نحن نشكر مايكل Dübbert ، أوليفر هوفمان ، هانز Scharstein ، سايدو يان آن وWosnitza لتقديم المساعدة التقنية الممتازة. وأيد هذه الدراسة من قبل منحة DFG بو 857 / 8،10 إلى AB

<ol>
	<li>B&#228;ssler, U., Wegener, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+output+of+the+denervated+thoracic+ventral+nerve+cord+in+the+stick+insect+Carausius+morosus.">Motor output of the denervated thoracic ventral nerve cord in the stick insect Carausius morosus.</a> J. Exp. Biol. 105, 127-147 (1983).</li><li>B&#246;hm, H., Messai, E., Heinzel, H. -. 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M., Levy, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organization+of+a+complex+movement:+fixed+and+variable+components+of+the+cockroach+escape+behavior.">Organization of a complex movement: fixed and variable components of the cockroach escape behavior.</a> J Comp Physiol A. 163, 317-328 (1988).</li><li>Camhi, J. M., Nolen, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+of+the+escape+system+of+cockroaches+during+walking.">Properties of the escape system of cockroaches during walking.</a> J Comp Physiol A. 142, 339-346 (1981).</li><li>Clarac, F., Libersat, F., Pfl&#252;ger, H. J., Rathmayer, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+pattern+analysis+in+the+shore+crab+(Carcinus+maenas)+walking+freely+in+water+and+on+land.">Motor pattern analysis in the shore crab (Carcinus maenas) walking freely in water and on land.</a> J Exp Biol. 133, 395-414 (1987).</li><li>Duch, C., Pfl&#252;ger, H. 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+patterns+for+horizontal+and+upside+down+walking+and+vertical+climbing+in+the+locust.">Motor patterns for horizontal and upside down walking and vertical climbing in the locust.</a> J Exp Biol. 198, 1963-1976 (1995).</li><li>Fermi, G., Reichhardt, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optomotorische+Reaktionen+der+Fliege+Musca+domestica.+Abh&#228;ngigkeit+der+Reaktion+von+der+Wellenl&#228;nge,+der+Geschwindigkeit,+dem+Kontrast+und+der+mittleren+Leuchtdichte+bewegter+periodischer+Muster.">Optomotorische Reaktionen der Fliege Musca domestica. 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Physiologie. 40, 556-592 (1958).</li><li>Jander, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Untersuchungen zum Mechanismus und zur zentralnerv&#246;sen Steuerung des Kurvenlaufs bei Stabheuschrecken (Carausius morosus) [dissertation]. , (1982).</li><li>Jander, R., Volk-Henrichs, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Das+strauch-spezifische+Perzeptorsystem+der+Stabheuschrecke+(Carausius+morosus).">Das strauch-spezifische Perzeptorsystem der Stabheuschrecke (Carausius morosus).</a> Zeitschr. f. vergl. Physiologie. 70, 425-447 (1970).</li><li>Kirschfeld, K., Zettler, F., Weiler, 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+resolution+of+lens+and+compound+eyes.">The resolution of lens and compound eyes.</a> Neural Principles in Vision. , 354-370 (1976).</li><li>Kutsch, W., Schwarz, G., Fischer, H., Kautz, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wireless+transmission+of+muscle+potentials+during+free+flight+of+a+locust.">Wireless transmission of muscle potentials during free flight of a locust.</a> J Exp Biol. 185, 367-373 (1993).</li><li>Scharstein, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+universal+projector+for+optomotor+stimulation.">A universal projector for optomotor stimulation.</a> , (1985).</li><li>Schlegtendal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beitrag+zum+Farbensinn+der+Arthropoden.">Beitrag zum Farbensinn der Arthropoden.</a> Zeitschr. f. vergl. Physiologie. 20, 573-581 .</li><li>Schmidt, J., Grund, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rhythmic+activity+in+a+motor+axon+induced+by+axotomy.">Rhythmic activity in a motor axon induced by axotomy.</a> Neuroreport. 14, 1267-1271 (2003).</li><li>Shankar, P. N., Kumar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+Determination+of+the+Kinematic+Viscosity+of+Glycerol-Water+Mixtures.">Experimental Determination of the Kinematic Viscosity of Glycerol-Water Mixtures.</a> Proc. Mathematical and Physical Sci. 444, 573-581 (1994).</li><li>Tryba, A. K., 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=Multi-Joint+coordination+during+walking+and+foothold+searching+in+the+Blaberus+cockroach+I.+Kinematics+and+electromyograms.">Multi-Joint coordination during walking and foothold searching in the Blaberus cockroach I. Kinematics and electromyograms.</a> J Neurophysiol. 83, 3323-3336 .</li><li>Tryba, A. K., 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=Multi-Joint+coordination+during+walking+and+foothold+searching+in+the+Blaberus+cockroach.+II.+Extensor+motor+neuron+pattern.">Multi-Joint coordination during walking and foothold searching in the Blaberus cockroach. II. Extensor motor neuron pattern.</a> J Neurophysiol. 83, 3337-3350 .</li><li>Tsuchida, Y., Hama, N., Takahata, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+optical+telemetry+system+for+underwater+recording+of+electromyogram+and+neuronal+activity+from+non-tethered+crayfish.">An optical telemetry system for underwater recording of electromyogram and neuronal activity from non-tethered crayfish.</a> J Neurosci Methods. 137, 103-109 (2004).</li><li>Wang, H., Ando, N., Kanzaki, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Active+control+of+free+flight+maneuvers+in+a+hawkmoth,+Agrius+convolvuli.">Active control of free flight maneuvers in a hawkmoth, Agrius convolvuli.</a> J Exp Biol. 211, 423-432 (2008).</li></ol>

