TAGLN，THBS2，TNNT2 </td><td>血管及心脏</td></tr><tr><td> &gt; 50倍</td><td> CCL19，CD74 一 ，DEFB1，Lrrc21，MGL1，MRC1，MSLN，PLa2g5，Prg4，RT1，RT1-DA-BB </td><td>免疫系统</td></tr><tr><td> &gt; 50倍</td><td> ADH1，Aebp1的，ALDH1A2，GSTM2 </td><td>维甲酸与脂质处理</td></tr><tr><td> &gt; 50倍</td><td> CDH1，COL3A1，COL1A2，Colec12，Cpxm2，氯丙嗪，DCN，EMP3，GPC3，Nupr1，OMD，PCOLCE Slamf9，Tmem27，Tspan8 </td><td>细胞外基质和细胞 - 细胞连接处</td></tr><tr><td> &gt; 50倍</td><td> AQP1，Asgr1，Kcnj13，Slc5a5，Slc6a13，Slc6a20，Slc22a6，Sned1，TTR </td><td>离子和溶质稳态</td></tr><tr><td> &gt; 50倍</td><td> Alx3，CDKN1C，FOXC2，IGFBP2，IFITM1，Ifitm2，Nkx6-1，OSR1，Prrx2，SFRP1，TBX15。 Upk1b，WISP2，Wnt6 </td><td>发展与转录调控</td></tr><tr><td> &gt; 50倍</td><td> Gpha2，Mfap5，Mpzl2 Plac8的，Scgb1c1，Sostdc1，Steap1 </td><td>未知及杂项</td></tr><tr><td> 30〜50倍</TD&gt; <td> ANXA1 一个 ，Angpt2，C6 一 ，GJB2，Ptgis，Thbd，TIMP1 </td><td>血管及心脏</td></tr><tr><td> 30〜50倍</td><td> Casp12，CCL2，CCR1，CD14，CXCL10，Ifitm3，Klra5，Lgals1，LGALS3，Ms4a4a，MS4A7，PLSCR1，SPP1，XCL1 </td><td>免疫系统</td></tr><tr><td> 30〜50倍</td><td> COL6A3，Cpxm1，Efemp1，FMOD，MGP，NID2 </td><td>细胞外基质和细胞 - 细胞连接处</td></tr><tr><td> 30〜50倍</td><td> CP，，SCT Cubn，GCGR，S100A6，Scn7a：Slc4a5，Slc16a11，Slco1a5 </td><td>离子和溶质平衡</td></tr><tr><td> 30〜50倍</td><td> CFD，Ch25h，CRABP2，Rarres2 </td><td>维甲酸与脂质处理</td></tr><tr><td> 30〜50倍</td><td> BMP6，Casp12，DAB2，FOLR1，IGF2，MSX1，Tbx18 TWIST1，Wnt5b </td><td>发展与转录调控</td></tr><tr><td> 30〜50倍</td><td> Cela3b，Cln6，C1qtnf7，Copz2，Dhrs7c，Gng11，Prss23时，Vim，SRPX </td><td>未知及杂项</td></tr><tr><td> 15〜30倍</td><td> ADM，Angpt1，血管生成素样蛋白，BGN，CKLF 一 ，Cnn1，Cox8h，CTSK，F13a1，GJA5，KLF4，液氧，LYZ，Myl9，PROCR，PROS1，RT1巴，Serpinb10，SERPING1，SERPINF1，TGM2，Tnmd，Trim63，TXNIP 一 ，Vamp5，VTN </td><td>血管及心脏</td></tr><tr><td> 15〜30倍</td><td> 阿达一 ，BST2，CCL6，CD40，CD68，CTSC，磷酸二铵，Faim3，Fkbp9，FXYD5，FMO1，GLIPR1，IER3，Ifi47，Igsf6，硕士，Nfatc4，RT1-DB1，Serpinb1a，Tir4，Tubb6 </td><td>免疫系统</td></tr><tr><td> 15〜30倍</td><td> Adamtsl4，COL1A1，Crb3的，DPT，Egfl3，FBN1，FBLN1，Fbln5，FN1，ITGB4，LAMA2，Lgals3bp，LOXL1，Mfap4，MMP14，Mmp23，PPIC，SERPINH1，TIMP3 </td><td>细胞外基质和细胞 - 细胞连接处</td></tr><tr><td> 15〜30倍</td><td> CYBRD1，SELENBP1，Slc2a4，Slc9的A2，Slc13a3，Slc16a4，Slc22a8，SLC22A18 </td><td>离子和溶质平衡</td></tr><tr><td> 15〜30倍</td><td> Agpat2，Bdh2，Cyp26b1，Lpar3，LTB4DH，OLR1，PON3，RBP1，RBP4 </td><td>维甲酸与脂质处理</td></tr><tr><td> 15〜30倍</td><td> ATF3，DKK4，EYA2，Ifitm7，Ltbp1，Mustn1，NR2F2，Ptrf，Sphk1，TGFBI，Tcea3 Tcfap2b，Wnt2b，而Wnt5a，ZIC1 </td><td>发展与转录调控</td></tr><tr><td> 15〜30倍</td><td> C1R Adamts12，Adamtsl3，Crispld2，Crygn，CYP1B1，DSE，ENPEP，ENPP2，Epn3，FLNA，FMO3，GNMT，GPRC5C，HSPB1，Klc3，KRT18，KRT19，MDK，Mesp1，Ms4a6a，Ms4a6b，Ms4a11，NET1，Pdlim2， PLP2 Phactr2，，Ppp1r3b，Pqlc3，RIN3，Spag11，SULT1A1，Tmem106a，WFIKKN2 </td><td>未知及杂项</td></tr></tbody></table> *表中的基因都必须是超过15倍以上表达在纹状体和pariet的人皮层和5倍以上的背景水平。这两个比率（MAV /纹状体MAV /顶叶皮层），每个基因被用于分组。 一个基因被发现在血管内皮细胞，但也有可能在介导的免疫反应中发挥了重要作用。 表1。了15倍*或更增加了表达的MAV相比，纹状体和顶叶皮层的基因。

