私たちの活動は、NIHの助成金NS048447とAG017216to KDによってサポートされています。里六は、オリジナルのプロトコルの開発中に、彼の監督と支援のために博士ヘイッキTanilaを（クオピオ大学、クオピオ、フィンランド）に感謝いたします。

ガラスキャピラリーチューブを引っ張る <ol><li>ガラスキャピラリーチューブはサターインストゥルメント社（ホウケイ酸ガラス、B100 - 75 - 10）から購入しています。 </li><li> 300で設定された熱指数と330で設定された圧力の指数で、マイクロピペットプラー迸るサターP - 87キャピラリーチューブを引き出します。 </li><li>テーパー先端が約0.5 mmの内径を持つように、はさみでガラスキャピラリーチューブの先端を切り取ります。 </li></ol> CSFのための大槽穿刺法 CSF試料は、大槽（図1）1以前に発表されたメソッドを使用してから取得されます。 <img src="/files/ftp_upload/old/960_2008_10_13_2/960_Figure-01.png" alt="図1" width="432" height="184" /> 図1 1。マウスは、ケタミン（100mg/kg）とキシラジン（10mg/kgより少ない）、腹腔内投与によってanesthesizedされています。麻酔導入の時間の間に、マウスは37℃のインキュベーター内で保持されます。 2。首の皮膚を剃毛され、そしてマウスは、加温パッドの直接接触を持つ定位固定装置に発生しやすく配置されます。加熱パッドから発生する熱が体温の変化に応じて調整されるように直腸温度プローブを直腸に挿入されます。ヘッドは、ヘッドアダプタで固定されています。手術部位は7 0％エタノール（3回繰り返す） に続いて、10％ポビドンヨードでswabbedされ、そして皮膚の矢状切開は、後頭部に劣って行われます。 3。解剖顕微鏡下で、皮下組織や筋肉が（M.二腹のcervicisとmの頭直背大手 ）鉗子で鈍的切開で区切られています。 microretractorsのペアは離れて筋肉を保持するために使用されます。 4。マウスは頭部がボディとほぼ135 Oの角度を形成するように定められている。 5。解剖顕微鏡下で、大槽の硬膜は、延髄と主要な血管（動脈背棘）、およびCSF領域が（図2）表示するかを介して輝くと明確な逆三角形として表示されます。 <img src="/files/ftp_upload/old/960_2008_10_13_3/960_Figure-02.png" alt="図2" width="304" height="307" /> 図2 6。硬膜は、滅菌綿棒で乾燥したブロットされています。背動脈の棘（図2）に横方向の硬膜を介してチテルナマグナにキャピラリーチューブを貫通する。キャピラリーチューブの挿入に抵抗の顕著な変化に続いて、CSFは、キャピラリー管に流れる。 7。慎重にキャピラリーチューブを取り外し、および1mm内径を持つポリエチレンチューブを介して3 mlシリンジに接続してください。事前にマーク0.5ミリリットルエッペンドルフチュー​​ブにCSFを注入し、直ちにドライアイスでチューブを凍結し、-80℃の冷凍庫にそれを転送する。 8 CSFのサンプリングの後、筋肉が再整列されており、皮膚が（4-0、エチコン、ジョンソン＆ジョンソン）縫合されています。 0.9％塩化ナトリウムの約1mlは、デハイドレーションを防止するために皮下注射される。マウスは、それが回復するまで体温を維持するためにインキュベーターに保たれ、マウスの体重は1日、および手術後1週間を監視されます。 結果と考察 我々は、検出可能なプラズマ汚染することなくマウスからのCSFのシリアルサンプリングするための信頼性の高いプロトコルを説明。 1。全体の手順は、通常、マウスあたり10分（麻酔を含む）を取ります。 CSF得られたの体積は、マウス系統に依存しています。 P301L（JNPL3）マウス3が約10μL-15μlの収率を得たながら、我々は2を使用したPS / APPダブルTgマウスでは、平均的な量は、約5μL（3〜7μL）です。シリアルサンプリングの場合は、7-8μlの最大は、安全に2-3ヶ月の間隔でそれぞれの時間を撮影することができます。 2。手術中は、それは大槽の硬膜を十分に露出することができるように定位固定フレーム上で適切にマウスの頭と体を配置することが重要です。アポ4の免疫ブロットによりCSFのサンプルで監視できる血漿タンパク質、からの汚染を防ぐためにキャピラリチューブを硬膜を貫通するときに慎重に血管を避けてください。 3。組織の癒着は、以下のサンプリングのための素因出血によって難易度を増加させることができるように各手術時の組織損傷を最小限に抑えることは、重要です。 4。麻酔によって引き起こされる低体温が大幅に非常に急速に5を 、脳内のリン酸化タウのレベルに影響する可能性があるため、体温がうまく手術中に維持される必要があります。 5。抗体ELISAプロトコルと我々が使用した抗体は、Refoloらに詳細に記載された。6。 1:50-1:60に希釈したときにPS / APPマウスからの脳脊髄液の二液は、満足のいく結果が得られます。タウELISAの場合は、P301L（JNPL3）マウス4から5μlのCSFは十分です総タウの検出、またはタウ 蛍光体は、トレオニンでylated 231（キットはバイオソースから購入されています）。 6。この手法は、脳脊髄液の少量が必要なアッセイには十分となっている神経疾患、の他のマウスモデルに適用することができます。

