Name: behavioral assessment of manual dexterity in non-human primates
Uploaded: 2011-11-11T13:00:00
Duration: 16 min
Description: Watch this Scientific Journal Video about Behavioral Assessment of Manual Dexterity in Non-Human Primates at JoVE.com

著者は、教授MEシュワブ、博士P.フロイント博士A. Wyssさん、博士S.バシル、博士A.ミール、博士J.ブロッホ、博士JF黒髪、博士J.に感謝したいと思いますAebischer、博士A.メーサーロシュ、以前の実験と解析への貢献のための博士V. Goetschman。実験的なセットアップはアンドレガイヤール、バーナードAebischerとローランMonneyすることにより構築した。ヨーゼフCorpataux、ローラン偉そうとジャックメイラード：動物施設では、サルは、プロの動物の世話の監督下に置かれた。ヴェロニクモレ（またウェブマスター）、フランソワーズティンゲリー、クリスティン配達夫、モニカBennefeld、クリスティマルティとジョーゼットフィッシャー：行動テストと分析データのほか、組織は、検査技師の非常に貴重な貢献を行った。この作品は、いいえ31〜61857.00、310000から110005、31003A - 132465（EMR）、310030から118357、31003A - 10406​​1（TW）、310030を許可しない、スイス国立科学財団によってサポートされていました-120411（ABS）、PZ00P3_121646（ES）、ノバルティス財団、&quot;神経可塑性と修復&quot;に関する研究の能力のナショナルセンター（NCCR）。

