SN performed the experiments described above, performed the data analysis and drafted the manuscript. AD and JDM implemented the technique at our research facilities and aided in the data analysis. HC aided in performing the experiments. WJ, CK and DG assisted in implementing the afferent measurement technique in our lab, the data analysis and interpretation of the results. SF, JDM and BDW designed the study. All authors critically read and approved the final manuscript. SN is an aspirant of the Fund for Scientific Research (FWO), Flanders (11G7415N). This work was supported financially by the FWO (G028615N and G034113N).

<ol>
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The authors have nothing to disclose.

本論文ではプロトコルは、当社グループおよびその他3,4,7,8,12,20,21,31によって使用されるようなげっ歯類で腸間膜求心性神経活動を研究するために、再現実験室の技術が記載されています。プロトコル内の重要なステップは、キャピラリーに、周囲の脂肪組織を吸引することにより、吸引電極および器官槽からガラスキャピラリーの適切な「シール」に組織の迅速な単離、神経ストラ?...

感覚シグナリングおよび疼痛知覚は、求心性神経、脊髄神経細胞、昇順と降順促通と抑制性経路およびいくつかの異なる脳領域間の複雑な相互作用から生じる複雑なプロセスです。このように、これらのレベルの一つ以上における変化は、変化した感覚シグナリングおよび疾患状態における内臓痛をもたらすことができます。感覚シグナリング複数の技術のすべてのこれらのさまざまな側面を研究するために、単一細胞実験に至るまで開発されている（ 例えば、神経細胞上のカルシウムイメージング）は、全体の動物モデルに（ 例えば、内臓運動応答として行動反応）。この論文に記載された技術は、研究者は、具体的にげっ歯類において小腸または結腸の孤立したセグメントからin vitroでの求心性神経活動を評価することができます。要するに、孤立した胃腸のセグメントは、（通常は空腸または結腸）生理Kで灌流専用記録チャンバー内に搭載されていますrebsソリューション。内臓神経は自由解剖し、内臓や骨盤の求心性神経における求心性神経活動の登録を可能にする電極に接続されています。神経活動は、求心性放電1-6に対する効果を評価するために、基底にまたは直接記録チャンバー（serosally）に適用することができる管腔内圧力および/または薬理学的化合物の増加に応じて記録された、または腔内灌流（粘膜）を介してすることができます。注目すべきは、内臓神経はまた、求心性感覚神経に加えて、遠心性繊維とviscerofugal求心性神経が含まれています。 ex vivoで内臓神経の記録の主要な利点の一つは、研究者が、神経活動に局所的に適用される化合物の直接の効果を研究するために、1つを可能にする、中枢神経系からの変調または入力なしで神経活動を定量化することができるという事実です。さらに、 インビボでのアプローチを使用して必要であるとして重要なパラメータの監視は、（下記参照）、nはO長く関連する。 インビトロ内臓記録は最終的にはるかに少ない時間がかかり、そのin vivoでの対応を超えています。 このような粘膜なでるなどの他の刺激、フォン・フレイ毛を使用したプロービングまたはセグメントの延伸に応じて、求心性神経活動は、腸組織が釘付けとは対照的である（縦方向に開放された修正された実験で研究することができます前号7,8で説明したように、無傷のセグメントを使用して私たちのセットアップ）、。また、ごく最近になって、技術が再び縦方向にセグメント9を開いて、釘付け使用して、カルシウムイメージングを介した結腸壁自体に結腸求心性神経の活性化を研究するために説明しました。 このin vivoでの技術の代替バージョンは、脊髄への求心性のエントリの近くに神経活動を測定するの外に構成されています。要するに、鎮静させた動物は、腹臥位で電子を配置されています、椎弓切除術による関心のプロジェクトの求心性神経に腰仙脊髄をxposingパラフィンで満たされただけでなく、切開部の皮膚を使用して、白金双極電極10,11の上に背細根を立体裁断を構築します。この技術は、さらに、研究者が彼らの伝導速度に基づいて、繊維を特徴付けることができ、薄く有髄Aδ線維から無髄C線維を区別する。また、背側根は、排他的に前述の混合求心性と遠心性内臓神経とは対照的に、感覚求心性繊維を含んでいます。 2つの研究グループが独立して人間の切除標本12,13における結腸求心性神経活動を記録するファースト・イン・マン写本を公開している、単離された腸セグメントからのin vitroでの求心性神経放電を記録することは、人間の標本を使用しても行うことができます。この技術の実装がより容易にtranslatiにつながる可能性人道的な状態にネズミのデータの上に、と研究者が容易に増感知覚神経を標的とする薬剤を同定する可能性があります。求心性神経活動を特徴づけるの臨床的重要性だけでなく、法外な求心性神経活動を標的とする新しい治療薬の発見、精巧フィールド14-19で多くの専門家によって議論されてきました。 上記のインビトロ技術は、より一般的に求心性神経活動のin vivoでの測定で知られて補完します。 インビボでのニューロンの活性測定の間、神経活動は、目的のセグメントが識別され、続いて挿管し、そして液体よくパラフィン充填されている間鎮静動物において直接測定することができる齧歯類20の腹壁および皮膚を用いて構成されています。関心の求心性神経は、その後、神経活動のmeasuremenを可能にする、双極白金電極上に切断し、置かれ、識別されトン。この技術は、鎮静動物いえ生きで求心性神経活動を調節するために研究を可能にします。以下のような、1は、このような管腔膨満または化合物の静脈内投与などの干渉に応答するニューロンの活動を学ぶことができます。 トランスレーショナル研究は、最近、主にヒト由来の上清のアプリケーションに焦点を当てて（ 例えば 、結腸生検、栽培末梢血単核細胞などからの）空腸および/または結腸マウスの求心性神経21,22上。粘膜アプリケーションに対する漿膜の差動効果が求心性神経放電に研究することができるので、研究者は直接、臓器浴にまたは腸セグメントを灌流管腔内溶液中にいずれかの上清を適用することができます。このように、それは過敏性腸症候群の患者からの結腸粘膜生検supernatansは、マウス結腸求心性神経、モルモット粘膜下神経細胞とマウスの後根に過敏症を引き起こすことができることが示されました神経節ニューロン21,23,24。 最後に、神経活動を記録することは腸間膜に制限および/または骨盤神経細胞は、胃腸管を支配されていません。他の人が他の人が26-28と同様の膀胱求心性神経活動を特徴とし、膀胱から骨盤の求心性神経だけでなく、消化管が収束することを証明し、おそらく神経細胞が得られているのに対し、神経録音は、25膝関節を供給求心性神経で実行することができることを実証しましたクロストーク29。

