وأيد هذا البحث من المنح المقدمة من مجلس البحوث الطبية ، والعلوم الطبيعية والهندسة مجلس البحوث وكندا ، والشبكة الكندية السكتة الدماغية. كما أيد حركة اتشيه الحرة من قبل مؤسسة التراث ألبرتا للأبحاث الطبية.

درجة سلم المشي جهاز الاختبار تألف درجة سلم الأفقي المشي جهاز الاختبار من الجدران المصنوعة من الجانب زجاجي واضحة ودرجات المعادن (3 مم) ، والتي يمكن أن تضاف إلى خلق الأرض مع الحد الأدنى لمسافة 1 سم بين درجات (انظر الشكل رقم 1 ؛ ميتز وWhishaw ، 2003). كانت الجدران الجانبية 1 م و 19 سم طويلة تقاس عالية من ارتفاع درجات. وقد ارتقى في السلم 30 سم فوق سطح الأرض مع بدء قفص محايدة وملجأ (قفص المنزل) في نهاية المطاف. لأن الحيوانات اعتادوا خلال التدريب ، وكان الارتفاع من المحتمل أن الجهاز تسبب القلق. تم تعديل العرض من زقاق إلى حجم الحيوان ، بحيث كان حوالي 1 سم أوسع من حيوان لمنع الحيوان من الدوران في المكان. تم تعديل صعوبة المهمة من خلال تغيير موقف درجات المعادن. سمح لنمط منتظم من درجات الحيوانات لمعرفة مدى نمط عدة دورات تدريبية وتوقع موقف درجات (الشكل 1 ، نمط). منعت نمطا غير النظامية التي تم تغييرها من المحاكمة من محاكمة الحيوان من نمط التعلم (الشكل 1 ، خطة B). لترتيب العادية ، كانت متباعدة في درجات على فترات سم 2. لنمط غير النظامية ، وتباينت درجات على مسافة منهجية 1-5 سم. وقد استخدمت خمسة نماذج من أنماط درجة غير النظامية ، بحيث تم تطبيق نفس الأنماط لجميع الحيوانات لتوحيد صعوبة الاختبار وتعزيز قابليتها للمقارنة لنتائج (انظر النتائج). Videorecording تم وضع كاميرا (Canovision ، Canon المؤتمر الوطني العراقي) في زاوية طفيف بطني ، حتى يمكن تسجيل مواقف جميع الأطراف الأربعة في وقت واحد. تم تعيين سرعة مصراع الكاميرا في 500 -- 2000 س. حللت videorecordings باستخدام الإطار حسب الإطار تحليل في 30 و / ثانية. السلوكية والتدريب واختبار تحليل وقد تم تدريب هذه الحيوانات لعبور سلم من قفص محايدة للوصول إلى القفص وطنهم ، وبالتالي فإن قفص المنزل مع تتزاحم قدم التعزيز الإيجابي للمشي. عبرت جميع الحيوانات السلم في نفس الاتجاه. لم يعط المزيد من التعزيز لتحفيز الحيوانات لعبور السلم. وقد تم تدريب واختبار جميع الحيوانات خمس مرات في كل دورة. القدم بتسجيله خطأ تم إجراء التقييم النوعي للتنسيب وforelimb hindlimb باستخدام القدم بتسجيله خطأ النظام كما هو موضح سابقا (ميتز وWhishaw ، 2003). تم تحليلها من قبل التفتيش على تسجيلات الفيديو الإطار حسب الإطار. وقد تم تحليل سوى خطوات متتالية من كل الاطراف. لذا ، لم يكن أحرز الخطوة الأخيرة قبل انقطاع مشية ، مثل وقف أو خطأ القدم ، والخطوة الأولى بعد انقطاع. كما تم استبعاد مشاركة في دورة تصعيد أجريت في نهاية سلم من الدرجات. وسجل وضع الطرف من حيث وضع الطرف على نتوء درجة وأطرافه بين درجات عندما يغيب حدث. تم تصنيف أنواع موضع الأقدام أو مخلب على درجات باستخدام مقياس 7 - الفئة (انظر الشكل 2). تم تصنيف الأقدام أو مخلب التنسيب على درجة وفقا لموقعها والأخطاء التي وقعت في دقة التنسيب. (0) أعطيت إجمالي تفوت. 0 نقطة عند الطرف غاب بشكل كامل درجة ، لم يكن أي اتصال به ، وحدث السقوط. وعرف يسقطون أحد أطرافه بعمق في هبوط درجات بين وضعية الجسم والتوازن وتضطرب. (1) قسيمة ديب. انصب في البداية على درجة أطرافهم ، ثم انزلقت قبالة