<ol>
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Med. 12, 790-792 (2006).</li><li>Freund, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-Nogo-A+antibody+treatment+promotes+recovery+of+manual+dexterity+after+unilateral+cervical+lesion+in+adult+primates--re-examination+and+extension+of+behavioral+data.">Anti-Nogo-A antibody treatment promotes recovery of manual dexterity after unilateral cervical lesion in adult primates--re-examination and extension of behavioral data.</a> Eur. J. Neurosci. 29, 983-996 (2009).</li><li>Kaeser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Unilateral+Motor+Cortex+Lesion+on+Ipsilesional+Hand's+Reach+and+Grasp+Performance+in+Monkeys:+Relationship+With+Recovery+in+the+Contralesional+Hand.">Effects of Unilateral Motor Cortex Lesion on Ipsilesional Hand's Reach and Grasp Performance in Monkeys: Relationship With Recovery in the Contralesional Hand.</a> J. Neurophysiol. 103, 1603-1645 (2010).</li><li>Kaeser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+adult+cortical+cell+transplantation+enhances+functional+recovery+following+unilateral+lesion+of+motor+cortex+in+primates:+a+pilot+study.">Autologous adult cortical cell transplantation enhances functional recovery following unilateral lesion of motor cortex in primates: a pilot study.</a> Neurosurgery. 68, 1405-1417 (2011).</li><li>Bashir, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-term+effects+of+unilateral+lesion+of+the+primary+motor+cortex+(M1)+on+Ipsilesional+hand+dexterity+in+adult+macaque+monkeys.">Short-term effects of unilateral lesion of the primary motor cortex (M1) on Ipsilesional hand dexterity in adult macaque monkeys.</a> Brain Structure and Function. , (2011).</li><li>Hamadjida, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+anti-Nogo-A+treatment+on+the+reorganization+of+callosal+connectivity+of+the+premotor+cortical+areas+following+unilateral+lesion+of+primary+motor+cortex+(M1)+in+adult+macaque+monkeys.">Influence of anti-Nogo-A treatment on the reorganization of callosal connectivity of the premotor cortical areas following unilateral lesion of primary motor cortex (M1) in adult macaque monkeys.</a> , (2011).</li><li>Jenny, A. 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Neurosci. 3, 567-575 (1983).</li><li>Schmidlin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+plastic+changes+in+the+hand+representation+of+the+primary+motor+cortex+parallel+incomplete+recovery+from+a+unilateral+section+of+the+corticospinal+tract+at+cervical+level+in+monkeys.">Progressive plastic changes in the hand representation of the primary motor cortex parallel incomplete recovery from a unilateral section of the corticospinal tract at cervical level in monkeys.</a> Brain Research. 1017, 172-183 (2004).</li><li>Schmidlin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+the+hand+representation+in+the+ipsilateral+primary+motor+cortex+following+unilateral+section+of+the+corticospinal+tract+at+cervical+level+in+monkeys.">Reduction of the hand representation in the ipsilateral primary motor cortex following unilateral section of the corticospinal tract at cervical level in monkeys.</a> BMC Neuroscience. 6, 56-56 (2005).</li><li>Prescott, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Refinement+of+the+use+of+food+and+fluid+control+as+motivational+tools+for+macaques+used+in+behavioural+neuroscience+research:+report+of+a+Working+Group+of+the+NC3Rs.">Refinement of the use of food and fluid control as motivational tools for macaques used in behavioural neuroscience research: report of a Working Group of the NC3Rs.</a> J. Neurosci. Methods. 193, 167-188 (2010).</li><li>Darling, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volumetric+effects+of+motor+cortex+injury+on+recovery+of+dexterous+movements.">Volumetric effects of motor cortex injury on recovery of dexterous movements.</a> Exp. Neurol. 220, 90-108 (2009).</li><li>Darling, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimal+forced+use+without+constraint+stimulates+spontaneous+use+of+the+impaired+upper+extremity+following+motor+cortex+injury.">Minimal forced use without constraint stimulates spontaneous use of the impaired upper extremity following motor cortex injury.</a> Exp. Brain. Res. 202, 529-542 (2010).</li><li>McNeal, 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=Selective+long-term+reorganization+of+the+corticospinal+projection+from+the+supplementary+motor+cortex+following+recovery+from+lateral+motor+cortex+injury.">Selective long-term reorganization of the corticospinal projection from the supplementary motor cortex following recovery from lateral motor cortex injury.</a> J. Comp. 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現在の行動のタスクは、様々な治療法をテストすることを目的と頚髄の病変にまたは運動野の病変に関連した研究の文脈でこれまでに我々の研究室で検討されているが（14,15,17および参照してください<a href="http://www.unifr.ch/neuro/rouiller">http:/を/ www.unifr.ch /神経/ rouiller</a> &gt;その後、トップバーメニューの&quot;研究&quot;を選択&gt;&quot;モーターシステム&quot;&gt;&quot;損傷後の回復&quot;）手先の器用さも側面であるとして、彼らはまた、広範なアプリケーションがあるかもしれませんそのようなパーキンソン病（MPTPのサル）など、感覚脱afferentation影響を与える固有感覚および/ ​​またはタッチの感覚（特に視覚的なコントロールがない場合のテスト2）の場合は他の病態で検討すること。 手先の器用さに関係するとしてここで提案する行動テストは、前肢の遠位の動きのモータ制御を調査するために適しています。テストの特異性はによって示されている制御システムの関連コンポーネントを損なわない病変の場合に赤字の不在（​​2日間を除く）：確かに、病変の場合には、手の筋肉を制御する運動ニューロンよりも尾側に配置、何赤字はありませんでした。テスト1の適切さは、おそらく細胞療法の治療17に関連して、25％の機能回復の強化によって異なるポスト病変の回復曲線の2つの時点、で撮影したビデオシーケンスを比較することによって理解することができます。 初めにいくつかの初期の、比較的短い訓練の段階（一般的には2-3ヶ月持続する）のにもかかわらず、ここで提案する行動テストは、複雑な（例えば、条件付きの）タスクに比べて、相対的に&quot;自然&quot;と分かりやすいですそのために猿の訓練ほぼ一年以上かかることがあります。正の強化は、通常よりcomに使用される水の欠乏よりも倫理的観点について、機密性の低い固体の食品、、に基づいているプレックスは、タスク23。安定的かつ一貫性のある結果を得るために食べ物からサルを剥奪する必要はありません。作業中に受信したペレットは、行動のセッション（ただし、追加の食べ物は、前日の午後の終わりまで与えることができる猿が先行する夜の間に食べることはないと仮定すると）の日の食べ物への最初のアクセスを表します。それは、それぞれのサルの行動の日の同じ時間でセッションだけでなく、住宅室内のグループを形成する別のサルの間に同一の順序を尊重して行うことが重要です。サルは、外部の妨害事象に敏感であるため、行動のタスクは、隣接する部屋や研究室からの潜在的な外乱ノイズをマスクする、バックグラウンドミュージックの存在下で実施されるべきである。与えられたサルは、実験の全期間中（最大最後の日々の実験的なセッションへの最初の訓練から、数ヶ月の期間ではない年な場合）することが重要それである同じ実験者の監督の下で毎日置いた。 手先の器用さを定量化するために我々の研究室で数年前から使用されて現在の行動テストでは、、最近、24から28まで文献で ​​報告されている手先の器用さの他のテストに匹敵ある程度です。本報告書の暫定的な目標ですが、異なる研究室間のテストを（より良い比較のため）、標準化する重大な必要性は、しかしある。オンデマンドで、試験1〜4のここに示すようなセットアップは、それらを複製するために、対応する著者により提供することができるの詳細な特性。再生医療（脊髄または大脳皮質の病変からの回復）の問題を超えて、テストの現在のパレットが調査する、通常のヒト以外の霊長類の発達の問題（器用な動きの運動発達の例：時間の経過）に対処するために適している側性化の局面（手の嗜好/優位性）とcによって解読進化の質問へ人間の被験者を含む霊長類の異なる種の運動機能をomparing。それは、タスクのパフォーマンスに影響を与える可能性があるため、装置の寸法は、霊長類の数字&quot;のサイズ（太さと長さ）に応じて適応されるべきであることに注意してください。本研究では、テストは2.5から8歳までと、2.5〜8キロの重さ、カニクイザル猿で実施されました。人差し指の末節骨（先端）の周りは、私たちの研究に含まれているサルで22および25 mmの間であったのに対し、人差し指の長さ（ペレットを操作するために最初に使用）32〜35 mmまでの範囲、。以前の実験でテストとして、同じ把持テストは同様にアカゲザルに適しています。 全ての実験は、 実験動物の管理と利用 （1996年） のためにガイドにしたがって実施し、現地（スイス）の獣医当局によって承認された。猿上のすべての実験手順のだけでなく、動物施設における拘禁の条件は、我々の研究室から最近のレポートで詳細に記述された：リファレンス12-18を参照してください。