<table><tbody><tr><td>sodium chloride (NaCl)</td><td>VWR Chemicals</td><td>27,810,295</td><td>compound Krebs solution</td></tr><tr><td>potassium chloride (KCl)</td><td>Acros organics</td><td>196770010</td><td>compound Krebs solution</td></tr><tr><td>sodium dihydrogen phosphate (NaH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;)</td><td>VWR Chemicals</td><td>1,063,461,000</td><td>compound Krebs solution</td></tr><tr><td>sodium bicarbonate (NaHCO&lt;sub&gt;3&lt;/sub&gt;)</td><td>Merck</td><td>1,063,291,000</td><td>compound Krebs solution</td></tr><tr><td>magnesium sulfate (MgSO&lt;sub&gt;4&lt;/sub&gt;)</td><td>Merck</td><td>1,058,861,000</td><td>compound Krebs solution</td></tr><tr><td>calcium chloride (CaCl&lt;sub&gt;2&lt;/sub&gt;)</td><td>Merck</td><td>23,811,000</td><td>compound Krebs solution</td></tr><tr><td>D-glucose</td><td>VWR Chemicals</td><td>1011175P</td><td>compound Krebs solution</td></tr><tr><td>Distilled water</td><td></td><td></td><td>compound Krebs solution</td></tr><tr><td>PVC tubing</td><td>Scientific Laboratory Supplies</td><td></td><td>The intestinal segment should be mounted over PVC tubing</td></tr><tr><td>Silicone tubing</td><td>Scientific Laboratory Supplies</td><td></td><td>The rest of the tubing, ideally silicone-based - more easily dislodging of debris in the tubing</td></tr><tr><td>Silk thread</td><td>Pearsall Limited</td><td>10B15S220</td><td>Attachment of the segment over the PVC tubing</td></tr><tr><td>Syringe driver</td><td>Harvard Apparatus</td><td>55-2222</td><td>Intraluminal infusion of Krebs</td></tr><tr><td>Binocular - including 10X magnification in oculair</td><td>Zeiss STEMI 2000</td><td></td><td>Optimal visualization for the dissection of the afferent nerve</td></tr><tr><td>Homeothermic Blanket Control Unit</td><td>Harvard Apparatus</td><td>507214</td><td>Heating of the organ chamber</td></tr><tr><td>Custom made organ bath with Sylgard covered bottom</td><td></td><td></td><td></td></tr><tr><td>Spike2 software</td><td></td><td></td><td>Recording and analysis of the data</td></tr><tr><td>Insect pins, 500 pieces, stainless steel, diameter 0.2 mm</td><td>Austerlitz insect pins minutiens</td><td></td><td>Dissection of the afferent nerve</td></tr><tr><td>Tweezer Dumont #5 inox 11 cm</td><td>World Precision Instrument</td><td>500341</td><td>Dissection of the afferent nerve</td></tr><tr><td>Scissors, spring, 14 cm</td><td>World Precision Instrument</td><td>15905</td><td>Dissection of the afferent nerve</td></tr><tr><td>DB digitimer&amp;nbsp;</td><td>NL 108T2/10</td><td></td><td>pressure transducer</td></tr><tr><td>Micromanipulator</td><td>Narishige</td><td>M-3333</td><td>3D manipulation of the suction electrode</td></tr><tr><td>Micromanipulator</td><td>X-4 rotating block</td><td></td><td>3D manipulation of the suction electrode</td></tr><tr><td>Micromanipulator</td><td>GJ-8 magnetic stand</td><td></td><td>3D manipulation of the suction electrode</td></tr><tr><td>LightSource</td><td>Euromex Microscopes Holland EK-1</td><td></td><td>Optimal visualization for the dissection of the afferent nerve</td></tr><tr><td>CED 1401 Recording Apparatus</td><td></td><td></td><td>Recording of afferent nerve activity</td></tr><tr><td>Humbug 50/60 Hz Noise Eliminator</td><td>Quest Scientific Instruments</td><td></td><td>Elimination of background noise</td></tr><tr><td>Infusion Pump</td><td>Gibson Minipuls 2</td><td></td><td>Infusion of the organ chamber in which the segment is mounted</td></tr><tr><td>Microelectrode Holder Half Cells 1.5 mm</td><td>World Precision Instrument</td><td>MEH2SW</td><td>Suction electrode for isolation of the afferent fiber</td></tr><tr><td>Borosilicate Glass Capillaries, 300 pc; 1.5/0.84 OD/ID</td><td>World Precision Instrument</td><td>1B150-4</td><td>Capillary for the isolation of the afferent nerve</td></tr></tbody></table>

