eoe sub articles

EoE Sub Articles

<figure class='table' style='width:1367pt;'><table class='ck-table-resized'><colgroup><col style='width:24.14%;'><col style='width:27.07%;'><col style='width:16.02%;'><col style='width:32.77%;'></colgroup><tbody><tr><td style='height:14.5pt;'>3-channel two-twisted electrode</td><td>Invivo1, Plastic One, Roanoke, Virginia, USA</td><td>MS333/3-B/SPC</td><td>Material</td></tr><tr><td style='height:14.5pt;'>guide cannula</td><td>Agn Tho's, Lindig&#246;, Sweden</td><td>MAB 4.15.IC</td><td>Material</td></tr><tr><td style='height:14.5pt;'>Resin KK2 Plastik</td><td>Elettra Sport, Lecco, Italy</td><td>KK2</td><td>Material</td></tr><tr><td style='height:14.5pt;'>Super Attack gel Loctite</td><td>Henkel Italia Srl, Milano, Italy</td><td>2047420_71941</td><td>Material</td></tr><tr><td style='height:14.5pt;'>Imalgene-Ketamine</td><td>Merial, Toulouse, France</td><td>221300288 (AIC)</td><td>Solution</td></tr><tr><td style='height:14.5pt;'>Xylazine</td><td>Sigma, Milano, Italy</td><td>X1251</td><td>Material</td></tr><tr><td style='height:14.5pt;'>Isoflurane-Vet</td><td>Merial, Toulouse, France</td><td>103120022 (AIC)</td><td>Solution</td></tr><tr><td style='height:14.5pt;'>Altadol 50 mg/ ml - tramadol</td><td>Formevet, Milano, Italy</td><td>103703017 (AIC)</td><td>Solution</td></tr><tr><td style='height:14.5pt;'>Gentalyn 0.1% crm - gentamycine</td><td>MSD Italia, Roma, Italy</td><td>20891077 (AIC)</td><td>Material</td></tr><tr><td style='height:14.5pt;'>simplex rapid dental cement</td><td>Kemdent, Associated Dental Products Ltd, Swindon, United Kingdom</td><td>ACR811</td><td>Material</td></tr><tr><td style='height:14.5pt;'>GlasIonomer CX-Plus Cement</td><td>Shofu, Kyoto, Japan</td><td>PN1167</td><td>Material</td></tr><tr><td style='height:14.5pt;'>probe clip holder</td><td>Agn Tho's, Lindig&#246;, Sweden</td><td>p/n 100 5001</td><td>Equipment</td></tr><tr><td style='height:14.5pt;'>Histoacryl&#174; 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Sweden</td><td>MAB 4.15.1.Cu</td><td>Material</td></tr><tr><td style='height:14.5pt;'>microdialysis probe</td><td>Synaptech, Colorado Springs, Colorado, USA</td><td>S-8010</td><td>Material</td></tr><tr><td style='height:14.5pt;'>block heater</td><td>Grant Instruments, Cambridge, England</td><td>QBD2</td><td>Equipment</td></tr><tr><td style='height:14.5pt;'>stirrer</td><td>Cecchinato A, Aparecchi Scientifici, Mestre, Italy</td><td>711</td><td>Equipment</td></tr><tr><td style='height:14.5pt;'>infusion pump</td><td>Univentor, Zejtun, Malta</td><td>864</td><td>Equipment</td></tr><tr><td style='height:14.5pt;'>fine bore polythene tubing</td><td>Smiths Medical International Ltd., Keene, New Hampshire, USA</td><td>800/100/100/100</td><td>Material</td></tr><tr><td style='height:14.5pt;'>blue tubing adapters</td><td>Agn Tho's, Lindig&#246; Sweden</td><td>1002</td><td>Material</td></tr><tr><td style='height:14.5pt;'>red tubing adapters</td><td>Agn Tho's, Lindig&#246; Sweden</td><td>1003</td><td>Material</td></tr><tr><td style='height:14.5pt;'>2.5 mL syringe with 22G needle</td><td>Chemil, Padova, Italy</td><td>S02G22</td><td>Material</td></tr><tr><td style='height:14.5pt;'>vial cap</td><td>Cronus, Labicom, Olomouc, Czech Republic</td><td>VCA-1004TB-100</td><td>Material</td></tr><tr><td style='height:14.5pt;'>septum</td><td>Thermo Scientific, Rockwoood, Tennessee, USA</td><td>National C4013-60 8 mm TEF/SIL septum</td><td>Material</td></tr><tr><td style='height:14.5pt;'>glass insert with bottom spring</td><td>Supelco, Sigma, Milano, Italy</td><td>27400-U</td><td>Material</td></tr><tr><td style='height:14.5pt;'>autosampler vial</td><td>National Scientific, Thermo Fisher Scientific, Monza, Italy</td><td>C4013-2</td><td>Material</td></tr><tr><td style='height:14.5pt;'>Smartline manager 5000 system controller and degasser unit</td><td>Knauer, Berlin, Germany</td><td>V7602</td><td>Equipment</td></tr><tr><td style='height:14.5pt;'>Smartline 1000 quaternary gradient pump</td><td>Knauer, Berlin, Germany</td><td>V7603</td><td>Equipment</td></tr><tr><td style='height:14.5pt;'>Ringer solution</td><td>composition in mM: MgCl2&#160;0.85, KCl 2.7, NaCl 148, CaCl2&#160;1.2, 0.3% BSA</td><td>&#160;</td><td>Solution</td></tr><tr><td style='height:14.5pt;'>modified Ringer solution</td><td>composition in mM: MgCl2&#160;0.85, KCl 100, NaCl 50.7, CaCl2&#160;1.2, 0.3% BSA</td><td>&#160;</td><td>Solution</td></tr><tr><td style='height:14.5pt;'>saline</td><td>0.9% NaCl, ph adjusted to 7.0</td><td>&#160;</td><td>Solution</td></tr><tr><td style='height:14.5pt;'>sucrose solution</td><td>10% sucrose in distilled water</td><td>&#160;</td><td>Solution</td></tr></tbody></table></figure>