behavioral assessment of manual dexterity in non-human primates

皮質脊髄（CS）管は巧みに、小さなオブジェクトを操作するために絶妙な運動能力、主に霊長類1の特権の解剖学的なサポートです。その起源（モーター皮質領域の病変）で、またはその軌道（頸髄病変）に沿って、CSの投影システムに影響を与える病変の場合には、などのいくつかの四肢麻痺や片麻痺の患者に見られる手先の器用さの劇的な損失（手の麻痺が）、ある。このような病変の後、いくつかの自発的な機能回復があるが、それは成人では非常に限られている。様々な治療戦略は、現在提案されている（例えば、細胞治療、抑制性軸索成長の分子の中和、成長因子などのアプリケーション）、主にげっ歯類で開発されている。しかし、臨床応用する前に、それはしばしば、ヒト以外の霊長類における治療の実現可能性、有効性、およびセキュリティをテストすることをお勧めします。目標は、後の手先の器用さを復元するとき、これは特にそうです中枢神経系の病変は、げっ歯類の運動系の組織としての霊長類1,2のものとは異なっている。マカクザルは、例えば運動野や頚髄の病変から生じる赤字を反映するために、霊長類で手先の器用さを定量化するために適切な行動のモデルとして、ここで図示されている、自発的な機能回復の程度と、治療が適用される場合、評価を測定するどれだけそれが機能回復を高めることができます。 手先の器用さの行動評価は、大人のマカクザルの初期トレーニングが必要な4つの別個の、補完的な、達すると把握手動タスク（ペレットを把握するために精度のグリップを使用する）、に基づいています。動物の準備が実証だけでなく、セットアップの行動に関しては、位置決めされる。典型的なサルのパフォーマンスは、各タスクのために示されています。正確な手のマニピュレータを反映し、関連するパラメータの収集と分析るだけでなく、力の制御は、説明と代表的な結果で実証されています。これらのデータは、行動データが脊髄損傷のや運動野の病変の影響を調査し、治療が比較することにより、自発的機能回復を高めることがどの程度に悪用される方法を示す、より広い文脈で、配置されていますサルの異なるグループ（例えば治療を偽対扱わ）。行動テストの利点と制限について説明します。現在の行動のアプローチは、神経系の疾患2,3の文脈における非ヒト霊長類モデルの適切さを強調する以前の報告に沿ったものです。

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Behavioral Assessment of Manual Dexterity in Non-Human Primates

手先の器用さは、主に霊長類の特権であるため、行動タスクはマカクザルで開発されている。 4つは手の操作能力や力を測定、把握のタスクを達成し、把握、中枢神経系の損傷後の機能回復を確立するために、治療の効果をテストすることができます。

ヒト以外の霊長類における手先の器用さの行動評価

The corticospinal (CS) tract is the anatomical support of the exquisite motor ability to skillfully manipulate small objects, a prerogative mainly of ...

behavioral-assessment-of-manual-dexterity-in-non-human-primates

Research

JoVE Journal

Neuroscience

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

ラットのショ糖の渇望のインキュベーションを評価するための一般的な方法

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

神経科学

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, ...