in vitro recording of mesenteric afferent nerve activity in mouse jejunal and colonic segments

求心性神経は正常な生理機能に関する情報を伝えるだけでなく、中枢神経系に向かって周囲から乱れ恒常性と異なる器官系の病態生理学的プロセスを知らせるだけでなく。このように、腸間膜求心性神経の活性の増加または「感作」は、内臓過敏症と腹部の疼痛症候群の病態生理に重要な役割を割り当てられています。 腸間膜求心性神経活動は、専用の器官浴中に取り付けられ、そこから内臓神経は、研究者が直接消化管セグメントに隣接する神経活動を評価することを可能にする、単離されている単離された腸セグメントにおいてインビトロで測定することができます。活動は、セグメントの膨満または管腔内またはserosally配信薬理学的化合物の添加後の間に、標準化された状態で、ベースラインで記録することができます。この技術は、可能研究者は、簡単コントロール検体における末梢神経系を標的とする薬剤の効果を研究します。他にも、それは病気の間に変更されている方法ニューロン活動に関する重要な情報を提供します。 例えば、後根神経節、脊髄または中枢神経系（求心性ニューロンの発火活性を測定することのみシグナル伝達カスケードの複雑な神経で1つの中継局を構成し、研究者は他のレベルでの神経活動を見落とさないように心に留めなければならないことに留意すべきです）完全に健康と病気における複雑な神経生理学を解明するためです。 一般的に使用されるアプリケーションは、リポ多糖の投与に応答した神経活動の研究、および過敏性腸症候群の動物モデルでの求心性神経活動の研究が含まれます。より翻訳アプローチでは、単離されたマウスの腸セグメントは、IBS患者の結腸の上清に曝露することができます。また、修正この技術の最近のヒト結腸検体において適用可能であることが示されています。

空腸求心性神経活動は、ベースラインで測定し、応答してOF-1マウス9 8週齢の雄で膨満をランプします。水と通常の飼料をタップする無制限のアクセスを持つ動物は、標準化された条件（50％、12時間の明暗サイクル - - 22°C、湿度40ケージ当たり6匹、20）のグループに収容しました。マウスの空腸セグメントは0 mmHgの（自発的活動11.47±3.31悪童/秒を意味する）の管腔?...

Watch this Scientific Journal Video about In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments at JoVE.com

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity.

マウス空腸および結腸セグメントにおける腸間膜求心性神経活動のin vitro記録で

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the ...

in-vitro-recording-mesenteric-afferent-nerve-activity-mouse-jejunal

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

11.4K ビュー.  University of Antwerp. Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity.