الإعلان عن أي تضارب في المصالح.

وقد وصفت لدينا الإعداد التي تسمح للجيل يسببها بصريا من تحول السلوك وتصاريح لدرجة كبيرة ولدت من uncoupling neuronally نشاط المشي من الآثار السلبية التي نجمت عن تشريد الميكانيكية للأطراف الأخرى المشي على أرض الواقع. احتمال تدفق المعلومات بين الساقين من خلال الجهاز العصبي حول رد فعل القوات البرية أو الاتصال الرصغي ، من ناحية أخرى ، لا يزال ممكنا ، ويسمح للمجرب لدراسة تأثير مثل هذه المعلومات في إعداد مخفضة. المزايا الرئيسية لبرنامج الإعداد سطح زلق أن تشمل الحيوانات تظهر نزعة عالية جدا على المشي ، ويتعارض مع المشي أو التنقل على treadwheel ، الحيوان يمكن أن تؤدي جولة والحركات المرحلة الموقف في اتجاهات كل من المشي الطبيعية. بالإضافة إلى ذلك ، على درجة من الحرية للجميع الساقين تسمح للحيوان لأداء منحنى سواء كان المشي هو إعداد سليمة أو شبه سليمة. لأنه لا يمكن نقل سلبي الساقين ببساطة عن طريق حركة إلى الأمام من الحيوان أو حركة الركيزة تحتها ، ويعكس كل حركة الإخراج الحركي لتلك المحطة (كروز ، 1976 ؛ غراهام وWendler ، 1981). الإعداد هو مناسبة للغاية لتحقيق أساس العصبية من السلوكيات التكيفية مثل تحول إلى الأمام أو إلى الخلف مقابل المشي ، لأن المرء لا يستطيع الجمع بين التسجيلات الكهربية من النشاط الحركي مع تحليل الكينماتيكا حركة الأطراف. استخدمنا استجابة عصا الحشرات في optomotor لانتزاع المشي. ردود الحيوانات للنمط شريطية الدورية إظهار استعدادها لإجراء منحنى المشي في حين المربوطة على سطح زلق. الأكثر غرابة بالنسبة لنا ، والساقين واحد في التحضير الساق واحدة تظهر نوعيا نفس النمط تتحرك كما في الحيوان سليمة. وبالتالي لدينا سبب للاعتقاد بأن السيطرة على المشي يمكن أن منحنى الدالة إلى حد كبير من دون تنسيق المدخلات الحسية من الساقين المجاورة. سيكون من المهم لاختبار ما اذا كان في تجارب أخرى ويتأثر أيضا نشاط الخلايا العصبية الحركية في الساقين التحفظي من قبل نمط optomotor. ويمكن بسهولة الإعداد يمكن تعديل للسماح للدراسة من المهام الأخرى مثل مستقيم إلى الأمام وإلى الوراء المشي من خلال وضع شريط واحد في الجبهة من حيوان أو سحب بلطف هوائيات. القياس الدقيق للاتصال الأرض تسمح لنا لربط نشاط العضلات وموقف الساق. القرار الأوان لهذه الإشارات الكهربائية اتصال أفضل من 1ms ويقودنا الى نظرة جديدة في توقيت الانتقال من مرحلة إلى أرجوحة الموقف. القرار هو أسوأ بالنسبة للموقف في التأرجح تمر بمرحلة انتقالية بسبب التأخير في القص من زيوت التشحيم إجراء وعدم وجود حاجة لإكمال انطلاقه خلال مطل على سطح زلق. ومع ذلك ، فإن المعرفة حول الأرجوحة دقيقة للانتقال الموقف هو خطوة مفيدة على وجه الخصوص أولا إذا أردنا أن نفهم الآليات التي تتحكم في العضلات وتوقيت الأنشطة المنسقة للعضلات الساق في سياقات سلوكية مختلفة (انظر أيضا : Büschges وآخرون ، 2008 ؛. وBüschges Gruhn 2008). كمثال على ذلك ، استخدمنا مبعاد ومنقلة العضلات الورك من الساق المتوسطة ويرتبط نشاطها بالتحديد مع التحول من مرحلة إلى أرجوحة موقف بينما نحن في نفس الوقت رصد سياق السلوكية التي كانت تستخدم في ساقه. لهذا الغرض ، ونحن يسببها المشي وتسجيل نشاط العضلات بشكل مستمر. ويمكن أن تعطى الساق يكون داخل أو خارج ساق ، اعتمادا على اتجاه تحول. في منتصف الساق التنقل ، بوصفها ساق داخل في معنى وظيفي ، يمكن ملاحظة أن مبعاد ومنقلة العضلات يمكن العمل على حد سواء وظيفية بسبب موقف عضلات الساق ينتج متقطع خطوات إلى الوراء ، بالإضافة إلى الخطوات الموجهة إلى الأمام (انظر Fig.4 ). وتصحيح electromyograms (EMGs) من كل من العضلات وتطبيع للوقت من أسفل لمسة وحسبت كمون والتموج العضلات الأولى. ومن المثير للاهتمام ، والإختفاء من كلا العضلات فيما يتعلق برفع الاقلاع وتهبط ، تعتمد على وظيفة العضلات مثل هذا التأرجح كل موقف أو العضلات (انظر Fig.4) وليس على العضلات نفسها ، وتظهر تغيرات طفيفة في توقيت بدء النشاط. معظم التفسيرات لتغيير الدولة من الأرجوحة لنفترض أن الموقف من إشارات حسية الزناد الاتصال الرصغي بدء الموقف. ويمكن الآن مسألة مثيرة للاهتمام عن كيفية جلب الإختفاء قصيرة بين الأسفل بلمسة وتنشيط العضلات في حوالي عصا الحشرات ، وعلى ما هي المعلومات الحسية التي تعتمد معالجتها مع الإعداد المعدلة. باختصار ، نحن تظهر الإعداد سطح زلق الذي يسمح لنا للحصول على التوالي ، والمشي بشكل صحيح في منحنى عصا الحشرات ثابتة. الكينماتيكا ، والنشاط العضلي وتوقيت الجفن الأسفل بلمسة وانطلاقه يمكن رصدها ومترابطة في ج two سلوكية مختلفةontexts في نفس الوقت. هذا يعطينا أداة ممتازة لدراسة العلاقة بين النشاط مفصلة العضلات والسياق السلوكية للساق واحدة وكذلك في الحيوانات سليمة والآليات الكامنة.