عندما حمل وزن الجسم وتسبب في سقوط. (2) زلة طفيف. انزلق وضعت الأطراف على درجة قبالة عندما تحمل الوزن ، ولكن لم يسفر عن سقوط ولا يقطع دورة المشي. في هذه الحالة ، كانت الحيوانات قادرة على الحفاظ على التوازن ومواصلة مشية منسقة. (3) استبدال. أطرافهم وضعت على الدرجة ، ولكن قبل أن تحمل الوزن ورفع سرعان ما وضعت على آخر درجة. (4) التصحيح ، والتي تهدف لأحد أطرافه درجة ، ولكن لم يوضع بعد ذلك على درجة أخرى دون لمس الأول. بدلا من ذلك ، تم تسجيلها على درجة من 4 إذا وضعت على أحد أطرافه ، وكان يدق بسرعة تعديل أوضاعها في حين تبقى على نفس الدرجة. (5) موضع الجزئي وضعت الأطراف على درجة مع أي أرقام أو المعصم من forelimb أو كعب أو أصابع hindlimb. (6) موضع الصحيح. midportion وضعت من النخيل من أحد أطرافه على درجة بدعم الوزن الكامل. عندما وقعت أخطاء مختلفة في الوقت نفسه ، سجلت أدنى الدرجات. على سبيل المثال ، إذا تم تسجيلها لأول مرة وضعت قدميك على درجة ثم وضعت على واحد آخر في نفس الخطوة (درجة 3) ، وتراجع بعد ذلك وقعت في الفترات الفاصلة بين درجات (درجة 1) ، وعلى درجة من 1. WHان السقوط حدث ، فقط تم تصنيف أطرافهم الشروع في الخطأ واحرز أيا من الأطراف الأخرى حتى الحيوان قد أعادت جميع الأطراف. وبلغ متوسط ​​درجات خطأ من خمس تجارب لتحليلها. قدم تحليل دقة التنسيب (عدد أخطاء) وقد أحصى عدد من الأخطاء في كل عبور. تم تحديد الأخطاء على أساس نظام تسجيل خطأ في القدم. تم تعريف خطأ لأن كل الأطراف التي تلقت التنسيب على درجة من النقاط 0 ، 1 أو 2 ، أي خطأ يمثل أي نوع من زلة قدم أو يغيب الإجمالي. تم تسجيل عدد من الأخطاء وعدد من الخطوات من أجل كل طرف على حدة. من هذه المعطيات ، تم احتساب متوسط ​​عدد من الأخطاء في خطوة وبلغ متوسط ​​مدة خمس محاكمات. كانت المعلمة الثانية الكمية تحليل متوسط ​​الوقت اللازم لعبور على طول المهمة السلم. بدأ قياس الوقت بعد ان وضعت الحيوان في سلم وبدأ المشي على طول منه. لم تكن مدرجة في الوقت الذي يقضيه في حيوان توقف في القياس. Forepaw يسجل رقما تم تسجيل درجة forepaw أرقام عندما وضعت بشكل صحيح مع مخلب midportion به على درجة (أي درجة 6 في نظام خطأ التهديف القدم). تم تصنيف الدرجة التي يمكن أن تكون الأرقام من forepaw استعرضوا حول درجة في مقياس من ثلاث نقاط (الشكل 2). تم تطبيق رصيده الى الموقف بدعم الوزن الأقصى ، أي كان وضعه في forelimb عموديا على درجة. اعطيت درجات على النحو التالي : (0) سيارة مغلقة في زاوية 90 درجة تقريبا. (1) سيارة مغلقة في زاوية 45 درجة تقريبا. (2) سيارة استعرضوا تماما حول درجة. وبلغ متوسط ​​درجات من خمس خطوات واستخدامها في التحليل. الرقم الأساطير <img src="/files/ftp_upload/old/1204_2009_1_20_6/Fig 1 Grid.png" alt="الشكل 1" /> الشكل 1. أشارت درجة سلم المهرة المشي جهاز اختبار في طريقة العرض الأمامي الوحشي ، مع مقاييس محددة للجهاز. واستخدمت نمطين درجة مختلفة في هذه الدراسة : نمط منتظم وترتيب درجة وباء خطة ، وعدم انتظام درجة الترتيب. الترتيب عدم انتظام درجة تتراوح بين محاكمات متتالية. <img src="/files/ftp_upload/old/1204_2009_1_20_7/Fig 2 (angle lines).png" alt="الشكل 2" /> التين. 2. الصور الفوتوغرافية التي تصور الممثل الفئات الثلاث من الدرجة أرقام. علما زاوية الأرقام فيما يتعلق forelimb عندما دعما الوزن الكامل.

