Name: a technique for serial collection of cerebrospinal fluid from the cisterna magna in mouse
Uploaded: 2008-11-10T00:00:00.000+00:00
Duration: 8 min 8 s
Description: Watch this Scientific Journal Video about A Technique for Serial Collection of Cerebrospinal Fluid from the Cisterna Magna in Mouse at JoVE.com

Il nostro lavoro è supportato dal NIH concede NS048447 e AG017216to KD. Li Liu desidera ringraziare il Dott. Heikki Tanila (Università di Kuopio, Kuopio, Finlandia) per la sua supervisione e supporto durante lo sviluppo del protocollo originale.

Tirare il tubo capillare di vetro <ol><li> Il tubo capillare di vetro viene acquistato dallo strumento Sutter Inc (vetro borosilicato, B100-75-10). </li><li> Tirare i tubi capillari su un Sutter P-87 Flaming micropipetta estrattore, con l&#39;indice di calore fissato a 300 e l&#39;indice di pressione fissato a 330. </li><li> Tagliare la punta del tubo capillare di vetro con le forbici, in modo che la punta conica ha un diametro interno di circa 0,5 mm. </li></ol> Cisterna magna tecnica di puntura di CSF Campioni di CSF sono prese dal cisterna magna (Figura 1) utilizzando un metodo che è stato pubblicato in precedenza 1. <img src="/files/ftp_upload/old/960_2008_10_13_2/960_Figure-01.png" alt="Figura 1" width="432" height="184" /> Figura 1 1. I topi sono anesthesized da ketamina (100mg/kg) e xilazina (10mg/kg), somministrato per via intraperitoneale. Durante il periodo di induzione dell&#39;anestesia, i topi sono conservati in un incubatore o 37 ° C. 2. La pelle del collo è rasata, e il mouse viene posto prono sullo strumento stereotassico con il contatto diretto di una piastra elettrica. Una sonda di temperatura rettale viene inserito nel retto modo che il calore prodotto dal pad di riscaldamento viene regolata in risposta ai cambiamenti della temperatura corporea. La testa è assicurato con le schede di testa. Il sito chirurgico è tamponata con iodio povidone al 10%, seguita da 7 0% di etanolo (ripetere 3 volte), e un&#39;incisione sagittale della pelle è fatta inferiore all&#39;occipite. 3. Sotto il microscopio dissezione, il tessuto sottocutaneo e dei muscoli (m. biventer del collo e m. retto capitis dorsale principale) sono separati per via smussa con una pinza. Un paio di microretractors è utilizzato per tenere i muscoli a pezzi. 4. Il mouse è stabilito in modo che la testa forma un angolo di quasi 135 ° con il corpo. 5. Sotto il microscopio dissezione, la dura madre della cisterna magna appare come un triangolo scintillante e chiara inverso attraverso il quale il midollo allungato e di un vaso sanguigno principale (arteria spinale dorsale), e lo spazio CSF ​​sono visibili (Figura 2). <img src="/files/ftp_upload/old/960_2008_10_13_3/960_Figure-02.png" alt="Figura 2" width="304" height="307" /> Figura 2 6. La dura madre è asciugate con tampone di cotone sterile. Penetrare il tubo capillare in magna Citerna attraverso la dura madre, laterale alla arteria spinale dorsale (Figura 2). A seguito di un notevole cambiamento nella resistenza per l&#39;inserimento del tubo capillare, il QCS scorre nel tubo capillare. 7. Rimuovere con attenzione il tubo capillare, e collegarlo a una siringa da 3 ml con un tubo di polietilene che ha un diametro interno 1 mm. Iniettare il liquido cerebrospinale in un pre-segnato 0,5 ml di tubo Eppendorf, e congelare immediatamente la provetta in ghiaccio secco e poi trasferirlo in un congelatore C ° -80. 8. Dopo il campionamento CSF, i muscoli sono nuovamente allineati, e la pelle viene suturata (4-0, Ethicon, Johnson &amp; Johnson). Circa 1 ml di NaCl 0,9% viene iniettata per via sottocutanea per evitare de-idratazione. Il mouse è conservato presso l&#39;incubatore per mantenere la temperatura corporea fino a che non recupera, il peso del mouse viene monitorata 1 giorno, e 1 settimana dopo l&#39;intervento chirurgico. Risultati e discussione Abbiamo descritto un protocollo altamente affidabile per il campionamento di serie del CSF da topi, senza contaminazioni plasma rilevabili. 1. L&#39;intera procedura richiede solitamente 10 min per il mouse (incluso anestesia). Il volume di CSF ottenuti dipende dal ceppo di topi. Nella PS / APP topi doppio Tg abbiamo usato 2, il volume medio è di circa 5 microlitri (3-7 mL), mentre la (JNPL3) topi P301L 3 danno una resa di circa 10 microlitri-15 microlitri. Per il campionamento di serie, un massimo di 7-8 microlitri può essere tranquillamente assunto di volta in volta ad un intervallo di 2-3 mesi. 2. Durante l&#39;intervento, è importante posizionare la testa e il corpo del mouse correttamente sul telaio stereotassico in modo che la dura madre della cisterna magna può essere esposto a sufficienza. Accuratamente evitare i vasi sanguigni quando penetra la dura madre con il tubo capillare per evitare la contaminazione da proteine ​​plasmatiche, che possono essere monitorate nel campione di liquido cerebrospinale di immunoblotting per ApoB 4. 3. Riducendo al minimo il danno ai tessuti durante ogni intervento chirurgico è importante, come conglutination tessuto può aumentare la difficoltà di sanguinamento predisponendo per il campionamento successivo. 4. La temperatura corporea deve essere ben mantenuto durante l&#39;intervento chirurgico a causa ipotermia causata da anestesia possono influenzare significativamente i livelli di tau fosforilata nel cervello molto rapidamente 5. 5. L&#39;Ab ELISA protocollo e gli anticorpi che abbiamo utilizzato sono stati descritti in dettaglio nel Refolo et al. 6. Due ml di liquor dal PS / APP topi darà risultati soddisfacenti se diluito 1:50-1:60. Per l&#39;ELISA tau, 4-5 CSF microlitri da P301L (JNPL3) topi sarà sufficiente per larilevazione del totale tau, o tau fosforo ylated a treonina 231 (I kit sono acquistati da Bio-Source). 6. Questa tecnica può essere applicata ad altri modelli murini di patologie neurologiche, per la quale un piccolo volume di CSF è soddisfacente per le analisi desiderata.

