المؤلفين أشكر نيك كاثمان للاقتراحات ومساعدة في التحضير للمخطوطة. وقد تم تطوير هذه التقنية بالتعاون مع العمل بدعم من AFOSR تحت منحة FA9550-10-1-0054 والمؤسسة الوطنية للعلوم تحت المنحة رقم IOS-1120305 لRER.

<ol>
	<li>Strauss, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+central+complex+and+the+genetic+dissection+of+locomotor+behaviour.">The central complex and the genetic dissection of locomotor behaviour.</a> Curr. Opin. Neurobiol. 12, 633-638 (2002).</li><li>Pick, S., Strauss, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Goal-driven+behavioral+adaptations+in+gap-climbing+Drosophila.">Goal-driven behavioral adaptations in gap-climbing Drosophila.</a> Curr. Biol. 15, 1473-1478 (2005).</li><li>Triphan, T., Poeck, B., Neuser, K., Strauss, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+targeting+of+motor+actions+in+climbing+Drosophila.">Visual targeting of motor actions in climbing Drosophila.</a> Curr. Biol. 20, 663-668 (2010).</li><li>Heinze, S., Gotthardt, S., Homberg, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transformation+of+polarized+light+information+in+the+central+complex+of+the+locust.">Transformation of polarized light information in the central complex of the locust.</a> J. Neuorosci. 29, 11783-11793 (2009).</li><li>Heinze, S., Homberg, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maplike+representation+of+celestial+E-vector+orientations+in+the+brain+of+an+insect.">Maplike representation of celestial E-vector orientations in the brain of an insect.</a> Science. 315, 995-997 (2007).</li><li>Heinze, S., Homberg, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroarchitecture+of+the+central+complex+of+the+desert+locust:+Intrinsic+and+columnar+neurons.">Neuroarchitecture of the central complex of the desert locust: Intrinsic and columnar neurons.</a> J. Comp. Neurol. 511, 454-478 (2008).</li><li>Heinze, S., Homberg, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Linking+the+input+to+the+output:+new+sets+of+neurons+complement+the+polarization+vision+network+in+the+locust+central+complex.">Linking the input to the output: new sets of neurons complement the polarization vision network in the locust central complex.</a> J. Neurosci. 29, 4911-4921 (2009).</li><li>Heinze, S., Reppert, 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=Sun+compass+integration+of+skylight+cues+in+migratory+monarch+butterflies.">Sun compass integration of skylight cues in migratory monarch butterflies.</a> Neuron. 69, 345-358 (2011).</li><li>Brill, M. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parallel+processing+via+a+dual+olfactory+pathway+in+the+honeybee.">Parallel processing via a dual olfactory pathway in the honeybee.</a> J Neurosci. 33, 2443-2456 (2013).</li><li>Guo, P., Ritzmann, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+activity+in+the+central+complex+of+the+cockroach+brain+is+linked+to+turning+behaviors.">Neural activity in the central complex of the cockroach brain is linked to turning behaviors.</a> J. Exp. Biol. 216, 992-1002 (2013).</li><li>Ritzmann, R. E., Ridgel, A. L., Pollack, 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=Multi-unit+recording+of+antennal+mechanosensitive+units+in+the+central+complex+of+the+cockroach,+Blaberus+discoidalis.">Multi-unit recording of antennal mechanosensitive units in the central complex of the cockroach, Blaberus discoidalis.</a> J. Comp. Physiol. A. 194, 341-360 (2008).</li><li>Buzs&#225;ki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+recording+of+neuronal+ensembles.">Large-scale recording of neuronal ensembles.</a> Nature Neurosci. 7.5, 446-445 (2004).</li><li>Huston, S. J., Jayaraman, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studying+sensorimotor+integration+in+insects.">Studying sensorimotor integration in insects.</a> Curr. Opin. Neurobiol. 21, 527-534 (2011).</li><li>Bender, J. A., Pollack, A. J., Ritzmann, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+activity+in+the+central+complex+of+the+insect+brain+is+linked+to+locomotor+changes.">Neural activity in the central complex of the insect brain is linked to locomotor changes.</a> Curr. Biol. 20, 921-926 (2010).</li><li>Harrison, R. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wireless+Neural/EMG+telemetry+systems+for+small+freely+moving+animals.">Wireless Neural/EMG telemetry systems for small freely moving animals.</a> IEEE. 5, 103-111 (2011).</li><li>Straw, A. D., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motmot,+an+open-source+toolkit+for+realtime+video+acquisition+and+analysis.">Motmot, an open-source toolkit for realtime video acquisition and analysis.</a> Source Code Biol. Med. 4, 5 (2009).</li><li>Tyrer, N. M., Shaw, M. K., Altman, J. S., Strausfeld, N. J., Miller, 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=.">.</a> Neuroanatomical Techniques. Insect Nervous System. , (1980).</li><li>Daly, K., Wright, G., Smith, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+features+of+odorants+systematically+influence+slow+temporal+responses+across+clusters+of+coordinated+antennal+lobe+units+in+the+moth,+Manduca+sexta.">Molecular features of odorants systematically influence slow temporal responses across clusters of coordinated antennal lobe units in the moth, Manduca sexta.</a> J. Neurophsyiol. 92, 236-254 (2004).</li><li>Branson, K., Robie, A., Bender, J., Perona, P., Dickinson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+ethomics+in+large+groups+of+Drosophila.">High-throughput ethomics in large groups of Drosophila.</a> Nat Methods. 6, 451-457 (2009).</li></ol>

