This work was supported by Interdisziplin&#228;res Zentrum f&#252;r Klinische Forschung (IZKF), University Clinics W&#252;rzburg, Germany (project N-215).

（：55.2から2531.01 11分の76承認番号）の動物実験は、動物保護ガイドラインと欧州共同体理事会のガイドラインに従ってウンターフランケンのヴュルツブルク大学と法的な州当局によって承認されました。すべての努力は、使用する動物の痛みや不快感を最小限にするために行われました。 注：他の場所18に記載の電極の移植を行いました。 1.シリンダー試験（図2） <ol><li> 0.1％酢酸溶液でシリンダーを洗浄することにより、透明なプラスチックガラスシリンダー（19センチメートル：40センチ、直径、高さ）を準備します。 </li><li>実験の日、各ラットの識別番号とカードを準備します。 </li><li>シリンダーの後ろに90°の角度で二つのミラーを配置します。 </li><li>カメラとシリンダとの間の距離は足の素晴らしい眺めを可能にするように、シリンダーの前にカメラを置きます。 </li><li>輸送箱にラットを置きます。 注：動物はストレスを避けるために、試験前に実験者によって処理されるべきです。 </li><li>輸送ボックスを使用して、シリンダにホームケージからラットを輸送。 </li><li>シリンダ（ 図3）にラットを置きます。 <ol><li>常に活動の概日違いを回避するために、1日の同じ時間にすべての行動試験を行います。動物がケーブルによって刺激発生器に接続されている場合は、ケーブルが実験中にねじれがないことを確認してください。 </li></ol></li><li>カメラの「録画」ボタンを押してください。カメラに実験およびラットの識別番号の実際の日付でカードを表示します。記録を開始します。 </li><li> 5分後、シリンダーから動物を削除し、輸送ボックスを使用して、ホームケージに戻します。 </li><li> 0.1％酢酸溶液でシリンダーを清掃してください。 </li><li>左右の足の壁に接触（パーセントで足の使用）と同様にRをカウントすることにより、記録した映像から足の使用を評価しますイヤリング（またはシリンダ壁に支持することなく、後足の上に立って）。シリンダテストは、適切なソフトウェアによって自動的に評価することができます。 注：健康なラットは均等に両方の足を使用しています。 hemiparkinsonianラットは、より少ない程度の病変による影響を受けた足を使用しています。 </li></ol> 2.階段テスト（図4） <ol><li>獲得フェーズ<ol><li>前訓練への一日は、階段の試験に用いたペレットを動物に慣れます。 <ol><li>オプション：食餌制限（自由摂食レベル16の90％で、体重を維持するための標準的な実験用飼料10〜15 g）を使用し、動物のモチベーションを高めるために。しかし、これは、正のトレーニング効果を達成するために必須ではありません。この研究は、食事制限なしで行いました。 </li></ol></li><li>透明なプラスチックのガラスの階段ボックスを準備し（高さ：34.5センチメートル、長さ：35.5センチメートル、幅12センチ、幅の狭い室6 cm）の0.1％の交流でボックスを清掃することによりETIC酸溶液。注：階段ボックスが狭い区画内の隆起したプラットフォームと2階段で二区画-ボックスです。狭い区画内の階段上の左のステップは、右のステップは右足で、唯一の左の足に到達することができます。 注：標準の階段ボックスは、それがケーブルを介して刺激されたラットを用いた実験のために使用される場合には、蓋なしで高いボックスを使用し、蓋を二つの区画で構成されています。 </li><li>階段を削除し、8 45 mgのペレットと各手順の井戸を埋めます。 </li><li>階段を挿入し、高めのプラットフォーム上で8つの追加ペレットを置きます。 </li><li>輸送箱にラットを置きます。 </li><li>輸送ボックスを使用して階段ボックスにホームケージからラットを輸送。 </li><li>階段ボックス（ 図5）にラットを置きます。 </li><li> 5分後、階段ボックスから動物を削除し、輸送ボックスを使用して、ホームケージに戻します。 </li><li>どのように多くの注意ペレットは、左右の階段からプラットフォームと（最終的に）から食べました。 </li><li> 8 45 mgのペレットと各ステップ上のウェルを充填することにより、階段を補充します。 </li><li> 0.1％酢酸溶液で階段ボックスをきれいにし、プラットフォーム上で追加のペレットを配置します。 </li><li>この手順（アクイジション・フェーズ）行の3日間を繰り返します。 注：記載された全ての実験は、雄のSprague Dawleyラットで実施しました。異なるトレーニングモジュールの所要時間は異なる系統、性別およびベンダーのラットで異なることができます。 </li></ol></li><li>自由選択のテスト<ol><li> 0.1％酢酸溶液で階段ボックスを清掃してください。 </li><li>階段を削除し、8 45 mgのペレットと各手順の井戸を埋めます。 </li><li>輸送箱にラットを置きます。 </li><li>輸送ボックスを使用して階段ボックスにホームケージからラットを輸送。 </li><li>階段ボックスにラットを置きます。 </li><li> 5分後、階段から動物を削除しますケースボックスと輸送ボックスを使用して、ホームケージにそれを戻します。 左右の階段から食べたどのように多くのペレットに注意してください。 </li><li>注：動物がまだペレットを把握に問題がある場合は、それらを簡単に達することができるプラットフォーム上でいくつかの詳細を追加します。 </li><li> 8 45 mgのペレットと各ステップ上のウェルを充填することにより、階段を補充します。 </li><li>次の動物のための0.1％酢酸溶液で階段ボックスを清掃してください。 </li><li>この手順（自由選択相）行の3日間を繰り返します。 注：提示された結果は、モジュール間の休止期間なく行わ訓練によって得ました。いくつかのグループは、トレーニングプロセスをサポートするため、連結のための休止日を好みます。 </li></ol></li><li>強制選択テスト<ol><li> 0.1％酢酸溶液で階段ボックスを清掃してください。 </li><li>階段を削除し、8（モジュールの最初の3日間）または4（連続したT​​HRと左側階段上の各手順の井戸を埋めますモジュールのEE日）45 mgのペレット。 <ol><li>減損が発生する側の強制選択テストを実行します。 注：私たちは右半球にパーキンソン病変を行うため、選択的に左足を訓練します。 </li></ol></li><li>輸送箱にラットを置きます。 </li><li>輸送ボックスを使用して階段ボックスにホームケージからラットを輸送。 </li><li>階段ボックスにラットを置きます。 </li><li> 5分後、階段ボックスから動物を削除し、輸送ボックスを使用して、ホームケージに戻します。 </li><li>左の階段から食べたどのように多くのペレットに注意してください。 </li><li> 8または4 45mgのペレット（ペレットの数は、トレーニング日に依存します）との各ス​​テップ上のウェルを充填することにより、階段を補充します。 </li><li>次の動物のための0.1％酢酸溶液で階段ボックスを清掃してください。 </li><li>この手順を繰り返し（強制選択相）行の6日間。 </li></ol></li><li>データ収集<ol> &lt;2日連続で強制選択モジュール（左の階段上の各ウェル中の4ペレット）について記載したように李&gt;は、実験を実行します。 2日間の平均値として成功率（食べたペレットの数）を計算します。 </li></ol></li></ol>

