فإن الكتاب أود أن أشكر الدكتور ديانا آدكنز الدكتور Woodlee مارتي ، الأزواج كول ، مونيكا مالدونادو A. ، كين جاكي ، والدكتور تيم Schallert للمساعدة في تطوير اختبار التعامل مع الشعيرية ، ولو تشو للمساعدة في تطوير Capellini التعامل مع اختبار. بدعم من المنح المقدمة من المعاهد الوطنية للصحة ، NINDS (NS05689) ، والمعاهد الوطنية للصحة ، NIA (1F31AG034032 - 01).

<ol>
	<li>Allred, R. P., Adkins, D. L., Woodlee, M. T., Husbands, L. C., Maldonado, M. A., Kane, J. R., Schallert, T., 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=The+Vermicelli+Handling+Test:+a+simple+quantitative+measure+of+dexterous+forepaw+function+in+rats.">The Vermicelli Handling Test: a simple quantitative measure of dexterous forepaw function in rats.</a> J. Neurosci. Methods. 170, 229-244 (2008).</li><li>Baird, A. L., Meldrum, A., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+staircase+test+of+skilled+reaching+in+mice.">The staircase test of skilled reaching in mice.</a> Brain Res. Bull. 54, 243-250 (2001).</li><li>Ballermann, M., Metz, G. A., McKenna, J. E., Klassen, F., 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=The+pasta+matrix+reaching+task:+a+simple+test+for+measuring+skilled+reaching+distance,+direction,+and+dexterity+in+rats.">The pasta matrix reaching task: a simple test for measuring skilled reaching distance, direction, and dexterity in rats.</a> J. Neurosci. Methods. 106, 39-45 (2001).</li><li>Castro, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+performance+in+rats.+The+effects+of+pyramidal+tract+section.">Motor performance in rats. The effects of pyramidal tract section.</a> Brain Res. 44, 313-323 (1972).</li><li>Farr, T. 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=Quantitative+and+qualitative+impairments+in+skilled+reaching+in+the+mouse+(Mus+musculus)+after+a+focal+motor+cortex+stroke.">Quantitative and qualitative impairments in skilled reaching in the mouse (Mus musculus) after a focal motor cortex stroke.</a> Stroke. 33, 1869-1875 (2002).</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/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+staircase+test+:+a+measure+of+independent+forelimb+reaching+and+grasping+abilities+in+rats.">The staircase test : a measure of independent forelimb reaching and grasping abilities in rats.</a> J. Neurosci. Methods. 36, 219-228 (1991).</li><li>Peterson, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+handedness+in+the+rat.">Mechanism of handedness in the rat.</a> Comp Psych Monographs. 9, 1-67 (1934).</li><li>Tennant, K. A., Adkins, D. L., Donlan, N. A., Asay, A. L., Thomas, N., Kleim, J. A., 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=The+effect+of+age+on+motor+skill+learning+and+organization+of+the+forelimb+motor+cortical+representation+in+C57BL/6+mice.">The effect of age on motor skill learning and organization of the forelimb motor cortical representation in C57BL/6 mice.</a> Society for Neuroscience Annual Meeting abstract. , 783-784 (2009).</li><li>Tennant, K. A., 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=Sensorimotor+behavioral+effects+of+endothelin-1+induced+small+cortical+infarcts+in+C57BL/6+mice.">Sensorimotor behavioral effects of endothelin-1 induced small cortical infarcts in C57BL/6 mice.</a> J. Neurosci. Methods. 181, 18-26 (2009).</li><li>Whishaw, I. Q., Coles, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Varieties+of+paw+and+digit+movement+during+spontaneous+food+handling+in+rats:+postures,+bimanual+coordination,+preferences,+and+the+effect+of+forelimb+cortex+lesions.">Varieties of paw and digit movement during spontaneous food handling in rats: postures, bimanual coordination, preferences, and the effect of forelimb cortex lesions.</a> Behav. Brain Res. 77, 135-148 (1996).</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 (1997).</li><li>Whishaw, I. Q., Dringenberg, H. C., 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=Spontaneous+forelimb+grasping+in+free+feeding+by+rats:+motor+cortex+aids+limb+and+digit+positioning.">Spontaneous forelimb grasping in free feeding by rats: motor cortex aids limb and digit positioning.</a> Behav. Brain Res. 48, 112-125 (1992).</li><li>Whishaw, I. Q., O'Connor, W. T., Dunnett, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+contributions+of+motor+cortex,+nigrostriatal+dopamine+and+caudate-putamen+to+skilled+forelimb+use+in+the+rat.">The contributions of motor cortex, nigrostriatal dopamine and caudate-putamen to skilled forelimb use in the rat.</a> Brain. 109, 805-843 (1986).</li></ol>

الإعلان عن أي تضارب في المصالح.