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Sci. 17, 227-230 (2002).</li><li>Johnston, M., Zakharov, A., Koh, L., Armstrong, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+injection+of+Microfil+reveals+connections+between+cerebrospinal+fluid+and+nasal+lymphatics+in+the+non-human+primate.">Subarachnoid injection of Microfil reveals connections between cerebrospinal fluid and nasal lymphatics in the non-human primate.</a> Neuropathol. Appl. Neurobiol. 31, 632-640 (2005).</li><li>Johnston, 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=Evidence+of+connections+between+cerebrospinal+fluid+and+nasal+lymphatic+vessels+in+humans,+non-human+primates+and+other+mammalian+species.">Evidence of connections between cerebrospinal fluid and nasal lymphatic vessels in humans, non-human primates and other mammalian species.</a> Cerebrospinal Fluid Res. 1, 2 (2004).</li><li>Kulik, T., 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=Regulation+of+cerebral+vasculature+in+normal+and+ischemic+brain.">Regulation of cerebral vasculature in normal and ischemic brain.</a> Neuropharmacol. 55, 281-288 (2008).</li><li>Bowyer, J. F., Ali, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+doses+of+methamphetamine+that+cause+disruption+of+the+blood-brain+barrier+in+limbic+regions+produce+extensive+neuronal+degeneration+in+mouse+hippocampus.">High doses of methamphetamine that cause disruption of the blood-brain barrier in limbic regions produce extensive neuronal degeneration in mouse hippocampus.</a> Synapse. 60, 521-532 (2006).</li><li>Sharma, H. S., Westman, J., Nyberg, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiology+of+brain+edema+and+cell+changes+following+hyperthermic+brain+injury.">Pathophysiology of brain edema and cell changes following hyperthermic brain injury.</a> Prog. Brain Res. 115, 351-412 (1998).</li><li>Sharma, H. S., Sjoquist, P. O., Ali, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drugs+of+abuse-induced+hyperthermia,+blood-brain+barrier+dysfunction+and+neurotoxicity:+neuroprotective+effects+of+a+new+antioxidant+compound+H-290/51.">Drugs of abuse-induced hyperthermia, blood-brain barrier dysfunction and neurotoxicity: neuroprotective effects of a new antioxidant compound H-290/51.</a> 13, 1903-1923 (2007).</li><li>Ho, E. L., Josephson, S. A., Lee, H. S., Smith, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebrovascular+complications+of+methamphetamine+abuse.">Cerebrovascular complications of methamphetamine abuse.</a> Neurocrit. Care. 10, 295-305 (2009).</li><li>Ohta, K., 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=Delayed+ischemic+stroke+associated+with+methamphetamine+use.">Delayed ischemic stroke associated with methamphetamine use.</a> J. Emerg. Med. 28, 165-167 (2005).</li><li>Brenner, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropsychological+and+neuroimaging+findings+in+traumatic+brain+injury+and+post-traumatic+stress+disorder.">Neuropsychological and neuroimaging findings in traumatic brain injury and post-traumatic stress disorder.</a> Dialogues Clin. Neurosci. 13, 311-323 (2011).</li><li>Case, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflicted+traumatic+brain+injury+in+infants+and+young+children.">Inflicted traumatic brain injury in infants and young children.</a> Brain Pathol. 18, 571-582 (2008).</li><li>Maas, A. I., 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=Prognostic+value+of+computerized+tomography+scan+characteristics+in+traumatic+brain+injury:+results+from+the+IMPACT+study.">Prognostic value of computerized tomography scan characteristics in traumatic brain injury: results from the IMPACT study.</a> J. Neurotrauma. 24, 303-314 (2007).</li></ol>