<ol><li>Fasano, A., Lozano, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deep+brain+stimulation+for+movement+disorders%3A+2015+and+beyond%5BTitle%5D)+AND+%22Current+opinion+in+neurology%22%5BJournal%5D)">Deep brain stimulation for movement disorders: 2015 and beyond.</a> Current opinion in neurology. , (2015).</li>
<li>McIntyre, C. C., Savasta, M., Kerkerian-Le Goff, L., Vitek, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Uncovering+the+mechanism%28s%29+of+action+of+deep+brain+stimulation%3A+activation%2C+inhibition%2C+or+both%5BTitle%5D)+AND+%22Clinical+neurophysiology+%3A+official+journal+of+the+International+Federation+of+Clinical+Neurophysiology%22%5BJournal%5D)">Uncovering the mechanism(s) of action of deep brain stimulation: activation, inhibition, or both.</a> Clinical neurophysiology : official journal of the International Federation of Clinical Neurophysiology. 115, 1239-1248 (2004).</li>
<li>Deniau, J. M., Degos, B., Bosch, C., Maurice, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deep+brain+stimulation+mechanisms%3A+beyond+the+concept+of+local+functional+inhibition%5BTitle%5D)+AND+%22The+European+journal+of+neuroscience%22%5BJournal%5D)">Deep brain stimulation mechanisms: beyond the concept of local functional inhibition.</a> The European journal of neuroscience. 32, 1080-1091 (2010).</li>
<li>Modolo, J., Legros, A., Thomas, A. W., Beuter, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Model-driven+therapeutic+treatment+of+neurological+disorders%3A+reshaping+brain+rhythms+with+neuromodulation%5BTitle%5D)+AND+%22Interface+focus%22%5BJournal%5D)">Model-driven therapeutic treatment of neurological disorders: reshaping brain rhythms with neuromodulation.</a> Interface focus. 1, 61-74 (2011).</li>
<li>Groppa, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Physiological+and+anatomical+decomposition+of+subthalamic+neurostimulation+effects+in+essential+tremor%5BTitle%5D)+AND+%22Brain+%3A+a+journal+of+neurology%22%5BJournal%5D)">Physiological and anatomical decomposition of subthalamic neurostimulation effects in essential tremor.</a> Brain : a journal of neurology. 137, 109-121 (2014).</li>
<li>Reich, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Short+pulse+width+widens+the+therapeutic+window+of+subthalamic+neurostimulation%5BTitle%5D)+AND+%22Annals+of+clinical+and+translational+neurology%22%5BJournal%5D)">Short pulse width widens the therapeutic window of subthalamic neurostimulation.</a> Annals of clinical and translational neurology. 2, 427-432 (2015).</li>
<li>Blandini, F., Armentero, M. T., Martignoni, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+6-hydroxydopamine+model%3A+news+from+the+past%5BTitle%5D)+AND+%22Parkinsonism+%26+related+disorders%22%5BJournal%5D)">The 6-hydroxydopamine model: news from the past.</a> Parkinsonism & related disorders. 14, Suppl 2 124-129 (2008).</li>
<li>Bove, J., Perier, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neurotoxin-based+models+of+Parkinson%27s+disease%5BTitle%5D)+AND+%22Neuroscience%22%5BJournal%5D)">Neurotoxin-based models of Parkinson's disease.</a> Neuroscience. 211, 51-76 (2012).</li>
<li>Metz, G. A., Tse, A., Ballermann, M., Smith, L. K., Fouad, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+unilateral+6-OHDA+rat+model+of+Parkinson%27s+disease+revisited%3A+an+electromyographic+and+behavioural+analysis%5BTitle%5D)+AND+%22The+European+journal+of+neuroscience%22%5BJournal%5D)">The unilateral 6-OHDA rat model of Parkinson's disease revisited: an electromyographic and behavioural analysis.</a> The European journal of neuroscience. 22, 735-744 (2005).</li>
<li>Miklyaeva, E. I., Castaneda, E., Whishaw, I. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skilled+reaching+deficits+in+unilateral+dopamine-depleted+rats%3A+impairments+in+movement+and+posture+and+compensatory+adjustments%5BTitle%5D)+AND+%22The+Journal+of+neuroscience+%3A+the+official+journal+of+the+Society+for+Neuroscience%22%5BJournal%5D)">Skilled reaching deficits in unilateral dopamine-depleted rats: impairments in movement and posture and compensatory adjustments.</a> The Journal of neuroscience : the official journal of the Society for Neuroscience. 14, 7148-7158 (1994).</li>