ビデオ: In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行性蛍光標識は、特定された神経細胞から標的細胞外シングルユニット記録を可能にします<em生体内で&gt;</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

神経科学

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

ローカスの神経活動を特徴づけるためにマルチユニット記録方法（<em&gt; Schistocercaアメリカーナ</em&gt;）嗅覚回路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

げっ歯類におけるランニングホイール活動の概日リズムの記録および分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

In Vivo Electrophysiological Measurements on Mouse Sciatic Nerves

Electrophysiological studies allow a rational classification of various neuromuscular diseases and are of help, together with neuropathological techniques, in the understanding of the underlying pathophysiology1. Here we describe a method to perform electrophysiological studies on mouse sciatic nerves in vivo.
The animals are anesthetized with isoflurane in order to ensure analgesia for the tested mice and undisturbed working environment during the measurements that take about 30 min/animal. A constant body temperature of 37 &#176;C is maintained by a heating plate and continuously measured by a rectal thermo probe2. Additionally, an electrocardiogram (ECG) is routinely recorded during the measurements in order to continuously monitor the physiological state of the investigated animals.
Electrophysiological recordings are performed on the sciatic nerve, the largest nerve of the peripheral nervous system (PNS), supplying the mouse hind limb with both motoric and sensory fiber tracts. In our protocol, sciatic nerves remain in situ and therefore do not have to be extracted or exposed, allowing measurements without any adverse nerve irritations along with actual recordings. Using appropriate needle electrodes3 we perform both proximal and distal nerve stimulations, registering the transmitted potentials with sensing electrodes at gastrocnemius muscles. After data processing, reliable and highly consistent values for the nerve conduction velocity (NCV) and the compound motor action potential (CMAP), the key parameters for quantification of gross peripheral nerve functioning, can be achieved.

In Vivo Electrophysiological Measurements on Mouse Sciatic Nerves

Measurements of nerve conduction properties in vivo exemplify a powerful tool to characterize various animal models of neuromuscular diseases. Here, we present an easy and reliable protocol by which electrophysiological analysis on sciatic nerves of anesthetized mice can be performed.

マウス坐骨神経に生体内電気生理学的測定値の

Electrophysiological studies allow a rational classification of various neuromuscular diseases and are of help, together with neuropathological ...

In vitro Functional Characterization of Mouse Colorectal Afferent Endings

This video demonstrates in detail an in vitro single-fiber electrophysiological recording protocol using a mouse colorectum-nerve preparation. The approach allows unbiased identification and functional characterization of individual colorectal afferents. Extracellular recordings of propagated action potentials (APs) that originate from one or a few afferent (i.e., single-fiber) receptive fields (RFs) in the colorectum are made from teased nerve fiber fascicles. The colorectum is removed with either the pelvic (PN) or lumbar splanchnic (LSN) nerve attached and opened longitudinally. The tissue is placed in a recording chamber, pinned flat and perfused with oxygenated Krebs solution. Focal electrical stimulation is used to locate the colorectal afferent endings, which are further tested by three distinct mechanical stimuli (blunt probing, mucosal stroking and circumferential stretch) to functionally categorize the afferents into five mechanosensitive classes. Endings responding to none of these mechanical stimuli are categorized as mechanically-insensitive afferents (MIAs). Both mechanosensitive and MIAs can be assessed for sensitization (i.e., enhanced response, reduced threshold, and/or acquisition of mechanosensitivity) by localized exposure of RFs to chemicals (e.g., inflammatory soup (IS), capsaicin, adenosine triphosphate (ATP)). We describe the equipment and colorectum&#8211;nerve recording preparation, harvest of colorectum with attached PN or LSN, identification of RFs in the colorectum, single-fiber recording from nerve fascicles, and localized application of chemicals to the RF. In addition, challenges of the preparation and application of standardized mechanical stimulation are also discussed.

In vitro Functional Characterization of Mouse Colorectal Afferent Endings

This video demonstrates a protocol for conducting single-fiber electrophysiological recordings on an in vitro mouse colorectum-nerve preparation.

マウス大腸求心性終末の体外機能解析では、

This video demonstrates in detail an in vitro single-fiber electrophysiological recording protocol using a mouse colorectum-nerve preparation. The ...

