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

この作品は、韓国科学技術機関プログラム (プロジェクト号 2E26190 と 2E26170) とフロンティア科学人間 (RGY0089/2012 年) によって支えられました。

説明のすべてのメソッドは、動物愛護と韓国科学技術研究所の使用委員会によって承認されている。
 1 マイクロ ドライブと電極の準備 <ol><li>マイクロ ドライブを組立。。
	<ol><li>マイクロ ドライブ (スライダー、ボディ、およびシェル) の部分は印刷 14 高解像度 3 D プリンターを使用しています。部品には欠陥がない確認します</li>。
		<...

<ol>
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S., Sokoloff, G., Tiriac, A., Del Rio-Bermudez, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+valuable+and+promising+method+for+recording+brain+activity+in+behaving+newborn+rodents.">A valuable and promising method for recording brain activity in behaving newborn rodents.</a> Dev Psychobiol. 57 (4), 506-517 (2015).</li><li>Haiss, F., Butovas, S., Schwarz, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+miniaturized+chronic+microelectrode+drive+for+awake+behaving+head+restrained+mice+and+rats.">A miniaturized chronic microelectrode drive for awake behaving head restrained mice and rats.</a> J Neurosci Methods. 187 (1), 67-72 (2010).</li><li>Dombeck, D. A., Khabbaz, A. N., Collman, F., Adelman, T. L., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+large-scale+neural+activity+with+cellular+resolution+in+awake,+mobile+mice.">Imaging large-scale neural activity with cellular resolution in awake, mobile mice.</a> Neuron. 56 (1), 43-57 (2007).</li><li>Villette, V., Malvache, A., Tressard, T., Dupuy, N., Cossart, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Internally+Recurring+Hippocampal+Sequences+as+a+Population+Template+of+Spatiotemporal+Information.">Internally Recurring Hippocampal Sequences as a Population Template of Spatiotemporal Information.</a> Neuron. 88 (2), 357-366 (2015).</li><li>Ziv, Y., 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=Long-term+dynamics+of+CA1+hippocampal+place+codes.">Long-term dynamics of CA1 hippocampal place codes.</a> Nat Neurosci. 16 (3), 264-266 (2013).</li><li>Danielson, N. 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=Distinct+Contribution+of+Adult-Born+Hippocampal+Granule+Cells+to+Context+Encoding.">Distinct Contribution of Adult-Born Hippocampal Granule Cells to Context Encoding.</a> Neuron. 90 (1), 101-112 (2016).</li><li>Stark, E., Koos, T., Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diode+probes+for+spatiotemporal+optical+control+of+multiple+neurons+in+freely+moving+animals.">Diode probes for spatiotemporal optical control of multiple neurons in freely moving animals.</a> J Neurophysiol. 108 (1), 349-363 (2012).</li><li>Wu, F., 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=An+implantable+neural+probe+with+monolithically+integrated+dielectric+waveguide+and+recording+electrodes+for+optogenetics+applications.">An implantable neural probe with monolithically integrated dielectric waveguide and recording electrodes for optogenetics applications.</a> J Neural Eng. 10 (5), 056012 (2013).</li></ol>

神経の慢性的な録音は海馬場所フィールドなど神経プロセスを理解することにとって重要です。慢性的なシリコン プローブ implantantation を実行へのアプローチがで電極パッケージを回復する比較的簡単だという事実によって他の方法7,18,19,20から自分自身を区別します。実験終了。場所細胞が通常...

シリコン プローブの電気生理学的記録、彼らは組織の損傷や彼らが密集サイト1、録音の正確なレイアウトを提示する最小限に抑えるシャープなプロファイル設計されているという事実を含むいくつかの利点を提示します。 2,3,4。種、人間3,5,6、多様なアプローチ1,7を含む様々 なシステムの研究に使用されます。まだの反復使用は、コスト、脆弱性、および慢性的な実験のための便利なメソッドが8を欠けている事実のためまだ比較的限られています。3 D 印刷技術の最近の進歩は、これらのデリケートな電極の取り扱いを簡単にできるようにヘッド プレート マイクロ ドライブなどのデバイスのカスタム設計に可能にしました。最初のステップで構築し、慢性的なシリコン プローブ14術の開発したツールのセットを使用する方法を説明します。
場所細胞は通常、迷路の実行している自由に動く動物を用いて調べた、間最近彼らも調べた仮想環境15とトレッドミル apparatii9 (図 1 a)。これらの実験方法は、動物の頭-拘束可能、optrode9,10、11パッチ ・ クランプの16、2 光子励起顕微鏡15の活用利点を提供します。動物の行動と環境手がかりの12の拡張コントロールを提供することに加えて、簡単テクニック。2 番目のステップでは、マウスをトレーニングとトレッドミル装置の場所細胞の活動を記録の手順を紹介します。