<li>Li, X. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High-frequency+stimulation+of+the+subthalamic+nucleus+restores+neural+and+behavioral+functions+during+reaction+time+task+in+a+rat+model+of+Parkinson%27s+disease%5BTitle%5D)+AND+%22Journal+of+neuroscience+research%22%5BJournal%5D)">High-frequency stimulation of the subthalamic nucleus restores neural and behavioral functions during reaction time task in a rat model of Parkinson's disease.</a> Journal of neuroscience research. 88, 1510-1521 (2010).</li>
<li>Darbaky, Y., Forni, C., Amalric, M., Baunez, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High+frequency+stimulation+of+the+subthalamic+nucleus+has+beneficial+antiparkinsonian+effects+on+motor+functions+in+rats%2C+but+less+efficiency+in+a+choice+reaction+time+task%5BTitle%5D)+AND+%22The+European+journal+of+neuroscience%22%5BJournal%5D)">High frequency stimulation of the subthalamic nucleus has beneficial antiparkinsonian effects on motor functions in rats, but less efficiency in a choice reaction time task.</a> The European journal of neuroscience. 18, 951-956 (2003).</li>
<li>Fang, X., Sugiyama, K., Akamine, S., Namba, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Improvements+in+motor+behavioral+tests+during+deep+brain+stimulation+of+the+subthalamic+nucleus+in+rats+with+different+degrees+of+unilateral+parkinsonism%5BTitle%5D)+AND+%22Brain+research%22%5BJournal%5D)">Improvements in motor behavioral tests during deep brain stimulation of the subthalamic nucleus in rats with different degrees of unilateral parkinsonism.</a> Brain research. 1120, 202-210 (2006).</li>
<li>Lindemann, C., Krauss, J. K., Schwabe, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deep+brain+stimulation+of+the+subthalamic+nucleus+in+the+6-hydroxydopamine+rat+model+of+Parkinson%27s+disease%3A+effects+on+sensorimotor+gating%5BTitle%5D)+AND+%22Behavioural+brain+research%22%5BJournal%5D)">Deep brain stimulation of the subthalamic nucleus in the 6-hydroxydopamine rat model of Parkinson's disease: effects on sensorimotor gating.</a> Behavioural brain research. 230, 243-250 (2012).</li>
<li>Schallert, T., Fleming, S. M., Leasure, J. L., Tillerson, J. L., Bland, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CNS+plasticity+and+assessment+of+forelimb+sensorimotor+outcome+in+unilateral+rat+models+of+stroke%2C+cortical+ablation%2C+parkinsonism+and+spinal+cord+injury%5BTitle%5D)+AND+%22Neuropharmacology%22%5BJournal%5D)">CNS plasticity and assessment of forelimb sensorimotor outcome in unilateral rat models of stroke, cortical ablation, parkinsonism and spinal cord injury.</a> Neuropharmacology. 39, 777-787 (2000).</li>
<li>Montoya, C. P., Campbell-Hope, L. J., Pemberton, K. D., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+%27staircase+test%27%3A+a+measure+of+independent+forelimb+reaching+and+grasping+abilities+in+rats%5BTitle%5D)+AND+%22Journal+of+neuroscience%22%5BJournal%5D)">The 'staircase test': a measure of independent forelimb reaching and grasping abilities in rats.</a> Journal of neuroscience. 36, 219-228 (1991).</li>
<li>Kloth, V., Klein, A., Loettrich, D., Nikkhah, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Colour-coded+pellets+increase+the+sensitivity+of+the+staircase+test+to+differentiate+skilled+forelimb+performances+of+control+and+6-hydroxydopamine+lesioned+rats%5BTitle%5D)+AND+%22Brain+research+bulletin%22%5BJournal%5D)">Colour-coded pellets increase the sensitivity of the staircase test to differentiate skilled forelimb performances of control and 6-hydroxydopamine lesioned rats.</a> Brain research bulletin. 70, 68-80 (2006).</li>
<li>Fluri, F., Volkmann, J., Kleinschnitz, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microelectrode+guided+implantation+of+electrodes+into+the+subthalamic+nucleus+of+rats+for+long-term+deep+brain+stimulation%5BTitle%5D)+AND+%22JoVE%22%5BJournal%5D)">Microelectrode guided implantation of electrodes into the subthalamic nucleus of rats for long-term deep brain stimulation.</a> JoVE. , (2015).</li>
<li>Paxinos, G., Watson, C. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aPaxinos%2C+G+%22The+rat+brain+in+stereotactic+coordinates%22">The rat brain in stereotactic coordinates</a>. , (2008).</li>