الكتاب تعلن أي تضارب في المصالح.

في حين قدمت الدراسات السابقة الكهربية على CC أو مناطق أخرى من الدماغ الحشرات لنا نظرة ثاقبة لسيطرة مركزية من السلوك، وأجريت معظمها في الاستعدادات إما مقيدة أو تلك المربوطة 9،11 10،14. ونتيجة لذلك، فإن الحيوان التجربة الحسية والحالة الفسيولوجية يمكن أن تكون مختلفة ...

توضح هذه المقالة نظام ناجح للتسجيل من الخلايا العصبية داخل المجمع المركزي (CC) من صرصور، discoidalis Blaberus، كما يمشي الحشرة في الساحة ويتعامل مع الأشياء التي تسبب ليستدير، نفق تحت، أو تسلق العقبات. كما يمكن توصيل الأسلاك إلى مشجعا لاستحضار النشاط في neuropil المحيطة بها مع التغيرات السلوكية يترتب على ذلك. على مدى العقد الماضي قد وجهت اهتماما كبيرا في الأدوار التي تقوم بها مناطق الدماغ المختلفة في السيطرة على سلوك الحشرات. وقد وجهت الكثير من هذا التركيز نحو neuropils الدماغ خط الوسط التي يشار إليها مجتمعة ب مجمع المركزية (CC). وقد تم إحراز تقدم نتيجة لأصناف واسعة من التقنيات التي تستهدف أسئلة حول دور CC في السلوك. وتتراوح تلك التقنيات من التلاعب العصبية الوراثية، في المقام الأول في ذبابة الفاكهة، إلى جانب behaviتحليل عن طريق الفم 1-3، إلى تقنيات الكهربية التي ترصد النشاط العصبي داخل CC ومحاولة لربط هذا النشاط إلى المعايير ذات الصلة سلوكيا. وتشمل تقنيات الكهربية داخل الخلايا من الخلايا العصبية تسجيل الفردية المحددة 4-9 وتسجيل خارج الخلية، وغالبا مع قناة تحقيقات متعددة 10،11. هذه التقنيات هما مجانية. تسجيل داخل الخلايا مع أقطاب حادة أو التصحيح خلية كاملة يوفر بيانات مفصلة جدا على الخلايا العصبية التي تم تحديدها، ولكن يقتصر على واحد أو اثنين من الخلايا في آن واحد، يتطلب حركة محدودة أو معدومة، ويمكن الحفاظ لفترات قصيرة نسبيا من الزمن. تسجيلات خارج الخلية يمكن بسهولة انشاء، لا تتطلب ضبط النفس، ويمكن الحفاظ على لساعات. مع tetrodes متعددة القنوات والقطع العنقودية، ويمكن تحليل أعداد كبيرة نسبيا من الخلايا العصبية في وقت واحد 9،12. بينما patc خلية كاملةح وقد استخدمت بنجاح في الحشرات المربوطة 13، ونحن نشعر أن هناك أيضا حاجة إلى التقنيات التي تسمح لنا لتسجيل النشاط العصبي في الدماغ لفترات طويلة من الوقت في التصرف بحرية الحشرات لأنها تعامل مع العوائق التي تحول دون حركة إلى الأمام. الحاجة لتسجيل مثل هذه التحركات الحشرات ويرتد صعودا وهبوطا دفعنا نحو أساليب تسجيل خارج الخلية. لقد كان لدينا تسجيل نجاح