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<td>2-component epoxy glue</td>
<td>&nbsp;</td>
<td>UHU plus, UHU GmbH, Germany</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>glass screens (diameter 130mm)</td>
<td>&nbsp;</td>
<td>Marata screens, Linos Photonics, G&#246;ttingen, Germany</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>dental cement</td>
<td>&nbsp;</td>
<td>ProTemp II, 3M ESPE, Seefeld, Germany</td>
<td>3M Id : 70-2011-0358-0 Catalog Number : 46430</td>
<td>Available through 3M (<a href="http://www.3m.com/">http://www.3m.com/</a>)or dental suppliers</td>
</tr>
<tr>
<td>fluorescent pigments </td>
<td>&nbsp;</td>
<td>Dr. Kremer Farbm&#252;hle, Aichstetten, Germany</td>
<td>Cat.#s: i.e 56200 Fluorescent Pigment Golden Yellow 56350 Fluorescent Pigment Flame Red</td>
<td><a href="http://kremer-pigmente.de/en">http://kremer-pigmente.de/en</a> or <a href="http://www.kremerpigments.com/
">http://www.kremerpigments.com/</a>
</td>
</tr>
<tr>
<td>histoacrylic glue </td>
<td>&nbsp;</td>
<td>3M Vetbond, St.Paul, MN, USA</td>
<td>&nbsp;</td>
<td>supplier: WPI</td>
</tr>
<tr>
<td>coated copper wire</td>
<td>&nbsp;</td>
<td>Elektrisola Eckernhagen</td>
<td>&nbsp;</td>
<td><a href="http://www.elektrisola.com/">http://www.elektrisola.com/</a></td>
</tr>
<tr>
<td>electrode cream </td>
<td>&nbsp;</td>
<td>Marquette Hellige, Freiburg, Germany</td>
<td>&nbsp;</td>
<td>Product is now discontinued, we suggest for example: <a href="www.grasstechnologies.com">www.grasstechnologies.com</a></td>
</tr>
<tr>
<td>pulse generator </td>
<td>&nbsp;</td>
<td>Model MS501, electronics workshop, Zoological Institute, Cologne, Germany</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>lock-in-amplifier </td>
<td>&nbsp;</td>
<td>electronics workshop, Zoological Institute, Cologne, Germany</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>AD converter </td>
<td>&nbsp;</td>
<td>Micro 1401k II, CED, Cambridge, UK</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>preamplifier </td>
<td>&nbsp;</td>
<td>electronics workshop, Zoological Institute, Cologne, Germany</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>high speed video camera </td>
<td>&nbsp;</td>
<td>Marlin F-033C, Allied Vision Technologies, Stadtroda, Germany</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>UV LED arrays </td>
<td>&nbsp;</td>
<td>30-50V DC, electronics workshop, Zoological Institute, Cologne, Germany</td>
<td>&nbsp;</td>
<td>&lambda;390-395nm 
Luminance 24cd</td>
</tr>