<li>Nikkhah, G., Rosenthal, C., Hedrich, H. J., Samii, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differences+in+acquisition+and+full+performance+in+skilled+forelimb+use+as+measured+by+the+%27staircase+test%27+in+five+rat+strains%5BTitle%5D)+AND+%22Behavioural+brain+research%22%5BJournal%5D)">Differences in acquisition and full performance in skilled forelimb use as measured by the 'staircase test' in five rat strains.</a> Behavioural brain research. 92, 85-95 (1998).</li>
<li>Angelov, S. D., Dietrich, C., Krauss, J. K., Schwabe, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+Deep+Brain+Stimulation+in+Rats+Selectively+Bred+for+Reduced+Prepulse+Inhibition%5BTitle%5D)+AND+%22Brain+stimulation%22%5BJournal%5D)">Effect of Deep Brain Stimulation in Rats Selectively Bred for Reduced Prepulse Inhibition.</a> Brain stimulation. , (2014).</li>
<li>de Haas, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Wireless+implantable+micro-stimulation+device+for+high+frequency+bilateral+deep+brain+stimulation+in+freely+moving+mice%5BTitle%5D)+AND+%22Journal+of+neuroscience+methods%22%5BJournal%5D)">Wireless implantable micro-stimulation device for high frequency bilateral deep brain stimulation in freely moving mice.</a> Journal of neuroscience methods. 209, 113-119 (2012).</li>
<li>Heo, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fully+Implantable+Deep+Brain+Stimulation+System+with+Wireless+Power+Transmission+for+Long-term+Use+in+Rodent+Models+of+Parkinson%27s+Disease%5BTitle%5D)+AND+%22Journal+of+Korean+Neurosurgical+Society%22%5BJournal%5D)">Fully Implantable Deep Brain Stimulation System with Wireless Power Transmission for Long-term Use in Rodent Models of Parkinson's Disease.</a> Journal of Korean Neurosurgical Society. 57, 152-158 (2015).</li>
<li>Gut, N. K., Winn, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deep+brain+stimulation+of+different+pedunculopontine+targets+in+a+novel+rodent+model+of+parkinsonism%5BTitle%5D)+AND+%22J.+Neurosci%22%5BJournal%5D)">Deep brain stimulation of different pedunculopontine targets in a novel rodent model of parkinsonism.</a> J. Neurosci. 35, 4792-4803 (2015).</li>
<li>Whishaw, I. Q., Gorny, B., Foroud, A., Kleim, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Long-Evans+and+Sprague-Dawley+rats+have+similar+skilled+reaching+success+and+limb+representations+in+motor+cortex+but+different+movements%3A+some+cautionary+insights+into+the+selection+of+rat+strains+for+neurobiological+motor+research%5BTitle%5D)+AND+%22Behavioural+brain+research%22%5BJournal%5D)">Long-Evans and Sprague-Dawley rats have similar skilled reaching success and limb representations in motor cortex but different movements: some cautionary insights into the selection of rat strains for neurobiological motor research.</a> Behavioural brain research. 145, 221-232 (2003).</li>
<li>Honndorf, S., Lindemann, C., Tollner, K., Gernert, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Female+Wistar+rats+obtained+from+different+breeders+vary+in+anxiety-like+behavior+and+epileptogenesis%5BTitle%5D)+AND+%22Epilepsy+research%22%5BJournal%5D)">Female Wistar rats obtained from different breeders vary in anxiety-like behavior and epileptogenesis.</a> Epilepsy research. 94, 26-38 (2011).</li>
<li>Jadavji, N. M., Metz, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sex+differences+in+skilled+movement+in+response+to+restraint+stress+and+recovery+from+stress%5BTitle%5D)+AND+%22Behavioural+brain+research%22%5BJournal%5D)">Sex differences in skilled movement in response to restraint stress and recovery from stress.</a> Behavioural brain research. 195, 251-259 (2008).</li>
<li>Kucker, S., Tollner, K., Piechotta, M., Gernert, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Kindling+as+a+model+of+temporal+lobe+epilepsy+induces+bilateral+changes+in+spontaneous+striatal+activity%5BTitle%5D)+AND+%22Neurobiology+of+disease%22%5BJournal%5D)">Kindling as a model of temporal lobe epilepsy induces bilateral changes in spontaneous striatal activity.</a> Neurobiology of disease. 37, 661-672 (2010).</li>
<li>Smith, L. K., Metz, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dietary+restriction+alters+fine+motor+function+in+rats%5BTitle%5D)+AND+%22Physiology+%26+behavior%22%5BJournal%5D)">Dietary restriction alters fine motor function in rats.</a> Physiology & behavior. 85, 581-592 (2005).</li>
</ol>