جيدة في الأعمال التحضيرية ضبط النفس مع المتاحة تجاريا تحقيقات السيليكون 11 16 قنوات، ولكن صغر حجم كبير حتى الصراصير يعني أن تحقيقات يجب أن يتم تنظيمها قبالة الجسم. التي، إلى جانب حساسية للأسنان التحقيق، جعلها غير مناسبة لإعداد المشي الحر. في اثنين من المشاريع السابقة، استخدمنا حزم من الأسلاك غرامة تشكيل tetrode لإنجاز تسجيل خصائص مماثلة ولكن في ترتيب أكثر قوة. سمحت هذه الباقات tetrode لنا لتسجيل من الصراصير المربوطة ود تتصل النشاط حدة CC للتغيرات في سرعة المشي 14 وتحول السلوك الناتج من الاتصال antennal مع قضيب 10. كما مفيدة كما كانت هذه الاستعدادات المربوطة وستظل، فإنها تفعل تقديم بعض القيود. أولا، السلوكيات التي يمكن أن تؤدي الحشرة محدودة لطائرة واحدة. وهذا هو، ونحن يمكن أن تثير بسهولة التغيرات في سرعة المشي أو تحول، ولكن كانت الإجراءات التسلق والأنفاق غير ممكن، على الأقل مع حبل ترتيب نموذجي. الثاني، الاستعدادات المربوطة لدينا هي &quot;حلقة مفتوحة&quot;. وهذا يعني أنها لا تسمح للحركة العادية المتعلقة ردود الفعل على النظام. وهكذا، كما تحولت صرصور على حبل لدينا، لم يتم تغييرها العالم البصرية وفقا لذلك. فمن الممكن لبناء نظم حلقة مغلقة حبل لتقديم هذا النوع من ردود الفعل. ومع ذلك، فهي محدودة بسبب تعقيد البرمجة والأجهزة البيئة البصرية المحاكاة. Neverthelesق، شعرنا أننا يمكن أن تحسن على طرقنا تسجيل المربوطة القائمة من خلال تسجيل من الحيوان لأنه مشى بحرية في الساحة أو المسار واجه الأشياء كما تفعل في محيطها الطبيعي. على الرغم من أن الأنظمة اللاسلكية لتسجيل نشاط المخ 15 سيكون مثاليا، النظم الحالية لديها قيود في عدد من القنوات تسجيل، والوقت من الحصول على البيانات، وعمر البطارية والوزن. ونحن، بالتالي، اختارت في محاولة للتكيف لدينا نظام تسجيل المربوطة لاستخدامها في التحرك بحرية الاستعدادات. كما تصبح أفضل الأنظمة اللاسلكية المتاحة، وهذه التقنية يمكن تكييفه بسهولة لمثل هذه الأجهزة. نظام الموضح في هذه المقالة هو خفيفة الوزن، ويعمل بشكل جيد جدا، ويبدو أن يكون لها تأثير ضار يذكر على سلوك صرصور و. مع رخيصة كاميرا عالية السرعة والكتلة البرمجيات القطع، والنشاط في الخلايا العصبية في الدماغ الفردية يمكن أن تكون ذات صلة الحركة. نحن هنا وصف preparأوجه من الأسلاك tetrode وترسيخها على الدماغ إلى الحشرة وكذلك تقنيات التسجيل للنشاط والحركة الكهربائية وكيف أن هذه البيانات لا يمكن أن تتحقق معا لتحليلها لاحقا.