我们什么都没有透露。这里的工作不一定代表美国食品和药物管理局（FDA）的意见。

技术方面的MAV和脉络丛解剖至关重要的是在剥离的MAV有大脑“淹没”的大部分时间在生理盐水或钠的磷酸盐缓冲盐水。 MAV和皮质脱水剥离时，将导致在MAV坚持紧紧皮质层一，这与MAV和/或造成无法删除这些坚持MAV部分，从皮质的皮质组织，收获量会增加。剥离时，耐心是必要的。同样，如果删除的MAV从一个半球皮质剥离时，遇到了一些阻力，切换到另一半球，回去以后遇到反抗的一侧。这种方法确实需要一些练习来完善，并需要约5〜15“实践”解剖之前，实现均匀一致的收获的MAV组织的。开始的解剖和圣雷莫的方法，在脑底动脉环的MAV val且便于去除脑膜通过使用较大的血管为支持“脚手架”。 大部分血液最初在MAV血管后牺牲应在剥离喷出，和小于5％的RNA收获MAV应该是血源性。这是可能的，以除去几乎所有的血液灌流动物用50至70毫升的生理盐水中，用组织学技术之前，去除斩首和脑。但是，它是可视化的MAV解剖组织中要困难得多。三个最可能的“污染”的MAV组织来源是松果体，皮质层I组织和残余嗅球组织，但它是在该制剂中，也有可能得到垂体组织。在MAV松果体污染被检测到由MAV含有一个圆形的1至2毫米直径的球体是有色的暗红色。的主要功能松果体通常被认为是从MAV是完全不同的。它的主要功能是产生褪黑激素，调节各方面的昼夜节律9，和10脂质前体lipoxygenation。虽然该基因的低 ，调节脂肪氧化酶活性，被认为是目前在成人主要是在松果体10，我们的结果表明，它是在显着水平MAV也始终存在。残余在幅状的粉红色MAV组织具有约1​​至2毫米的致密组织，是很大括在在颜色苍白的的皮质或嗅球污染结果。这可以通过“浮动”的MAV的缓冲液的表面上，然后使用两个解剖钳除去较重的皮质或灯泡组织发现。 如果污染的MAV由我组织的皮质层，下丘脑组织或松果体存在，那么将有一个显着的几个基因着地表达比通常存在于一个“干净”解剖。污染的MAV松果体会导致高表达芳烷脘N-乙酰转移酶（AANAT），这是必要的褪黑激素合成。不幸的是，如果松果体污染过程中发生MAV夹层，MAV 液氧在水平将不能够被精确地确定。皮质组织的MAV的污染会导致神经元特异性基因，如hippocalcin（HPCA），小白蛋白（Pvalb）和/或神经元五聚的受体（Nptxr）中高表达。下丘脑组织的MAV污染会导致在高表达的生长激素1（GH1），阿黑皮素原（POMC）。事件，确实存在一定的污染，在收获组织，大部分这些基因可以被识别，而不是用在基因表达分析。这是特别重要的基因，存在于循环BLO外径，MAV或脉络膜丛的可能仍然存在。 影视之收获MAV和脉络丛的基因表达数据血管以及与相关的脑膜（MAV）和脉络丛的脑表面（表面）是必要的大脑血液流量和脑脊髓液（CSF）的动态平衡4,11，12，13，14，15，16，17。该组织的收获是适合生理和生化分析，MAV已被证明是敏感的损坏生产的安非他明和热疗1,2通过基因表达分析。这些结果是相称的，也被称为严重的高热和安非他明的脑血管一般18,19,20曝光。人体临床数据支持的信念，硬膜下血管容易损坏的安非他明和甲基安非他明21,22。