Name: an organotypic slice assay for high-resolution time-lapse imaging of neuronal migration in the postnatal brain
Uploaded: 2010-12-11T16:00:00
Description: Watch this Scientific Journal Video about An Organotypic Slice Assay for High-Resolution Time-Lapse Imaging of Neuronal Migration in the Postnatal Brain at JoVE.com

我々はビデオでプロトコルを語るためのダンマホーターに感謝。この作品は、NIHのグラント5R01NS062182、老化研究のためのアメリカ連合からの助成金、およびHTGに授与機関の資金によってサポートされています。

I.手続次のような手法は、滅菌のツールを使用して、層流フードで、無菌条件下で実行する必要があります。 器官型スライス用ガラスボトムディッシュの準備<ol><li>料理は、無菌環境で製造し、滅菌のツールを使用している必要があります。 </li><li>スライス培地の150μLドロップは（レシピを参照）培地中の気泡を避けるために注意して皿のガラス底?...

<ol>
	<li>Ghashghaei, H. T., Lai, C., Anton, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+migration+in+the+adult+brain:+are+we+there+yet.">Neuronal migration in the adult brain: are we there yet.</a> Nat. Rev. Neurosci. 8, 141-151 (2007).</li><li>Valiente, M., Marin, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+migration+mechanisms+in+development+and+disease.">Neuronal migration mechanisms in development and disease.</a> Curr. Opin. Neurobiol. 20, 68-78 (2010).</li><li>Rakic, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+of+the+neocortex:+a+perspective+from+developmental+biology.">Evolution of the neocortex: a perspective from developmental biology.</a> Nat. Rev. Neurosci. 10, 724-735 (2009).</li><li>Jaglin, X. H., Chelly, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tubulin-related+cortical+dysgeneses:+microtubule+dysfunction+underlying+neuronal+migration+defects.">Tubulin-related cortical dysgeneses: microtubule dysfunction underlying neuronal migration defects.</a> Trends Genet. 25, 555-566 (2009).</li><li>Carro, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+transcriptional+network+for+mesenchymal+transformation+of+brain+tumours.">The transcriptional network for mesenchymal transformation of brain tumours.</a> Nature. 463, 318-325 (2010).</li><li>Wu, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Directional+guidance+of+neuronal+migration+in+the+olfactory+system+by+the+protein+Slit.">Directional guidance of neuronal migration in the olfactory system by the protein Slit.</a> Nature. 400, 331-336 (1999).</li><li>Hu, H., Tomasiewicz, H., Magnuson, T., Rutishauser, U., U, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+polysialic+acid+in+migration+of+olfactory+bulb+interneuron+precursors+in+the+subventricular+zone.">The role of polysialic acid in migration of olfactory bulb interneuron precursors in the subventricular zone.</a> Neuron. 16, 735-743 (1996).</li><li>Shapiro, E. M., Gonzalez-Perez, O., Garcia-Verdugo, M. a. n. u. e. l., Alvarez-Buylla, J., &amp;amp, A., Koretsky, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetic+resonance+imaging+of+the+migration+of+neuronal+precursors+generated+in+the+adult+rodent+brain.">Magnetic resonance imaging of the migration of neuronal precursors generated in the adult rodent brain.</a> Neuroimage. , (2006).</li><li>Vreys, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI+visualization+of+endogenous+neural+progenitor+cell+migration+along+the+RMS+in+the+adult+mouse+brain:+validation+of+various+MPIO+labeling+strategies.">MRI visualization of endogenous neural progenitor cell migration along the RMS in the adult mouse brain: validation of various MPIO labeling strategies.</a> Neuroimage. 49, 2094-2103 (2010).</li><li>Davenne, M., Custody, C., Charneau, P., Lledo, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+of+migrating+neurons+in+the+mammalian+forebrain.">In vivo imaging of migrating neurons in the mammalian forebrain.</a> Chem. Senses. 30, 115-116 (2005).</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.">The subventricular zone en-face: wholemount staining and ependymal.</a> J. Vis. Exp. , (2010).</li><li>Shen, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+SVZ+stem+cells+lie+in+a+vascular+niche:+a+quantitative+analysis+of+niche+cell-cell+interactions.">Adult SVZ stem cells lie in a vascular niche: a quantitative analysis of niche cell-cell interactions.</a> Cell Stem Cell. 3, 289-300 (2008).</li><li>Tavazoie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+specialized+vascular+niche+for+adult+neural+stem+cells.">A specialized vascular niche for adult neural stem cells.</a> Cell Stem Cell. 3, 279-288 (2008).</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>Polleux, F. &. a. m. p. ;. a. m. p., Ghosh, 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+slice+overlay+assay:+a+versatile+tool+to+study+the+influence+of+extracellular+signals+on+neuronal.">The slice overlay assay: a versatile tool to study the influence of extracellular signals on neuronal.</a> Sci. STKE. , L9-L9 (2002).</li><li>Murase, S. &. a. m. p. ;. a. m. p., Horwitz, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deleted+in+colorectal+carcinoma+and+differentially+expressed+integrins+mediate+the+directional+migration+of+neural+precursors+in+the+rostral+migratory+stream.">Deleted in colorectal carcinoma and differentially expressed integrins mediate the directional migration of neural precursors in the rostral migratory stream.</a> J. Neurosci. 22, 3568-3579 (2002).</li><li>Suzuki, S. O. &. a. m. p. ;. a. m. p., Goldman, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+cell+populations+in+the+early+postnatal+subventricular+zone+take+distinct+migratory+pathways:+a+dynamic+study+of+glial+and+neuronal+progenitor+migration.">Multiple cell populations in the early postnatal subventricular zone take distinct migratory pathways: a dynamic study of glial and neuronal progenitor migration.</a> J. Neurosci. 23, 4240-4250 (2003).</li><li>Ghashghaei, H. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+neuregulin-ErbB4+interactions+on+the+proliferation+and+organization+of+cells+in+the+subventricular+zone.">The role of neuregulin-ErbB4 interactions on the proliferation and organization of cells in the subventricular zone.</a> Proc. Natl. Acad. Sci. U. S. A. 103, 1930-1935 (2006).</li><li>Khodosevich, K., Seeburg, P. H., Monyer, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Major+signaling+pathways+in+migrating+neuroblasts.">Major signaling pathways in migrating neuroblasts.</a> Front Mol. Neurosci. 2, 7-7 (2009).</li><li>Jacquet, B. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+neuronal+proliferation,+migration+and+differentiation+in+the+postnatal+brain+using+equine+infectious+anemia+virus-based+lentiviral+vectors.">Analysis of neuronal proliferation, migration and differentiation in the postnatal brain using equine infectious anemia virus-based lentiviral vectors.</a> Gene Ther. 16, 1021-1033 (2009).</li></ol>

