Çalışmalarımız NIH hibe NS048447 ve AG017216to KD tarafından desteklenmektedir. Li Liu orijinal protokol gelişimi sırasında Dr. Heikki Tanila (Kuopio Üniversitesi, Kuopio, Finlandiya) onun gözetimi ve desteği için teşekkür etmek istiyorum.

Çekme cam kapiller tüp <ol><li> Cam kapiller tüp Sutter Enstrüman A.Ş. (borosilikat cam, B100-75-10) satın alınmaktadır. </li><li> Kılcal tüpler çekin bir Sutter P-87, 300 ayarlanan ısı indeksi ve 330 olarak belirlenmiştir basınç indeksi, mikropipet çektirmesi Flaming. </li><li> Sivri ucu yaklaşık 0.5 mm iç çapa sahip, böylece cam kapiller tüp ucu makas ile kesin. </li></ol> CSF Cisterna magna ponksiyonu tekniği BOS örnekleri sisterna magna (Şekil 1), daha önce 1 yayımlandı bir yöntem kullanarak alınır. <img src="/files/ftp_upload/old/960_2008_10_13_2/960_Figure-01.png" alt="Şekil 1" width="432" height="184" /> Şekil 1 1. Fareler Ketamin (100mg/kg) ve xylazine (10mg/kg) intraperitoneal yolla anesthesized. Anestezi indüksiyonu sırasında, fareler 37 o C inkübatör tutulur. 2. Boyun derisi traş ve fare sonra bir ısıtma yastığı doğrudan temas ile stereotaksik alet eğilimli yerleştirilir. Isıtma yastığı tarafından üretilen ısı, vücut ısısı değişikliklerine yanıt olarak ayarlanır, böylece bir rektal ısı probu rektum içine yerleştirilir. Kafa kafa adaptörleri ile güvence altına alınmıştır. 7 0% etanol (3 kez tekrarlamanız), cerrahi sitesi% 10 povidon iyot ile temizlenmesi, ve oksiput aşağı cilt sagital bir kesi yapılır. 3. Diseksiyon mikroskobu altında, subkutan doku ve kasları (m. biventer cervicis ve m. rektus kapitis dorsalis majör) künt diseksiyon forseps ile ayrılır. Microretractors bir çift dışında kaslar tutmak için kullanılır. 4. Fare kafası, gövdesi ile yaklaşık 135 o açı oluşturur, böylece ortaya konulmaktadır . 5. Diseksiyon mikroskobu altında, sisterna magna duramater medulla oblongata ve büyük bir kan damarı (arteria dorsalis spinalis) ve BOS alanı (Şekil 2) görünür olan bir parlak ve net bir ters üçgen gibi görünür. <img src="/files/ftp_upload/old/960_2008_10_13_3/960_Figure-02.png" alt="Şekil 2" width="304" height="307" /> Şekil 2 6. Duramater steril pamuklu bir bez ile kuru lekelenen. Arteria dorsalis spinalis (Şekil 2) Lateral duramater, kılcal tüp aracılığıyla citerna magna içine nüfuz ederler. Kılcal boru ekleme direnç farkedilir bir değişikliğin ardından, BOS kılcal tüp içine akar. 7. Kılcal tüp dikkatlice çıkarın ve 1 mm iç çapı olan bir polietilen boru ile 3 ml şırınga bağlayın. BOS önceden işaretli 0.5 ml Eppendorf tüp içine enjekte edilir ve tüp dondurma ve kuru buz üzerinde hemen sonra -80 ° C derin dondurucu içine transfer. 8. BOS örnekleme sonra, kasların yeniden hizalanır ve cilt sütüre edilir (4-0, Ethicon, Johnson &amp; Johnson). Yaklaşık 1 ml% 0.9 NaCl de-hidrasyon önlemek için subkutan enjekte edilir. Fare kurtarır kadar vücut ısısını korumak için inkübatör tutulur; farenin ağırlığı 1 gün ve 1 hafta sonra ameliyat izlenir. Sonuçlar ve Tartışma Biz saptanabilir plazma kirlenme olmadan farelerden alınan BOS seri örnekleme için son derece güvenilir bir protokol nitelendirdi. 1. Tüm prosedür genellikle fare başına 10 dakika (anestezi dahil) alır. BOS elde hacmi fare suşları bağlıdır. P301L (JNPL3) farelerde 3 yaklaşık 10 ul-15 ul verimi verirken, biz 2 PS / APP çift Tg fareler ortalama hacmi, yaklaşık 5 ul (3 7 ul). Seri numune almak için, en fazla 7-8 ul bir güvenle 2-3 ay aralıklarla her zaman olabilir. 2. Ameliyat sırasında, sisterna magna duramater yeterince maruz böylece stereotaksik çerçeve düzgün fare baş ve vücut pozisyonu önemlidir. ApoB 4 immunoblotting BOS örnek izlenebilir plazma proteinlerine, kirlenmeyi önlemek için kapiller tüp ile duramater nüfuz kan damarlarını dikkatle kaçının. 3. Doku kaynama Aşağıdaki örnekleme için predispozan kanama zorluk artışı gibi her ameliyat sırasında doku hasarı en aza indirmek önemlidir. 4. Hipotermi anestezi nedeniyle önemli ölçüde beyin çok hızlı bir şekilde 5 fosforile tau düzeyleri etkileyebilir, çünkü vücut ısısı ameliyat sırasında iyi bir bakıma ihtiyacı vardır. 5. Kullanılan Ab ELISA protokolü ve antikorlar Refolo ark ayrıntılı şekilde tarif edildi. 6. İki PS / APP farelerden alınan BOS ul 1:50-1:60 seyreltilmiş zaman tatmin edici sonuçlar verecektir. Tau ELISA için 4-5 ul P301L (JNPL3) farelerden alınan BOS için yeterli olacaktırtoplam tau tau ​​tespit fosfor treonin az ylated 231 (Bio-Kaynak Kitleri satın alınır). 6. Bu teknik, küçük bir BOS hacmi istenilen deneyleri için tatmin edici olduğu nörolojik hastalıklar, diğer fare modelleri uygulanabilir.