同时，主要和次要的的脑vasculatures含ING在软脑膜层的脑膜，MAV的收获是潜在的高利率调查震荡的头部外伤23,24 25时。目前的基因的表达谱产生的MAV表示，可能是软脑膜和蛛网膜脑脊膜膜可能会发挥更大的作用比预期的调节CSF组成。在未来，通过解剖显微镜技术的使用，它可以是可能的，以进一步分离的组织，包括使得软脑膜和蛛网膜膜可以从较大的​​动脉血管本分离，并可能从每个其他的MAV。这将使更好的诠释了这3个组成部分的收获MAV的功能。

<table border="1"><tbody><tr><td> 的试剂/设备名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>生理盐水（盐水/9克，1升H 2 O） </td><td>在内部</td><td></td><td>解剖缓冲</td></tr><tr><td> 0.1 M磷酸钠pH值7.4 </td><td>在内部</td><td></td><td>解剖缓冲</td></tr><tr><td>骨咬骨钳</td><td> Roboz </td><td> RS-8300 </td><td>去除颅骨</td></tr><tr><td>钳弯尖（4“长，0.8毫米宽的尖） </td><td> Roboz </td><td> RS-5135或RS-5137 </td><td>解剖钳</td></tr><tr><td>钳直头（5.5“） </td><td> Roboz </td><td> RS-8124或RS-8104 </td><td>拇指敷料钳</td></tr></tbody></table>

a visual description of the dissection of the cerebral surface vasculature and associated meninges and the choroid plexus from rat brain

该视频演示显示的方法收获​​的两个最重要的血管结构，而不是居住在大脑中正确的，即支持前脑功能。它们是血管以及与相关的脑膜（MAV）和脉络膜丛这是必要的大脑血液流量和脑脊髓液（CSF）的稳态的脑表面（表面）。收获的组织是适合生理和生化分析，和MAV已被证明是敏感的损坏所产生安非他明和热疗1,2。同时，在MAV的主要和轻微脑vasculatures的，收获的是调查震荡，头部外伤时，潜在的高利率。在此演示文稿中的MAV解剖的软脑膜和蛛网膜少硬脑膜的脑膜及脑表面血管的主要和次要一些。脉络丛的解剖结构驻留在升抵押品脑室所描述的Oldfield和麦金利3,4,5,6。收获这两个组织所采用的方法，也有利于区域皮质组织缺乏脑膜及脑表面血管的收获，是兼容的收获大脑的其他组织，如纹状体，下丘脑，海马，等这两个组织需要的解剖从总5到10分钟。可以发现，如图所示，在此演示文稿中的基因表达水平的解剖MAV和脉络丛，在GSE23093（MAV）和GSE29733（脉络膜丛）在NCBI的GEO资料库。此数据已经，正在，用于帮助进一步了解MAV和脉络丛的功能和神经中毒的事件，如严重的高热和AMPH如何影响他们的功能。

