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 border="0" cellpadding="0" cellspacing="0" width="728">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:207px;">Nichrome wire&#160;</td>
			<td style="width:179px;">Sandvik Heating Technology</td>
			<td style="width:137px;">Kanthal RO-800</td>
			<td style="width:205px;">Use for tetrode</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biomedical polyethylene tubing</td>
			<td>A-M Systems</td>
			<td>800700</td>
			<td>Use for tetrode tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lynx-8</td>
			<td>Neuralynx</td>
			<td></td>
			<td>Use for multi-unit recording</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cheetah 32</td>
			<td>Neuralynx</td>
			<td></td>
			<td>Use for multi-unit recording</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">High speed camera</td>
			<td>Basler</td>
			<td>A602f</td>
			<td>Use fir video recording for walking experiments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">High speed camera</td>
			<td>Casio</td>
			<td>EX-FC150&#160;</td>
			<td>Use for video recording for climbing experiments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">WINanalyze</td>
			<td>Winanalyze</td>
			<td>version 1.4 3D</td>
			<td>Use for video tracking&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Matlab</td>
			<td>MathWorks</td>
			<td>MATLAB R2012b</td>
			<td>Use for TTL pulse generation and off-line 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

16.4K 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

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

الفلورسنت وصفها الوراء يسمح لخارج الخلية المستهدفة واحدة وحدة التسجيل من الخلايا العصبية التي تم تحديدها<em&gt; في الجسم الحي</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Electrophysiological Recording in the Brain of Intact Adult Zebrafish

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram recordings. This paper describes how an adult zebrafish can be immobilized, intubated, and used for in vivo electrophysiological experiments, allowing recording of neural activity. Immobilization of the adult requires a mechanism to deliver dissolved oxygen to the gills in lieu of buccal and opercular movement. With our technique, animals are immobilized and perfused with habitat water to fulfill this requirement. A craniotomy is performed under tricaine methanesulfonate (MS-222; tricaine) anesthesia to provide access to the brain. The primary electrode is then positioned within the craniotomy window to record extracellular brain activity. Through the use of a multitube perfusion system, a variety of pharmacological compounds can be administered to the adult fish and any alterations in the neural activity can be observed. The methodology not only allows for observations to be made regarding changes in neurological activity, but it also allows for comparisons to be made between larval and adult zebrafish. This gives researchers the ability to identify the alterations in neurological activity due to the introduction of various compounds at different life stages.

This paper describes how an adult zebrafish can be immobilized, intubated, and used for in vivo electrophysiological experiments to allow recordings and manipulation of neural activity in an intact animal.

تسجيل الكهربية في الدماغ سليمة من اسماك الزرد الكبار

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Recording Single Neurons&#039; Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

تسجيل إمكانات العمل احدة الخلايا العصبية &#39;من التنقل بحرية في جميع أنحاء الحمام ثلاث مراحل التعلم

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

تسجيل إيغ في التحرك بحرية الفئران حديثي الولادة باستخدام طريقة رواية

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful analysis of synaptic transmission modulation when performing field potential or intracellular recordings. This manuscript describes methodological aspects that might be helpful in improving experimental conditions for the maintenance of acute brain slices and for recording field excitatory postsynaptic potentials in a commercially available submersion chamber with an outflow-carbogenation unit. The outflow-carbogenation helps to stabilize the oxygen level in experiments that rely on the recycling of a small buffer reservoir to enhance the cost-efficiency of drug experiments. In addition, the manuscript presents representative experiments that examine the effects of different carbogenation modes and stimulation paradigms on the activity-dependent synaptic plasticity of synaptic transmission.

This protocol describes the stabilization of the oxygen level in a small volume of recycled buffer and methodological aspects of recording activity-dependent synaptic plasticity in submerged acute hippocampal slices.

تسجيل اللدونة متشابك في شرائح هيبوكامبال الحاد تحتفظ في صغيرة الحجم-إعادة التدوير، ونضح-، وغمر من نوع نظام الدائرة

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...

Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems neuroscience. One key component of this technique consists of the precise placement of multiple electrodes into desired brain regions and the maintenance of their stability. Here, we describe a protocol for the construction of a 3D-printable hyperdrive, which includes eighteen independently adjustable tetrodes, and is specifically designed for in vivo extracellular neural recording in freely behaving rats. The tetrodes attached to the microdrives can either be individually advanced into multiple brain regions along the track, or can be used to place an array of electrodes into a smaller area. The multiple tetrodes allow for simultaneous examination of action potentials from dozens of individual neurons, as well as local field potentials from populations of neurons in the brain during active behavior. In addition, the design provides for simpler 3D drafting software that can easily be modified for differing experimental needs.

We present the construction of a 3D-printable hyperdrive with eighteen independently adjustable tetrodes. The hyperdrive is designed to record brain activity in freely behaving rats over a period of several weeks.

بناء Hyperdrive تيترودي متعددة محسنة لتسجيل العصبية على نطاق واسع في التصرف الفئران

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

نظام Microdrive الهجين مع مسبار أوبتو السيليكون القابل ة للاسترداد وTetrode لتسجيل الكثافة العالية في الموقع المزدوج في الفئران المتحركة بحرية

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and smaller functional subgroups, it becomes paramount to record from projections and/or genetically-defined subpopulations of neurons. Fiber photometry is an accessible and powerful approach that can overcome these challenges. By combining optical and genetic methodologies, neural activity can be measured in deep brain structures by expressing genetically-encoded calcium indicators, which translate neural activity into an optical signal that can be easily measured. The current protocol details the components of a multi-fiber photometry system, how to access deep brain structures to deliver and collect light, a method to account for motion artifacts, and how to process and analyze fluorescent signals. The protocol details experimental considerations when performing single and dual color imaging, from either single or multiple implanted optic fibers.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

القياس الضوئي متعدد ألياف لتسجيل النشاط العصبي في الكائنات المتحركة بحريه

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...

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 ...

Recording Brain Activity with Ear-Electroencephalography

The c-grid (ear-electroencephalography, sold under the name cEEGrid) is an unobtrusive and comfortable electrode array that can be used for investigating brain activity after affixing around the ear. The c-grid is suitable for use outside of the laboratory for long durations, even for the whole day. Various cognitive processes can be studied using these grids, as shown by previous research, including research beyond the lab. To record high-quality ear-EEG data, careful preparation is necessary. In this protocol, we explain the steps needed for its successful implementation. First, how to test the functionality of the grid prior to a recording is shown. Second, a description is provided on how to prepare the participant and how to fit the c-grid, which is the most important step for recording high-quality data. Third, an outline is provided on how to connect the grids to an amplifier and how to check the signal quality. In this protocol, we list best practice recommendations and tips that make c-grid recordings successful. If researchers follow this protocol, they are comprehensively equipped for experimenting with the c-grid both in and beyond the lab.

Presented here is the procedure for using the c-grid (ear-electroencephalography, sold under the name cEEGrid) for recording brain activity in and beyond the lab for extended durations. This protocol describes how to set up these arrays and how to record brain activity using them.

تسجيل نشاط الدماغ باستخدام تخطيط كهربية الدماغ للأذن

The c-grid (ear-electroencephalography, sold under the name cEEGrid) is an unobtrusive and comfortable electrode array that can be used for investigating ...