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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. SE Induction
<ol>
	<li>Purchase 8-week-old male C57BL/6NHsd mice and weigh each mouse. Then, use a marker pen to mark the tails of all mice for their easy identification during SE induction.</li>
	<li>Calculate the amount of scopolamine methyl bromide (scopolamine; 2 mg/kg), terbutaline hemisulfate (terbutaline; 2 mg/kg), and pilocarpine hydrochloride (pilocarpine; 280 mg/kg) based on the mouse weight and add saline (0.9% NaCl, 10 mL/kg) to make solutions. 
	NOTE:&#160;For example, if the weight of the mouse is 25 g, the following amounts are applied: Scopolamine and terbutaline: 2 mg/kg * (25 g/1,000 g) = 0.05 mg, Saline: 10 mL/kg * (25 g/1,000 g) = 0.25 mL, Pilocarpine: 280 mg/kg * (25 g/1,000 g) = 7 mg, Saline: 10 mL/kg * (25 g/1,000 g) = 0.25 mL.</li>
	<li>Load the scopolamine and terbutaline mixture in a 1 mL syringe with a 30 G needle. Inject the solution intraperitoneally into each mouse, and then return the mice to their cages. At 30 min after scopolamine and terbutaline administration, inject pilocarpine solution into each mouse (i.p.; 1 mL syringe; 30 G needle). Immediately after pilocarpine injection, place the mice in an incubator (28&#8211;30&#176;C) for observation.</li>
	<li>Carefully monitor the behavior of the mice until SE is induced. If limbic motor seizures that correspond to stage 3 or higher according to Racine&#39;s scale are detected, record the time and monitor the mice to determine whether the seizures occur more frequently. Once continuous motor seizures last more than 2 min, place the mouse in a new cage at room temperature and keep monitoring for 3 h to determine whether their convulsive seizures continue and SE is induced. Euthanize the mice that failed to enter SE at about 2 h after pilocarpine injection. 
	NOTE:&#160;Racine&#39;s scale; stage 1, mouth and facial movement; stage 2, head nodding; stage 3, forelimb clonus; stage 4, rearing with forelimb clonus; stage 5, rearing and falling with forelimb clonus. If the mouse is not transferred to the cage at room temperature immediately after SE induction, the mouse can die due to hyperthermia.</li>
	<li>Terminate behavioral acute seizures at 3 h after SE onset by injecting diazepam solution (i.p.; 10 mg/kg; 1 mL syringe; 30 G needle). Make the diazepam solution by diluting 5 mg/mL of diazepam in 10% polyoxyl 35 hydrogenated castor oil by adding saline (i.p.; 10 mL/kg; 1 mL syringe; 30 G needle).&#160; 
	NOTE:&#160;10% polyoxyl 35 hydrogenated castor oil solution: 1 mL polyoxyl 35 hydrogenated castor oil solution + 9 mL saline. Store the solution at room temperature. For example, if the mouse body weight is 25 g, inject 250 &#181;L of diazepam solution: 50 &#181;L 5 mg/mL diazepam + 200 &#181;L 10% polyoxyl 35 hydrogenated castor oil solution = 250 &#181;L 1 mg/mL diazepam.</li>
	<li>For sham mice, perform an i.p. injection of the scopolamine methyl bromide and terbutaline hemisulfate mixture (i.p.; both 2 mg/kg, 10 mL/kg; 1 mL syringe; 30 G needle). 30 min later, inject the saline intraperitoneally (i.p.; 10 mL/kg; 1 mL syringe; 30 G needle). 
	NOTE:&#160;If the mouse body weight is 25 g, inject 250 &#181;L of saline instead of pilocarpine.</li>
	<li>After diazepam injection (i.p.; 1 mL syringe; 30 G needle), administer 1 mL of 5% dextrose per individual mouse (i.p.; 1 mL syringe; 26 G needle). 
	NOTE:&#160;1 mL of 5% dextrose injection provides energy source and hydration that can increase the survival rate.</li>
	<li>During post-care in the incubator (28&#8211;30 &#176;C), wipe off excessive secretions such as saliva, tears, and feces.</li>
	<li>At day 1 after SE induction, weigh the mice and keep them in the incubator (28&#8211;30 &#176;C) for one extra day. At day 2, after SE induction, weigh the mice and return them to their home cage. Provide moist chow to facilitate their recovery. 
	NOTE:&#160;In this experiment, the mortality rate for C57BL/6NHsd after SE termination was 8.57% on average (3 out of 35 mice tested).</li>
	<li>Measure daily body weight of the animals until 7 days post-pilocarpine and inject the mice with 5% dextrose (i.p.; 1 mL syringe; 26 G needle) when the body weight has not increased. Stop body weight monitoring when the mice start to regain body weight and consume moist chow without difficulty at 2 days after pilocarpine injection. 
	NOTE:&#160;If the body weight of a mouse did not increase until 7 days post SE, exclude the mouse from the experiment and euthanize by carbon dioxide. Depending on the background of the mouse, death can occasionally occur; however, in the case of C57BL/6NHsd, less than 1% of mice died in these experiments. With survival of more than 7 days after SE induction, the mice rarely die during the latent period.</li>
</ol>