<table><tbody><tr><td>Nichrome wire&amp;nbsp;</td><td>Sandvik Heating Technology</td><td>Kanthal RO-800</td><td>Use for tetrode</td></tr><tr><td>Biomedical polyethylene tubing</td><td>A-M Systems</td><td>800700</td><td>Use for tetrode tubing</td></tr><tr><td>Lynx-8</td><td>Neuralynx</td><td></td><td>Use for multiunit recording</td></tr><tr><td>Cheetah 32</td><td>Neuralynx</td><td></td><td>Use for multiunit recording</td></tr><tr><td>High speed camera</td><td>Basler</td><td>A602f</td><td>Use for video recording for walking experiments</td></tr><tr><td>High speed camera</td><td>Casio</td><td>EX-FC150</td><td>Use for video recording for climbing experiments</td></tr><tr><td>WINanalyze</td><td>Winanalyze</td><td>version 1.4 3D</td><td>Use for video tracking&amp;nbsp;</td></tr><tr><td>MATLAB</td><td>MathWorks</td><td>MATLAB R2012b</td><td>Use for TTL pulse generation and offline data analysis</td></tr></tbody></table>

extracellular wire tetrode recording in brain of freely walking insects

يتطلب اهتماما متزايدا في دور نشاط الدماغ في التحكم في المحركات الحشرات أن نكون قادرين على مراقبة النشاط العصبي في حين الحشرات أداء السلوك الطبيعي. وضعنا سابقا تقنية لزرع الأسلاك tetrode في المجمع المركزي للأدمغة الصراصير التي سمحت لنا لتسجيل نشاط الخلايا العصبية متعددة في نفس الوقت من حين تحولت صرصور المربوطة أو تغير سرعة المشي. في حين أن تقدما كبيرا، والأعمال التحضيرية المربوطة توفير الوصول إلى سلوكيات محدودة وغالبا ما تفتقر عمليات التقييم التي تحدث في التحرك بحرية الحيوانات. نقدم الآن نسخة معدلة من هذا الأسلوب الذي يسمح لنا لتسجيل من المجمع المركزي من التحرك بحرية الصراصير وهم يسيرون في الساحة والتعامل مع الحواجز من خلال تحويل، التسلق أو نفق. إلى جانب سرعة عالية الفيديو والقطع العنقودية، يمكننا الآن تتصل نشاط الدماغ لمعاملات شتى من حركة الحشرات يتصرف بحرية.

سجلنا النشاط العصبي من 50 وحدة من 27 CC في الاستعدادات لاجراء تجارب على الأقدام. ل15 من تلك التحضيرات (23 وحدة)، وأجريت أيضا تجارب التسلق. تتم تسمية الوحدات الفردية وفقا لأرقام إعداد وحدة (مثل وحدة 1-2 يشير إعداد 1، 2 وحدة). وتظهر لقطا?...

Watch this Scientific Journal Video about Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects at JoVE.com

Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

وضعنا سابقا تقنية لزرع الأسلاك tetrode في المجمع المركزي للأدمغة صرصور الذي يسمح لنا لرصد النشاط في وحدات فردية من الصراصير المربوطة. نحن هنا تقديم نسخة معدلة من هذا الأسلوب الذي يسمح لنا أيضا تسجيل نشاط المخ في تحريك الحشرات بحرية.