assessment of excitatory neurotransmitter levels in chronic epilepsy-induced rats

<a href='https://app.jove.com/v/58455/microdialysis-excitatory-amino-acids-during-eeg-recordings-freely'>Source: Soukupov&#225;, M., et al. Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats. J. Vis. Exp. (2018)</a>This video demonstrates the assessment of extracellular excitatory neurotransmitter levels in rats with chronic epilepsy induction. After implanting a microdialysis probe into the brain and confirming the absence of seizures using electroencephalography (EEG), extracellular fluid is collected to evaluate elevated neurotransmitter levels in epileptic rats. A high-potassium solution is then infused to sustain neuronal excitability and trigger an increased release of neurotransmitters, with samples collected to assess the potassium-induced peak in excitatory neurotransmitters.

<figure class='image image-style-align-center image_resized' style='display:table;margin-left:auto;margin-right:auto;width:50%;'><img style='aspect-ratio:2875/2449;' src='https://cloudfront.jove.com/files/ftp_upload/1/23312_Figure1_editor_1.jpg' alt='23312_Figure1.jpg' /></figure>Figure 1. Step-by-step preparation of the device to be implanted.&#160;(A) 3-channel electrode with 10 cm long grounding electrode in its protective sleeve on the left and guide cannula for microdialysis on the right needed to assemble the device. (B). The bare electrode and guide cannula in detail. The first step is to remove (C) and insert (D) the metal guide cannula from and into its plastic dummy few times to ease its release once implanted into the animal's head. The second step is to bend two times the twisted registering electrode to be aligned to dialysis guide cannula (E, F). (G) The electrode tip should be cut to be 0.5 mm longer than the tip of the metal guide cannula. (H) Check for the precision of the cut using the digital caliper. Subsequently, about 1 mm long silicon circlet should be used to fix the alignment of electrode to guide cannula foot (I). (J) The photograph showing how to ring the electrode and guide cannula shaft. The final step is to put a drop of resin or glue onto the guide cannula pedestal fixing the silicon circlet to it (K). (L) Assembled device ready to be sterilized.<figure class='image image-style-align-center image_resized' style='display:table;margin-left:auto;margin-right:auto;width:50%;'><img style='aspect-ratio:1139/559;' src='https://cloudfront.jove.com/files/ftp_upload/1/23312_Figure2_editor_1.jpg' alt='23312_Figure2.jpg' /></figure>Figure 2. Photographs of different types of devices for microdialysis-EEG in rats used (A) and the photograph of the probe clip holder (B) used to implant these devices.&#160;(A) The guide cannula (in green) is replaced by a microdialysis cannula typically 24 h before the experiment. The electrical connector of the device (first left was used for the recordings described in this manuscript) permits the attachment of wires that conduct electrical signals to amplifier and data collection equipment. The device is surgically attached to the skull of anesthetized rats and recordings may be obtained later without causing pain or discomfort in freely behaving rats.&#160;<figure class='image image-style-align-center image_resized' style='display:table;margin-left:auto;margin-right:auto;width:50%;'><img style='aspect-ratio:1363/351;' src='https://cloudfront.jove.com/files/ftp_upload/1/23312_Figure3_editor_1.jpg' alt='23312_Figure3.jpg' /></figure>Figure 3. Experimental design.&#160;The week before status epilepticus (SE) induction, the rats are implanted with the device. SE is induced by pilocarpine and animals (if not dialyzed and killed at 24 h after SE; the rats from acute group) are video monitored for 5 days (blue line), then video-EEG monitored to assess the seizure frequency and duration in their home cages (green line). For the microdialysis experiment, the epileptic and respective non-epileptic control rats are transferred to another EEG set up equipped with cylinder cages in 24 h before the microdialysis session and still video-EEG monitored (light green line). The vertical red lines represent the dialysis sessions at different time-points of epileptic disease development. The horizontal red lines represent the different groups of epileptic animals (and respective non-epileptic animals), where the arrow indicates the last day of the microdialysis and the day of animal's death.&#160;