The authors declare that they have no competing financial interests.

この記事では、シリンダおよび階段テストの詳細なトレーニングプロトコルについて説明します。後者が原因ラット16,17に到達業者に複雑な把持動作と細かい運動の動きを評価するために設計されています。成果の測定は、客観的な測定であるテスト、中に食べたペレットの数として表現さ​​れます。プロトコルは、パーキンソン病および他の運動疾患モデルのラットモデルにおい...

視床下核（STN）の脳深部刺激はパーキンソン病1および他の運動障害のための有効な治療選択肢です。基礎となるメカニズムはまだ十分に理解し、多因子であるが、主要な機能は、刺激電極2-4付近の軸索の繰り返し脱分極によって神経ネットワーク活動の変調です。高周波数（&gt; 100 Hz）で刺激が最も脳の目標及びDBSのほとんどの適応症のための有益な効果のために必要とされます。刺激量で覆われている他の繊維、の不注意による同時活性化から深部脳刺激の結果の副作用は、錐体路などのさまざまな機能を、補助します。したがって、副作用素子5,6の同時活性化を回避しつつ、優先的に有益な神経要素を活性化する刺激パラメータを開発することが望ましいです。神経生理学は、このような微細なTUNIを提供することがありますが、DBSのオプションngを、科学の進歩は、プログラミング戦略は、主に患者の「試行錯誤」により評価し、市販のDBSデバイスの限られたプログラミングオプションによって制限されているため、過去20年の間に最小限されてではなく、神経生理学的知見を使用していますかつ体系フルパラメータ空間を探索する実験的な設定を定義しました。 DBS研究の翻訳障害を克服するために、我々は、臨床探査の前にパーキンソン症候群の齧歯類モデルにおける代替刺激パラメータをスクリーニングするためのプロトコルを提案しています。ラットにおける片側パーキンソン病は、右の内側前脳束7,8に6-ヒドロキシドーパミン注射を使用してモデル化されます。得られた病変が、hemiparkinsonianとしてさらに説明、低用量のアポモルフィン注射後の回転スコアの評価によってアポモルフィン試験で評価し、チロシンヒドロキシラーゼimmunohistによって死後に確認されましたochemistry。低死亡率および罹患率を保有しながら、この方法は、適用が容易で再現性の高いです。結果として得られる運動障害は、7,8非常に離散的です。動物は、自発的な探査や複雑な把握行動9,10の両方の間に反対側の左足のわずかな障害を呈します。 脳深部刺激プロトコルテストの有効性を評価するためには、モータ性能の迅速かつ信頼性の変化を測定できるように、別の神経刺激設定で時間をかけて繰り返すことができる必要とされます。いくつかのグループが非常に変動し、一貫性のない成果11-14ラット11で運動機能を評価するために、異なる刺激のアプローチと異なるテストを提案しました。これは、妥当性と相補性を予測する高で一連のテストを選択することが私たちを余儀なくされました。さらに、深部脳刺激条件下でのモータ結果の評価のために、試験は、ANIによって行うことができた好ましMALSは、刺激発生器にケーブルを介して接続されています。これらの目的のために、我々は足の使用の非対称性のための1つの試験や熟練到達するための1つの試験からなる当社の試験電池を設置しました。研究デザインは、 図1に示されています。 自発的な足の使用のために我々は、垂直探査中の足の使用のために広く使用されている試験であるShallert 15によって記述円筒試験を行いました。動物のいかなるトレーニングは必要ありません。より複雑な把持動作の評価のために我々はモントーヤ16に従って階段テストを確立しました。私たちのプロトコルはKloth 17に応じて変更されます。ラットを試験箱からペレットに到達するには12日間の期間のために訓練されています。トレーニング期間の後に試験を食べたペレットの数として記載成功率をカウントすることにより、複雑な把持動作を測定するために適用することができます。記事では、BEH階段ボックス内の詳細なトレーニングだけでなく、両方の性能を提示しますナイーブ、hemiparkinsonianと脳深部刺激条件の下でavioralテスト。

<table><tbody><tr><td>Staircase box without lid</td><td>Glas Keil, Germany</td><td></td><td>custom made</td></tr><tr><td>Cylinder box</td><td>Glas Keil, Germany</td><td></td><td>custom made</td></tr><tr><td>Dustless precision pellets, 45 mg</td><td>Bio Serv</td><td>F0021</td><td></td></tr></tbody></table>

a novel approach to assess motor outcome of deep brain stimulation effects in the hemiparkinsonian rat: staircase and cylinder test

視床下核の脳深部刺激は、パーキンソン病のための有効な治療選択肢です。私たちの研究室では、hemiparkinsonian（一方的な病変）ラットで異なる神経刺激パターンをスクリーニングするためのプロトコルを確立しました。それは、右の内側前脳束に6-ヒドロキシドーパミン（6-OHDA）を注入視床下核への慢性刺激電極を移植し、ケーブルに結合した外部神経刺激の24時間の期間の終了時にモータの成果を評価することによって、一方的なパーキンソン病斑を作成で構成されてい。刺激は、定電流刺激を行いました。振幅は、副作用のために、個々の閾値を下回る20％に設定しました。モータアウトカム評価がShallertにし、モントーヤに応じて階段試験で到達業者の評価によって応じ円筒試験における自発的な足の使用の評価によって行われました。このプロトコルを詳細に階段ボックスの訓練を説明し、Cylinderテスト、ならびにhemiparkinsonianラットの両方での使用。階段テストは細かい運動技能障害のために、より敏感であるように思われると神経刺激の間に変化するより高い感度を示すため、両方のテストの使用が必要です。一方的なパーキンソンモデルと2行動試験の組み合わせは、標準化された方法で異なる刺激パラメータの評価を可能にします。

すべての動物は、ドーパミン作動性病変と電極の位置の両方の死後組織学的検査を受けました。 STN内部の正しい電極配置（ 図6）と完全なドーパミン作動性病変（&gt;黒質におけるドーパミン作動性ニューロンの90％の損失）と動物のみが結果セクション（ 図7）に含まれていました。 病変条件で?...

Watch this Scientific Journal Video about A Novel Approach to Assess Motor Outcome of Deep Brain Stimulation Effects in the Hemiparkinsonian Rat: Staircase and Cylinder Test at JoVE.com

A Novel Approach to Assess Motor Outcome of Deep Brain Stimulation Effects in the Hemiparkinsonian Rat: Staircase and Cylinder Test

Deep brain stimulation (DBS) is an effective treatment option for Parkinson's disease. We established a study design to screen novel stimulation paradigms in rats. The protocol describes the use of the staircase test and cylinder test for motor outcome assessment in DBS treated hemiparkinsonian rats.