<tr>
<td>Digitalizing software Spike2 </td>
<td>&nbsp;</td>
<td>Vers.5.05, CED, Cambridge, UK</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>motion tracking software </td>
<td>&nbsp;</td>
<td>(WINanalyze, Vers.1.9, Mikromak service, Berlin, Germany</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

studying the neural basis of adaptive locomotor behavior in insects

دراسة الأساس العصبي للسلوك المشي ، واحدة غالبا ما يواجه المشكلة أنه من الصعب الفصل بين الانتاج المنتجة neuronally التنقل من تلك الحركات التي تنتج المحطة من القوات السلبية والتفاعل مع الآخرين من خلال الساقين اتصال مشتركة مع الركيزة. إذا كنا نريد أن نفهم ، التي هي جزء من حركة معينة هي التي ينتجها الجهاز العصبي الناتج المحرك ، وتحليل حركية حركات التنقل ، وبالتالي ، تحتاج إلى أن تستكمل مع التسجيلات الكهربية من النشاط الحركي. غالبا ما تقتصر على تسجيل نشاط الخلايا العصبية في العضلات أو حيوان يتصرف من المعدات الكهربية التي يمكن أن تحد من هذا الحيوان في قدرته على التحرك مع درجة أكبر عدد ممكن من الحرية. يمكن أن يكون هذا إما تجنب زرع أقطاب كهربائية باستخدام وبعد هذه الخطوة ثم الحيوان على حبل طويل (أي Clarac وآخرون ، 1987 ؛. دوتش وفلوغر ، 1995 ؛ بوم وآخرون ، 1997 ؛. Gruhn وRathmayer ، 2002) أو عن طريق يحيل البيانات باستخدام أجهزة القياس عن بعد (Kutsch وآخرون ، 1993 ؛ فيشر وآخرون ، 1996 ؛. Tsuchida وآخرون 2004 ؛. حماة وآخرون ، 2007 ؛. انغ وآخرون ، 2008). كل من هذه الأساليب الأنيقة ، والتي تستخدم بنجاح في جو من المفصليات ، يثبت في كثير من الأحيان من الصعب تطبيقها في الحشرات الصغيرة التي إما سيرا على الأقدام بسهولة الحصول على شباكها في حبل طويل أو تعوقها وزن الجهاز القياس عن بعد والبطاريات والخمسين. بالإضافة إلى ذلك ، في جميع هذه الحالات ، فإنه لا يزال من المستحيل التمييز بين الخلايا العصبية على أساس محض للتنقل ، والآثار التي تبذلها اقتران الميكانيكية بين الساقين المشي من خلال الركيزة. أحد الحلول لهذه المشكلة هو لإجراء التجارب في الحيوانات المربوطة التي هي خالية من السير في المكان وعلق العمل محليا ، على سبيل المثال على سطح زلق ، الذي يزيل بفعالية الميكانيكا الاتصال معظم الأرض. وقد استخدمت هذه الدراسة الى ردود الهروب (Camhi وNolen ، 1981 ؛ Camhi وليفي ، 1988) ، تحول (Tryba وRitzman ، 2000A ، ب ؛. Gruhn وآخرون ، 2009a) ، والمشي إلى الخلف (غراهام وإبستين ، 1985) أو تغييرات في السرعة (Gruhn وآخرون ، 2009b) وأنه يسمح للمجرب بسهولة إلى الجمع بين داخل وخارج الخلية مع الفيزيولوجيا تحليلات الحركية (Gruhn وآخرون ، 2006). نستخدم سطح زلق الإعداد للتحقيق في توقيت عضلات الساق في الحشرات عصا يتصرف فيما يتعلق لمس أسفل وانطلاقه في إطار نماذج سلوكية مختلفة مثل المشي قدما المستقيمة والمنحنية في الاستعدادات سليمة وتخفيضها.

