<ol>
	<li>Boeckh, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elektrophysiologische+Untersuchungen+an+einzelnen+Geruchsrezeptoren+auf+der+Antenne+des+TotengrAbers+(Necrophorus+Coleoptera).">Elektrophysiologische Untersuchungen an einzelnen Geruchsrezeptoren auf der Antenne des TotengrAbers (Necrophorus Coleoptera).</a> Z. Vergl. Physiol. 46, 212-248 (1962).</li><li>Schneider, D., Hecker, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zur+Elektrophysiologic+der+Antenne+des+Seidenspinners+Bombyx+mori+bei+Reizung+mit+angereicherten+Extrakten+des+Sexuallockstoffes.">Zur Elektrophysiologic der Antenne des Seidenspinners Bombyx mori bei Reizung mit angereicherten Extrakten des Sexuallockstoffes.</a> Z. Naturforschg. 11b, 121-124 (1956).</li><li>Touhara, K., Vosshall, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensing+odorants+and+pheromones+with+chemosensory+receptors.">Sensing odorants and pheromones with chemosensory receptors.</a> Annu Rev Physiol. 71, 307-332 (2009).</li><li>Sato, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insect+olfactory+receptors+are+heteromeric+ligand-gated+ion+channels.">Insect olfactory receptors are heteromeric ligand-gated ion channels.</a> Nature. 452, 1002-1006 (2008).</li><li>Wicher, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+odorant+receptors+are+both+ligand-gated+and+cyclic-nucleotide-activated+cation+channels.">Drosophila odorant receptors are both ligand-gated and cyclic-nucleotide-activated cation channels.</a> Nature. 452, 1007-1011 (2008).</li><li>Hallem, E. A., Ho, M. G., Carlson, J. 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+molecular+basis+of+odor+coding+in+the+Drosophila+antenna.">The molecular basis of odor coding in the Drosophila antenna.</a> Cell. 117, 965-979 (2004).</li><li>Hallem, E. A., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coding+of+odors+by+a+receptor+repertoire.">Coding of odors by a receptor repertoire.</a> Cell. 125, 143-160 (2006).</li><li>Boeckh, J., Kaissling, K. E., Schneider, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insect+olfactory+receptors.">Insect olfactory receptors.</a> Cold Spring Harb Symp Quant Biol. 30, 263-280 (1965).</li><li>de Bruyne, M., Foster, K., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odor+coding+in+the+Drosophila+antenna.">Odor coding in the Drosophila antenna.</a> Neuron. 30, 537-552 (2001).</li><li>Lu, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odor+coding+in+the+maxillary+palp+of+the+malaria+vector+mosquito+Anopheles+gambiae.">Odor coding in the maxillary palp of the malaria vector mosquito Anopheles gambiae.</a> Curr Biol. 17, 1533-1544 (2007).</li><li>Hallem, E. A., Fox, A. N., Zwiebel, L. J., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfaction:+mosquito+receptor+for+human-sweat+odorant.">Olfaction: mosquito receptor for human-sweat odorant.</a> Nature. 427, 212-213 (2004).</li><li>Couto, A., Alenius, M., Dickson, B. J. M. o. l. e. c. u. l. a. r. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=anatomical,+and+functional+organization+of+the+Drosophila+olfactory+system.">anatomical, and functional organization of the Drosophila olfactory system.</a> Curr. Biol. 15, 1535-1547 (2005).</li><li>Fishilevich, E., Vosshall, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+and+functional+subdivision+of+the+Drosophila+antennal+lobe.">Genetic and functional subdivision of the Drosophila antennal lobe.</a> Curr Biol. 15, 1548-1553 (2005).</li></ol>

تستخدم إشارات شمية بواسطة الكائنات الحية لتحديد مصادر الغذاء والاصحاب المحتملة ، والحيوانات المفترسة. الخلايا العصبية الحسية الشمية (OSNs) هي المركز الأول بين التتابع مؤثرات الخارجية وأعلى المراكز في الدماغ حيث يتم معالجة المزيد من المعلومات. في melanogaster ذبابة الفاكهة والأنوفيلة الغامبية ، OSNs يمكن الحصول عليها بسهولة ، ويمكن رصد نشاطهم الكهربائية بينما يحفزه نفث رائحة. أوضح تسجيل sensillum واحد (SSR) تقنية في هذا الفيديو قد استخدمت على نطاق واسع لتسجيل ودراسة من OSNs ردودهم الكهربائية لعدد كبير من odorants 6 ، 7. جعلت من المستقبلات الشمية deorphanization (ORS) 6 ، (11) ورسم خرائط لنسب الأرجحية في مواقع محددة على ذبابة الفاكهة هوائي 9 ، 12 ، 13 SSR التقنية أداة قوية لتحليل الخصائص الكهربية من نسب الأرجحية محددة في الجسم الحي ، وأول خطوة لفهم الكيفية التي ترجمت في العالم الخارجي الشمية إلى إشارات كهربائية من خلال OSNs وينظر إليها في نهاية المطاف من قبل الحيوانات.