<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>

Abbiamo descritto un protocollo altamente affidabile per il campionamento di serie del CSF da topi, senza contaminazioni plasma rilevabili. 1. L&#39;intera procedura richiede solitamente 10 min per il mouse (incluso anestesia). Il volume di CSF ottenuti dipende dal ceppo di topi. Nella PS / APP topi doppio Tg abbiamo used2, il volume medio è di circa 5 microlitri (3-7 mL), mentre la (JNPL3) topi P301L 3 danno una resa di circa 10 microlitri-15 microlitri. Per il campionamento di serie, un massimo di 7-8 microlitri può essere tranq...

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

La malattia di Alzheimer (AD) è una malattia neurodegenerativa progressiva che è patologicamente caratterizzata dalla deposizione extracellulare del peptide β-amiloide (Aβ) e l&#39;accumulo di intraneuronale iperfosforilata proteina tau. A causa del liquido cerebrospinale (CSF) è in contatto diretto con lo spazio extracellulare del cervello, fornisce un riflesso dei cambiamenti biochimici nel cervello in risposta a processi patologici. CSF di pazienti dC mostra una diminuzione del 42 aminoacidi forma di Aβ (Aβ42), e aumenta in totale iperfosforilata tau e tau, anche se i meccanismi responsabili di questi cambiamenti non sono ancora pienamente compreso. Transgenici (Tg) modelli murini di AD forniscono un&#39;eccellente opportunità per indagare come e perché i livelli di Aβ o tau nel liquido cerebrospinale cambiare con la progressione della malattia. Qui, dimostrano una raffinata tecnica di puntura cisterna magna per il campionamento CSF ​​dal mouse. Questa tecnica di campionamento estremamente delicata permette seriale campioni di liquido cerebrospinale di ottenere dal mouse stesso a intervalli di 2-3 mesi, che riduce sensibilmente l&#39;effetto confondente della tra-mouse variabilità nei livelli di Aβ o tau, che consente di rilevare alterazioni sottili nel tempo. In combinazione con Aβ e tau ELISA, questa tecnica sarà utile per studi volti a indagare il rapporto tra i livelli di CSF Aβ42 e tau, e il loro metabolismo nel cervello in modelli murini dC. Studi nei topi Tg potrebbe fornire la convalida importante per il potenziale di Aβ CSF o livelli di tau per essere utilizzati come marcatori biologici per monitorare la progressione della malattia, e per monitorare l&#39;effetto degli interventi terapeutici. Come i topi possono essere sacrificati e il cervello possono essere esaminati per cambiamenti biochimici o istologici, i meccanismi alla base dei cambiamenti CSF può essere meglio valutata. Questi dati sono suscettibili di essere informativo per l&#39;interpretazione dei cambiamenti umani CSF dC.