Assessment of Dendritic Arborization in the Dentate Gyrus of the Hippocampal Region in Mice

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity and extent of dendritic arborization correlates well with the synaptic plasticity in these cells. A reliable method for assessment of dendritic arborization is needed to characterize the deleterious effects of neurological disorders on these structures and to determine the effects of therapeutic interventions. However, quantification of these structures has proven to be a formidable task given their complex and dynamic nature. Fortunately, sophisticated imaging techniques can be paired with conventional staining methods to assess the state of dendritic arborization, providing a more reliable and expeditious means of assessment. Below is an example of how these imaging techniques were paired with staining methods to characterize the dendritic arborization in wild type mice. These complementary imaging methods can be used to qualitatively and quantitatively assess dendritic arborization that span a rather wide area within the hippocampal region.

We describe two methods for visualization and quantification of dendritic arborization in the hippocampus of mouse models: real-time and extended depth of field imaging. While the former method allows sophisticated topographical tracing and quantification of the extent of branching, the latter allows speedy visualization of the dendritic tree.

マウスの海馬領域の歯状回における樹状樹枝状分岐の評価

Dendritic arborization has been shown to be a reliable marker for examination of structural and functional integrity of neurons. Indeed, the complexity ...

Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich structures suggest alterations in synaptic integrity and connectivity. Here we provide a detailed protocol for culturing primary rat cortical neurons, Phalloidin staining for F-actin puncta, and subsequent quantification techniques. First, the frontal cortex of E18 rat embryos are dissociated into low-density cell culture, then the neurons grown in vitro for at least 12-14 days. Following experimental treatment, the cortical neurons are stained with AlexaFluor 488 Phalloidin (to label the dendritic F-actin puncta) and microtubule-associated protein 2 (MAP2; to validate the neuronal cells and dendritic integrity). Finally, specialized software is used to analyze and quantify randomly selected neuronal dendrites. F-actin rich structures are identified on second order dendritic branches (length range 25-75 &#181;m) with continuous MAP2 immunofluorescence. The protocol presented here will be a useful method for investigating changes in dendritic synapse structures subsequent to experimental treatments.

Quantification of Filamentous Actin F-actin Puncta in Rat Cortical Neurons

Filamentous actin (F-actin) plays an important role in spinogenesis, synaptic plasticity, and synaptic stability. Quantification of F-actin puncta is therefore a useful tool to study the integrity of synaptic structures. This protocol describes the procedures of quantifying F-actin puncta labeled with Phalloidin in low-density primary cortical neuronal cultures.

ラット皮質ニューロンにおける繊維状アクチン（F-アクチン）涙点の定量化

Filamentous actin protein (F-actin) plays a major role in spinogenesis, synaptic plasticity, and synaptic stability. Changes in dendritic F-actin rich ...

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in regulating neuronal growth and development. However, the mechanisms underlying Drd1&#945; receptor abnormalities mediating behavioral responses and modulating working memory function are still unclear. Using a novel RNA in situ hybridization assay, the current study identified dopamine Drd1&#945; receptor and tyrosine hydroxylase (TH) RNA expression from DA-related circuitry in the nucleus accumbens (NAc) area and substantia nigra region (SNR), respectively. Drd1&#945; expression in the NAc shows a &#34;discrete dot&#34; staining pattern. Clear sex differences in Drd1&#945; expression were observed. In contrast, TH shows a &#34;clustered&#34; staining pattern. Regarding TH expression, female rats displayed a higher signal expression per cell relative to male animals. The methods presented here provide a novel in situ hybridization technique for investigating changes in dopamine system dysfunction during the progression of central nervous system diseases.

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Identification of dopamine D1-alpha receptor in the nucleus accumbens is critical for clarifying D1 receptor dysfunction during a central nervous system disease. We performed a novel RNA in situ hybridization assay to visualize single RNA molecules in a specific brain area.