في هذا الفيديو ، ونحن لشرح كيفية وتجرى اختبارات التعامل مع الشعيرية وCapellini في الفئران والجرذان ، على التوالي. هذه التجارب هي وسيلة بسيطة والكمية لدراسة مخلب وظيفة أرقام في الحيوانات العادية ، بعد الأضرار التي لحقت أنظمة السيارات من الدماغ ، أو أثناء الشيخوخة. البطيء تحليل شريط فيديو للمخلب والحركات أرقام تستخدم للتعامل مع قطع صغيرة من المعكرونة غير مطبوخة تسمح للباحث لتحديد التعديلات التي تجري مع كل مخلب ، وكذلك مراقبة والسلوكيات الشاذة التي يمكن أن ينظر إليها في أعقاب الأضرار التي يسببها الجهاز العصبي المركزي بشكل تجريبي. ويمكن القيام جلسات الاختبار مرارا وتكرارا في أيام مختلفة ومعظم الحيوانات استكمال جميع المحاكمات من جلسة في غضون دقائق قليلة. وبالتالي ، فإن الاختبار هو مناسبة لإدراجها في أكبر اختبار البطارية أو التصميم التجريبي. ويمكن أيضا أن تستخدم هذه الاختبارات في دراسات السيارات مهارة التعلم (شو وآخرون ، 2009) ، ولكن هنا نقدم بروتوكولا للتغيرات في فحص مهارة أنشئت على مر الزمن. حاليا ، اختبارات التوصل الماهرة هي الاختبارات الأكثر استخداما من وظيفة forelimb حاذق في الفئران (على سبيل المثال ، Whishaw وآخرون ، 1986 ؛. Ballermann وآخرون ، 2001 ؛. مونتويا وآخرون ، 1991.) والفئران (فار &amp; Whishaw ، 2002 ؛ تينانت وجونز ، 2009 ؛ بيرد وآخرون ، 2001). اختبارات التوصل تصريح اختبار معزولة من كل أطرافه ، في حين أن التعامل مع المعكرونة مهمة باليدين. ومع ذلك ، فإن التجارب تتشابه في حساسيتها للضعف في وظيفة forepaw المهرة واكتساب المهارة بالنسبة للاختبارات تتضمن مرونة في التمثيل forelimb من القشرة الحركية (شو وآخرون ، 2009). في تجربتنا ، والتوصل إلى اختبارات أكثر تعقيدا وشاقة لتنفيذ ، وبالنسبة لبعض الأسئلة التجريبية ، والتجارب الحالية قد توفر بديلا أكثر فعالية. ومع ذلك ، خلافا للاختبارات الموصوفة هنا ، تم التوصل إلى اختبارات المهارة في استخدام لعدة عقود (بيترسون ، 1934) وأنها تضم الآن مجموعة متنوعة من التدابير حساسة للغاية والمعلمات الاختبار (Ballermann وآخرون ، 2001 ؛. فار &amp; Whishaw ، 2002 ؛ مونتويا وآخرون ، 1991 ؛. Whishaw وآخرون ، 1992). في المقابل ، فإن التعامل مع الشعيرية والاختبارات Capellini تمتد من الأوصاف حديثة نسبيا للسلوك مناولة الأغذية البارع من الفئران (Whishaw وكولز ، 1996). في كل من الإدارة والتحليل ، ويمكن أن تكون هذه الاختبارات مما لا شك فيه مزيد من الصقل والموسعة ، على سبيل المثال ، إلى أكثر تحديدا كميا غرامة أنماط الحركة أرقام واليدين وقياس تحركات منسقة في الفم واللسان والفك.

the vermicelli and capellini handling tests: simple quantitative measures of dexterous forepaw function in rats and mice