خارج الخلية سلك Tetrode تسجيل في الدماغ من المشي بحرية الحشرات

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform ...

extracellular-wire-tetrode-recording-brain-freely-walking

Research

JoVE Journal

Neuroscience

16.5K Views.  Case Western Reserve University.  وضعنا سابقا تقنية لزرع الأسلاك tetrode في المجمع المركزي للأدمغة صرصور الذي يسمح لنا لرصد النشاط في وحدات فردية من الصراصير المربوطة. نحن هنا تقديم نسخة معدلة من هذا الأسلوب الذي يسمح لنا أيضا تسجيل نشاط المخ في تحريك الحشرات بحرية. 

Video: Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

Early Metamorphic Insertion Technology for Insect Flight Behavior Monitoring

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on insects during their metamorphic development. Here, the implants are fused within the structure and function of the neuromuscular system as a result of metamorphic tissue remaking. The implants emerge with the insect where the development of tissue around the electronics during pupal development results in a bioelectrically and biomechanically enhanced tissue interface. This relatively more reliable and stable interface would be beneficial for many researchers exploring the neural basis of the insect locomotion with alleviated traumatic effects caused during adult stage insertions. In this article, we implant our electrodes into the indirect flight muscles of Manduca sexta. Located in the dorsal-thorax, these main flight powering dorsoventral and dorsolongitudinal muscles actuate the wings and supply the mechanical power for up and down strokes. Relative contraction of these two muscle groups has been under investigation to explore how the yaw maneuver is neurophysiologically coordinated. To characterize the flight dynamics, insects are often tethered with wires and their flight is recorded with digital cameras. We also developed a novel way to tether Manduca sexta on a magnetically levitating frame where the insect is connected to a commercially available wireless neural amplifier. This set up can be used to limit the degree of freedom to yawing &#8220;only&#8221; while transmitting the related electromyography signals from dorsoventral and dorsolongitudinal muscle groups.

We present a novel surgical procedure to implant electrodes in Manduca sexta during its early metamorphic stages. This technique allows mechanically stable and electrically reliable coupling with the neuromuscular tissue to study flight neurophysiology dynamics. We also present a novel magnetic levitation platform for tethered studies of insect yaw.

المتحولة الإدراج المبكر لتكنولوجيا الطيران سلوك الحشرات الرصد

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on ...

Behavior

Flat-floored Air-lifted Platform: A New Method for Combining Behavior with Microscopy or Electrophysiology on Awake Freely Moving Rodents

It is widely acknowledged that the use of general anesthetics can undermine the relevance of electrophysiological or microscopical data obtained from a living animal&#8217;s brain. Moreover, the lengthy recovery from anesthesia limits the frequency of repeated recording/imaging episodes in longitudinal studies. Hence, new methods that would allow stable recordings from non-anesthetized behaving mice are expected to advance the fields of cellular and cognitive neurosciences. Existing solutions range from mere physical restraint to more sophisticated approaches, such as linear and spherical treadmills used in combination with computer-generated virtual reality. Here, a novel method is described where a head-fixed mouse can move around an air-lifted mobile homecage and explore its environment under stress-free conditions. This method allows researchers to perform behavioral tests (e.g., learning, habituation or novel object recognition) simultaneously with two-photon microscopic imaging and/or patch-clamp recordings, all combined in a single experiment. This video-article describes the use of the awake animal head fixation device (mobile homecage), demonstrates the procedures of animal habituation, and exemplifies a number of possible applications of the method.

This method creates a tangible, familiar environment for the mouse to navigate and explore during microscopic imaging or single-cell electrophysiological recordings, which require firm fixation of the animal&#8217;s head.

مسطحة طوابق منصة رفع الهواء و: طريقة جديدة للجمع بين السلوك مع الميكروسكوب أو الكهربية على استيقظ تتحرك بحرية القوارض

It is widely acknowledged that the use of general anesthetics can undermine the relevance of electrophysiological or microscopical data obtained from a ...