<table><tbody><tr><td>C57BL/6</td><td>Envigo</td><td>C57BL/6NHsd</td><td></td></tr><tr><td>Scopolamine methyl nitrate</td><td>Sigma</td><td>S2250</td><td>Make 10X stock</td></tr><tr><td>Terbutaline hemisulfate salt</td><td>Sigma</td><td>T2528</td><td>Make 10X stock</td></tr><tr><td>Pilocarpine hydrochloride</td><td>Sigma</td><td>P6503</td><td></td></tr><tr><td>Diazepam</td><td>SAMJIN</td><td></td><td></td></tr><tr><td>Castor oil (Kolliphor EL)</td><td>Sigma</td><td>C5135</td><td>Polyoxyl 35 hydrogenated castor oil</td></tr><tr><td>Saline</td><td>Daihan pharm. Co.</td><td></td><td></td></tr><tr><td>5% Dextrose</td><td>Daihan pharm. Co.</td><td></td><td></td></tr><tr><td>1 mL syringe</td><td>Sung shim medial Co.&amp;nbsp; Ltd.</td><td></td><td></td></tr><tr><td>26 guage needle</td><td>Sung shim medial Co.&amp;nbsp; Ltd.</td><td></td><td>26 G * 13 mm (1/2&quot;)</td></tr><tr><td>30 guage needle</td><td>Sung shim medial Co.&amp;nbsp; Ltd.</td><td></td><td>30 G * 13 mm (1/2&quot;)</td></tr></tbody></table>

systemic pilocarpine treatment: mouse model of temporal lobe epilepsy

Source: Kim, J. E. and Cho, K. O. <a href="https://www.jove.com/video/56831/the-pilocarpine-model-temporal-lobe-epilepsy-eeg-monitoring-using">The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice</a>. J. Vis. Exp. (2018).
This video presents the procedure for inducing status epilepticus (SE) in mice by systemic administration of pilocarpine to model temporal lobe epilepsy.

Systemic Pilocarpine Treatment: Mouse Model of Temporal Lobe Epilepsy

Source: Kim, J. E. and Cho, K. O. The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice. J. Vis. Exp. ...

To begin, administer an injection of scopolamine and terbutaline solution to minimize the peripheral side effects of pilocarpine. Next, administer an injection of pilocarpine solution to induce status epilepticus or SE. Pilocarpine will alter the brain&#39;s neuronal excitability.
Immediately after that, place the mouse in a warm incubator and monitor for induction of SE. Look for head nodding and rhythmic movements of the front legs or any behavior of stage 3 or higher on the Racine&#39;s scale. Then, place the mouse into a room temperature incubator and continue to monitor the animal&#39;s behavior.
Next, administer an injection of diazepam solution to end SE. Lastly, administer an injection of dextrose solution to aid with the animal&#39;s recovery. Following SE induction, the animal enters a seizure-free latent period until the onset of spontaneous recurrent seizures.

systemic-pilocarpine-treatment-mouse-model-of-temporal-lobe-epilepsy

Universidad San Sebastian

Research

JoVE Encyclopedia of Experiments

Biology

Rodent Models

Adult

1.6K Views. المصدر: Kim ، J. E. and Cho ، K. O. نموذج بيلوكاربين لصرع الفص الصدغي ومراقبة مخطط كهربية الدماغ باستخدام نظام القياس الإشعاعي عن بعد في الفئران. J. Vis. Exp. (2018). يعرض هذا الفيديو إجراء إحداث حالة الصرع (SE) في الفئران عن طريق الإعطاء الجهازي للبيلوكاربين لنمذجة صرع الفص الصدغي. 