وقد كشفت الأوصاف السابقة للسلوك الأكل القوارض التي تستخدمها الحركات forepaw منسقة لمعالجة قطع الطعام. لقد قدمنا ​​على هذا العمل لوضع مقياس كمي بسيط من البراعة forepaw التي تعتبر حساسة للعاهات lateralized والتغيرات التي تعتمد على سن. القوارض تعلم forepaw ماهرا والحركات أرقام لإدارة قطع معكرونة رقيقة ، والتي تستهلك بفارغ الصبر. لقد وصفنا من قبل وسائل لقياس الشعيرية المناولة في الفئران وتبين أن التدابير هي حساسة جدا لالعاهات الناتجة عن forelimb الآفات الدماغية من جانب واحد ، وسط انسداد الشريان الدماغي استنفاد الدوبامين من جانب واحد مخططي [الريد ، RP ، آدكنز ، DL ، Woodlee ، MT ، الأزواج ، LC ، مالدونادو MA ، كين ، JR ، Schallert ، T. &amp; جونز ، TA اختبار التعامل مع الشعيرية : وهو قياس كمية بسيطة من وظيفة forepaw حاذق في الفئران.<em&gt; J. Neurosci. طرق</em&gt; 170 ، 229-244 (2008)]. هنا نقدم بروتوكول أكثر تفصيلا لهذا الاختبار في الفئران ومقارنتها مع النسخة التي وضعت حديثا عن الفئران ، واختبار التعامل مع Capellini. وشريط فيديو الجرذان والفئران أثناء التعامل مع أطوال قصيرة من الشعيرية والمعكرونة غير مطبوخة أو capellini ، على التوالي ، مع كاميرا لتحسين وضعه وجهة نظر الحركات مخلب. بطء تشغيل الفيديو الاقتراح يسمح لتحديد التعديلات forepaw ، على النحو المحدد أي إزالة متميزة واستبدال مخلب ، أو أي عدد من الأرقام ، على قطعة المعكرونة بعد تناول الطعام تبدأ. وبلغ متوسط ​​التعديلات Forepaw للقطعة الواحدة خلال محاكمات في كل دورة الاختبار. اختبار التحليل الكمي المتكررة تصاريح حساسة للتغيرات في البراعة forepaw مع مرور الوقت. موصوفة بروتوكولات للاختبار قبل التعود والممارسة المناولة ، فضلا عن إجراءات لوصف أنماط التعامل مع غير نمطية. لأن الفئران والجرذان أداء اختبارات التعامل مع المعكرونة بشكل مختلف قليلا ، ويسلط الضوء على أنواع محددة في إدارة الخلافات وسجل من هذه الاختبارات. كان كل استخدام الحيوانات وفقا للبروتوكولات المعتمدة من قبل جامعة تكساس في أوستن ، ورعاية الحيوان اللجنة الاستخدام.

Watch this Scientific Journal Video about The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice at JoVE.com

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice

والشعيرية والتعامل مع الاختبارات Capellini من البراعة forepaw الاستفادة من الميل الطبيعي للالقوارض لمعالجة المواد الغذائية باستخدام forepaw ماهرا والحركات أرقام. وأثناء التعامل مع الحيوانات شريط فيديو قصير من خيوط المعكرونة الجافة غير مطبوخة. بطء تشغيل الفيديو الاقتراح يسمح لتقدير حجم التعديلات forepaw.

والشعيرية والتعامل مع الاختبارات Capellini : تدابير كمية بسيطة من وظيفة forepaw حاذق في الجرذان والفئران

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended ...

the-vermicelli-capellini-handling-tests-simple-quantitative-measures

Research

JoVE Journal

Neuroscience

20.8K Views.  University of Texas at Austin. والشعيرية والتعامل مع الاختبارات Capellini من البراعة forepaw الاستفادة من الميل الطبيعي للالقوارض لمعالجة المواد الغذائية باستخدام forepaw ماهرا والحركات أرقام. وأثناء التعامل مع الحيوانات شريط فيديو قصير من خيوط المعكرونة الجافة غير مطبوخة. بطء تشغيل الفيديو الاقتراح...