<ol>
	<li>Liu, 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=Longitudinal+observation+on+CSF+Abeta42+levels+in+young+to+middle-aged+amyloid+precursor+protein/presenilin-1+doubly+transgenic+mice.">Longitudinal observation on CSF Abeta42 levels in young to middle-aged amyloid precursor protein/presenilin-1 doubly transgenic mice.</a> Neurobiol Dis. 17, 516 (2004).</li><li>Holcomb, 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=Accelerated+Alzheimer-type+phenotype+in+transgenic+mice+carrying+both+mutant+amyloid+precursor+protein+and+presenilin+1+transgenes.">Accelerated Alzheimer-type phenotype in transgenic mice carrying both mutant amyloid precursor protein and presenilin 1 transgenes.</a> Nat Med. 4, 97 (1998).</li><li>Lewis, 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=Neurofibrillary+tangles,+amyotrophy+and+progressive+motor+disturbance+in+mice+expressing+mutant+(P301L)+tau+protein.">Neurofibrillary tangles, amyotrophy and progressive motor disturbance in mice expressing mutant (P301L) tau protein.</a> Nat Genet. 25, 402 (2000).</li><li>DeMattos, R. B., 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=Plaque-associated+disruption+of+CSF+and+plasma+amyloid-beta+(Abeta)+equilibrium+in+a+mouse+model+of+Alzheimer's+disease.">Plaque-associated disruption of CSF and plasma amyloid-beta (Abeta) equilibrium in a mouse model of Alzheimer's disease.</a> J Neurochem. 81, 229 (2002).</li><li>Planel, E., 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=Anesthesia+leads+to+tau+hyperphosphorylation+through+inhibition+of+phosphatase+activity+by+hypothermia.">Anesthesia leads to tau hyperphosphorylation through inhibition of phosphatase activity by hypothermia.</a> J Neurosci. 27, 3090 (2007).</li><li>Refolo, 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=A+cholesterol-lowering+drug+reduces+beta-amyloid+pathology+in+a+transgenic+mouse+model+of+Alzheimer's+disease.">A cholesterol-lowering drug reduces beta-amyloid pathology in a transgenic mouse model of Alzheimer's disease.</a> Neurobiol Dis. 8, 890 (2001).</li></ol>

我々は、検出可能なプラズマ汚染することなくマウスからのCSFのシリアルサンプリングするための信頼性の高いプロトコルを説明。 1。全体の手順は、通常、マウスあたり10分（麻酔を含む）を取ります。 CSF得られたの体積は、マウス系統に依存しています。 P301L（JNPL3）マウス3が約10μL-15μlの収率を得た一方、PS / APPダブルTgマウスではused2、平均容積は、約5μL（3〜7μL）です。シリアルサンプリング?...

a technique for serial collection of cerebrospinal fluid from the cisterna magna in mouse 