RMSの神経細胞の移動は、嗅球1の出生後の神経新生の重要なコンポーネントです。 RMSを介しての移行は、脳の表面への接平面で発生します。接線方向に移行する神経芽細胞が放射状にその前駆細胞のソースの場所だけでなく、彼らの最終的な神経の製品1、2、3の分岐運命に基づいて細胞の移行とは別個のものである。出生後のRMSでの接線方向に移行し細胞の比較的純粋な人口...

an organotypic slice assay for high-resolution time-lapse imaging of neuronal migration in the postnatal brain

既存の神経回路に前駆細胞、神経芽細胞の遊走、だけでなく、新たな神経細胞の分化と統合の増殖：出生後の脳における神経新生は、3つの生物学的事象のメンテナンスに依存します。嗅球における出生後の神経新生のために、これらのイベントは3つの解剖学的に別個のドメイン内で分離されています：増殖は主に神経芽細胞は吻側渡り鳥ストリーム（RMS）を横断し、新しい神経細胞が分化し、移行、側脳室の上衣ゾーン（SEZ）で発生嗅球（OB）内に統合する。三つのドメインは、はっきりと生物学的事象の各々を調節する、細胞の分子、および生理学的メカニズムを研究する理想的なプラットフォームとして機能する。このホワイトペーパーでは、細胞外の条件が密接に神経芽を移行するための生体内環境で模倣した生後脳組織に最適化された器官スライスアッセイを、説明しています。私たちは、アッセイは、RMS内神経芽細胞の、均一な指向、そしてスピーディな動きを提供することを示している。このアッセイは、異なる遺伝的背景の上にマウスからのクロス移植のアプローチを利用することにより、ニューロン移動の細胞自律的で、非自律的規制の研究のために非常に適したものとなる。

Watch this Scientific Journal Video about An Organotypic Slice Assay for High-Resolution Time-Lapse Imaging of Neuronal Migration in the Postnatal Brain at JoVE.com

An Organotypic Slice Assay for High-Resolution Time-Lapse Imaging of Neuronal Migration in the Postnatal Brain

このプロトコルは、吻側渡り鳥ストリームにおける出生後の脳および神経芽細胞の遊走の高解像度のタイムラプスイメージングに最適化された器官スライスのアッセイを説明します。

出生後の脳における神経細胞の移動の高分解能タイムラプスイメージングのための器官型スライスアッセイ

Neurogenesis in the postnatal brain depends on maintenance of three biological events: proliferation of progenitor cells, migration of neuroblasts, as ...