Assessment of Excitatory Neurotransmitter Levels in Chronic Epilepsy-Induced Rats

Source: Soukupov&#225;, M., et al. Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats. J. Vis. Exp. (2018)This video ...

Take a control rat and a chronic epilepsy-induced rat implanted with a guide cannula and an electroencephalogram, or EEG, electrode in the hippocampus.Epilepsy induction increases excitatory neurotransmitter release, causing ion influx into postsynaptic cells, which generates continuous excitatory potentials. This leads to synchronized bursts of electrical activity, termed seizures, recorded using the electrode.Insert a microdialysis probe with a semi-permeable membrane into the cannula.Tether the rat to the cage of an EEG recording system.Infuse a physiological buffer into the brain tissue via the probe inlet.Ensure the absence of seizure and collect extracellular fluid via the probe outlet to assess neurotransmitter levels.An elevated basal level in the epileptic rat indicates an epilepsy-mediated increase.Infuse a high potassium buffer and recollect samples.&#160;The increased potassium prolongs neuronal excitability, triggering continuous neurotransmitter release.A temporary spike in neurotransmitter levels in both the epileptic and control rats indicates a high potassium-induced increase.

assessment-excitatory-neurotransmitter-levels-chronic-epilepsy

Nanyang Technological University

Research

JoVE Encyclopedia of Experiments

Neuroscience

Neuropathology

Epilepsy and Seizure Disorders

8 Aufrufe. Source: Soukupová, M., et al. Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats. J. Vis. Exp. (2018) This video demonstrates the assessment of extracellular excitatory neurotransmitter levels in rats with chronic epilepsy induction. After implanting a microdialysis probe into the brain and confirming the absence of seizures using electroencephalography (EEG), extracellular fluid is collected to evaluate elevated neurotransmitter levels in epileptic rats. A ...

Serie: Assessment of Excitatory Neurotransmitter Levels in Chronic Epilepsy-Induced Rats - Konzept

In Vivo Microdialysis Method to Collect Large Extracellular Proteins from Brain Interstitial Fluid with High-molecular Weight Cut-off Probes

In vivo microdialysis is a powerful technique to collect ISF from awake, freely-behaving animals based on a dialysis principle. While microdialysis is an established method that measures relatively small molecules including amino acids or neurotransmitters, it has been recently used to also assess dynamics of larger molecules in ISF using probes with high molecular weight cut off membranes. Upon using such probes, microdialysis has to be run in a push-pull mode to avoid pressure accumulated inside of the probes. This article provides step-by-step protocols including stereotaxic surgery and how to set up microdialysis lines to collect proteins from ISF. During microdialysis, drugs can be administered either systemically or by direct infusion into ISF. Reverse microdialysis is a technique to directly infuse compounds into ISF. Inclusion of drugs in the microdialysis perfusion buffer allows them to diffuse into ISF through the probes while simultaneously collecting ISF. By measuring tau protein as an example, the author shows how its levels are altered upon stimulating neuronal activity by reverse microdialysis of picrotoxin. Advantages and limitations of microdialysis are described along with the extended application by combining other in vivo methods.

