Name: implantation of chronic silicon probes and recording of hippocampal place cells in an enriched treadmill apparatus
Uploaded: 2017-10-11T13:00:00
Description: Watch this Scientific Journal Video about Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus at JoVE.com

Эта работа была поддержана Корейский институт науки и технологии институциональной программы (проекты № 2E26190 и 2E26170) и программу человека пограничной науки (RGY0089/2012).

животное уход и использование Комитетом Корейский институт науки и технологии были утверждены все методы, описанные.
 1. Подготовка микро привод и электрод <ol><li>Сборка микро привода.
	<ol><li>Печать части микро привода (слайдер, тело и оболочки) 14 с выс?...

<ol>
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Хронический записи активности нейронов имеет решающее значение для понимания нервные процессы такие как гиппокампа место поля. Наш подход для выполнения хронических кремния зонд implantantation отличается от других методов7,18,19,<sup class="...

Кремния датчики представляют ряд преимуществ для электрофизиологических записей, включая тот факт, что они разработаны с острыми профили, минимизации повреждения тканей и что они представляют точную разметку из плотно упакованных записи сайтов1, 2,3,4. Они используются для изучения различных систем в разных видов, в том числе люди3,5,6, разнообразные подходы1,7. Тем не менее их повторного использования все еще сравнительно ограничено из-за их стоимости, хрупкость и тот факт, что удобные методы для хронических экспериментов хватает8. Последние достижения в технологии 3D печати сделали возможным индивидуальное проектирование устройств, таких как микро диски и головы пластины позволяют облегчить обработку этих деликатных электродов. В качестве первого шага мы опишем, как построить и использовать набор инструментов, которые мы разработали для имплантации хронической кремния зонды14.
В то время как место клетки обычно изучаются с использованием свободно перемещающихся животных в лабиринты, недавно они были также расследование в виртуальных средах15 и беговая дорожка apparatii9 (рис. 1A). Эти экспериментальные методы предлагают преимущество, что животные могут быть голова сдержанный, использования 2-Фотон Микроскоп15, патч хомут16и optrode9,10,11 методы проще, помимо усиления контроля на поведение животных и окружающей среды сигналы12. В качестве второго шага мы представим процедуры подготовки мышей и записи активности клеток место в аппарат беговой дорожки.

<table>
	<tbody>
		<tr>
			<td>Silicon Probe</td>
			<td>Neuronexus</td>
			<td>Buzsabi32</td>
			<td>Recording electrode</td>
		</tr>
		<tr>
			<td>Recording system</td>
			<td>Intantech</td>
			<td>RHD2132/RHD2000</td>
			<td></td>
		</tr>
		<tr>
			<td>3D printer</td>
			<td>Asiga</td>
			<td>Pico Plus 27</td>
			<td>High resolution printer for micro-drive</td>
		</tr>
		<tr>
			<td>3D printer</td>
			<td>Stratasys</td>
			<td>Mojo</td>
			<td>Lower resolution printer for hat components</td>
		</tr>
		<tr>
			<td>Stereotaxic apparatus</td>
			<td>Kopf</td>
			<td>Model 963</td>
			<td></td>
		</tr>
		<tr>
			<td>Binocular microscope</td>
			<td>Leica</td>
			<td>M60</td>
			<td></td>
		</tr>
		<tr>
			<td>Treadmill apparatus</td>
			<td></td>
			<td></td>
			<td>We build them</td>
		</tr>
	</tbody>
</table>

implantation of chronic silicon probes and recording of hippocampal place cells in an enriched treadmill apparatus