Video: The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response is a vision-based, reflexive behavior that has been observed in all vertebrates tested. Tadpole eyes are small so the tail flip response was used as alternative measure, which requires a trained technician to record the subtle response. We developed an alternative behavior assay based on the fact that tadpoles prefer to swim on the white side of a tank when placed in a tank with both black and white sides. The assay presented here is an inexpensive, simple alternative that creates a response that is easily measured. The setup consists of a tripod, webcam and nested testing tanks, readily available in most Xenopus laboratories. This article includes a movie showing the behavior of tadpoles, before and after severing the optic nerve. In order to test the function of one eye, we also include representative results of a tadpole in which each eye underwent retinal axotomy on consecutive days. Future studies could develop an automated version of this assay for testing the vision of many tadpoles at once.

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Xenopus laevis tadpoles prefer swimming on the white side of a black/white tank. This behavior is guided by their vision. Based on this behavior, we present a simple assay to test the visual function of tadpoles.

والفحص السلوكية بسيطة لاختبار وظيفة البصرية في<em&gt; القيطم المورق</em

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

طريقة بسيطة وغير مكلفة لتحديد الباردة الحساسية والتكيف في الفئران

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Imaging Serotonergic Fibers in the Mouse Spinal Cord Using the CLARITY/CUBIC Technique

Long descending fibers to the spinal cord are essential for locomotion, pain perception, and other behaviors. The fiber termination pattern in the spinal cord of the majority of these fiber systems have not been thoroughly investigated in any species. Serotonergic fibers, which project to the spinal cord, have been studied in rats and opossums on histological sections and their functional significance has been deduced based on their fiber termination pattern in the spinal cord. With the development of CLARITY and CUBIC techniques, it is possible to investigate this fiber system and its distribution in the spinal cord, which is likely to reveal previously unknown features of serotonergic supraspinal pathways. Here, we provide a detailed protocol for imaging the serotonergic fibers in the mouse spinal cord using the combined CLARITY and CUBIC techniques. The method involves perfusion of a mouse with a hydrogel solution and clarification of the tissue with a combination of clearing reagents. Spinal cord tissue was cleared in just under two weeks, and the subsequent immunofluorescent staining against serotonin was completed in less than ten days. With a multi-photon fluorescent microscope, the tissue was scanned and a 3D image was reconstructed using Osirix software.

Supraspinal projections are important for pain perception and other behaviors, and serotonergic fibers are one of these fiber systems. The present study focused on the application of the combined CLARITY/CUBIC protocol to the mouse spinal cord in order to investigate the termination of these serotonergic fibers.

التصوير ألياف هرمون السيروتونين في الحبل الشوكي الماوس عن طريق وضوح / تقنية مكعب

Long descending fibers to the spinal cord are essential for locomotion, pain perception, and other behaviors. The fiber termination pattern in the spinal ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

وبسيطة خطوة واحدة بروتوكول تشريح لجبل لالجامع إعداد الكبار<em&gt; ذبابة الفاكهة</em&gt; أدمغة

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and Neuropathological Assessments

Experimental autoimmune neuritis (EAN) is a well-appreciated experimental model of autoimmune peripheral demyelinating diseases. EAN disease is induced by immunizing mice with neurogenic peptides to direct an inflammatory attack toward components of the peripheral nervous system (PNS). Recent advances have enabled the induction of EAN in the relatively resistant C57BL/6 mouse line using myelin protein zero (P0)106-125 or P0180-199 peptides delivered in adjuvant combined with the injection of pertussis toxin. The ability to induce EAN in the C57BL/6 strain allows for the use of the numerous genetic tools that exist on this mouse background, and thus allows the sophisticated study of disease pathogenesis and interrogation of the mechanistic action of novel therapeutics in combination with transgenic approaches. In this study, we demonstrate a simple approach to successfully induce EAN using the P0180-199 peptide in C57BL/6 mice. We also outline a protocol for the assessment of functional deficits that occur in this model, accompanied by an array of neuropathological features. Thus, this model is a powerful experimental model to study the pathogenesis of human peripheral demyelinating neuropathies, and to determine the efficacy of potential therapies that aim to promote myelin repair and protect against nerve damage in autoimmune neuritis.