In Vivo Microdialysis Method to Collect Large Extracellular Proteins from Brain Interstitial Fluid with High-molecular Weight Cut-off Probes

In vivo microdialysis has enabled collection of molecules present in brain interstitial fluid (ISF) from awake, freely-behaving animals. In order to analyze relatively large molecules in ISF, the current article specifically focuses on the microdialysis protocol using probes with high molecular weight cut off membranes.

In Vivo Mikrodialyse-Methode, um große extrazellulären Proteine Sammeln von Gehirn interstitielle Flüssigkeit mit hochmolekularen Gewicht Cut-Off-Sonden

In vivo microdialysis is a powerful technique to collect ISF from awake, freely-behaving animals based on a dialysis principle. While microdialysis is an ...

Forschung

JoVE Journal

Neurowissenschaften

Real-Time Dynamic Collection of Hippocampal Extracellular Fluid from Conscious Rats Using a Microdialysis System

A variety of central nervous system (CNS) diseases are associated with changes in the composition of hippocampal extracellular fluid (HECF). However, difficulty in obtaining HECF in real time from conscious rats has long restricted the evaluation of CNS disease progression and the effectiveness of ethnomedicine therapy. Encouragingly, a brain microdialysis technique can be used for continuous sampling with the advantages of dynamic observation, quantitative analysis, and a small sampling size. This allows the monitoring of changes in the extracellular fluid content for compounds from traditional herbs and their metabolites in the brain of living animals. The aim of this study was thus to accurately implant a cerebrospinal fluid microdialysis probe into the hippocampal region of Sprague Dawley (SD) rats with a three-dimensional brain stereotaxic apparatus, cutting off molecular weights greater than 20 kDa. The high-quality HECF was then obtained from conscious rats using a microdialysis sampling control system with an adjustable sampling rate from 2.87 nL/min - 2.98 mL/min. In conclusion, our protocol provides an efficient, rapid, and dynamic method to obtain HECF in awake rats with the help of microdialysis technology, which provides us with unlimited possibilities to further explore the pathogenesis of CNS-related diseases and evaluate drug efficacy.

The protocol here provides a detailed real-time dynamic sampling of extracellular fluid from the hippocampus of awake rats using a microdialysis system.

Dynamische Echtzeit-Entnahme von extrazellulärer Flüssigkeit aus dem Hippocampus von bewussten Ratten mit Hilfe eines Mikrodialysesystems

A variety of central nervous system (CNS) diseases are associated with changes in the composition of hippocampal extracellular fluid (HECF). However, ...

Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique

Dynamic analysis of blood components is of great importance in understanding cardiovascular diseases and their related diseases, such as myocardial infarction, arrhythmia, atherosclerosis, cardiogenic pulmonary edema, pulmonary embolism, and cerebral embolism. At the same time, it is urgent to break through the continuous heart blood sampling technique in live rats to evaluate the effectiveness of distinctive ethnic medicine therapy. In this study, a blood microdialysis probe was implanted in the right jugular vein of rats in a precise and noninvasive surgical procedure. Cardiac blood samples were then collected at a rate of 2.87 nL/min to 2.98 mL/min by connecting to an online microdialysis sample collection system. Even more momentously, the acquired blood samples can temporarily be stored in microdialysis containers at 4 &#176;C. The microdialysis-based online continuous blood collection program from rat heart has greatly guaranteed the quality of blood samples, advancing and invigorating the scientific rationality of the research on systemic cardiovascular diseases and evaluating ethnomedicine therapy from the perspective of hematology.

Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique

The present protocol describes a simple and efficient method for the real-time and dynamic collection of rat heart blood using the microdialysis technique.

Dynamische kontinuierliche Blutentnahme aus Rattenherzen mittels nicht-invasiver Mikrodialysetechnik

Dynamic analysis of blood components is of great importance in understanding cardiovascular diseases and their related diseases, such as myocardial ...

Assessment of Excitatory Neurotransmitter Levels in Chronic Epilepsy-Induced Rats

Transkript

Assessment of Excitatory Neurotransmitter Levels in Chronic Epilepsy-Induced Rats

In Vivo Mikrodialyse-Methode, um große extrazellulären Proteine Sammeln von Gehirn interstitielle Flüssigkeit mit hochmolekularen Gewicht Cut-Off-Sonden

Dynamische Echtzeit-Entnahme von extrazellulärer Flüssigkeit aus dem Hippocampus von bewussten Ratten mit Hilfe eines Mikrodialysesystems

Dynamische kontinuierliche Blutentnahme aus Rattenherzen mittels nicht-invasiver Mikrodialysetechnik