<ol>
	<li>Chu, C. J., Jones, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+structural+plasticity+in+cortex+heterotopic+to+focal+sensorimotor+cortical+damage.">Experience-dependent structural plasticity in cortex heterotopic to focal sensorimotor cortical damage.</a> Exp Neurol. 166, 403-414 (2000).</li><li>Emerick, A. J., Kartje, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+recovery+and+anatomical+plasticity+in+adult+rats+after+cortical+lesion+and+treatment+with+monoclonal+antibody+IN-1.">Behavioral recovery and anatomical plasticity in adult rats after cortical lesion and treatment with monoclonal antibody IN-1.</a> Behav Brain Res. 152, 315-325 (2004).</li><li>Faraji, J., Metz, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sequential+bilateral+striatal+lesions+have+additive+effects+on+single+skilled+limb+use+in+rats.">Sequential bilateral striatal lesions have additive effects on single skilled limb use in rats.</a> Behav Brain Res. 177, 195-204 (2007).</li><li>Farr, T. D., Liu, L., Colwell, K. L., Whishaw, I. Q., Metz, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bilateral+alteration+in+stepping+pattern+after+unilateral+motor+cortex+injury:+a+new+test+strategy+for+analysis+of+skilled+limb+movements+in+neurological+mouse+models.">Bilateral alteration in stepping pattern after unilateral motor cortex injury: a new test strategy for analysis of skilled limb movements in neurological mouse models.</a> J Neurosci Methods. 153, 104-113 (2006).</li><li>Kleim, J. A., Barbay, S., Nudo, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+reorganization+of+the+rat+motor+cortex+following+motor+skill+learning.">Functional reorganization of the rat motor cortex following motor skill learning.</a> J Neurophysiol. 80, 3321-3325 (1998).</li><li>Merkler, D., Metz, G. A., Raineteau, O., Dietz, V., Schwab, M. E., Fouad, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Locomotor+recovery+in+spinal+cord-injured+rats+treated+with+an+antibody+neutralizing+the+myelin-associated+neurite+growth+inhibitor+Nogo-A.">Locomotor recovery in spinal cord-injured rats treated with an antibody neutralizing the myelin-associated neurite growth inhibitor Nogo-A.</a> J Neurosci. 21, 3665-3673 (2001).</li><li>McVea, D. A., Pearson, K. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stepping+of+the+forelegs+over+obstacles+establishes+long-lasting+memories+in+cats.">Stepping of the forelegs over obstacles establishes long-lasting memories in cats.</a> Curr Biol. 17, R621-R623 (2007).</li><li>Metz, G. A. S., Dietz, V., Schwab, M. E., Meent, H. v. a. n. d. e. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+unilateral+pyramidal+tract+section+on+hindlimb+motor+performance+in+the+rat.">The effects of unilateral pyramidal tract section on hindlimb motor performance in the rat.</a> Behav Brain Res. 95, 37-46 (1998).</li><li>Metz, G. A. S., Dietz, V., Schwab, M. E., Fouad, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+testing+of+motor+function+in+spinal+cord+injured+rats.">Efficient testing of motor function in spinal cord injured rats.</a> Brain Res. 883, 165-177 (2000).</li><li>Metz, G. A., Whishaw, I. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug-induced+rotation+intensity+in+unilateral+dopamine-depleted+rats+is+not+correlated+with+end+point+or+qualitative+measures+of+forelimb+or+hindlimb+motor+performance.">Drug-induced rotation intensity in unilateral dopamine-depleted rats is not correlated with end point or qualitative measures of forelimb or hindlimb motor performance.</a> Neuroscience. 111, 325-336 (2002).</li><li>Miklyaeva, E. I., Castaneda, E., Whishaw, I. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skilled+reaching+deficits+in+unilateral+dopamine-depleted+rats:+impairments+in+movement+and+posture+and+compensatory+adjustments.">Skilled reaching deficits in unilateral dopamine-depleted rats: impairments in movement and posture and compensatory adjustments.</a> J Neurosci. 14, 7148-7158 (1994).</li><li>Muir, G. D., Whishaw, I. Q., Kolb, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Locomotion in The Behavior of the laboratory rat. , 150-161 (2005).</li><li>Muir, G. D., Whishaw, I. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ground+reaction+forces+in+locomoting+hemi-parkinsonian+rats:+a+definitive+test+for+impairments+and+compensations.">Ground reaction forces in locomoting hemi-parkinsonian rats: a definitive test for impairments and compensations.</a> Exp Brain Res. 126, 307-314 (1999).</li><li>Muir, G. D., Whishaw, I. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Red+nucleus+lesions+impair+overground+locomotion+in+rats:+a+kinetic+analysis.">Red nucleus lesions impair overground locomotion in rats: a kinetic analysis.</a> Eur J Neurosci. 12, 1113-1122 (2000).</li><li>Riek-Burchardt, M., Henrich-Noack, P., Metz, G. A., Reymann, K. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+chronic+sensorimotor+impairments+in+the+ladder+rung+walking+task+in+rats+with+endothelin-1-induced+mild+focal+ischemia.">Detection of chronic sensorimotor impairments in the ladder rung walking task in rats with endothelin-1-induced mild focal ischemia.</a> J Neurosci Methods. 137, 227-233 (2004).</li><li>Ploughman, M., Attwood, Z., White, N., Dor&#233;, J. J., Corbett, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endurance+exercise+facilitates+relearning+of+forelimb+motor+skill+after+focal+ischemia.">Endurance exercise facilitates relearning of forelimb motor skill after focal ischemia.</a> Eur J Neurosci. 25, 3453-3460 (2007).</li><li>Thallmair, M., Metz, G. A. S., Z'Graggen, W. J., Kartje, G. L., Schwab, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+recovery+parallels+enhanced+plasticity+of+the+lesioned+adult+CNS+by+antibodies+to+neurite+growth+inhibitors.">Functional recovery parallels enhanced plasticity of the lesioned adult CNS by antibodies to neurite growth inhibitors.</a> Nature Neurosci. 1, 124-131 (1998).</li><li>Meyenburg, J. v. o. n., Brosamle, C., Metz, G. A., Schwab, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regeneration+and+sprouting+of+chronically+injured+corticospinal+tract+fibers+in+adult+rats+promoted+by+NT-3+and+the+mAb+IN-1,+which+neutralizes+myelin-associated+neurite+growth+inhibitors.">Regeneration and sprouting of chronically injured corticospinal tract fibers in adult rats promoted by NT-3 and the mAb IN-1, which neutralizes myelin-associated neurite growth inhibitors.</a> Exp Neurol. 154, 583-594 (1998).</li><li>Whishaw, I. Q., Coles, B. L., Pellis, S. M., Miklyaeva, E. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impairments+and+compensation+in+mouth+and+limb+use+in+free+feeding+after+unilateral+dopamine+depletions+in+a+rat+analog+of+human+Parkinson's+disease.">Impairments and compensation in mouth and limb use in free feeding after unilateral dopamine depletions in a rat analog of human Parkinson's disease.</a> Behav Brain Res. 84, 167-177 (1997a).</li><li>Whishaw, I. Q., Woodward, N. C., Miklyaeva, E., Pellis, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+limb+use+by+control+rats+and+unilateral+DA-depleted+rats+in+the+Montoya+staircase+test:+movements,+impairments+and+compensatory+strategies.">Analysis of limb use by control rats and unilateral DA-depleted rats in the Montoya staircase test: movements, impairments and compensatory strategies.</a> Behav Brain Res. 89, 167-177 (1997b).</li><li>Whishaw, I. Q., Gorny, B., Sarna, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paw+and+limb+use+in+skilled+and+spontaneous+reaching+after+pyramidal+tract,+red+nucleus+and+combined+lesions+in+the+rat:+behavioral+and+anatomical+dissociations.">Paw and limb use in skilled and spontaneous reaching after pyramidal tract, red nucleus and combined lesions in the rat: behavioral and anatomical dissociations.</a> Behav Brain Res. 93, 167-183 (1998).</li><li>Z'Graggen, W. J., Metz, G. A. S., Kartje, G. L., Schwab, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+recovery+and+enhanced+ccrtico-fugal+plasticity+in+the+adult+rat+after+unilateral+pyramidal+tract+section+and+blockade+of+myelin-associated+neurite+growth+inhibitors.">Functional recovery and enhanced ccrtico-fugal plasticity in the adult rat after unilateral pyramidal tract section and blockade of myelin-associated neurite growth inhibitors.</a> J Neurosci. 18, 4744-4757 (1998).</li></ol>