Важным условием для понимания функции мозга является определение поведения и соотносит клеточной активности. Кремния зонды являются передовыми электроды для крупномасштабных электрофизиологических записи нейронной активности, но по-прежнему развиты процедуры их хронических имплантации. Активность клеток гиппокампа место, как известно, коррелируют с позиции животного в окружающей среде, но основные механизмы по-прежнему неясны. Исследовать место клетки, здесь мы описываем набор методов, которые варьируются от изготовления устройств для хронических кремния зонд имплантаты для мониторинга активности поле место в аппарат кий обогащенный беговой дорожки. Микро привод и шляпу строятся установки и крепления вместе 3D-печатных пластиковых деталей. Зонд кремния монтируется на микро привода, уборка и покрыты краской. Первая операция выполняется исправить шляпу на череп мыши. Небольшие достопримечательности изготавливаются и придает ремня беговой дорожке. Мышь обучены выполнять голова исправлена на беговой дорожке. Вторая операция выполняется для имплантата кремния зонд в гиппокампе, после которого широкополосного электрофизиологических сигналов записываются. Наконец кремния зонд оправился и очищен для повторного использования. Анализ активности клеток место в беговой дорожке показывает разнообразие механизмов поля, изложением преимущество подхода.

Мышь был сначала обучить работать на два метра длиной пояса лишенный сигналы (рис. 1 c). После имплантации электродов, новый ремень той же длины, но представляя 3 пары подсказки был установлен на беговой дорожке, чтобы генерировать аллоцентрическом прост?...

Watch this Scientific Journal Video about Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus at JoVE.com

Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus

Мы описываем различные шаги имплантата хронических кремниевых датчиков и записать место клетки мышей, которые работает голова исправлена на Кий обогащенный беговая дорожка аппарат.

Имплантация хронической кремниевых датчиков и записи клеток гиппокампа место в аппарат обогащенного беговая дорожка

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced ...