アルツハイマー病（AD）は、病理学的にβ-アミロイドペプチド（Aβ）と過リン酸化タウタンパク質の神経細胞内蓄積の細胞外沈着によって特徴づけられる進行性神経変性疾患である。脳脊髄液（CSF）は脳の細胞外空間と直接接触しているので、それは病理学的プロセスに対応して脳内の生化学的変化を反映しています。これらの変化の原因のメカニズムはまだ完全には理解されていないものの、AD患者のCSFは、42アミノ酸Aβ（Aβ42）の形態、および総タウと過リン酸化タウの増加の減少を示しています。 ADのトランスジェニック（Tg）マウスモデルでは、どのように、なぜAβやタウのレベルCSFの変化で病気の進行状況に応じて調査する絶好の機会を提供しています。ここで、我々は、マウスからの脳脊髄液のサンプリングのための洗練された大槽穿刺のテクニックを示しています。この非常に穏やかなサンプリング手法は、シリアルCSFのサンプルが大幅にそれが可能な時間をかけて微妙な変化を検出すること、Aβまたはタウのレベルでの間に、マウスの変動の交絡影響を最小化する2〜3カ月ごとに同一のマウスから取得することを可能とする。 AβとタウELISAとの組み合わせで、この手法は、ADのマウスモデルにおける脳内のCSFAβ42とタウのレベルとの関係、およびそれらの代謝を調査するために設計された研究に有用となります。 Tgマウスの研究では、監視の病気の進行のための生物学的マーカーとして使用されるCSFのAβやタウのレベルの潜在的なまでに重要な検証を提供することができる、そして治療的介入の効果を監視する。マウスを屠殺することができ、脳の生化学的または組織学的変化を調べることができるように、CSFの変化のメカニズムは良く評価することができる。これらのデータは、ヒトADの脳脊髄液の変化の解釈のために有益であると思われる。

Watch this Scientific Journal Video about A Technique for Serial Collection of Cerebrospinal Fluid from the Cisterna Magna in Mouse at JoVE.com

A Technique for Serial Collection of Cerebrospinal Fluid from the Cisterna Magna in Mouse

ADのトランスジェニック（Tg）マウスモデルでは、病気は人間の患者で進行するにつれてどのように、なぜCSFの変化でAβやタウのレベルを調査するために素晴らしい機会を提供しています。ここで、我々は、マウスからシリアルCSFのサンプリングのための洗練された大槽穿刺のテクニックを示しています。

大槽マウスでの脳脊髄液のシリアル収集のためのテクニック

Alzheimer's disease (AD) is a progressive neurodegenerative disease that is pathologically characterized by extracellular deposition of &#946;-amyloid ...

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Biology

A Technique for Serial Collection of Cerebrospinal Fluid from the Cisterna Magna in Mouse 

101.5K ビュー.  Columbia University.  ADのトランスジェニック（Tg）マウスモデルでは、病気は人間の患者で進行するにつれてどのように、なぜCSFの変化でAβやタウのレベルを調査するために素晴らしい機会を提供しています。ここで、我々は、マウスからシリアルCSFのサンプリングのための洗練された大槽穿刺のテクニックを示しています。 

ビデオ: A Technique for Serial Collection of Cerebrospinal Fluid from the Cisterna Magna in Mouse

Isolation of Cerebrospinal Fluid from Rodent Embryos for use with Dissected Cerebral Cortical Explants

The CSF is a complex fluid with a dynamically varying proteome throughout development and in adulthood. During embryonic development, the nascent CSF differentiates from the amniotic fluid upon closure of the anterior neural tube. CSF volume then increases over subsequent days as the neuroepithelial progenitor cells lining the ventricles and the choroid plexus generate CSF. The embryonic CSF contacts the apical, ventricular surface of the neural stem cells of the developing brain and spinal cord. CSF provides crucial fluid pressure for the expansion of the developing brain and distributes important growth promoting factors to neural progenitor cells in a temporally-specific manner. To investigate the function of the CSF, it is important to isolate pure samples of embryonic CSF without contamination from blood or the developing telencephalic tissue. Here, we describe a technique to isolate relatively pure samples of ventricular embryonic CSF that can be used for a wide range of experimental assays including mass spectrometry, protein electrophoresis, and cell and primary explant culture. We demonstrate how to dissect and culture cortical explants on porous polycarbonate membranes in order to grow developing cortical tissue with reduced volumes of media or CSF. With this method, experiments can be performed using CSF from varying ages or conditions to investigate the biological activity of the CSF proteome on target cells.