<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

脳の機能を理解するための重要な前提条件は挙動の同定と細胞活性の相関します。シリコン プローブ、神経活動の大規模な電気生理学的記録用電極を高度な彼らの慢性的な注入の手順がまだ未発達。環境で動物の位置に関連付ける海馬場所細胞の活動が知られているが、根本的なメカニズムはまだ明確ではないです。場所細胞を調べるためには、ここで我々 は慢性的なシリコン プローブのデバイスの作製からキュー濃縮トレッドミル装置の場所フィールド活動の監視にインプラント技術のセットをについて説明します。マイクロ ドライブと帽子は、フィッティングとプラスチック部品の 3 D プリントを一緒に固定して構築されます。シリコン プローブ マイクロ ドライブにマウントされているが、洗浄、染料でコーティングします。最初手術を実行して、マウス頭蓋の帽子を修正します。目印を作製し、トレッドミルのベルトに接続されています。頭固定を実行するマウスを訓練トレッドミルで。2 番目の手術が次のブロード バンド電気生理学的信号を記録、海馬におけるシリコン プローブをインプラントに実行されます。最後に、シリコン プローブを回収し、再利用のためきれいです。トレッドミルの場所細胞の活動の分析では、アプローチのメリットをまとめた場所フィールド メカニズムの多様性を明らかにします。

マウスは最初キュー (図 1) に欠けている 2 メートルの長いベルト上で、訓練されました。次の電極注入、同じ長さの新しいベルトが、キューの 3 つのペアを提示は、呈示空間表現12,14を生成するために、トレッドミルにインストールされました。広帯域信号が 250 チャンネル録音システム (アンプ基板?...

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

我々 は、慢性シリコン プローブをインプラントしてキュー濃縮トレッドミル装置の実行の頭固定されているマウスの場所細胞を記録するための多様な手順を説明します。

慢性的なシリコン プローブと豊かなトレッドミル装置海馬場所細胞の記録

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.

implantation-chronic-silicon-probes-recording-hippocampal-place-cells

Research

JoVE Journal

Neuroscience

Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of behaviors from the spiking activity of its neurons. One of the most effective methods to monitor spiking activity from a large number of neurons in multiple local neuronal circuits simultaneously is by using silicon electrode arrays1-3.
Action potentials produce large transmembrane voltage changes in the vicinity of cell somata. These output signals can be measured by placing a conductor in close proximity of a neuron. If there are many active (spiking) neurons in the vicinity of the tip, the electrode records combined signal from all of them, where contribution of a single neuron is weighted by its 'electrical distance'. Silicon probes are ideal recording electrodes to monitor multiple neurons because of a large number of recording sites (+64) and a small volume. Furthermore, multiple sites can be arranged over a distance of millimeters, thus allowing for the simultaneous recordings of neuronal activity in the various cortical layers or in multiple cortical columns (Fig. 1). Importantly, the geometrically precise distribution of the recording sites also allows for the determination of the spatial relationship of the isolated single neurons4. Here, we describe an acute, large-scale neuronal recording from the left and right forelimb somatosensory cortex simultaneously in an anesthetized rat with silicon probes (Fig. 2).

Extracellular recordings of neuronal activity using silicon probes in the anesthetized rat will be described. This technique allows information to be obtained across multiple brain areas from more than 100 neurons simultaneously. It provides information with single cell resolution about neuronal ensembles dynamics in multiple local circuits.

麻酔ラットのシリコンプローブで大規模なニューロンアンサンブルの録音

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

神経科学

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

The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery. Here we provide useful information for surgeons who are learning the process of implanting electrode systems. We demonstrate the implantation procedure of both a penetrating and a surface electrode. The surgical process is described from start to finish, including detailed descriptions of each step throughout the procedure. It should also be noted that this video guide is focused towards procedures conducted in rodent models and other small animal models. Modifications of the described procedures are feasible for other animal models.

We provide useful information for surgeons who are learning the process of implanting chronic neural recording electrodes. Techniques for both penetrating and surface electrode systems are described in a rodent animal model.

単一ユニット活動とElectrocorticographic信号を記録するための慢性神経電極の外科的移植

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

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

動作げっ歯類の可動シリコンプローブによるニューロンの大規模なレコーディング

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

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of neural circuits underlying a variety of natural behaviors, including navigation1 , decision making 2,3, and the generation of complex motor sequences4,5,6. Advances in precision machining has allowed for the fabrication of light-weight devices suitable for chronic recordings in small animals, such as mice and songbirds. The ability to adjust the electrode position with small remotely controlled motors has further increased the recording yield in various behavioral contexts by reducing animal handling.6,7
 Here we describe a protocol to build an ultra-light motorized microdrive for long-term chronic recordings in small animals. Our design evolved from an earlier published version7, and has been adapted for ease-of use and cost-effectiveness to be more practical and accessible to a wide array of researchers. This proven design 8,9,10,11 allows for fine, remote positioning of electrodes over a range of ~ 5 mm and weighs less than 750 mg when fully assembled. We present the complete protocol for how to build and assemble these drives, including 3D CAD drawings for all custom microdrive components.

The design, fabrication and assembly of an ultra-light motorized microdrive is described. The device provides a cost-effective and easy-to-use solution for chronic recordings of single units in small behaving animals.