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

Transgenici (Tg) modelli murini di AD forniscono un&#39;eccellente opportunità per indagare come e perché i livelli di Aβ o tau nel liquido cerebrospinale cambiare il progredire della malattia nei pazienti umani. Qui, dimostrano una raffinata tecnica di puntura cisterna magna per il campionamento CSF ​​seriale dal mouse.

Una tecnica per la raccolta seriale del liquido cerebrospinale dalla Magna Cisterna in Mouse

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 

101.4K Views.  Columbia University.  Transgenici (Tg) modelli murini di AD forniscono un'eccellente opportunità per indagare come e perché i livelli di Aβ o tau nel liquido cerebrospinale cambiare il progredire della malattia nei pazienti umani. Qui, dimostrano una raffinata tecnica di puntura cisterna magna per il campionamento CSF ​​seriale dal mouse. 

Video: 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.

Isolamento del liquido cerebrospinale da embrioni roditori per l&#39;uso con sezionato espianti cerebrali corticali

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

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Neuroscienze

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.

Un metodo migliore per la raccolta di liquido cerebrospinale da topi anestetizzati

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.

Impianto di cannula in Cisterna Magna di roditori

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.

In vivo Imaging del trasporto del fluido cerebrospinale attraverso il teschio del topo intatto utilizzando la macroscopia di fluorescenza

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.

Misurazione quantitativa delle proteine intratecalliamente sintetizzate nei topi

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.

Induzione di cellule leptomeningeali Modifica tramite iniezione intracisternale

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.

Approccio chirurgico stereotassico per microiniettare il tronco encefalico caudale e il midollo spinale cervicale superiore tramite la Cisterna Magna nei topi

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. ...

Prelievo di liquido cerebrospinale e sangue dai ratti con registrazione EEG concomitante

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.

Autore in primo piano: Ottenere campioni di sangue e CSF di alta qualità per la scoperta di biomarcatori dell'epilessia 

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.

Campionamento del liquido cerebrospinale e del sangue dalla vena caudale laterale nei ratti durante le registrazioni EEG

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

Una tecnica per la raccolta seriale del liquido cerebrospinale dalla Magna Cisterna in Mouse

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Isolamento del liquido cerebrospinale da embrioni roditori per l'uso con sezionato espianti cerebrali corticali

Un metodo migliore per la raccolta di liquido cerebrospinale da topi anestetizzati

Impianto di cannula in Cisterna Magna di roditori

In vivo Imaging del trasporto del fluido cerebrospinale attraverso il teschio del topo intatto utilizzando la macroscopia di fluorescenza

Misurazione quantitativa delle proteine intratecalliamente sintetizzate nei topi

Induzione di cellule leptomeningeali Modifica tramite iniezione intracisternale

Approccio chirurgico stereotassico per microiniettare il tronco encefalico caudale e il midollo spinale cervicale superiore tramite la Cisterna Magna nei topi

Campionamento del liquido cerebrospinale e del sangue dalla vena caudale laterale nei ratti durante le registrazioni EEG

Campionamento del liquido cerebrospinale e del sangue dalla vena caudale laterale nei ratti durante le registrazioni EEG

Campionamento del liquido cerebrospinale e del sangue dalla vena caudale laterale nei ratti durante le registrazioni EEG