医学

Video Imaging and Spatiotemporal Maps to Analyze Gastrointestinal Motility in Mice

The enteric nervous system (ENS) plays an important role in regulating gastrointestinal (GI) motility and can function independently of the central nervous system. Changes in ENS function are a major cause of GI symptoms and disease and may contribute to GI symptoms reported in neuropsychiatric disorders including autism. It is well established that isolated colon segments generate spontaneous, rhythmic contractions known as Colonic Migrating Motor Complexes (CMMCs). A procedure to analyze the enteric neural regulation of CMMCs in ex vivo preparations of mouse colon is described. The colon is dissected from the animal and flushed to remove fecal content prior to being cannulated in an organ bath. Data is acquired via a video camera positioned above the organ bath and converted to high-resolution spatiotemporal maps via an in-house software package. Using this technique, baseline contractile patterns and pharmacological effects on ENS function in colon segments can be compared over 3-4 hr. In addition, propagation length and speed of CMMCs can be recorded as well as changes in gut diameter and contraction frequency. This technique is useful for characterizing gastrointestinal motility patterns in transgenic mouse models (and in other species including rat and guinea pig). In this way, pharmacologically induced changes in CMMCs are recorded in wild type mice and in the Neuroligin-3R451C mouse model of autism. Furthermore, this technique can be applied to other regions of the GI tract including the duodenum, jejunum and ileum and at different developmental ages in mice.

This article describes a video imaging technique and high-resolution spatiotemporal mapping to identify changes in the neural regulation of colonic motility in adult mice. Subtle effects on gastrointestinal (GI) function can be detected using this approach in isolated tissue preparations to advance our understanding of GI disease.

マウスにおける胃腸の運動を分析するためにビデオ画像と時空間マップ

The enteric nervous system (ENS) plays an important role in regulating gastrointestinal (GI) motility and can function independently of the central ...

In Vitro Characterization of the Electrophysiological Properties of Colonic Afferent Fibers in Rats

Dysfunction of the colonic sensory nerves has been implicated in the pathophysiology of several common conditions, including functional and inflammatory bowel diseases and diabetes. Here, we describe a protocol for the in vitro characterization of the electrophysiological properties of colonic afferents in rats. The colorectum, with the intact pelvic ganglion (PG) attached, is removed from the rat; superfused with carbogenated Krebs solution in the recording chamber; and cannulated at the oral and anal ends to allow for distension. A fine nerve bundle emanating from the PG is identified, and the multiunit afferent nerve activity is recorded using a suction electrode. Distension of the colonic segment elicits gradual increases in multiunit discharge. A principal component analysis is conducted to differentiate the low-threshold, the high-threshold, and the wide-dynamic range afferent fibers. Chemical sensitivity of colonic afferents can be studied through the bath or intraluminal administration of test compounds. This protocol can be modified for application to other species, such as mice and guinea pigs, and to study the differences in the electrophysiological properties of thoracolumbar/hypogastric and lumbosacral/pelvic afferents of the descending colon in normal and pathological conditions.

In Vitro Characterization of the Electrophysiological Properties of Colonic Afferent Fibers in Rats

Abnormal sensory function underlies visceral pain and other symptoms of functional and inflammatory bowel diseases. A protocol for the electrophysiological recording of the colonic afferent nerves in an ex vivo rat colorectum preparation is presented here.

体外ラットの結腸求心性線維の電気生理学的特性の評価

Dysfunction of the colonic sensory nerves has been implicated in the pathophysiology of several common conditions, including functional and inflammatory ...

Electrophysiological Recording from Colonic Afferent Nerves in an Ex Vivo Rat Colon

Source: Meng, Y. et al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/56090">In Vitro Characterization of the Electrophysiological Properties of Colonic Afferent Fibers in Rats.</a> J. Vis. Exp. (2017)
This video demonstrates a method to record the activity of colonic afferent nerves in an ex-vivo rat colon, cannulated at both ends and infused with saline to increase intraluminal pressure. As pressure increases, the nerve signal rises, indicating its stimulation.

Ex Vivoラット結腸における結腸求心性神経からの電気生理学的記録

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
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JoVE Encyclopedia of Experiments

Neurophysiology

Electrophysiology Techniques

Recording of Mesenteric Afferent Nerve Activity in an Ex Vivo Mouse Jejunum Segment

Source: Nullens, S., et al. <a href="https://appjove.unicartagenaproxy.elogim.com/v/54576">In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments.</a> J. Vis. Exp. (2016).
This video shows the process of recording electrical signals from mesenteric afferent nerves in a dissected mouse jejunal segment to study the effects of tissue pressure and movement changes on afferent nerve activity.

マウス空腸および結腸セグメントにおける腸間膜求心性神経活動のin vitro記録で

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逆行性蛍光標識は、特定された神経細胞から標的細胞外シングルユニット記録を可能にします

ローカスの神経活動を特徴づけるためにマルチユニット記録方法（