小動物の慢性神経録音用超軽量電動マイクロドライブの設計およびアセンブリ

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

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

急性海馬スライスにおけるシナプス誘発アストログリアとニューロン活動の二重の電気生理学的記録

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

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard for analyzing patterns of synaptic connectivity, but paired electrophysiological recordings can be both cumbersome and experimentally limiting. The development of optogenetics has introduced an elegant method to stimulate neurons and circuits, both in vitro1 and in vivo2,3. By exploiting cell-type specific promoter activity to drive opsin expression in discrete neuronal populations, one can precisely stimulate genetically defined neuronal subtypes in distinct circuits4-6. Well described methods to stimulate neurons, including electrical stimulation and/or pharmacological manipulations, are often cell-type indiscriminate, invasive, and can damage surrounding tissues. These limitations could alter normal synaptic function and/or circuit behavior. In addition, due to the nature of the manipulation, the current methods are often acute and terminal. Optogenetics affords the ability to stimulate neurons in a relatively innocuous manner, and in genetically targeted neurons. The majority of studies involving in vivo optogenetics currently use a optical fiber guided through an implanted cannula6,7; however, limitations of this method include damaged brain tissue with repeated insertion of an optical fiber, and potential breakage of the fiber inside the cannula. Given the burgeoning field of optogenetics, a more reliable method of chronic stimulation is necessary to facilitate long-term studies with minimal collateral tissue damage. Here we provide our modified protocol as a video article to complement the method effectively and elegantly described in Sparta et al.8 for the fabrication of a fiber optic implant and its permanent fixation onto the cranium of anesthetized mice, as well as the assembly of the fiber optic coupler connecting the implant to a light source. The implant, connected with optical fibers to a solid-state laser, allows for an efficient method to chronically photostimulate functional neuronal circuitry with less tissue damage9 using small, detachable, tethers. Permanent fixation of the fiber optic implants provides consistent, long-term in vivo optogenetic studies of neuronal circuits in awake, behaving mice10 with minimal tissue damage.

The development of optogenetics now provides the means to precisely stimulate genetically defined neurons and circuits, both in vitro and in vivo. Here we describe the assembly and implantation of a fiber optic for chronic photostimulation of brain tissue.

脳組織の慢性Optogenetic刺激のための光ファイバ注入

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

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

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

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

の記録細胞内の電圧応答のための電気生理学的手法<em&gt;ショウジョウバエ</em&gt;光刺激に感光体と介在ニューロン<em&gt;インビボ</em

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

Quantification of Endosome and Lysosome Motilities in Cultured Neurons Using Fluorescent Probes

In the brain, membrane trafficking systems play important roles in regulating neuronal functions, such as neuronal morphology, synaptic plasticity, survival, and glial communications. To date, numerous studies have reported that defects in these systems cause various neuronal diseases. Thus, understanding the mechanisms underlying vesicle dynamics may provide influential clues that could aid in the treatment of several neuronal disorders. Here, we describe a method for quantifying vesicle motilities, such as motility distance and rate of movement, using a software plug-in for the ImageJ platform. To obtain images for quantification, we labeled neuronal endosome-lysosome structures with EGFP-tagged vesicle marker proteins and observed the movement of vesicles using a time-lapse microscopy. This method is highly useful and simplify measuring vesicle motility in neurites, such as axons and dendrites, as well as in the soma of both neurons and glial cells. Furthermore, this method can be applied to other cell lines, such as fibroblasts and endothelial cells. This approach could provide a valuable advancement of our understanding of membrane trafficking.

The investigation of membrane trafficking is crucial for understanding neuronal functions. Here, we introduce a method for quantifying vesicle motility in neurons. This is a convenient method that can be adapted to the quantification of membrane trafficking in the nervous system.

蛍光プローブを用いた培養ニューロンにおけるエンドソームおよびリソソームの動力学の定量

In the brain, membrane trafficking systems play important roles in regulating neuronal functions, such as neuronal morphology, synaptic plasticity, ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a closed-loop event-triggered manner. Although many studies are focusing on the possible benefits and side-effects of TES in healthy and pathologic brains, there are still many fundamental open questions regarding the mechanism of action of the stimulation. Therefore, there is a clear need for a robust and reproducible method to test the acute and the chronic effects of TES in rodents. TES can be combined with regular behavioral, electrophysiological, and imaging techniques to investigate neuronal networks in vivo. The implantation of transcranial stimulation electrodes does not impose extra constraints on the experimental design while it offers a versatile, flexible tool to manipulate brain activity. Here we provide a detailed, step-by-step protocol to fabricate and implant transcranial stimulation electrodes to influence brain activity in a temporally constrained manner for months.

This detailed protocol describes transcranial stimulation electrode placement on the temporal bone in order to investigate the short- and long-term effects of transcranial electrical stimulation in freely moving rats.

慢性的な経頭蓋電気刺激およびラットの皮質内の記録

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

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

一般的なセットの全皮質のシェットランドシープドッグ配列の慢性注入

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

慢性的なシリコン プローブと豊かなトレッドミル装置海馬場所細胞の記録

慢性的なシリコンプローブと豊かなトレッドミル装置海馬場所細胞の記録