Hemiparkinsonianラットにおける脳深部刺激効果のモーターの成果を評価するための新しいアプローチ：階段室とシリンダーテスト

Deep brain stimulation of the subthalamic nucleus is an effective treatment option for Parkinson&#39;s disease. In our lab we established a protocol to ...

a-novel-approach-to-assess-motor-outcome-deep-brain-stimulation

Research

JoVE Journal

Behavior

14.7K ビュー.  University Hospital Wuerzburg. Deep brain stimulation (DBS) is an effective treatment option for Parkinson's disease. We established a study design to screen novel stimulation paradigms in rats. The protocol describes the use of the staircase test and cylinder test for motor outcome assessment in DBS treated hemiparkinsonian rats.

ビデオ: A Novel Approach to Assess Motor Outcome of Deep Brain Stimulation Effects in the Hemiparkinsonian Rat: Staircase and Cylinder Test

Assessment of Sensorimotor Function in Mouse Models of Parkinson's Disease

Sensitive and reliable behavioral outcome measures are essential to the evaluation of potential therapeutic treatments in preclinical trials for many neurodegenerative diseases. In Parkinson's disease, sensorimotor tests sensitive to varying degrees of nigrostriatal dysfunction are fundamental for testing the efficacy of potential therapeutics. Reliable and quite elegant sensorimotor measures exist for rats, however many of these tests measure sensorimotor asymmetry within the rat and are not entirely suitable for the newer genetic mouse models of PD. We have put together a battery of sensorimotor tests inspired by the sensitive tests in rats and adapted for mice. The test battery highlighted in this study is chosen for a) its sensitivity in a wide variety of mouse models of PD, b) its ease in implementing into a study, and c) its low expense. These tests have proven useful in characterizing novel genetic mouse models of PD as well as in testing potential disease-modifying therapies.

In Parkinson's disease and movement disorders in general, sensitive and reliable behavioral assays are essential for testing novel potential therapeutics. Here, we describe a manageable battery of sensorimotor tests for mice that are sensitive to varying degrees of injury to the nigrostriatal system and useful for preclinical studies.

パーキンソン病のマウスモデルにおける感覚運動機能の評価

Sensitive and  reliable behavioral outcome measures are essential to the evaluation of  potential therapeutic treatments in preclinical trials for many  ...

行動学

Assessing Forelimb Function after Unilateral Cervical SCI using Novel Tasks: Limb Step-alternation, Postural Instability and Pasta Handling

Cervical spinal cord injury (cSCI) can cause devastating neurological deficits, including impairment or loss of upper limb and hand function. A majority of the spinal cord injuries in humans occur at the cervical levels. Therefore, developing cervical injury models and developing relevant and sensitive behavioral tests is of great importance. Here we describe the use of a newly developed forelimb step-alternation test after cervical spinal cord injury in rats. In addition, we describe two behavioral tests that have not been used after spinal cord injury: a postural instability test (PIT), and a pasta-handling test. All three behavioral tests are highly sensitive to injury and are easy to use. Therefore, we feel that these behavioral tests can be instrumental in investigating therapeutic strategies after cSCI.

Three new behavioral tests (forelimb step-alternation, postural instability test, pasta handling test) for evaluating forelimb function after cervical spinal cord injury in rodents are described.

新規タスクを使用した片側子宮頸SCI後前肢の機能を評価する：四肢ステップ交替、姿勢の不安定やパスタの取り扱い

Cervical spinal cord injury (cSCI) can cause devastating neurological deficits, including impairment or loss of upper limb and hand function. A majority ...

Use of Rotorod as a Method for the Qualitative Analysis of Walking in Rat

High speed videoanalysis of the details of movement can provide a source of information about qualitative aspects of walking movements. When walking on a rotorod, animals remain in approximately the same place making repetitive movements of stepping. Thus the task provides a rich source of information on the details of foot stepping movements. Subjects were hemi-Parkinson analogue rats, produced by injection of 6-hydroxydopamine (6-OHDA) into the right nigrostriatal bundle to deplete nigrostriatal dopamine (DA). The present report provides a video analysis illustration of animals previously were filmed from frontal, lateral, and posterior views as they walked (15). Rating scales and frame-by-frame replay of the video records of stepping behavior indicated that the hemi-Parkinson rats were chronically impaired in posture and limb use contralateral to the DA-depletion. The contralateral limbs participated less in initiating and sustaining propulsion than the ipsilateral limbs. These deficits secondary to unilateral DA-depletion show that the rotorod provides a use task for the analysis of stepping movements.

The rotorod test is used to assess motor status in the walking movement of hemi-Parkinson analogue rats.

ラットの歩行の質的分析のための方法としてRotorodの使用

High speed videoanalysis of the details of movement can provide a source of information about qualitative aspects of walking movements. When walking on a ...

生物学

Behavioral Assessments of Spontaneous Locomotion in a Murine MPTP-induced Parkinson's Disease Model

Parkinson&#8217;s disease (PD) is a common neurodegenerative disorder disease, causing the phenomenon of shaking, rigidity, slowness of movement and dementia. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) can lead to some Parkinson&#8217;s-like symptoms by destroying dopaminergic neurons in the substantia nigra of the brain. It has been thus used to establish PD models in various animal studies. Here, mice receive MPTP injections (20 mg/kg/day) for seven days and the behavioral tests are performed on the eighth day. This model is adapted efficiently in the study of PD. The behavioral tests here include the cylinder test and the open field test. The cylinder experiment is used to detect the animals&#8217; ability to lift their front paws when put into a different environment. As the PD model mice show arching&#8212;the mouse arches its back&#8212;the number of paw liftings decrease. This test is easy to execute. The open field test is used to detect the amount of time the mice spend on running, walking, and remaining immobile. We analyze animals&#8217; movements in open field using software and obtain data. Lastly, we use L-DOPA, one of the most commonly used PD drugs, as one example to show how to apply this model to the study of PD drugs. Our results indicate that MPTP neurotoxicity induces motor deficit which can be mitigated by L-DOPA.