検出技術その場で革新的な RNA 側坐齧歯動物の核内ドーパミン D1-アルファ受容体の同定

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.

刺激間隔の齧歯動物の一時的な処理の評価のための力

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from parallel fibers (PF) to Purkinje cells (PC) is considered the basis for motor learning, and deficiencies of both LTD and motor learning are observed in various gene-manipulated animals. Common motor learning sets, such as adaptation of the optokinetic reflex (OKR), the vestibular-ocular reflex (VOR), and rotarod test were used for evaluation of motor learning ability. However, results obtained from the GluA2-carboxy terminus modified knock-in mice demonstrated normal adaptation of the VOR and the OKR, despite lacking PF-LTD. In that report, induction of LTD was only attempted using one type of stimulation protocol at room temperature. Thus, conditions to induce cerebellar LTD were explored in the same knock-in mutants using various protocols at near physiological temperature. Finally, we found stimulation protocols, by which LTD could be induced in these gene-manipulated mice. In this study, a set of protocols are proposed to evaluate LTD-induction, which will more accurately allow examination of the causal relationship between LTD and motor learning. In conclusion, experimental conditions are crucial when evaluating LTD in gene-manipulated mice.

In some gene-manipulated animals, using a single protocol may fail to induce LTD in cerebellar Purkinje cells, and there may be a discrepancy between LTD and motor learning. Multiple protocols are necessary to assess LTD-induction in gene-manipulated animals. Standard protocols are shown.

成人小脳スライスにおける長期うつ病誘導の評価

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...

Ballistic Labeling of Pyramidal Neurons in Brain Slices and in Primary Cell Culture

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of pyramidal neurons and dendritic spines, a ballistic labeling technique can be utilized. In the present protocol, pyramidal neurons are labeled with DilC18(3) dye and analyzed using neuronal reconstruction software to assess neuronal morphology and dendritic spines. To investigate neuronal structure, dendritic branching analysis and Sholl analysis are performed, allowing researchers to draw inferences about dendritic branching complexity and neuronal arbor complexity, respectively. The evaluation of dendritic spines is conducted using an automatic assisted classification algorithm integral to the reconstruction software, which classifies spines into four categories (i.e., thin, mushroom, stubby, filopodia). Furthermore, an additional three parameters (i.e., length, head diameter, and volume) are also chosen to assess alterations in dendritic spine morphology. To validate the potential of wide application of the ballistic labeling technique, pyramidal neurons from in vitro cell culture were successfully labeled. Overall, the ballistic labeling method is unique and useful for visualizing neurons in different brain regions in rats, which in combination with sophisticated reconstruction software, allows researchers to elucidate the possible mechanisms underlying neurocognitive dysfunction.

We present a protocol to label and analyze pyramidal neurons, which is critical for evaluating potential morphological alterations in neurons and dendritic spines that may underlie neurochemical and behavioral abnormalities.

脳スライスと一次細胞培養における錐体ニューロンの弾道標識

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of ...

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered tissue samples. Traditional immunolabeling procedures require cutting the sample into thin sections, which restricts the ability to label and examine intact structures. However, if brain tissue can remain intact during processing, structures and circuits can remain intact for the analysis. Previously established clearing methods take significant time to completely clear the tissue, and the harsh chemicals can often damage sensitive antibodies. The iDISCO method quickly and completely clears tissue, is compatible with many antibodies, and requires no special lab equipment. This technique was initially validated for the use in mice tissue, but the current protocol adapts this method to image hemispheres of control and transgenic rat brains. In addition to this, the present protocol also makes several adjustments to preexisting protocol to provide clearer images with less background staining. Antibodies for Iba-1 and tyrosine hydroxylase were validated in the HIV-1 transgenic rat and in F344/N control rats using the present hydrophobic tissue clearing method. The brain is an interwoven network, where structures work together more often than separately of one another. Analyzing the brain as a whole system as opposed to a combination of individual pieces is the greatest benefit of this whole brain clearing method.

Here we present a hydrophobic tissue clearing method that allows for the viewing of target molecules as part of intact brain structures. This technique has now been validated for F344/N control and HIV-1 transgenic rats of both sexes.

ラット脳組織の疎水性組織除去法

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered ...

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

エコHIV脳感染症のラットモデル

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...