The ventricular cerebrospinal fluid (CSF) bathes the neuroepithelial and cerebral cortical progenitor cells during early brain development in the embryo. Here we describe the method developed to isolate ventricular CSF from rodent embryos of different ages in order to investigate its biological function. In addition, we demonstrate our cerebral cortical explant dissection and culture technique that allows for explant growth with minimal volumes of culture medium or CSF.

解剖大脳皮質植で使用するための齧歯類の胚から脳脊髄液の分離

The CSF is a complex fluid with a dynamically varying proteome throughout development and in adulthood. During embryonic development, the nascent CSF ...

神経科学

Minimally Invasive Cisterna Magna Injection Model for Leptomeningeal Metastasis Studies in Mice

Leptomeningeal metastasis (LM), the spread of cancer cells into the cerebrospinal fluid (CSF)-filled leptomeninges, is a rare yet devastating complication of advanced solid tumors. Patients with LM often have a poor prognosis, with survival measured in weeks to months. Developing in vivo models that accurately replicate the complexities of LM is essential for understanding its cellular and pathological mechanisms and evaluating potential therapies. Murine LM models are typically created through intra-cardiac, carotid artery, or cisterna magna injection of tumor cells. However, intra-cardiac or carotid injections often result in substantial extracranial and brain tumor burden, complicating bioluminescent imaging and leading to mortality unrelated to LM. Meanwhile, conventional cisterna magna injection requires invasive procedures, such as skin incision and muscle dissection, making it both traumatic and resource intensive. Here, we describe a minimally invasive procedure for tumor cell injection into the leptomeningeal space through the cisterna magna without the need for a skin incision. This approach reduces extracranial tumor formation, minimizes surgical trauma, and shortens the time and postoperative care required compared to other surgical methods. Importantly, it consistently induces LM with minimal brain parenchyma infiltration, as confirmed by two-photon microscopy and histological analysis. This streamlined approach offers an efficient and reliable model for studying LM in preclinical research.

The protocol describes a method for injecting tumor cells through the percutaneous puncture route into cisterna magna that robustly induces leptomeningeal metastasis in mice, reducing trauma and extracranial tumor burden.

マウスにおける軟髄膜転移研究のための低侵襲槽マグナ注射モデル

Leptomeningeal metastasis (LM), the spread of cancer cells into the cerebrospinal fluid (CSF)-filled leptomeninges, is a rare yet devastating complication ...

がん研究

An Improved Method for Collection of Cerebrospinal Fluid from Anesthetized Mice

The cerebrospinal fluid (CSF) is a valuable body fluid for analysis in neuroscience research. It is one of the fluids in closest contact with the central nervous system and thus, can be used to analyze the diseased state of the brain or spinal cord without directly accessing these tissues. However, in mice it is difficult to obtain from the cisterna magna due to its closeness to blood vessels, which often contaminate samples. The area for CSF collection in mice is also difficult to dissect to and often only small samples are obtained (maximum of 5-7 &#181;L or less). This protocol describes in detail a technique that improves on current methods of collection to minimize contamination from blood and allow for the abundant collection of CSF (on average 10-15 &#181;L can be collected). This technique can be used with other dissection methods for tissue collection from mice, as it does not impact any tissues during CSF extraction. Thus, the brain and spinal cord are not affected with this technique and remain intact. With greater CSF sample collection and purity, more analyses can be used with this fluid to further aid neuroscience research and better understand diseases affecting the brain and spinal cord.

This protocol describes an improved technique for the abundant collection of cerebrospinal fluid (CSF) with no contamination from blood. With greater sample collection and purity, more analyses can be performed using CSF to further our understanding of diseases that affect the brain and spinal cord.