<ol><li>Liu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Longitudinal+observation+on+CSF+Abeta42+levels+in+young+to+middle-aged+amyloid+precursor+protein%2Fpresenilin-1+doubly+transgenic+mice%5BTitle%5D)+AND+%22Neurobiol+Dis%22%5BJournal%5D)">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/pubmed?term=((Accelerated+Alzheimer-type+phenotype+in+transgenic+mice+carrying+both+mutant+amyloid+precursor+protein+and+presenilin+1+transgenes%5BTitle%5D)+AND+%22Nat+Med%22%5BJournal%5D)">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/pubmed?term=((Neurofibrillary+tangles%2C+amyotrophy+and+progressive+motor+disturbance+in+mice+expressing+mutant+%28P301L%29+tau+protein%5BTitle%5D)+AND+%22Nat+Genet%22%5BJournal%5D)">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/pubmed?term=((Plaque-associated+disruption+of+CSF+and+plasma+amyloid-beta+%28Abeta%29+equilibrium+in+a+mouse+model+of+Alzheimer%27s+disease%5BTitle%5D)+AND+%22J+Neurochem%22%5BJournal%5D)">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/pubmed?term=((Anesthesia+leads+to+tau+hyperphosphorylation+through+inhibition+of+phosphatase+activity+by+hypothermia%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">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/pubmed?term=((A+cholesterol-lowering+drug+reduces+beta-amyloid+pathology+in+a+transgenic+mouse+model+of+Alzheimer%27s+disease%5BTitle%5D)+AND+%22Neurobiol+Dis%22%5BJournal%5D)">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>

Biz saptanabilir plazma kirlenme olmadan farelerden alınan BOS seri örnekleme için son derece güvenilir bir protokol nitelendirdi. 1. Tüm prosedür genellikle fare başına 10 dakika (anestezi dahil) alır. BOS elde hacmi fare suşları bağlıdır. P301L (JNPL3) farelerde 3 ul ul-10 15 verim verirken biz used2 PS / APP çift Tg fareler, ortalama hacmi, yaklaşık 5 ul (3 7 ul). Seri numune almak için, en fazla 7-8 ul bir güvenle 2-3 ay aralıklarla her zaman olabilir. 2. Ameliyat sırasında, sisterna magna duramater...