ليس لدينا شيء في الكشف عنها.

هذه الدراسة بتقديم درجة سلم المشي المهمة بمثابة اختبار جديد لتقييم مهارة المشي والتنسيب أطرافهم وأعضائهم التنسيق. لدرجة سلم المشي مهمة التمييز بين الاضطرابات يسمح خفية من وظيفة الحركة من خلال الجمع بين التحليل الكمي والنوعي من المشي المهرة. ويمكن استخدام الفيديو ل?...

the ladder rung walking task: a scoring system and its practical application.

التقدم في تطوير نماذج حيوانية ل/ السكتة الدماغية وإصابات الحبل الشوكي ، ومرض الاعصاب الأخرى يتطلب اختبارات حساسية عالية لوضع جوانب بارزة من وظيفة الحركة ، وتحديد دقيق حتى فقدان القدرة على الحركة. لتعزيز فعالية ودقة من الاختبار ، وينبغي اتخاذ تدابير تسمح الاختبارات النوعية والكمية وظيفة المحرك وتكون حساسة للتغيرات في الأداء أثناء فترات الانتعاش. هذه الدراسة توضح مهمة جديدة لتقييم مهارة المشي في الفئران لقياس كل وظيفة forelimb hindlimb في نفس الوقت. مطلوبة الحيوانات على المشي على طول سلم الأفقية التي تباعد للدرجات هو متغير ويتغير بشكل دوري. التغيرات في درجة التباعد منع الحيوانات من مكان التعلم المطلق والنسبي للدرجات والتقليل بذلك من قدرة الحيوانات على تعويض عن العاهات من خلال التعلم. بالإضافة إلى ذلك ، تغيير التباعد بين درجات يسمح لاستخدامها في اختبار مرارا وتكرارا في الدراسات طويلة الأجل. ووصف الطرق لوصف كل من كمية ونوعية الأداء الصدارة وhindlimb على حد سواء ، بما في ذلك وضع الأطراف ، يخطو ، التنسيق. وعلاوة على ذلك ، يشار إلى استخدام استراتيجيات التعويضية التي كتبها عثرات أو خطوات تعويضية ردا على الطرف الآخر في غير موضعها ل.

Watch this Scientific Journal Video about The Ladder Rung Walking Task: A Scoring System and its Practical Application. at JoVE.com

The Ladder Rung Walking Task: A Scoring System and its Practical Application.

لدرجة سلم المشي المهمة اختبارا جديدا لتقييم وقياس مهارة المشي على حد سواء وforelimb hindlimb وضع في تصعيد ، وبين أطرافه التنسيق.

وسلم المشي رونغ العمل : نظام تسجيل النقاط وتطبيقها العملي.

Progress in the development of animal models for/stroke, spinal cord injury, and other neurodegenerative disease requires tests of high sensitivity to ...

the-ladder-rung-walking-task-scoring-system-its-practical

Research

JoVE Journal

Biology

25.8K Views.  University of Lethbridge. لدرجة سلم المشي المهمة اختبارا جديدا لتقييم وقياس مهارة المشي على حد سواء وforelimb hindlimb وضع في تصعيد ، وبين أطرافه التنسيق.

Video: The Ladder Rung Walking Task: A Scoring System and its Practical Application.

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.

استخدام Rotorod كأسلوب للتحليل النوعي من المشي في الجرذ

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

Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants

The lens plays an important role in the development of the optic cup[1,2]. Using the zebrafish as a model organism, questions regarding lens development can be addressed. The zebrafish is useful for genetic studies due to several advantageous characteristics, including small size, high fecundity, short lifecycle, and ease of care. Lens development occurs rapidly in zebrafish. By 72 hpf, the zebrafish lens is functionally mature [3]. Abundant genetic and molecular resources are available to support research in zebrafish. In addition, the similarity of the zebrafish eye to those of other vertebrates provides basis for its use as an excellent animal model of human defects[4-7]. Several zebrafish mutants exhibit lens abnormalities, including high levels of cell death, which in some cases leads to a complete degeneration of lens tissues [8]. To determine whether lens abnormalities are due to intrinsic causes or to defective interactions with the surrounding tissues, transplantation of a mutant lens into a wild-type eye is performed. Using fire-polished metal needles, mutant or wild-type lenses are carefully dissected from the donor animal, and transferred into the host. To distinguish wild-type and mutant tissues, a transgenic line is used as the donor. This line expresses membrane-bound GFP in all tissues, including the lens. This transplantation technique is an essential tool in the studies of zebrafish lens mutants.

Lens development involves interactions with other tissues. Several zebrafish eye mutants are characterized by an abnormally small lens size. Here we demonstrate a lens transplantation experiment to determine whether this phenotype is due to intrinsic causes or defective interactions with tissues that surround the lens.