Watch this Scientific Journal Video about A Visual Description of the Dissection of the Cerebral Surface Vasculature and Associated Meninges and the Choroid Plexus from Rat Brain at JoVE.com

A Visual Description of the Dissection of the Cerebral Surface Vasculature and Associated Meninges and the Choroid Plexus from Rat Brain

该视频演示了一种方法收获的两个最重要的的高度血管结构，支持前脑功能。它们是血管以及与相关的脑膜（MAV）和脉络膜丛这是必要的大脑血液流量和脑脊髓液（CSF）的稳态的脑表面（表面）。

从大鼠脑的解剖脑表面血管及相关脑膜和脉络丛的可视描述，

This video presentation was created to show a method of harvesting the two most important highly vascular structures, not residing within the brain ...

a-visual-description-dissection-cerebral-surface-vasculature

Research

JoVE Journal

Neuroscience

47.9K 次观看.  National Center for Toxicological Research. 该视频演示了一种方法收获的两个最重要的的高度血管结构，支持前脑功能。它们是血管以及与相关的脑膜（MAV）和脉络膜丛这是必要的大脑血液流量和脑脊髓液（CSF）的稳态的脑表面（表面）。

视频: A Visual Description of the Dissection of the Cerebral Surface Vasculature and Associated Meninges and the Choroid Plexus from Rat Brain

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable clinical state; in the development of animal models of focal ischemia, however, achieving reproducibility of experimentally induced infarct volume is essential. The rat is a widely used animal model for stroke due to its relatively low animal husbandry costs and to the similarity of its cranial circulation to that of humans2,3. In humans, the middle cerebral artery (MCA) is most commonly affected in stroke syndromes and multiple methods of MCA occlusion (MCAO) have been described to mimic this clinical syndrome in animal models. Because recanalization commonly occurs following an acute stroke in the human, reperfusion after a period of occlusion has been included in many of these models. In this video, we demonstrate the transient endovascular suture MCAO model in the spontaneously hypertensive rat (SHR). A filament with a silicon tip coating is placed intraluminally at the MCA origin for 60 minutes, followed by reperfusion. Note that the optimal occlusion period may vary in other rat strains, such as Wistar or Sprague-Dawley. Several behavioral indicators of stroke in the rat are shown. Focal ischemia is confirmed using T2-weighted magnetic resonance images and by staining brain sections with 2,3,5-triphenyltetrazolium chloride (TTC) 24 hours after MCAO.

Surgical induction of ischemic brain damage in the rat is a widely used model for stroke research. Here we demonstrate the induction of focal cerebral ischemia by occlusion of the middle cerebral artery. Visualization of the resulting infarct by histological staining and magnetic resonance imaging is also shown.

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

科研

神经科学

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to monitor the disease course in the follow-up period. Changes in the VEP parameters closely correlate with pathological damage in the optic nerve. This protocol provides a detailed description about the rodent model of optic nerve microinjection, in which a partial demyelination lesion is produced in the optic nerve. VEP recording techniques are also discussed. Using skull implanted electrodes, we are able to acquire reproducible intra-session and between-session VEP traces. VEPs can be recorded on individual animals over a period of time to assess the functional changes in the optic nerve longitudinally. The optic nerve demyelination model, in conjunction with the VEP recording protocol, provides a tool to investigate the disease processes associated with demyelination and remyelination, and can potentially be employed to evaluate the effects of new remyelinating drugs or neuroprotective therapies.