Watch this Scientific Journal Video about Studying the Neural Basis of Adaptive Locomotor Behavior in Insects at JoVE.com

Studying the Neural Basis of Adaptive Locomotor Behavior in Insects

نحن تصف طريقة لتسجيل النشاط الحركي والتي تزامنت مع إشارة الاتصال الرصغي سجلت كهربائيا في حشرة المربوطة ، والمشي على سطح زلق. ويستخدم هذا لدراسة الأساس العصبي للسلوك التكيف تحت تأثير انخفاض التفاعل الميكانيكية بين الساقين من خلال الركيزة.

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 ...

studying-the-neural-basis-of-adaptive-locomotor-behavior-in-insects

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

12.7K Views.  University of Cologne. نحن تصف طريقة لتسجيل النشاط الحركي والتي تزامنت مع إشارة الاتصال الرصغي سجلت كهربائيا في حشرة المربوطة ، والمشي على سطح زلق. ويستخدم هذا لدراسة الأساس العصبي للسلوك التكيف تحت تأثير انخفاض التفاعل الميكانيكية بين الساقين من خلال الركيزة.

Video: Studying the Neural Basis of Adaptive Locomotor Behavior in Insects

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

تسجيلات Sensillum واحدة في الحشرات ذبابة الفاكهة السوداء البطن و الأنوفيلة الغامبية</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

Studying the Integration of Adult-born Neurons

Neurogenesis occurs in adult mammalian brains in the sub-ventricular zone (SVZ) of the lateral ventricle and in the sub-granular zone (SGZ) of the hippocampal dentate gyrus throughout life. Previous reports have shown that adult hippocampal neurogenesis is associated with diverse brain disorders, including epilepsy, schizophrenia, depression and anxiety (1). Deciphering the process of normal and aberrant adult-born neuron integration may shed light on the etiology of these diseases and inform the development of new therapies.
SGZ adult neurogenesis mirrors embryonic and post-natal neuronal development, including stages of fate specification, migration, synaptic integration, and maturation. However, full integration occurs over a prolonged, 6-week period. Initial synaptic input to adult-born SGZ dentate granule cells (DGCs) is GABAergic, followed by glutamatergic input at 14 days (2). The specific factors which regulate circuit formation of adult-born neurons in the dentate gyrus are currently unknown.
Our laboratory uses a replication-deficient retroviral vector based on the Moloney murine leukemia virus to deliver fluorescent proteins and hypothesized regulatory genes to these proliferating cells. This viral technique provides high specificity and resolution for analysis of cell birth date, lineage, morphology, and synaptogenesis.
A typical experiment often employs two or three viruses containing unique label, transgene, and promoter elements for single-cell analysis of a desired developmental process in vivo. The following protocol describes a method for analyzing functional newborn neuron integration using a single green (GFP) or red (dTomato) fluorescent protein retrovirus and patch-clamp electrophysiology.

A way to study the integration of newborn dentate granule cells in adult animals is described. This technique uses an engineered retrovirus to label newborn neurons, followed by electrophysiological recordings to determine in vivo functional integration.