麻酔下のマウスから脳脊髄液のコレクションのための改良法

The cerebrospinal fluid (CSF) is a valuable body fluid for analysis in neuroscience research. It is one of the fluids in closest contact with the central ...

Cannula Implantation into the Cisterna Magna of Rodents

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to the skull or the brain parenchyma. In anesthetized rodents, the exposure of the dura mater by blunt dissection of the neck muscles allows the insertion of a cannula into the cisterna magna (CM). The cannula, composed either by a fine beveled needle or borosilicate capillary, is attached via a polyethylene (PE) tube to a syringe. Using a syringe pump, molecules can then be injected at controlled rates directly into the CM, which is continuous with the subarachnoid space. From the subarachnoid space, we can trace CSF fluxes by convective flow into the perivascular space around penetrating arterioles, where solute exchange with the interstitial fluid (ISF) occurs. CMc can be performed for acute injections immediately following the surgery, or for chronic implantation, with later injection in anesthetized or awake, freely moving rodents. Quantitation of tracer distribution in the brain parenchyma can be performed by epifluorescence, 2-photon microscopy, and magnetic resonance imaging (MRI), depending on the physico-chemical properties of the injected molecules. Thus, CMc in conjunction with various imaging techniques offers a powerful tool for assessment of the glymphatic system and CSF dynamics and function. Furthermore, CMc can be utilized as a conduit for fast, brain-wide delivery of signaling molecules and metabolic substrates that could not otherwise cross the blood brain barrier (BBB).

Here we describe a protocol to perform cisterna magna cannulation (CMc), a minimally invasive way to deliver tracers, substrates and signaling molecules into the cerebrospinal fluid (CSF). Combined with different imaging modalities, CMc enables glymphatic system and CSF dynamics assessment, as well as brain-wide delivery of various compounds.

齧歯動物の槽注入カニューレ

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to ...

In Vivo Imaging of Cerebrospinal Fluid Transport through the Intact Mouse Skull using Fluorescence Macroscopy

Cerebrospinal fluid (CSF) flow in rodents has largely been studied using ex vivo quantification of tracers. Techniques such as two-photon microscopy and magnetic resonance imaging (MRI) have enabled in vivo quantification of CSF flow but they are limited by reduced imaging volumes and low spatial resolution, respectively. Recent work has found that CSF enters the brain parenchyma through a network of perivascular spaces surrounding the pial and penetrating arteries of the rodent cortex. This perivascular entry of CSF is a primary driver of the glymphatic system, a pathway implicated in the clearance of toxic metabolic solutes (e.g., amyloid-&#946;). Here, we illustrate a new macroscopic imaging technique that allows real-time, mesoscopic imaging of fluorescent CSF tracers through the intact skull of live mice. This minimally-invasive method facilitates a multitude of experimental designs and enables single or repeated testing of CSF dynamics. Macroscopes have high spatial and temporal resolution and their large gantry and working distance allow for imaging while performing tasks on behavioral devices. This imaging approach has been validated using two-photon imaging and fluorescence measurements obtained from this technique strongly correlate with ex vivo fluorescence and quantification of radio-labeled tracers. In this protocol, we describe how transcranial macroscopic imaging can be used to evaluate glymphatic transport in live mice, offering an accessible alternative to more costly imaging modalities.

Transcranial optical imaging allows wide-field imaging of cerebrospinal fluid transport in the cortex of live mice through an intact skull.

蛍光マクロスコピーを用いた無傷マウス頭蓋骨を介した脳脊髄液輸送の生体内イメージング

Cerebrospinal fluid (CSF) flow in rodents has largely been studied using ex vivo quantification of tracers. Techniques such as two-photon microscopy and ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the CSF protein composition delivers crucial information in basic neuroscience research as well as neurological diseases. One caveat is that proteins measured in CSF may derive from both intrathecal synthesis and transudation from serum, and protein analysis of CSF can only determine the sum of these two components. To discriminate between protein transudation from the blood and intrathecally produced proteins in animal models as well as in humans, CSF protein profiling measurements using conventional protein analysis tools must include the calculation of the albumin CSF/serum quotient (Qalbumin), a marker of the integrity of the blood-brain interface (BBI), and the protein index (Qprotein/Qalbumin), an estimate of intrathecal protein synthesis. This protocol illustrates the entire procedure, from CSF and blood collection to quotients and indices calculations, for the quantitative measurement of intrathecal protein synthesis and BBI impairment in mouse models of neurological disorders.