We describe establishment of a murine model for Parkinson's disease using MPTP, and behavioral assessments using cylinder and open field tests to measure motor function. We then use L-DOPA as one example to show how to apply this model in the study of PD drugs.

マウス MPTP 誘発パーキンソン病モデルにおける自発運動の行動評価

Parkinson&#8217;s disease (PD) is a common neurodegenerative disorder disease, causing the phenomenon of shaking, rigidity, slowness of movement and ...

Controlling Parkinson's Disease With Adaptive Deep Brain Stimulation

Adaptive deep brain stimulation (aDBS) has the potential to improve the treatment of Parkinson&#39;s disease by optimizing stimulation in real time according to fluctuating disease and medication state. In the present realization of adaptive DBS we record and stimulate from the DBS electrodes implanted in the subthalamic nucleus of patients with Parkinson&#39;s disease in the early post-operative period. Local field potentials are analogue filtered between 3 and 47 Hz before being passed to a data acquisition unit where they are digitally filtered again around the patient specific beta peak, rectified and smoothed to give an online reading of the beta amplitude. A threshold for beta amplitude is set heuristically, which, if crossed, passes a trigger signal to the stimulator. The stimulator then ramps up stimulation to a pre-determined clinically effective voltage over 250 msec and continues to stimulate until the beta amplitude again falls down below threshold. Stimulation continues in this manner with brief episodes of ramped DBS during periods of heightened beta power.
Clinical efficacy is assessed after a minimum period of stabilization (5 min) through the unblinded and blinded video assessment of motor function using a selection of scores from the Unified Parkinson&#39;s Rating Scale (UPDRS). Recent work has demonstrated a reduction in power consumption with aDBS as well as an improvement in clinical scores compared to conventional DBS. Chronic aDBS could now be trialed in Parkinsonism.

Adaptive deep brain stimulation (aDBS) is effective for Parkinson&#8217;s disease, improving symptoms and reducing power consumption compared to conventional deep brain stimulation (cDBS). In aDBS we track a local field potential biomarker (beta oscillatory amplitude) in real time and use this to control the timing of stimulation.

アダプティブ脳深部刺激とパーキンソン病の防除

Adaptive deep brain stimulation (aDBS) has the potential to improve the treatment of Parkinson&#39;s disease by optimizing stimulation in real time ...

医学

Unilateral Pyramidotomy of the Corticospinal Tract in Rats for Assessment of Neuroplasticity-inducing Therapies

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has great importance for distal muscle control in humans and, to a lesser extent, in rodents. A unilateral cut of one pyramid results in loss of CST innervation of the spinal cord mainly on the contralateral side of the spinal cord leading to transient motor disability in the forelimbs and sustained loss of dexterity. Ipsilateral projections of the corticospinal tract are minor. We have refined our surgical method to increase the chances of lesion completeness. We describe postsurgical care. Deficits on the Montoya staircase pellet reaching test and the horizontal ladder test shown here are detected up to 8 weeks postinjury. Deficits on the cylinder rearing test are only detected transiently. Therefore, the cylinder test may only be suitable for detection of short term recovery. We show how, electrophysiologically and anatomically, one may assess lesions and plastic changes. We also describe how to analyse fibers from the uninjured CST sprouting across the midline into the deprived areas. It is challenging to obtain &#62;90% complete lesions consistently due to the proximity to the basilar artery in the medulla oblongata and survival rates can be low. Alternative surgical approaches and behavioural testing are described in this protocol. The pyramidotomy model is a good tool for assessing neuroplasticity-inducing treatments, which increase sprouting of intact fibers after injury.

The corticospinal tract, one of the major sensorimotor tracts, can be lesioned unilaterally in the rodent brainstem in order to test neuroplasticity-inducing therapies for the central nervous system. This surgical procedure (&#8220;pyramidotomy&#8221;) and postoperative assessments are described in this protocol.

神経可塑性を誘導する治療法の評価のためのラットの皮質脊髄路の片側Pyramidotomy

The corticospinal tract (CST) can be completely severed unilaterally in the medullary pyramids of the rodent brainstem. The CST is a motor tract that has ...

神経科学

Motor and Hippocampal Dependent Spatial Learning and Reference Memory Assessment in a Transgenic Rat Model of Alzheimer's Disease with Stroke

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk factor for AD, stroke has also been implicated as a risk factor and an exacerbating factor. The co-morbidity of stroke and AD results in worsened stroke-related motor control and AD-related cognitive deficits when compared to each condition alone. To model the combined condition of stroke and AD, a novel transgenic rat model of AD, with a mutated form of amyloid precursor protein (a key protein involved in the development of AD) incorporated into its DNA, is given a small unilateral striatal stroke.
For a model with the combination of both stroke and AD, behavioral tests that assess stroke-related motor control, locomotion and AD-related cognitive function must be implemented. The cylinder task involves a cost-efficient, multipurpose apparatus that assesses spontaneous forelimb motor use. In this task, a rat is placed in a cylindrical apparatus, where the rat will spontaneously rear and contact the wall of the cylinder with its forelimbs. These contacts are considered forelimb motor use and quantified during video analysis after testing. Another cost-efficient motor task implemented is the beam-walk task, which assesses forelimb control, hindlimb control and locomotion. This task involves a rat walking across a wooden beam allowing for the assessment of limb motor control through analysis of forelimb slips, hindlimb slips and falls. Assessment of learning and memory is completed with Morris water maze for this behavioral paradigm. The protocol starts with spatial learning, whereby the rat locates a stationary hidden platform. After spatial learning, the platform is removed and both short-term and long-term spatial reference memory is assessed. All three of these tasks are sensitive to behavioral differences and completed within 28 days for this model, making this paradigm time-efficient and cost-efficient.