Focal demyelination is induced in the optic nerve using lysolecithin microinjection. Visual evoked potentials are recorded via skull electrodes implanted over the visual cortex to examine the signal conduction along the visual pathway in vivo. This protocol details the surgical procedures underlying electrode implantation and optic nerve microinjection.

在实验视神经脱髓鞘模型大鼠视觉诱发电位记录

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

Observation of the Ciliary Movement of Choroid Plexus Epithelial Cells Ex Vivo

The choroid plexus is located in the ventricular wall of the brain, the main function of which is believed to be production of cerebrospinal fluid. Choroid plexus epithelial cells (CPECs) covering the surface of choroid plexus tissue harbor multiple unique cilia, but most of the functions of these cilia remain to be investigated. To uncover the function of CPEC cilia with particular reference to their motility, an ex vivo observation system was developed to monitor ciliary motility during embryonic, perinatal and postnatal periods. The choroid plexus was dissected out of the brain ventricle and observed under a video-enhanced contrast microscope equipped with differential interference contrast optics. Under this condition, a simple and quantitative method was developed to analyze the motile profiles of CPEC cilia for several hours ex vivo. Next, the morphological changes of cilia during development were observed by scanning electron microscopy to elucidate the relationship between the morphological maturity of cilia and motility. Interestingly, this method could delineate changes in the number and length of cilia, which peaked at postnatal day (P) 2, while the beating frequency reached a maximum at P10, followed by abrupt cessation at P14. These techniques will enable elucidation of the functions of cilia in various tissues. While related techniques have been published in a previous report1, the current study focuses on detailed techniques to observe the motility and morphology of CPEC cilia ex vivo.

Observation of the Ciliary Movement of Choroid Plexus Epithelial Cells Ex Vivo

In this study, a detailed light microscopic technique was optimized for real-time observation and analysis of the motion of CPEC cilia ex vivo together with an electron microscopic method for ultrastructural analysis.

脉络丛上皮细胞的纤毛运动的观察<em&gt;离体</em

The choroid plexus is located in the ventricular wall of the brain, the main function of which is believed to be production of cerebrospinal fluid. ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful analysis of synaptic transmission modulation when performing field potential or intracellular recordings. This manuscript describes methodological aspects that might be helpful in improving experimental conditions for the maintenance of acute brain slices and for recording field excitatory postsynaptic potentials in a commercially available submersion chamber with an outflow-carbogenation unit. The outflow-carbogenation helps to stabilize the oxygen level in experiments that rely on the recycling of a small buffer reservoir to enhance the cost-efficiency of drug experiments. In addition, the manuscript presents representative experiments that examine the effects of different carbogenation modes and stimulation paradigms on the activity-dependent synaptic plasticity of synaptic transmission.

This protocol describes the stabilization of the oxygen level in a small volume of recycled buffer and methodological aspects of recording activity-dependent synaptic plasticity in submerged acute hippocampal slices.

小容积回收-灌注-浸没式室系统中维持的急性海马切片的突触可塑性记录

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...

Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System

During cerebral cortex development, progenitor cells undergo several rounds of symmetric and asymmetric cell divisions to generate new progenitors or postmitotic neurons. Later, some progenitors switch to a gliogenic fate, adding to the astrocyte and oligodendrocyte populations. Using time-lapse video-microscopy of primary cerebral cortex cell cultures, it is possible to study the cellular and molecular mechanisms controlling the mode of cell division and cell cycle parameters of progenitor cells. Similarly, the fate of postmitotic cells can be examined using cell-specific fluorescent reporter proteins or post-imaging immunocytochemistry. More importantly, all these features can be analyzed at the single-cell level, allowing the identification of progenitors committed to the generation of specific cell types. Manipulation of gene expression can also be performed using viral-mediated transfection, allowing the study of cell-autonomous and non-cell-autonomous phenomena. Finally, the use of fusion fluorescent proteins allows the study of symmetric and asymmetric distribution of selected proteins during division and the correlation with daughter cells fate. Here, we describe the time-lapse video-microscopy method to image primary cerebral cortex murine cells for up to several days and analyze the mode of cell division, cell cycle length and fate of newly generated cells. We also describe a simple method to transfect progenitor cells, which can be applied to manipulate genes of interest or simply label cells with reporter proteins.