Elevated spinal fluid protein levels can either be the result of diffusion of plasma protein across an altered blood-brain barrier or intrathecal synthesis. An optimized testing protocol is presented in this article that helps to discriminate both cases and provides quantitative measurements of intrathecally synthesized proteins.

マウスにおけるインテコール内合成タンパク質の定量測定

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

Induction of Leptomeningeal Cells Modification Via Intracisternal Injection

The protocol outlined here describes how to safely and manually inject solutions through the cisterna magna while eliminating the risk of damage to the underlying parenchyma. Previously published protocols recommend using straight needles that should be lowered to a maximum of 1-2 mm from the dural surface. The sudden drop in resistance once the dural membrane has been punctured makes it difficult to maintain the needle in a steady position. Our method, instead, employs a needle bent at the tip that can be stabilized against the occipital bone of the skull, thus preventing the syringe from penetrating into the tissue after perforation of the dural membrane. The procedure is straightforward, reproducible, and does not cause long-lasting discomfort in the operated animals. We describe the intracisternal injection strategy in the context of genetic fate mapping of vascular leptomeningeal cells. The same technique can, furthermore, be utilized to address a wide range of research questions, such as probing the role of leptomeninges in neurodevelopment and the spreading of bacterial meningitis, through genetic ablation of genes putatively implicated in these phenomena. Additionally, the procedure can be combined with an automatized infusion system for a constant delivery and used for tracking cerebrospinal fluid movement via injection of fluorescently labelled molecules.

We describe an intracisternal injection that employs a needle bent at the tip that can be stabilized to the skull, thus eliminating the risk of damage to the underlying parenchyma. The approach can be used for genetic fate mapping and manipulations of leptomeningeal cells and for tracking cerebrospinal fluid movement.

インタビセナーリナーリ注射によるレプトメニンゲ細胞改変の誘導

The protocol outlined here describes how to safely and manually inject solutions through the cisterna magna while eliminating the risk of damage to the ...

Stereotaxic Surgical Approach to Microinject the Caudal Brainstem and Upper Cervical Spinal Cord via the Cisterna Magna in Mice

Stereotaxic surgery to target brain sites in mice is commonly guided by skull landmarks. Access is then obtained via burr holes drilled through the skull. This standard approach can be challenging for targets in the caudal brainstem and upper cervical cord due to specific anatomical challenges as these sites are remote from skull landmarks, leading to imprecision. Here we outline an alternative stereotaxic approach via the cisterna magna that has been used to target discrete regions of interest in the caudal brainstem and upper cervical cord. The cisterna magna extends from the occipital bone to the atlas (i.e., the second vertebral bone), is filled with cerebrospinal fluid, and is covered by dura mater. This approach provides a reproducible route of access to select central nervous system (CNS) structures that are otherwise hard to reach due to anatomical barriers. Furthermore, it allows for direct visualization of brainstem landmarks in close proximity to the target sites, increasing accuracy when delivering small injection volumes to restricted regions of interest in the caudal brainstem and upper cervical cord. Finally, this approach provides an opportunity to avoid the cerebellum, which can be important for motor and sensorimotor studies.

Stereotaxic Surgical Approach to Microinject the Caudal Brainstem and Upper Cervical Spinal Cord via the Cisterna Magna in Mice

Stereotaxic surgery to target brain sites in mice commonly involves access through the skull bones and is guided by skull landmarks. Here we outline an alternative stereotaxic approach to target the caudal brainstem and upper cervical spinal cord via the cisterna magna that relies on direct visualization of brainstem landmarks.