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" (or "startle") response during the adult stage is mediated by the multi-component neuronal circuit called the Giant Fiber System (GFS). The comparative large size of the neurons, their distinctive morphology and simple connectivity make the GFS an attractive model system for studying neuronal circuitry. The GFS pathway is composed of two bilaterally symmetrical Giant Fiber (GF) interneurons whose axons descend from the brain along the midline into the thoracic ganglion via the cervical connective. In the mesothoracic neuromere (T2) of the ventral ganglia the GFs form electro-chemical synapses with 1) the large medial dendrite of the ipsilateral motorneuron (TTMn) which drives the tergotrochanteral muscle (TTM), the main extensor for the mesothoracic femur/leg, and 2) the contralateral peripherally synapsing interneuron (PSI) which in turn forms chemical (cholinergic) synapses with the motorneurons (DLMns) of the dorsal longitudinal muscles (DLMs), the wing depressors. The neuronal pathway(s) to the dorsovental muscles (DVMs), the wing elevators, has not yet been worked out (the DLMs and DVMs are known jointly as indirect flight muscles - they are not attached directly to the wings, but rather move the wings indirectly by distorting the nearby thoracic cuticle) (King and Wyman, 1980; Allen et al., 2006). The di-synaptic activation of the DLMs (via PSI) causes a small but important delay in the timing of the contraction of these muscles relative to the monosynaptic activation of TTM (~0.5 ms) allowing the TTMs to first extend the femur and propel the fly off the ground. The TTMs simultaneously stretch-activate the DLMs which in turn mutually stretch-activate the DVMs for the duration of the flight. The GF pathway can be activated either indirectly by applying a sensory (e.g."air-puff" or "lights-off") stimulus, or directly by a supra-threshold electrical stimulus to the brain (described here). In both cases, an action potential reaches the TTMs and DLMs solely via the GFs, PSIs, and TTM/DLM motoneurons, although the TTMns and DLMns do have other, as yet unidentified, sensory inputs. Measuring "latency response" (the time between the stimulation and muscle depolarization) and the "following to high frequency stimulation" (the number of successful responses to a certain number of high frequency stimuli) provides a way to reproducibly and quantitatively assess the functional status of the GFS components, including both central synapses (GF-TTMn, GF-PSI, PSI-DLMn) and the chemical (glutamatergic) neuromuscular junctions (TTMn-TTM and DLMn-DLM). It has been used to identify genes involved in central synapse formation and to assess CNS function.

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

The Giant Fiber System is a simple neuronal circuit of adult Drosophila melanogaster containing the largest neurons in the fly. We describe the protocol for monitoring synaptic transmission through this pathway by recording post synaptic potentials in dorsal longitudinal (DLM) and tergotrochanteral (TTM) muscles following direct stimulation of the Giant Fiber interneurons.

تسجيلات الكهربية من الألياف المسار عملاق د. melanogaster</em

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" ...