Video: علاج بيلوكاربين الجهازي: نموذج الفأر لصرع الفص الصدغي - Concept

Simultaneous Video-EEG-ECG Monitoring to Identify Neurocardiac Dysfunction in Mouse Models of Epilepsy

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can potentially increase risk of sudden unexpected death in epilepsy (SUDEP). Electroencephalography (EEG) and electrocardiography (ECG) are widely used clinical diagnostic tools to monitor for abnormal brain and cardiac rhythms in patients. Here, a technique to simultaneously record video, EEG, and ECG in mice to measure behavior, brain, and cardiac activities, respectively, is described. The technique described herein utilizes a tethered (i.e., wired) recording configuration in which the implanted electrode on the head of the mouse is hard-wired to the recording equipment. Compared to wireless telemetry recording systems, the tethered arrangement possesses several technical advantages such as a greater possible number of channels for recording EEG or other biopotentials; lower electrode costs; and greater frequency bandwidth (i.e., sampling rate) of recordings. The basics of this technique can also be easily modified to accommodate recording other biosignals, such as electromyography (EMG) or plethysmography for assessment of muscle and respiratory activity, respectively. In addition to describing how to perform the EEG-ECG recordings, we also detail methods to quantify the resulting data for seizures, EEG spectral power, cardiac function, and heart rate variability, which we demonstrate in an example experiment using a mouse with epilepsy due to Kcna1 gene deletion. Video-EEG-ECG monitoring in mouse models of epilepsy or other neurological disease provides a powerful tool to identify dysfunction at the level of the brain, heart, or brain-heart interactions.

Here, we present a protocol to record brain and heart bio signals in mice using simultaneous video, electroencephalography (EEG), and electrocardiography (ECG). We also describe methods to analyze the resulting EEG-ECG recordings for seizures, EEG spectral power, cardiac function, and heart rate variability.

متزامنة الفيديو-EEG-ECG الرصد لتحديد الخلل نيوروكاردياك في نماذج الماوس من الصرع

In epilepsy, seizures can evoke cardiac rhythm disturbances such as heart rate changes, conduction blocks, asystoles, and arrhythmias, which can ...

JoVE Journal

Genetics

Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test interventions at various stages of the disease process. These animal models are particularly difficult to implement in immature rodents, a classic model organism for laboratory study of these disorders. Recording continuous EEG in young animal models of seizures and other neurological disorders presents a technical challenge due to the small physical size of young rodents and their dependence on the dam prior to weaning. Therefore, there is not only a clear need for improving pre-clinical research that will better identify those therapies suitable for translation to the clinic but also a need for new devices capable of recording continuous EEG in immature rodents. Here, we describe the technology behind and demonstrate the use of a novel miniature telemetry system, specifically engineered for use in immature rats or mice, which is also effective for use in adult animals.

Here we demonstrate the use of a wireless enabling technology for electroencephalogram (EEG) in neonatal rodent models of human disease. With telemetry, there are no encumbering connections, thus allowing natural behaviors.

على المدى الطويل رصد EEG المستمر في القوارض الصغيرة نماذج من الأمراض البشرية عن طريق نظام عصر الارسال اللاسلكي

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test ...

Neuroscience

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

نموذج Pilocarpine صرع الفص الصدغي ورصد EEG باستخدام نظام راديوتيليميتري في الفئران

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Systemic Pilocarpine Treatment: Mouse Model of Temporal Lobe Epilepsy

Transcript

Systemic Pilocarpine Treatment: Mouse Model of Temporal Lobe Epilepsy