To investigate the co-morbid Alzheimer&#39;s disease (AD) and stroke condition in a novel model, three behavior tasks are described that assess both motor control and cognitive behaviors. These tasks include the beam-walk task, cylinder task and Morris water maze.

モーターと脳卒中とアルツハイマー病のトランスジェニックラットモデルにおける海馬依存性の空間学習とリファレンスメモリ評価

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk ...

Rodent Behavioral Testing to Assess Functional Deficits Caused by Microelectrode Implantation in the Rat Motor Cortex

Medical devices implanted in the brain hold tremendous potential. As part of a Brain Machine Interface (BMI) system, intracortical microelectrodes demonstrate the ability to record action potentials from individual or small groups of neurons. Such recorded signals have successfully been used to allow patients to interface with or control&#160;computers, robotic limbs, and their own limbs. However, previous animal studies have shown that a microelectrode implantation in the brain not only damages the surrounding tissue but can also result in functional deficits. Here, we discuss a series of behavioral tests to quantify potential motor impairments following the implantation of intracortical microelectrodes into the motor cortex of a rat. The methods for open field grid, ladder crossing, and grip strength testing provide valuable information regarding the potential complications resulting from a microelectrode implantation. The results of the behavioral testing are correlated with endpoint histology, providing additional information on the pathological outcomes and impacts of this procedure on the adjacent tissue.

We have shown that a microelectrode implantation in the motor cortex of rats causes immediate and lasting motor deficits. The methods proposed herein outline a microelectrode implantation surgery and three rodent behavioral tasks to elucidate potential changes in the fine or gross motor function due to implantation-caused damage to the motor cortex.

ラット皮質運動野の微小電極注入による機能的障害を評価するためにテスト行動の齧歯動物

Medical devices implanted in the brain hold tremendous potential. As part of a Brain Machine Interface (BMI) system, intracortical microelectrodes ...

生物工学

The 6-hydroxydopamine Rat Model of Parkinson's Disease

Motor symptoms of Parkinson's disease (PD)-bradykinesia, akinesia, and tremor at rest-are consequences of the neurodegeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc) and dopaminergic striatal deficit. Animal models have been widely used to simulate human pathology in the laboratory. Rodents are the most used animal models for PD due to their ease of handling and maintenance. Moreover, the anatomy and molecular, cellular, and pharmacological mechanisms of PD are similar in rodents and humans. The infusion of the neurotoxin, 6-hydroxydopamine (6-OHDA), into a medial forebrain bundle (MFB) of rats reproduces the severe destruction of dopaminergic neurons and simulates PD symptoms. This protocol demonstrates how to perform the unilateral microinjection of 6-OHDA in the MFB in a rat model of PD and shows the motor deficits induced by 6-OHDA and predicted dopaminergic lesions through the stepping test. The 6-OHDA causes significant impairment in the number of steps performed with the contralateral forelimb.

The 6-hydroxydopamine (6-OHDA) model has been used for decades to advance the understanding of Parkinson's Disease. In this protocol, we demonstrate how to perform unilateral nigrostriatal lesions in the rat by injecting 6-OHDA in the medial forebrain bundle, assess motor deficits, and predict lesions using the stepping test.

パーキンソン病の6-ヒドロキシドーパミンラットモデル

Motor symptoms of Parkinson's disease (PD)-bradykinesia, akinesia, and tremor at rest-are consequences of the neurodegeneration of dopaminergic neurons in ...

The Mouse Stroke Unit Protocol with Standardized Neurological Scoring for Translational Mouse Stroke Studies

The filament model of middle artery occlusion (fMCAo) is perhaps the most translational mouse stroke model, allowing for controlled ischemia with intravascular reperfusion/recanalization. However, it lacks alignment with current clinical advances for stroke care (e.g., Stroke Units), usually employs subjective or vague neurological scoring among laboratories, and exhibits high acute-phase mortality. Here, we address these limitations with validated video-guided protocols. We present the mouse Stroke Unit (mSU) protocol with instructional videos and a decision algorithm (Risk Stratification Score), bridging the gap between clinical and mouse stroke modeling. To increase accuracy and sensitivity of stroke neurological scoring, we present for the first time a video-standardized format of the focal Experimental Stroke Scale (fESS) and prove its value up to 6 months post-stroke. Additionally, protocols for mice Ladder-rung test, as well as the known Cylinder test, for unbiased, quantitative assessment of limbs&#180; motor function are presented. Results highlight mSU&#39;s translational efficacy. Focal ESS (fESS) excels over other known scales in detecting focal stroke deficits, capturing recovery, and maintaining sensitivity for up to 6 months post-stroke. Ladder-rung and Cylinder tests objectively quantify and monitor fore- and hind-limb motor deficits, long-term. In summary, integrating mSU, fESS, and motor function tests provides a robust framework for clinically relevant stroke investigations. Our protocols improve the translational value in mouse stroke research.

This article presents a protocol for executing the highly translational and independently proven mouse Stroke Unit (mSU) concept in large-stroke mice. It also provides a standardized protocol for executing the focal experimental stroke scale in mice, which evaluates stroke focal deficits detected even long-term.