Research

JoVE Journal

Neuroscience

The Organotypic Hippocampal Slice Culture Model for Examining Neuronal Injury

神経損傷を調べるための器官型海馬スライス培養モデル

Organotypic hippocampal slice culture is an in vitro method to examine mechanisms of neuronal injury in which the basic architecture and composition of ...

Time-lapse Live Imaging of Clonally Related Neural Progenitor Cells in the Developing Zebrafish Forebrain

開発ゼブラフィッシュ前脳におけるクローン関連の神経前駆細胞のタイムラプスライブイメージング

Precise patterns of division, migration and differentiation of neural progenitor cells are crucial for proper brain development and function1,2. To ...

Multi-electrode Array Recordings of Neuronal Avalanches in Organotypic Cultures

器官培養における神経雪崩のマルチ電極アレイの録音

The cortex is spontaneously active, even in the absence of any particular input or motor output.  During development, this activity is important for the ...

Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor

遺伝的にコードされたカルシウムセンサーを発現する鋤鼻器のスライス標本のイメージング神経細胞応答

The vomeronasal organ (VNO) detects chemosensory signals that carry information about the social, sexual and reproductive status of the individuals within ...

High-resolution Functional Magnetic Resonance Imaging Methods for Human Midbrain

人間の中脳の高解像度機能的磁気共鳴画像法

Functional MRI (fMRI) is a widely used tool for non-invasively measuring correlates of human brain activity. However, its use has mostly been focused upon ...

Time-lapse Imaging of Neuroblast Migration in Acute Slices of the Adult Mouse Forebrain

成体マウスの前脳の急性スライスにおける神経芽細胞の移行のタイムラプスイメージング

There is a substantial body of evidence indicating that new functional neurons are constitutively generated from an endogenous pool of neural stem cells ...

In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

生体産後エレクトロポレーションおよびマウスの急性脳スライスにおける神経芽細胞移動のタイムラプスイメージング中

The subventricular zone (SVZ) is one of the main neurogenic niches in the postnatal brain. Here, neural progenitors proliferate and give rise to ...

Genetic Manipulation of Cerebellar Granule Neurons In Vitro and In Vivo to Study Neuronal Morphology and Migration

Genetic Manipulation of Cerebellar Granule Neurons In Vitro and In Vivo to Study Neuronal Morphology and Migration

小脳顆粒ニューロンの遺伝子操作<em&gt;体外</em&gt;と<em&gt;インビボ</em&gt;ニューロンの形態と移行を研究するために、

Developmental events in the brain including neuronal morphogenesis and migration are highly orchestrated processes. In vitro and in vivo analyses allow ...

High-resolution In Vivo Manual Segmentation Protocol for Human Hippocampal Subfields Using 3T Magnetic Resonance Imaging

High-resolution In Vivo Manual Segmentation Protocol for Human Hippocampal Subfields Using 3T Magnetic Resonance Imaging

高解像度<em&gt;インビボ</em&gt; 3T磁気共鳴画像を用いたヒト海馬サブフィールドのためのマニュアルセグメンテーションプロトコル

The human hippocampus has been broadly studied in the context of memory and normal brain function and its role in different neuropsychiatric disorders has ...

Time-lapse Confocal Imaging of Migrating Neurons in Organotypic Slice Culture of Embryonic Mouse Brain Using In Utero Electroporation

Time-lapse Confocal Imaging of Migrating Neurons in Organotypic Slice Culture of Embryonic Mouse Brain Using In Utero Electroporation

胚性マウス脳の器官型スライス培養における移行ニューロンの経時的共焦点イメージング<em&gt;ウテロの</em&gt;エレクトロポレーション

In utero electroporation is a rapid and powerful approach to study the process of radial migration in the cerebral cortex of developing mouse embryos. It ...