دراسة دمج الكبار المولد الخلايا العصبية

Neurogenesis occurs in adult mammalian brains in the sub-ventricular zone (SVZ) of the lateral ventricle and in the sub-granular zone (SGZ) of the ...

High-resolution Live Imaging of Cell Behavior in the Developing Neuroepithelium

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central nervous system (CNS). Although much is known about the molecular mechanisms that pattern the spinal cord and elicit neuronal differentiation1, 2, we lack a deep understanding of these early events at the level of cell behavior. It is thus critical to study the behavior of neural progenitors in real time as they undergo neurogenesis.
In the past, real-time imaging of early embryonic tissue has been limited by cell/tissue viability in culture as well as the phototoxic effects of fluorescent imaging. Here we present a novel assay for imaging such tissue for long periods of time, utilizing a novel ex vivo slice culture protocol and wide-field fluorescence microscopy (Fig. 1). This approach achieves long-term time-lapse monitoring of chick embryonic spinal cord progenitor cells with high spatial and temporal resolution.
This assay may be modified to image a range of embryonic tissues3, 4 In addition to the observation of cellular and sub-cellular behaviors, the development of novel and highly sensitive reporters for gene activity (for example, Notch signaling5) makes this assay a powerful tool with which to understand how signaling regulates cell behavior during embryonic development.

Imaging embryonic tissue in real-time is challenging over long periods of time. Here we present an assay for monitoring cellular and sub-cellular changes in chick spinal cord for long periods with high spatial and temporal resolution. This technique can be adapted for other regions of the nervous system and developing embryo.

عالية الدقة التصوير لايف من سلوك خلية في الظهارة العصبية النامية

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central ...

Co-analysis of Brain Structure and Function using fMRI and Diffusion-weighted Imaging

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when studying the brain, as the functional role that a brain area fulfills may be largely defined by its connections to other brain areas. In this report, we describe a novel, non-invasive approach for relating brain structure and function using magnetic resonance imaging (MRI). This approach, a combination of structural imaging of long-range fiber connections and functional imaging data, is illustrated in two distinct cognitive domains, visual attention and face perception. Structural imaging is performed with diffusion-weighted imaging (DWI) and fiber tractography, which track the diffusion of water molecules along white-matter fiber tracts in the brain (Figure 1). By visualizing these fiber tracts, we are able to investigate the long-range connective architecture of the brain. The results compare favorably with one of the most widely-used techniques in DWI, diffusion tensor imaging (DTI). DTI is unable to resolve complex configurations of fiber tracts, limiting its utility for constructing detailed, anatomically-informed models of brain function. In contrast, our analyses reproduce known neuroanatomy with precision and accuracy. This advantage is partly due to data acquisition procedures: while many DTI protocols measure diffusion in a small number of directions (e.g., 6 or 12), we employ a diffusion spectrum imaging (DSI)1, 2 protocol which assesses diffusion in 257 directions and at a range of magnetic gradient strengths. Moreover, DSI data allow us to use more sophisticated methods for reconstructing acquired data. In two experiments (visual attention and face perception), tractography reveals that co-active areas of the human brain are anatomically connected, supporting extant hypotheses that they form functional networks. DWI allows us to create a "circuit diagram" and reproduce it on an individual-subject basis, for the purpose of monitoring task-relevant brain activity in networks of interest.

We describe a novel approach for simultaneous analysis of brain function and structure using magnetic resonance imaging (MRI). We assess brain structure with high-resolution diffusion-weighted imaging and white-matter fiber tractography. Unlike standard structural MRI, these techniques allow us to directly relate anatomical connectivity to functional properties of brain networks.

شارك في تحليل بنية الدماغ والدالة باستخدام الرنين المغناطيسي الوظيفي وانتشارها المرجحة التصوير

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when ...