This report outlines a simple approach to successfully induce experimental autoimmune neuritis (EAN) using the myelin protein zero (P0)180-199 peptide in combination with Freund&#39;s complete adjuvant and pertussis toxin. We present a sophisticated paradigm capable of accurately assessing the extent of functional deficits and neuropathology that occur in this EAN.

نهج بسيط للحث على العصب الذاتية التجريبية في الفئران C57BL/6 للتقييمات الفنية ولاثيل

Experimental autoimmune neuritis (EAN) is a well-appreciated experimental model of autoimmune peripheral demyelinating diseases. EAN disease is induced by ...

Intracerebroventricular Treatment with Resiniferatoxin and Pain Tests in Mice

The transient receptor potential vanilloid type 1 (TRPV1), a thermosensitive cation channel, is known to trigger pain in the peripheral nerves. In addition to its peripheral function, its involvement in brain functions has also been suggested. Resiniferatoxin (RTX), an ultrapotent TRPV1 agonist, has been known to induce long-term desensitization of TRPV1, and this desensitization has been an alternative approach for investigating the physiological relevance of TRPV1-expressing cells. Here we describe a protocol for intracerebroventricular (i.c.v.) treatment with RTX in mice. Procedures are described for testing nociception to peripheral TRPV1 stimulation (RTX test) and mechanical stimulation (tail pressure test) then follow. Although the nociceptive responses of mice that had been administered RTX i.c.v. were comparable to those of the control groups, RTX-i.c.v.-administered mice were insensitive to the analgesic effect of acetaminophen, suggesting that i.c.v. RTX treatment can induce supraspinal-selective TRPV1 desensitization. This mouse model can be used as a convenient experimental system for studying the role of TRPV1 in brain/supraspinal function. These techniques can also be applied to studies of the central actions of other drugs.

The transient receptor potential vanilloid type 1 (TRPV1) in the supraspinal region has been suggested to play some roles in the brain function. Described here is a protocol for intracerebroventricular injection of resiniferatoxin for supraspinal TRPV1 desensitization in mice. Procedures for some pain tests are also presented.

العلاج البطيني داخل المنظار مع Resiniferatoxin واختبارات الألم في الفئران

The transient receptor potential vanilloid type 1 (TRPV1), a thermosensitive cation channel, is known to trigger pain in the peripheral nerves. In ...

Using Optical Coherence Tomography and Optokinetic Response As Structural and Functional Visual System Readouts in Mice and Rats

Optical coherence tomography (OCT) is a fast, non-invasive, interferometric technique allowing high-resolution retinal imaging. It is an ideal tool for the investigation of processes of neurodegeneration, neuroprotection and neuro-repair involving the visual system, as these often correlate well with retinal changes. As a functional readout, visually evoked compensatory eye and head movements are commonly used in experimental models involving the visual function. Combining both techniques allows a quantitative in vivo investigation of structure and function, which can be used to investigate the pathological conditions or to evaluate the potential of novel therapeutics. A great benefit of the presented techniques is the possibility to perform longitudinal analyses allowing the investigation of dynamic processes, reducing variability and cuts down the number of animals needed for the experiments. The protocol described aims to provide a manual for acquisition and analysis of high quality retinal scans of mice and rats using a low cost customized holder with an option to deliver inhalational anesthesia. Additionally, the proposed guide is intended as an instructional manual for researchers using optokinetic response (OKR) analysis in rodents, which can be adapted to their specific needs and interests.

A detailed protocol for the assessment of structural and visual readouts in rodents by optical coherence tomography and optokinetic response is presented. The results provide valuable insights for ophthalmologic as well as neurologic research.

استخدام التصوير المقطعي التماسك البصري والاستجابة أوبتوكينيتيك كقراءات النظام المرئي الهيكلية والوظيفية في الفئران والجرذان

Optical coherence tomography (OCT) is a fast, non-invasive, interferometric technique allowing high-resolution retinal imaging. It is an ideal tool for ...