Live imaging is a powerful tool to study cellular behaviors in real time. Here, we describe a protocol for time-lapse video-microscopy of primary cerebral cortex cells that allows a detailed examination of the phases enacted during the lineage progression from primary neural stem cells to differentiated neurons and glia.

生活成像的脑皮层细胞使用二维培养系统

During cerebral cortex development, progenitor cells undergo several rounds of symmetric and asymmetric cell divisions to generate new progenitors or ...

A Volumetric Method for Quantification of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor determining patient outcome and is therefore frequently taken as a study endpoint. However, in small animal studies on SAH, quantification of cerebral vasospasm is a major challenge. Here, an ex vivo method is presented that allows quantification of volumes of entire vessel segments, which can be used as an objective measure to quantify cerebral vasospasm. In a first step, endovascular casting of the cerebral vasculature is performed using a radiopaque casting agent. Then, cross-sectional imaging data are acquired by micro computed tomography. The final step involves 3-dimensional reconstruction of the virtual vascular tree, followed by an algorithm to calculate center lines and volumes of the selected vessel segments. The method resulted in a highly accurate virtual reconstruction of the cerebrovascular tree shown by a diameter-based comparison of anatomical samples with their virtual reconstructions. Compared with vessel diameters alone, the vessel volumes highlight the differences between vasospastic and non-vasospastic vessels shown in a series of SAH and sham-operated mice.

The goal of this article is to present a method that allows a 3-dimensional reconstruction of the cerebrovascular tree in mice after micro computed tomography and determination of volumes of entire vessel segments that can be used to quantify cerebral vasospasm in murine models of subarachnoid hemorrhage.

小鼠蛛网膜下腔出血模型脑血管痉挛量化的容积法

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor ...

Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral vascular effects. Impact (i.e., non-blast) traumatic brain injury (TBI) is known to decrease pressure autoregulation in the cerebral vasculature in both humans and experimental animals. The hypothesis that blast-induced traumatic brain injury (bTBI), like impact TBI, results in impaired cerebral vascular reactivity was tested by measuring myogenic dilatory responses to reduced intravascular pressure in rodent middle cerebral arterial (MCA) segments from rats subjected to mild bTBI using an Advanced Blast Simulator (ABS) shock tube. Adult, male Sprague-Dawley rats were anesthetized, intubated, ventilated and prepared for Sham bTBI (identical manipulation and anesthesia except for blast injury) or mild bTBI. Rats were randomly assigned to receive Sham bTBI or mild bTBI followed by sacrifice 30 or 60 min post-injury. Immediately after bTBI, righting reflex (RR) suppression times were assessed, euthanasia at the time points post-injury was completed, the brain was harvested and the individual MCA segments were collected, mounted and pressurized. As the intraluminal pressure perfused through the arterial segments was reduced in 20 mmHg increments from 100 to 20 mmHg, MCA diameters were measured and recorded. With decreasing intraluminal pressure, MCA diameters steadily increased significantly above baseline in the Sham bTBI groups while MCA dilator responses were significantly reduced (p &#60; 0.05) in both bTBI groups as evidenced by the impaired, smaller MCA diameters recorded for the bTBI groups. In addition, RR suppression in the bTBI groups was significantly (p &#60; 0.05) higher than in the Sham bTBI groups. MCA&#39;s collected from the Sham bTBI groups exhibited typical vasodilatory properties to decreases in intraluminal pressure while MCA&#39;s collected following bTBI exhibited significantly impaired myogenic vasodilatory responses to reduced pressure that persisted for at least 60 min after bTBI.