Studying the Neural Basis of Adaptive Locomotor Behavior in Insects

Studying the neural basis of walking behavior, one often faces the problem that it is hard to separate the neuronally produced stepping output from those leg movements that result from passive forces and interactions with other legs through the common contact with the substrate. If we want to understand, which part of a given movement is produced by nervous system motor output, kinematic analysis of stepping movements, therefore, needs to be complemented with electrophysiological recordings of motor activity. The recording of neuronal or muscular activity in a behaving animal is often limited by the electrophysiological equipment which can constrain the animal in its ability to move with as many degrees of freedom as possible. This can either be avoided by using implantable electrodes and then having the animal move on a long tether (i.e. Clarac et al., 1987; Duch &amp; Pfl&uuml;ger, 1995; B&ouml;hm et al., 1997; Gruhn &amp; Rathmayer, 2002) or by transmitting the data using telemetric devices (Kutsch et al, 1993; Fischer et al., 1996; Tsuchida et al. 2004; Hama et al., 2007; Wang et al., 2008). Both of these elegant methods, which are successfully used in larger arthropods, often prove difficult to apply in smaller walking insects which either easily get entangled in the long tether or are hindered by the weight of the telemetric device and its batteries. In addition, in all these cases, it is still impossible to distinguish between the purely neuronal basis of locomotion and the effects exerted by mechanical coupling between the walking legs through the substrate. One solution for this problem is to conduct the experiments in a tethered animal that is free to walk in place and that is locally suspended, for example over a slippery surface, which effectively removes most ground contact mechanics. This has been used to study escape responses (Camhi and Nolen, 1981; Camhi and Levy, 1988), turning (Tryba and Ritzman, 2000a,b; Gruhn et al., 2009a), backward walking (Graham and Epstein, 1985) or changes in velocity (Gruhn et al., 2009b) and it allows the experimenter easily to combine intra- and extracellular physiology with kinematic analyses (Gruhn et al., 2006).
We use a slippery surface setup to investigate the timing of leg muscles in the behaving stick insect with respect to touch-down and lift-off under different behavioral paradigms such as straight forward and curved walking in intact and reduced preparations.

We describe a method to record motor activity, timed to the electrically recorded tarsal contact signal in a tethered insect, walking on a slippery surface. This is used to study the neural basis of adaptive behavior under reduced influence of mechanical interaction between legs through the substrate.

دراسة الأساس العصبي للسلوك الحركي التكيفية في الحشرات

Studying the neural basis of walking behavior, one often faces the problem that it is hard to separate the neuronally produced stepping output from those ...

Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of behaviors from the spiking activity of its neurons. One of the most effective methods to monitor spiking activity from a large number of neurons in multiple local neuronal circuits simultaneously is by using silicon electrode arrays1-3.
Action potentials produce large transmembrane voltage changes in the vicinity of cell somata. These output signals can be measured by placing a conductor in close proximity of a neuron. If there are many active (spiking) neurons in the vicinity of the tip, the electrode records combined signal from all of them, where contribution of a single neuron is weighted by its 'electrical distance'. Silicon probes are ideal recording electrodes to monitor multiple neurons because of a large number of recording sites (+64) and a small volume. Furthermore, multiple sites can be arranged over a distance of millimeters, thus allowing for the simultaneous recordings of neuronal activity in the various cortical layers or in multiple cortical columns (Fig. 1). Importantly, the geometrically precise distribution of the recording sites also allows for the determination of the spatial relationship of the isolated single neurons4. Here, we describe an acute, large-scale neuronal recording from the left and right forelimb somatosensory cortex simultaneously in an anesthetized rat with silicon probes (Fig. 2).

Extracellular recordings of neuronal activity using silicon probes in the anesthetized rat will be described. This technique allows information to be obtained across multiple brain areas from more than 100 neurons simultaneously. It provides information with single cell resolution about neuronal ensembles dynamics in multiple local circuits.

تسجيل على نطاق واسع اطقم متعلق بالخلايا العصبية مع مجسات للسيليكون في الجرذ خدر

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

جهاز لاسلكي، واجهة ثنائية الاتجاه للتسجيل في فيفو وحفز النشاط العصبي في التصرف بحرية الفئران

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

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

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

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

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

طرق تسجيل متعددة الوحدات لوصف النشاط العصبي في الجراد (<em&gt; الجراد الصحراوي أمريكانا</em&gt;) الدوائر الشم

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

Neural Circuit Recording from an Intact Cockroach Nervous System

The cockroach ventral nerve cord preparation is a tractable system for neuroethology experiments, neural network modeling, and testing the physiological effects of insecticides. This article describes the scope of cockroach sensory modalities that can be used to assay how an insect nervous system responds to environmental perturbations. Emphasis here is on the escape behavior mediated by cerci to giant fiber transmission in Periplaneta americana. This in situ preparation requires only moderate dissecting skill and electrophysiological expertise to generate reproducible recordings of neuronal activity. Peptides or other chemical reagents can then be applied directly to the nervous system in solution with the physiological saline. Insecticides could also be administered prior to dissection and the escape circuit can serve as a proxy for the excitable state of the central nervous system. In this context the assays described herein would also be useful to researchers interested in limb regeneration and the evolution of nervous system development for which P. americana is an established model organism.