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

Alzheimer hastalığı (AH) patolojik, beta-amiloid peptid (Aβ) ve hyperphosphorylated tau proteininin intraneuronal birikimi ekstrasellüler birikimi ile karakterize ilerleyici bir nörodejeneratif hastalıktır. Beyin omurilik sıvısı (BOS) beynin ekstraselüler boşluk ile doğrudan temas halinde olduğundan, patolojik süreçler yanıt olarak beyindeki biyokimyasal değişikliklerin bir yansıması sağlar. AD hastalardan alınan BOS, bu değişiklikler için sorumlu olan mekanizmalar hala tam olarak anlaşılmış değildir ama 42 amino asit formu Aβ (Aβ42) ve total tau ve hyperphosphorylated tau artışlar bir azalma gösterir. (Tg) AD transgenik fare modelleri Aβ veya tau düzeyleri BOS değişiklik neden ve nasıl hastalık ilerledikçe araştırmak için mükemmel bir fırsat sağlar. Burada, fare BOS örnekleme için rafine bir sisterna magna ponksiyonu tekniği göstermektedir. Bu son derece nazik bir örnekleme tekniği seri BOS örnekleri zamanla ince değişiklikleri tespit etmek mümkün kılan büyük ölçüde, fare Aβ veya tau düzeyleri arasında değişkenlik karıştırıcı etkisini en aza indirir 2-3 aylık aralıklarla aynı fare elde edilecek sağlar. Aβ ve tau ELISA ile birlikte, bu tekniğin AD fare modelleri, beyin, BOS Aβ42 ve tau düzeyleri arasındaki ilişki ve onların metabolizmasını araştırmak için tasarlanmış çalışmalar için yararlı olacaktır. Tg fareler üzerinde yapılan çalışmalar, hastalığın ilerlemesi izlenmesi için biyolojik belirteç olarak kullanılmak üzere BOS Aβ veya tau düzeyleri potansiyel olarak önemli bir doğrulama sağlayabilir ve tedavi edici girişimlerin etkisi izlemek için. , Farelerin feda edilebilir ve beyinlerinde biyokimyasal ve histolojik değişiklikler için incelenecektir gibi, BOS değişikliklerin altında yatan mekanizmaları daha iyi değerlendirilebilir. Bu veriler insan AD BOS değişiklikleri yorumlanması için bilgilendirici olması muhtemeldir.

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

(Tg) AD transgenik fare modelleri, nasıl ve neden Aβ veya tau düzeyleri BOS değişikliği insan hastalarda hastalık ilerledikçe araştırmak için mükemmel bir fırsat sağlar. Burada, fare seri BOS örnekleme için rafine bir sisterna magna ponksiyonu tekniği göstermektedir.

Cisterna Magna Fare Beyin Omurilik Sıvısı Seri Toplama Tekniği

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

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

101.5K Görüntüleme.  Columbia University.  (Tg) AD transgenik fare modelleri, nasıl ve neden Aβ veya tau düzeyleri BOS değişikliği insan hastalarda hastalık ilerledikçe araştırmak için mükemmel bir fırsat sağlar. Burada, fare seri BOS örnekleme için rafine bir sisterna magna ponksiyonu tekniği göstermektedir. 

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.

Dissected Serebral Kortikal Eksplantlarından ile kullanım için Kemirgen Embriyolar arasından Beyin Omurilik Sıvısı İzolasyonu

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

Nörobilim

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.

Beyin omurilik sıvısı imzalat fareler gelen topluluğu için geliştirilmiş bir yöntem

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.

Kanül implantasyon Sisterna Magna kemirgenler içine

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.

İçinde vivo beyin omurilik akışkan taşınması floresan Makroskopisi kullanarak bozulmamış fare kafatası aracılığıyla görüntüleme

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.

Farelerde İntrathecally Sentezlenmiş Proteinlerin Kantitatif Ölçümü

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 ...