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety of brain disorders1,2. To better understand the mechanisms by which brain circuits react to an animal's experience requires the ability to monitor the experience-dependent molecular changes in a given set of neurons, over a prolonged period of time, in the live animal. While experience and associated neural activity is known to trigger gene expression changes in neurons1,2, most of the methods to detect such changes do not allow repeated observation of the same neurons over multiple days or do not have sufficient resolution to observe individual neurons3,4. Here, we describe a method that combines in vivo two-photon microscopy with a genetically encoded fluorescent reporter to track experience-dependent gene expression changes in individual cortical neurons over the course of day-to-day experience. 
One of the well-established experience-dependent genes is Activity-regulated cytoskeletal associated protein (Arc)5,6. The transcription of Arc is rapidly and highly induced by intensified neuronal activity3, and its protein product regulates the endocytosis of glutamate receptors and long-term synaptic plasticity7. The expression of Arc has been widely used as a molecular marker to map neuronal circuits involved in specific behaviors3. In most of those studies, Arc expression was detected by in situ hybridization or immunohistochemistry in fixed brain sections. Although those methods revealed that the expression of Arc was localized to a subset of excitatory neurons after behavioral experience, how the cellular patterns of Arc expression might change with multiple episodes of repeated or distinctive experiences over days was not investigated. 
In vivo two-photon microscopy offers a powerful way to examine experience-dependent cellular changes in the living brain8,9. To enable the examination of Arc expression in live neurons by two-photon microscopy, we previously generated a knock-in mouse line in which a GFP reporter is placed under the control of the endogenous Arc promoter10. This protocol describes the surgical preparations and imaging procedures for tracking experience-dependent Arc-GFP expression patterns in neuronal ensembles in the live animal. In this method, chronic cranial windows were first implanted in Arc-GFP mice over the cortical regions of interest. Those animals were then repeatedly imaged by two-photon microscopy after desired behavioral paradigms over the course of several days. This method may be generally applicable to animals carrying other fluorescent reporters of experience-dependent molecular changes4.

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

Experience-dependent molecular changes in neurons are essential for the brain's ability to adapt in response to behavioral challenges. An in vivo two-photon imaging method is described here that allows the tracking of such molecular changes in individual cortical neurons through genetically encoded reporters.

في الجسم الحي ثنائي الفوتون التصوير من الخبرة التي تعتمد على التغيرات الجزيئية في الخلايا العصبية القشرية

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases.

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity.&#160; Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity.&#160;

تحديد العفوي الحركي آخر في<em&gt; ذبابة الفاكهة السوداء البطن</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy and cheap methods to distinguish between individuals in a trial. A standard technique for this is CO2 anaesthesia and then labelling or wing clipping each fly. However, this is invasive and has been shown to affect behavior. Other techniques have used coloration to identify flies. This article presents a simple and non-invasive method for labelling Drosophila that allows them to be individually identified within experiments, using food coloring. This method is used in trials where two males compete to mate with a female. Dyeing allowed quick and easy identification. There was, however, some difference in the strength of the coloration across the three species tested. Data is presented showing the dye has a lower impact on mating behavior than CO2 in Drosophila melanogaster. The impact of CO2 anaesthesia is shown to depend on the species of Drosophila, with D. pseudoobscura and D. subobscura showing no impact, whereas D. melanogaster males had reduced mating success. The dye method presented is applicable to a wide range of experimental designs.

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

This protocol describes a simple, cost effective way to individually identify Drosophila or other insects. Demonstration data investigating mating success across three species of Drosophila show that this method is comparable or better than the use of CO2 anaesthesia.

الصباغة الحشرات لفحوصات السلوكية: في سلوك التزاوج من تخدير<em&gt; ذبابة الفاكهة</em

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

A Simple Flight Mill for the Study of Tethered Flight in Insects

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have shown that flight mills are valuable tools for the experimental study of insect flight behavior, allowing researchers to examine how factors such as age, host plants, or population source can influence an insects' propensity to disperse. Flight mills allow researchers to measure components of flight such as speed and distance flown. Lack of detailed information about how to build such a device can make their construction appear to be prohibitively complex. We present a simple and relatively inexpensive flight mill for the study of tethered flight in insects. Experimental insects can be tethered with non-toxic adhesives and revolve around an axis by means of a very low friction magnetic bearing. The mill is designed for the study of flight in controlled conditions as it can be used inside an incubator or environmental chamber. The strongest points are the very simple electronic circuitry, the design that allows sixteen insects to fly simultaneously allowing the collection and analysis of a large number of samples in a short time and the potential to use the device in a very limited workspace. This design is extremely flexible, and we have adjusted the mill to accommodate different species of insects of various sizes.

Flight in insects is influenced by a number of factors and the propensity to disperse is an important variable in understanding insect ecology and biological control strategies. We describe the construction and use of a simple, relatively inexpensive, and flexible flight mill for measuring parameters of tethered flight in insects.