マウスのCisterna Magna を介して 尾側脳幹および上部頸椎脊髄をマイクロインジェクションする定位的外科的アプローチ

Stereotaxic surgery to target brain sites in mice is commonly guided by skull landmarks. Access is then obtained via burr holes drilled through the skull. ...

脳波記録の同時実施によるラットからの脳脊髄液および血液の回収

Sampling Cerebrospinal Fluid and Blood from Lateral Tail Vein in Rats During EEG Recordings

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many experimental contexts to measure molecules of interest, such as hormones, growth factors, proteins, or small non-coding RNAs. A specific example is the sampling of biological liquids in the research of biomarkers for epilepsy. In these studies, it is desirable to compare the levels of molecules in cerebrospinal fluid (CSF) and in plasma, by withdrawing CSF and plasma in parallel and considering the time distance of the sampling from and to seizures. The combined CSF and plasma sampling, coupled with video-EEG monitoring in epileptic animals, is a promising approach for the validation of putative diagnostic and prognostic biomarkers. Here, a procedure of combined CSF withdrawal from cisterna magna and blood sampling from the lateral tail vein in epileptic rats that are continuously video-EEG monitored is described. This procedure offers significant advantages over other commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, and reduced time of anesthesia. Additionally, it can be used to obtain CSF and plasma samples in both tethered and telemetry EEG recorded rats, and it may be used repeatedly across multiple days of experiment. By minimizing the stress due to sampling by shortening isoflurane anesthesia, measures are expected to reflect more accurately the true levels of investigated molecules in biofluids. Depending on the availability of an appropriate analytical assay, this technique may be used to measure the levels of multiple, different molecules while performing EEG recording at the same time.

著者スポットライト:てんかんバイオマーカー探索のための高品質なCSFおよび血液サンプルの取得 

The protocol shows repeated cerebrospinal fluid and blood collections from epileptic rats performed in parallel with continuous video-electroencephalogram (EEG) monitoring. These are instrumental for exploring possible links between changes in various body fluid molecules and seizure activity.

脳波記録中のラットの側尾静脈からの脳脊髄液および血液のサンプリング

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many ...

大槽マウスでの脳脊髄液のシリアル収集のためのテクニック

要約

さらに動画を探す

解剖大脳皮質植で使用するための齧歯類の胚から脳脊髄液の分離

マウスにおける軟髄膜転移研究のための低侵襲槽マグナ注射モデル

麻酔下のマウスから脳脊髄液のコレクションのための改良法

齧歯動物の槽注入カニューレ

蛍光マクロスコピーを用いた無傷マウス頭蓋骨を介した脳脊髄液輸送の生体内イメージング

マウスにおけるインテコール内合成タンパク質の定量測定

インタビセナーリナーリ注射によるレプトメニンゲ細胞改変の誘導

マウスのCisterna Magna を介して尾側脳幹および上部頸椎脊髄をマイクロインジェクションする定位的外科的アプローチ

脳波記録中のラットの側尾静脈からの脳脊髄液および血液のサンプリング

脳波記録中のラットの側尾静脈からの脳脊髄液および血液のサンプリング

大槽マウスでの脳脊髄液のシリアル収集のためのテクニック

要約

さらに動画を探す

解剖大脳皮質植で使用するための齧歯類の胚から脳脊髄液の分離

マウスにおける軟髄膜転移研究のための低侵襲槽マグナ注射モデル

麻酔下のマウスから脳脊髄液のコレクションのための改良法

齧歯動物の槽注入カニューレ

蛍光マクロスコピーを用いた無傷マウス頭蓋骨を介した脳脊髄液輸送の生体内イメージング

マウスにおけるインテコール内合成タンパク質の定量測定

インタビセナーリナーリ注射によるレプトメニンゲ細胞改変の誘導

マウスのCisterna Magna を介して 尾側脳幹および上部頸椎脊髄をマイクロインジェクションする定位的外科的アプローチ

脳波記録中のラットの側尾静脈からの脳脊髄液および血液のサンプリング

脳波記録中のラットの側尾静脈からの脳脊髄液および血液のサンプリング

マウスのCisterna Magna を介して尾側脳幹および上部頸椎脊髄をマイクロインジェクションする定位的外科的アプローチ