Here, we present a protocol to describe methods for ex vivo vascular reactivity determination following a primary blast traumatic brain injury (bTBI) using isolated, pressurized, rodent middle cerebral arterial (MCA) segments. bTBI induction is accomplished using a shock tube, also known as an Advanced Blast Simulator (ABS) device.

爆炸性神经损伤对压生啮鼠中脑动脉的影响

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to intracranial delivery, intra-arterial administration of NSCs is less invasive and produces a more diffuse distribution of NSCs within the brain parenchyma. Further, intra-arterial delivery allows the first-pass effect in the brain circulation, lessening the potential for trapping of cells in peripheral organs, such as liver and spleen, a complication associated with peripheral injections. Here, we detail the methodology, in both mice and rats, for delivery of NSCs through the common carotid artery (mouse) or external carotid artery (rat) to the ipsilateral hemisphere after an ischemic stroke. Using GFP-labeled NSCs, we illustrate the widespread distribution achieved throughout the rodent ipsilateral hemisphere at 1 d, 1 week and 4 weeks after postischemic delivery, with a higher density in or near the ischemic injury site. In addition to long-term survival, we show evidence of differentiation of GFP-labeled cells at 4 weeks. The intra-arterial delivery approach described here for NSCs can also be used for administration of therapeutic compounds, and thus has broad applicability to varied CNS injury and disease models across multiple species.

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

神经干细胞向大鼠和小鼠大脑的动脉内传递：对脑缺血的应用

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...

Measuring Post-Stroke Cerebral Edema, Infarct Zone and Blood-Brain Barrier Breakdown in a Single Set of Rodent Brain Samples

One of the most common causes of morbidity and mortality worldwide is ischemic stroke. Historically, an animal model used to stimulate ischemic stroke involves middle cerebral artery occlusion (MCAO). Infarct zone, brain edema and blood-brain barrier (BBB) breakdown are measured as parameters that reflect the extent of brain injury after MCAO. A significant limitation to this method is that these measurements are normally obtained in different rat brain samples, leading to ethical and financial burdens due to the large number of rats that need to be euthanized for an appropriate sample size. Here we present a method to accurately assess brain injury following MCAO by measuring infarct zone, brain edema and BBB permeability in the same set of rat brains. This novel technique provides a more efficient way to evaluate the pathophysiology of stroke.

This protocol describes a novel technique of measuring the three most important parameters of ischemic brain injury on the same set of rodent brain samples. Using only one brain sample is highly advantageous in terms of ethical and economic costs.

测量一组啮齿动物脑样本中的中风后脑水肿、梗塞区和血脑屏障破裂

One of the most common causes of morbidity and mortality worldwide is ischemic stroke. Historically, an animal model used to stimulate ischemic stroke ...

Extraction and Dissection of the Domesticated Pig Brain

Use of the pig as a preclinical and translatable animal model has been well-documented and accepted by research fields investigating cardiovascular systems, gastrointestinal systems, and nutrition, and the pig is increasingly being used as a large animal model in neuroscience. Furthermore, the pig is an accepted model to study neurodevelopment as it displays brain growth and development patterns similar to what occurs in humans. As a less common animal model in neuroscience, surgical and dissection procedures on pigs may not be as familiar or well-practiced among researchers. Therefore, a standardized visual protocol detailing consistent extraction and dissection methods may prove valuable for researchers working with the pig. The following video showcases a technique to remove the pig brain while keeping the cortex and brainstem intact and reviews methods to dissect several commonly investigated brain regions including the brainstem, cerebellum, midbrain, hippocampus, striatum, thalamus, and medial prefrontal cortex. The purpose of this video is to provide researchers with the tools and knowledge necessary to consistently perform a brain extraction and dissection on the four-week-old pig.

This protocol details the technique for removal of the pig brain in its entirety and dissection of several brain regions commonly studied in neuroscience.

驯养猪脑的提取和解剖

Use of the pig as a preclinical and translatable animal model has been well-documented and accepted by research fields investigating cardiovascular ...