زرع عدسة في اسماك الزرد وتطبيقاتها في تحليل المسوخ العين

The lens plays an important role in the development of the optic cup[1,2]. Using the zebrafish as a model organism, questions regarding lens development ...

Visualization of Caenorhabditis elegans Cuticular Structures Using the Lipophilic Vital Dye DiI

The cuticle of C. elegans is a highly resistant structure that surrounds the exterior of the animal1-4. The cuticle not only protects the animal from the environment, but also determines body shape and plays a role in motility4-6. Several layers secreted by epidermal cells comprise the cuticle, including an outermost lipid layer7.
Circumferential ridges in the cuticle called annuli pattern the length of the animal and are present during all stages of development8. Alae are longitudinal ridges that are present during specific stages of development, including L1, dauer, and adult stages2,9. Mutations in genes that affect cuticular collagen organization can alter cuticular structure and animal body morphology5,6,10,11. While cuticular imaging using compound microscopy with DIC optics is possible, current methods that highlight cuticular structures include fluorescent transgene expression12, antibody staining13, and electron microscopy1. Labeled wheat germ agglutinin (WGA) has also been used to visualize cuticular glycoproteins, but is limited in resolving finer cuticular structures14. Staining of cuticular surface using fluorescent dye has been observed, but never characterized in detail15. We present a method to visualize cuticle in live C. elegans using the red fluorescent lipophilic dye DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), which is commonly used in C. elegans to visualize environmentally exposed neurons. This optimized protocol for DiI staining is a simple, robust method for high resolution fluorescent visualization of annuli, alae, vulva, male tail, and hermaphrodite tail spike in C. elegans.

Visualization of Caenorhabditis elegans Cuticular Structures Using the Lipophilic Vital Dye DiI

We present a method to visualize cuticle in live C. elegans using the red fluorescent lipophilic dye DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), which is commonly used in C. elegans to visualize environmentally exposed neurons. With this optimized protocol, alae and annular cuticular structures are stained by DiI and observed using compound microscopy.

تصور انواع معينة ايليجانس</emالهياكل البشرة> استخدام الصبغة الحيوية محبة للدهون ، صلاح الغيدان

The cuticle of C. elegans is a highly resistant structure that surrounds the exterior of the animal1-4. The cuticle not only protects the animal from the ...

A Fluorescence-based Exonuclease Assay to Characterize DmWRNexo, Orthologue of Human Progeroid WRN Exonuclease, and Its Application to Other Nucleases

WRN exonuclease is involved in resolving DNA damage that occurs either during DNA replication or following exposure to endogenous or exogenous genotoxins. It is likely to play a role in preventing accumulation of recombinogenic intermediates that would otherwise accumulate at transiently stalled replication forks, consistent with a hyper-recombinant phenotype of cells lacking WRN. In humans, the exonuclease domain comprises an N-terminal portion of a much larger protein that also possesses helicase activity, together with additional sites important for DNA and protein interaction. By contrast, in Drosophila, the exonuclease activity of WRN (DmWRNexo) is encoded by a distinct genetic locus from the presumptive helicase, allowing biochemical (and genetic) dissection of the role of the exonuclease activity in genome stability mechanisms. Here, we demonstrate a fluorescent method to determine WRN exonuclease activity using purified recombinant DmWRNexo and end-labeled fluorescent oligonucleotides. This system allows greater reproducibility than radioactive assays as the substrate oligonucleotides remain stable for months, and provides a safer and relatively rapid method for detailed analysis of nuclease activity, permitting determination of nuclease polarity, processivity, and substrate preferences.

Exonucleases play critical roles in ensuring genome stability. Loss of WRN exonuclease function results in premature aging. Studying substrates and other requirements of the nuclease in vitro can help elucidate its role in vivo. Here we demonstrate a rapid and reproducible fluorescence-based assay to measure its nuclease activity.

WRN exonuclease is involved in resolving DNA damage that occurs either during DNA replication or following exposure to endogenous or exogenous genotoxins. ...

A Practical Guide to Phylogenetics for Nonexperts

Many researchers, across incredibly diverse foci, are applying phylogenetics to their research question(s). However, many researchers are new to this topic and so it presents inherent problems. Here we compile a practical introduction to phylogenetics for nonexperts.&#160;We outline in a step-by-step manner, a pipeline for generating reliable phylogenies from gene sequence datasets. We begin with a user-guide for similarity search tools via online interfaces as well as local executables. Next, we explore programs for generating multiple sequence alignments followed by protocols for using software to determine best-fit models of evolution. We then outline protocols for reconstructing phylogenetic relationships via maximum likelihood and Bayesian criteria&#160;and finally describe tools for visualizing phylogenetic trees. While this is not by any means an exhaustive description of phylogenetic approaches, it does provide the reader with practical starting information on key software applications commonly utilized by phylogeneticists. The vision for this article would be that it could serve as a practical training tool for researchers embarking on phylogenetic studies&#160;and also serve as an educational resource that could be incorporated into a classroom or teaching-lab.