This article describes the cockroach ventral nerve cord dissection and extracellular recordings from the cercal nerve and connectives. Evoked responses are generated by electrical stimulation of the cercal nerve or direct mechanical stimulation of the cerci.

العصبية الدائرة تسجيل من الصرصور الجهاز العصبي سليمة

The cockroach ventral nerve cord preparation is a tractable system for neuroethology experiments, neural network modeling, and testing the physiological ...

Simultaneous Long-term Recordings at Two Neuronal Processing Stages in Behaving Honeybees

In both mammals and insects neuronal information is processed in different higher and lower order brain centers. These centers are coupled via convergent and divergent anatomical connections including feed forward and feedback wiring. Furthermore, information of the same origin is partially sent via parallel pathways to different and sometimes into the same brain areas. To understand the evolutionary benefits as well as the computational advantages of these wiring strategies and especially their temporal dependencies on each other, it is necessary to have simultaneous access to single neurons of different tracts or neuropiles in the same preparation at high temporal resolution. Here we concentrate on honeybees by demonstrating a unique extracellular long term access to record multi unit activity at two subsequent neuropiles1, the antennal lobe (AL), the first olfactory processing stage and the mushroom body (MB), a higher order integration center involved in learning and memory formation, or two parallel neuronal tracts2 connecting the AL with the MB.&#160;The latter was chosen as an example and will be described in full. In the supporting video the construction and permanent insertion of flexible multi&#160;channel wire electrodes is demonstrated. Pairwise differential amplification of the micro&#160;wire electrode channels drastically reduces the noise and verifies that the source of the signal is closely related to the position of the electrode tip. The mechanical flexibility of the used wire electrodes allows stable invasive long&#160;term recordings over many hours up to days, which is a clear advantage compared to conventional extra&#160;and intracellular in vivo recording techniques.

Simultaneous extracellular long term recordings from two different brain neuropiles or two different anatomical tracts were established in honeybees. These recordings allow the investigation of temporal aspects of neuronal processing across different brain areas at the single neuron as well as at the ensemble level in a behaving animal.

تسجيلات في وقت واحد على المدى الطويل في مرحلتين العصبية في معالجة يتصرفون نحل العسل

In both mammals and insects neuronal information is processed in different higher and lower order brain centers. These centers are coupled via convergent ...

Wireless Electrophysiological Recording of Neurons by Movable Tetrodes in Freely Swimming Fish

The neural mechanisms governing fish behavior remain mostly unknown, although fish constitute the majority of all vertebrates. The ability to record brain activity from freely moving fish would advance research on the neural basis of fish behavior considerably. Moreover, precise control of the recording location in the brain is critical to studying coordinated neural activity across regions in fish brain. Here, we present a technique that records wirelessly from the brain of freely swimming fish while controlling for the depth of the recording location. The system is based on a neural logger associated with a novel water-compatible implant that can adjust the recording location by microdrive-controlled tetrodes. The capabilities of the system are illustrated through recordings from the telencephalon of goldfish.

A novel wireless technique for recording extracellular neural signals from the brain of freely swimming goldfish is presented. The recording device is composed of two tetrodes, a microdrive, a neural data logger, and a waterproof case. All parts are custom-made except for the data logger and its connector.

لاسلكيه الكهربائية تسجيل الخلايا العصبية من قبل Tetrodes المنقولة في بحريه السباحة الأسماك

The neural mechanisms governing fish behavior remain mostly unknown, although fish constitute the majority of all vertebrates. The ability to record brain ...

خارج الخلية سلك Tetrode تسجيل في الدماغ من المشي بحرية الحشرات

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