Direct Cannula Implantation in the Cisterna Magna of Pigs

The glymphatic system is a waste clearance system in the brain that relies on the flow of cerebrospinal fluid (CSF) in astrocyte-bound perivascular spaces and has been implicated in the clearance of neurotoxic peptides such as amyloid-beta. Impaired glymphatic function exacerbates disease pathology in animal models of neurodegenerative diseases, such as Alzheimer's, which highlights the importance of understanding this clearance system. The glymphatic system is often studied by cisterna magna cannulations (CMc), where tracers are delivered directly into the cerebrospinal fluid (CSF). Most studies, however, have been carried out in rodents. Here, we demonstrate an adaptation of the CMc technique in pigs. Using CMc in pigs, the glymphatic system can be studied at a high optical resolution in gyrencephalic brains and in doing so bridges the knowledge gap between rodent and human glymphatics.

This article presents a step-by-step protocol for the direct cannula implantation in the cisterna magna of pigs.

Domuzların Sarnıç Magnasında Doğrudan Kanın İmplantasyonu

The glymphatic system is a waste clearance system in the brain that relies on the flow of cerebrospinal fluid (CSF) in astrocyte-bound perivascular spaces ...

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.

Farelerde Cisterna Magna ile Kaudal Beyin Sapı ve Üst Servikal Omuriliğin Mikroenjekte Edilmesine Stereotaksik Cerrahi Yaklaşım

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. ...

Intraventricular Drug Delivery and Sampling for Pharmacokinetics and Pharmacodynamics Study

Although the blood-brain barrier (BBB) protects the brain from foreign entities, it also prevents some therapeutics from crossing into the central nervous system (CNS) to ameliorate diseases or infections. Drugs are administered directly into the CNS in animals and humans to circumvent the BBB. The present protocol describes a unique way of treating brain infections through intraventricular delivery of antibiotics, i.e., polymyxins, the last-line antibiotics to treat multi-drug resistant Gram-negative bacteria. A straightforward stereotaxic surgery protocol was developed to implant a guide cannula reaching into the lateral ventricle in rats. After a recovery period of 24 h, rats can be injected consciously and repeatedly through a cannula that is fitted to the guide. Injections can be delivered manually as a bolus or infusion using a microinjection pump to obtain a slow and controlled flow rate. The intraventricular injection was successfully confirmed with Evans Blue dye. Cerebrospinal fluid (CSF) can be drained, and the brain and other organs can be collected. This approach is highly amenable for studies involving drug delivery to the CNS and subsequent assessment of pharmacokinetic and pharmacodynamic activity.

<meta charset="utf8"></head><body>Farmakokinetik ve Farmakodinamik Çalışması için İntraventriküler İlaç Salımı ve Numune Alımı

Delivery of therapeutics directly into the central nervous system is one way of circumventing the blood-brain barrier. The present protocol demonstrates intracerebroventricular injection for subsequent collection of cerebrospinal fluid and bodily organs. This facilitates the investigation of drug pharmacokinetics and pharmacodynamics in animal models for developing new treatments.

Farmakokinetik ve Farmakodinamik Çalışması için İntraventriküler İlaç Salımı ve Numune Alımı

Although the blood-brain barrier (BBB) protects the brain from foreign entities, it also prevents some therapeutics from crossing into the central nervous ...

Eş Zamanlı EEG Kaydı ile Sıçanlardan Beyin Omurilik Sıvısı ve Kan Çekilmesi

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.

Yazar Spotlight: Epilepsi Biyobelirteç Keşfi için Yüksek Kaliteli BOS ve Kan Örnekleri Alma 

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.

EEG Kayıtları Sırasında Sıçanlarda Lateral Kuyruk Veninden Beyin Omurilik Sıvısı ve Kan Örneği Alınması

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

Cisterna Magna Fare Beyin Omurilik Sıvısı Seri Toplama Tekniği

Özet

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EEG Kayıtları Sırasında Sıçanlarda Lateral Kuyruk Veninden Beyin Omurilik Sıvısı ve Kan Örneği Alınması