Here we describe a step-by-step pipeline for generating reliable phylogenies from nucleotide or amino acid sequence datasets. This guide aims to serve researchers or students new to phylogenetic analysis.&#160;

دليل عملي لPhylogenetics لNonexperts

Many researchers, across incredibly diverse foci, are applying phylogenetics to their research question(s). However, many researchers are new to this ...

Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

The circadian clock regulates many aspects of life, including sleep, locomotor activity, and body temperature (BTR) rhythms1,2. We recently identified a novel Drosophila circadian output, called the temperature preference rhythm (TPR), in which the preferred temperature in flies rises during the day and falls during the night 3. Surprisingly, the TPR and locomotor activity are controlled through distinct circadian neurons3. Drosophila locomotor activity is a well&#160;known circadian behavioral output and has provided strong contributions to the discovery of many conserved mammalian circadian clock genes and mechanisms4. Therefore, understanding TPR will lead to the identification of hitherto unknown molecular and cellular circadian mechanisms. Here, we describe how to perform and analyze the TPR assay. This technique not only allows for dissecting the molecular and neural mechanisms of TPR, but also provides new insights into the fundamental mechanisms of the brain functions that integrate different environmental signals and regulate animal behaviors. Furthermore, our recently published data suggest that the fly TPR shares features with the mammalian BTR3. Drosophila are ectotherms, in which the body temperature is typically behaviorally regulated. Therefore, TPR is a strategy used to generate a rhythmic body temperature in these flies5-8. We believe that further exploration of Drosophila TPR will facilitate the characterization of the mechanisms underlying body temperature control in animals.

Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

We recently identified a novel Drosophila circadian output, temperature preference rhythm (TPR), in which the preferred temperature in flies rises during the day and falls during the night. TPR is regulated independently from another circadian output, locomotor activity. Here&#160;we describe the design and analysis of TPR in Drosophila.

تصميم وتحليل سلوك تفضيل درجة الحرارة وإيقاعها Circadian في Drosophila

The circadian clock regulates many aspects of life, including sleep, locomotor activity, and body temperature (BTR) rhythms1,2. We recently identified a ...

Reconstruction of 3-Dimensional Histology Volume and its Application to Study Mouse Mammary Glands

Histology volume reconstruction facilitates the study of 3D shape and volume change of an organ at the level of macrostructures made up of cells. It can also be used to investigate and validate novel techniques and algorithms in volumetric medical imaging and therapies. Creating 3D high-resolution atlases of different organs1,2,3 is another application of histology volume reconstruction. This provides a resource for investigating tissue structures and the spatial relationship between various cellular features. We present an image registration approach for histology volume reconstruction, which uses a set of optical blockface images. The reconstructed histology volume represents a reliable shape of the processed specimen with no propagated post-processing registration error. The Hematoxylin and Eosin (H&#38;E) stained sections of two mouse mammary glands were registered to their corresponding blockface images using boundary points extracted from the edges of the specimen in histology and blockface images. The accuracy of the registration was visually evaluated. The alignment of the macrostructures of the mammary glands was also visually assessed at high resolution.
This study delineates the different steps of this image registration pipeline, ranging from excision of the mammary gland through to 3D histology volume reconstruction. While 2D histology images reveal the structural differences between pairs of sections, 3D histology volume provides the ability to visualize the differences in shape and volume of the mammary glands.

We present an image registration approach for 3-dimensional (3D) histology volume reconstruction, which facilitates the study of the changes of an organ at the level of macrostructures made up of cells . Using this approach, we studied the 3D changes between wild-type and Igfbp7-null mammary glands.

إعادة بناء 3 الأبعاد حجم الأنسجة وتطبيقاتها لدراسة ماوس الغدد الثديية

Histology volume reconstruction facilitates the study of 3D shape and volume change of an organ at the level of macrostructures made up of cells. It can ...