The overall goal of this procedure is to implant chronic silicon probes in the mouse hippocampus, and to record place cell activity in mice that are running head-fixed in a cue-enriched treadmill. This method can help answer key questions about the underlying mechanisms of place cell activity. However, it could be applied to a wide range of other systems.The main advantage of silicon probes is that hundreds of place cells can be monitored simultaneously with temporal resolution of a single action potential, and with information about cells'especial configuration. Visual demonstration is critical, as it involves a series of manual steps that are delicate and need to be practiced. Begin probe preparation by carefully detaching the probe from the shipment enclosure.While working under a binocular microscope, use a flat alligator clip with rubber padding to grab the flex cable of the silicon probe and lift it from the enclosure while holding the enclosure with forceps. Fix the alligator clip to a manipulator, then use the manipulator to lay the silicon probe on the micro drive slider parallel to the direction of movement. Apply a drop of room temperature curing acrylic dental repair material to fix the probe to the slider.Correct the probe position if it is shifted. Fix the connector of the probe to the connector holder with dental cement. Then, fix the micro drive and probe assemblage on the probe cleaning device, which is equipped with two small rotating foam sponges.Adjust the gap using the manipulator. Soak the sponges with detergent. Then, monitor the cleaning process under a microscope while slowly and gently moving the probe up and down between the sponges.Begin by installing an anesthetized mouse in a stereotaxic apparatus. Place a nose cone to administer 1.5 to 2%isoflurane. Apply eye ointment to the eyes.Then, after shaving the scalp, clean the head of the animal with antiseptic. Inject buprenorphine under the scalp for analgesia. Then, check for a surgical plane of anesthesia by the absence of a reaction to a paw pinch.Next, cut and remove part of the parietal skin of the mouse head using fine scissors to expose the skull at its edges. Use saline and a hemostatic sponge to clean and control bleeding during the surgery. After removing the periosteum using a scraper tool, find bregma and lambda and the coronal and sagittal suture reference points on the skull.Adjust the head's angle along the sagittal access such that the bregma and lambda points are at the same height. Drill two holes in the skull for the reference and ground electrodes. The hole should be approximately one milometer caudal and one millimeter lateral to lambda.Insert the ground and reference electrodes into the holes. Then, apply ultraviolet light bunting dentin activator on the skull and expose to UV light for 45 to 60 seconds. Next, apply a layer of dental cement to the edges of the skull.Then, fix the head plate to a stereotaxic manipulator and position it above the skull. Slowly lower the head plate until it slightly touches the skull and apply dental cement at the junction with the skull. Let the dental cement cure for 15 minutes.After removing the nose cone, fix a connector holder and a cap to the head plate. Put the mouse in its cage after giving a subcutaneous injection of analgesic. Anesthetize the mouse as before and place it in the stereotaxic frame.After opening the skin and cleaning the skull with saline, measure the distance to the point of insertion and mark it. Drill the bone carefully until it becomes thin and transparent. Moisten and clean with saline while drilling.Use precision forceps to carefully remove the thinned bone and the dura mater. Keep the brain surface wet with saline all the time. Fix the micro drive and silicon probe assemblage to the stereotaxic manipulator.Bring the silicon probe to just above the craniotomy, then screw the silicon probe connector holder to the head plate. Connect the recording amplifier and electrodes. Shield the mouse with aluminum foil to protect from electromagnetic noise.Start the recording system to monitor the electrical activity of the brain. Slowly insert the silicon probe into the brain using the micromanipulator. Continuously check the electrical signals, the manipulator traveled distance, and the shanks of the probe to make sure they are penetrating in the brain.Unit activity is visible in the cortex while the white matter underneath is relatively silent. Unit activity reappears when the shanks touch the pyramidal layer of the hippocampus. From this point, retract the silicon probe 200 microns.Cover the surface of the brain with a mixture of bone wax and mineral oil, then fix the micro drive to the head plate using dental cement. After allowing the dental cement to cure for 15 minutes, detach the micro drive from the stereotaxic manipulator and put the head cap back on. Return the mouse to the cage after administering analgesia and monitor for any sign of pain.As the silicon probe needs to stabilize in the brain, allow the mouse to recover for an entire day before recording when the mouse runs on the treadmill. Install the anesthetized mouse in the stereotaxis apparatus by fixing the head plate. Remove the head cap.Bring the stereotaxic manipulator just above the micro drive. Fix the micro drive to the manipulator. Fix the micro drive to the micro drive handle by tightening the dedicated screws, and remove the screw connecting the micro drive to the shell part.Remove the screw connecting the shell and body of the micro drive. Under binocular microscope supervision, slowly pull up the micro drive and silicon probe assemblage with the stereotaxic manipulator, leaving the shell part behind. This is a color coded representation of place fields for a cell recorded on day one of treadmill running after probe implantation.Some cells showed repeated place fields correlated with the identity of the cues. This is the spike auto correlogram for the cell, and this image shows the spike wave forms for the cell example recorded on day one. This image shows the color coded representation of place fields for a cell recorded on day five after repeated exposure to the belt.Cells here showed a unique place field. Once mastered, each surgical procedure takes about one hour. Special care should be given to the craniotomy and the remitter removal to avoid any brain damage.Always keep the tools and surgical area clean and aseptic, and monitor the animal wellbeing during and after the surgeries. After watching this video, you should have a good understanding of how to implant chronic silicon probes in the hippocampus and how to record place cell activity.

Research

JoVE Journal

Neuroscience

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Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of ...

Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals

Хирургическая имплантация хронических Нейронные электроды для записи один вид деятельности группы и Electrocorticographic сигналы

The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery.  Here we provide useful information for ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

Крупномасштабные Запись нейронов Зонды Движимое кремния в Поведение грызунов

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals

Проектирование и монтаж Сверхлегкий моторизованная Microdrive при хронических нейронных Recordings у мелких животных

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Двойной Электрофизиологические Записи синаптически-вызванных астроглиального и нейрональных ответов, при остром срезов гиппокампа

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

Волоконно-оптические имплантации для хронической стимуляции Optogenetic ткани головного мозга

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Электрофизиологические Способ записи ответов внутриклеточный амперных<em&gt; Drosophila</em&gt; Фоторецепторов и Интернейроны к световой раздражитель<em&gt; В Vivo</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

Хронический транскраниальной электростимуляции и Intracortical запись в крыс

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Хронический имплантации целом корковых Electrocorticographic массива в общей игрунка

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Chronic Implantation of Multiple Flexible Polymer Electrode Arrays

Хроническая имплантация нескольких гибких полимерных электродов

Simultaneous recordings from large populations of individual neurons across distributed brain regions over months to years will enable new avenues of ...