A مطحنة الطيران بسيط لدراسة الطيران في الحشرات مربوط

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network dynamics in neuronal cultures. In contrast with more traditional methods, such as patch-clamping, which can only record activity from a single cell, MEAs can record simultaneously from multiple sites in a network, without requiring the arduous task of placing each electrode individually. Moreover, numerous control and stimulation configurations can be easily applied within the same experimental setup, allowing for a broad range of dynamics to be explored. One of the key dynamics of interest in these in vitro studies has been the extent to which cultured networks display properties indicative of learning. Mouse neuronal cells cultured on MEAs display an increase in response following training induced by electrical stimulation. This protocol demonstrates how to culture neuronal cells on MEAs; successfully record from over 95% of the plated dishes; establish a protocol to train the networks to respond to patterns of stimulation; and sort, plot, and interpret the results from such experiments. The use of a proprietary system for stimulating and recording neuronal cultures is demonstrated. Software packages are also used to sort neuronal units. A custom-designed graphical user interface is used to visualize post-stimulus time histograms, inter-burst intervals, and burst duration, as well as to compare the cellular response to stimulation before and after a training protocol. Finally, representative results and future directions of this research effort are discussed.

Mouse neuronal cells cultured on multi-electrode arrays display an increase in response following electrical stimulation. This protocol demonstrates how to culture neurons, how to record activity, and how to establish a protocol to train the networks to respond to patterns of stimulation.

زيادة تعتمد على الوقت في استجابة الشبكة لتحفيز الثقافات خلية الخلايا العصبية على صفائف الدقيقة الكهربائي

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

Advances in next-generation sequencing technologies contribute to the identification of (candidate) disease genes for movement disorders and other neurological diseases at an increasing speed. However, little is known about the molecular mechanisms that underlie these disorders. The genetic, molecular, and behavioral toolbox of Drosophila melanogaster makes this model organism particularly useful to characterize new disease genes and mechanisms in a high-throughput manner. Nevertheless, high-throughput screens require efficient and reliable assays that, ideally, are cost-effective and allow for the automatized quantification of traits relevant to these disorders. The island assay is a cost-effective and easily set-up method to evaluate Drosophila locomotor behavior. In this assay, flies are thrown onto a platform from a fixed height. This induces an innate motor response that enables the flies to escape from the platform within seconds. At present, quantitative analyses of filmed island assays are done manually, which is a laborious undertaking, particularly when performing large screens.
This manuscript describes the &#34;Drosophila Island Assay&#34; and &#34;Island Assay Analysis&#34; algorithms for&#160;high-throughput, automated data processing and quantification of island assay data. In the setup, a simple webcam connected to a laptop collects an image series of the platform while the assay is performed. The &#34;Drosophila Island Assay&#34; algorithm developed for the open-source software Fiji processes these image series and quantifies, for each experimental condition, the number of flies on the platform over time. The &#34;Island Assay Analysis&#34; script, compatible with the free software R, was developed to automatically process the obtained data and to calculate whether treatments/genotypes are statistically different. This greatly improves the efficiency of the island assay and makes it a powerful readout for basic locomotion and flight behavior. It can thus be applied to large screens investigating fly locomotor ability, Drosophila models of movement disorders, and drug efficacy.

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

The island assay is a relatively new, cost-effective assay that can be used to evaluate the basic locomotor behavior of Drosophila melanogaster. This manuscript describes algorithms for automatic data processing and objective quantification of island assay data, making this assay a sensitive, high-throughput readout for large genetic or pharmacological screens.

دراسة الأساس العصبي للسلوك الحركي التكيفية في الحشرات

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تسجيلات Sensillum واحدة في الحشرات ذبابة الفاكهة السوداء البطن و الأنوفيلة الغامبية

دراسة دمج الكبار المولد الخلايا العصبية

عالية الدقة التصوير لايف من سلوك خلية في الظهارة العصبية النامية

شارك في تحليل بنية الدماغ والدالة باستخدام الرنين المغناطيسي الوظيفي وانتشارها المرجحة التصوير

في الجسم الحي ثنائي الفوتون التصوير من الخبرة التي تعتمد على التغيرات الجزيئية في الخلايا العصبية القشرية

تحديد العفوي الحركي آخر في

الصباغة الحشرات لفحوصات السلوكية: في سلوك التزاوج من تخدير

A مطحنة الطيران بسيط لدراسة الطيران في الحشرات مربوط

زيادة تعتمد على الوقت في استجابة الشبكة لتحفيز الثقافات خلية الخلايا العصبية على صفائف الدقيقة الكهربائي

تحليل الفائق للسلوك الحركي في مقايسة الجزيرة المورفولوجية