Imaging Spatial Reorganization of a MAPK Signaling Pathway Using the Tobacco Transient Expression System

Visualization of dynamic signaling events in live cells has been a challenge. We expanded an established transient expression system, the biomolecular fluorescent complementation (BiFC) assay in tobacco epidermal cells, from testing protein-protein interaction to monitoring spatial distribution of signal transduction in plant cells. In this protocol, we used the BiFC assay to show that the interaction and the signaling between the Arabidopsis MAPKKK YODA and MAPK6 occur at the plasma membrane. When the scaffold protein BASL was co-expressed, the YODA-MAPK interaction redistributed and spatially co-polarized with CFP-BASL. This modified tobacco expression system allows for quick examination of signaling localization and dynamic changes (less than 4 days) and can accommodate multiple of fluorescent protein colors (at least three). We also presented detailed methods to quantify protein distribution (asymmetric spatial localization, or &#34;polarization&#34;) in tobacco cells. This advanced tobacco expression system has a potential to be used widely for quick testing of dynamic signaling events in live plant cells.

At the subcellular level, signaling events are dynamically modulated by developmental and environmental cues. Here we describe a protocol that employs the tobacco transient expression system to monitor dynamic protein-protein interaction and to disclose spatial organization of signal transduction in plant cells.

التصوير إعادة التنظيم المكاني لMAPK اشارة المسار باستخدام نظام التعبير عابر التبغ

Visualization of dynamic signaling events in live cells has been a challenge. We expanded an established transient expression system, the biomolecular ...

Agrobacterium-Mediated Genetic Transformation, Transgenic Production, and Its Application for the Study of Male Reproductive Development in Rice

Male sterility is an important agronomic trait for hybrid seed production that is usually characterized by functional defects in male reproductive organs/gametes. Recent advances in CRISPR-Cas9 genome editing technology allow for high editing efficacy and timesaving knockout mutations of endogenous candidate genes at specific sites. Additionally, Agrobacterium-mediated genetic transformation of rice is also a key method for gene modification, which has been widely adopted by many public and private laboratories. In this study, we applied CRISPR-Cas9 genome editing tools and successfully generated three male sterile mutant lines by targeted genome editing of OsABCG15 in a japonica cultivar. We used a modified Agrobacterium-mediated rice transformation method that could provide excellent means of genetic emasculation for hybrid seed production in rice. Transgenic plants can be obtained within 2&#8211;3 months and homozygous transformants were screened by genotyping using PCR amplification and Sanger sequencing. Basic phenotypic characterization of the male sterile homozygous line was performed by microscopic observation of the rice male reproductive organs, pollen viability analysis by iodine potassium iodide (I2-KI) staining semi-thin cross-sectioning of developing anthers.

Agrobacterium-Mediated Genetic Transformation, Transgenic Production, and Its Application for the Study of Male Reproductive Development in Rice

This work describes the use of CRISPR-Cas9 genome editing technology to knockout endogenous gene OsABCG15 followed by a modified Agrobacterium-mediated transformation protocol to produce a stable male-sterile line in rice.

البكتيريا الزراعية-بوساطة التحول الجيني، وإنتاج تحويل الجينات، وتطبيقها لدراسة التنمية الإنجابية للذكور في الأرز

Male sterility is an important agronomic trait for hybrid seed production that is usually characterized by functional defects in male reproductive ...

A Simple Dry Sectioning Method for Obtaining Whole-Seed-Sized Resin Section and Its Applications

The morphology, size and quantity of cells, starch granules&#160;and protein bodies in seed determine the weight and quality of seed. They are significantly different among different regions of seed. In order to view the morphologies of cells, starch granules and protein bodies clearly, and quantitatively analyze their morphology parameters accurately, the whole-seed-sized section is needed. Though the whole-seed-sized paraffin section can investigate the accumulation of storage materials in seeds, it is very difficult to quantitatively analyze the morphology parameters of cells and storage materials due to the low resolution of the thick section. The thin resin section has high resolution, but the routine resin sectioning method is not suitable to prepare the whole-seed-sized section of mature seeds with a large volume and high starch content. In this study, we present a simple dry sectioning method for preparing the whole-seed-sized resin section. The technique can prepare the cross and longitudinal whole-seed-sized sections of developing, mature, germinated, and cooked seeds embedded in LR White resin, even for large seeds with high starch content. The whole-seed-sized section can be stained with fluorescent brightener 28, iodine, and Coomassie brilliant blue R250 to specifically exhibit the morphology of cells, starch granules, and protein bodies clearly, respectively. The image obtained can also be analyzed quantitatively to show the morphology parameters of cells, starch granules, and protein bodies in different regions of seed.

This technique allows for the fast and simple preparation of whole-seed-sized resin section for the observation and analysis of cells, starch granules, and protein bodies in different regions of the seed.