Electrophysiological Investigations of Retinogeniculate and Corticogeniculate Synapse Function

The lateral geniculate nucleus is the first relay station for the visual information. Relay neurons of this thalamic nucleus integrate input from retinal ganglion cells and project it to the visual cortex. In addition, relay neurons receive top-down excitation from the cortex. The two main excitatory inputs to the relay neurons differ in several aspects. Each relay neuron receives input from only a few retinogeniculate synapses, which are large terminals with many release sites. This is reflected by the comparably strong excitation, the relay neurons receive, from retinal ganglion cells. Corticogeniculate synapses, in contrast, are simpler with few release sites and weaker synaptic strength. The two synapses also differ in their synaptic short-term plasticity. Retinogeniculate synapses have a high release probability and consequently display a short-term depression. In contrast, corticogeniculate synapses have a low release probability. Corticogeniculate fibers traverse the reticular thalamic nuclei before entering the lateral geniculate nucleus. The different locations of the reticular thalamic nucleus (rostrally from the lateral geniculate nucleus) and optic tract (ventro-laterally from the lateral geniculate nucleus) allow stimulating corticogeniculate or retinogeniculate synapses separately with extracellular stimulation electrodes. This makes the lateral geniculate nucleus an ideal brain area where two excitatory synapses with very different properties impinging onto the same cell type, can be studied simultaneously. Here, we describe a method to investigate the recording from relay neurons and to perform detailed analysis of the retinogeniculate and corticogeniculate synapse function in acute brain slices. The article contains a step-by-step protocol for the generation of acute brain slices of the lateral geniculate nucleus and steps for recording activity from relay neurons by stimulating the optic tract and corticogeniculate fibers separately.

Here, we present protocols for the preparation of acute brain slices containing the lateral geniculate nucleus and the electrophysiological investigation of retinogeniculate and corticogeniculate synapses function. This protocol provides an efficient way to study synapses with the high-release and low-release probability in the same acute brain slices.

التحقيقات الكهرولوجية من وظائف المشبك وCorticogeniculate

The lateral geniculate nucleus is the first relay station for the visual information. Relay neurons of this thalamic nucleus integrate input from retinal ...

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of sciatic nerve injury rodent models. In this protocol, we describe a novel kinematic analysis method that uses a three-dimensional (3D) motion capture apparatus for functional evaluations using a rat sciatic nerve crush injury model. First, the rat is familiarized with treadmill walking. Markers are then attached to the designated bone landmarks and the rat is made to walk on the treadmill at the desired speed. Meanwhile, the posterior limb movements of the rat are recorded using four cameras. Depending on the software used, marker tracings are created using both automatic and manual modes and the desired data are produced after subtle adjustments. This method of kinematic analysis, which uses a 3D motion capture apparatus, offers numerous advantages, including superior precision and accuracy. Many more parameters can be investigated during the comprehensive functional evaluations. This method has several shortcomings that require consideration: The system is expensive, can be complicated to operate, and may produce data deviations due to skin shifting. Nevertheless, kinematic analysis using a 3D motion capture apparatus is useful for performing functional anterior and posterior limb evaluations. In the future, this method may become increasingly useful for generating accurate assessments of various traumas and diseases.

We introduce a kinematic analysis method that uses a three-dimensional motion capture apparatus containing four cameras and data processing software for performing functional evaluations during fundamental research involving rodent models.

3D تحليل الحركية للتقييم الوظيفي في نموذج الفئران من عرق النسا الأعصاب سحق الإصابة

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the CSF protein composition delivers crucial information in basic neuroscience research as well as neurological diseases. One caveat is that proteins measured in CSF may derive from both intrathecal synthesis and transudation from serum, and protein analysis of CSF can only determine the sum of these two components. To discriminate between protein transudation from the blood and intrathecally produced proteins in animal models as well as in humans, CSF protein profiling measurements using conventional protein analysis tools must include the calculation of the albumin CSF/serum quotient (Qalbumin), a marker of the integrity of the blood-brain interface (BBI), and the protein index (Qprotein/Qalbumin), an estimate of intrathecal protein synthesis. This protocol illustrates the entire procedure, from CSF and blood collection to quotients and indices calculations, for the quantitative measurement of intrathecal protein synthesis and BBI impairment in mouse models of neurological disorders.

Elevated spinal fluid protein levels can either be the result of diffusion of plasma protein across an altered blood-brain barrier or intrathecal synthesis. An optimized testing protocol is presented in this article that helps to discriminate both cases and provides quantitative measurements of intrathecally synthesized proteins.

القياس الكمي للبروتينات التركيبية داخل الفئران

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...