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Paraffin oil</td>
<td>Odors</td>
<td>Fluka</td>
<td>76235</td>
<td>&nbsp;</td>
<tr>
<td>High purity odors (&gt;98%)</td>
<td>Odors</td>
<td>Sigma-Aldrich</td>
<td>&nbsp;</td>
<td>Methyl acetate 
#296996 
1-octen-3-ol 
#74950 </td>
</tr>
<tr>
<td>Filter paper strips</td>
<td>Odors</td>
<td>Fisherbrand</td>
<td>05-714-1</td>
<td>Chromatography paper</td>
</tr>
<tr>
<td>Connectors</td>
<td>Odors</td>
<td>Cole-Parmer</td>
<td>EW-06365-40</td>
<td>1/16x1/8"</td>
</tr>
<tr>
<td>Glass vials</td>
<td>Odors</td>
<td>Agilent Technologies</td>
<td>5182-0556</td>
<td>&nbsp;</td>
<tr>
<td>Air line plastic tubing</td>
<td>Odor Delivery</td>
<td>Python Products</td>
<td>500PAL</td>
<td>&nbsp;</td>
<tr>
<td>1 serological pipette</td>
<td>Odor Delivery</td>
<td>Corning</td>
<td>4101</td>
<td>10 mL</td>
</tr>
<tr>
<td>Plastic tubing</td>
<td>Odor Delivery</td>
<td>Cole-Parmer</td>
<td>EW-06418-0</td>
<td>0.050"x0.090"OD</td>
</tr>
<tr>
<td>Disposable borosilicate glass Pasteur pipettes</td>
<td>Odor Delivery</td>
<td>FisherBrand</td>
<td>13-678-20A</td>
<td>5-3/4 inches</td>
</tr>
<tr>
<td>Programmable stimulus controller</td>
<td>Odor Delivery</td>
<td>Syntech</td>
<td>CS-55</td>
<td>&nbsp;</td>
<tr>
<td>Anti-vibration table</td>
<td>Electrophysiology Equipment</td>
<td>TMC</td>
<td>63533</td>
<td>36&#x201D;Wx30&#x201D;Dx29&#x201D;H</td>
</tr>
<tr>
<td>Faraday cage</td>
<td>Electrophysiology Equipment</td>
<td>TMC</td>
<td>MI8133303</td>
<td>&nbsp;</td>
<tr>
<td>Inverted microscope</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>E600FN ECLIPSE</td>
<td>Recording microscope</td>
</tr>
<tr>
<td>10x and 100x objectives</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>10x Plan Fluor 100x L Plan</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting microscope</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>EZ645</td>
<td>electrode sharpening/insect prep microscope</td>
</tr>
<tr>
<td>Magnetic stands</td>
<td>Electrophysiology Equipment</td>
<td>Newport</td>
<td>MODEL 150</td>
<td>&nbsp;</td>
<tr>
<td>IDAC</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>IDAC-4</td>
<td>&nbsp;</td>
<tr>
<td>Acquisition software</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>Autospike</td>
<td>&nbsp;</td>
<tr>
<td>1 macromanipulator</td>
<td>Electrophysiology Equipment</td>
<td>NARISHIGE</td>
<td>MN-151</td>
<td>Joystick manipulator 
Used for positioning reference electrode
</td>
</tr>
<tr>
<td>1 micromanipulator</td>
<td>Electrophysiology Equipment</td>
<td>EXFO</td>
<td>PCS-6000</td>
<td>Used for positioning recording electrode</td>
</tr>
<tr>
<td>Crocodile clip</td>
<td>Electrophysiology Equipment</td>
<td>Pomona</td>
<td>AL-B-12-0</td>
<td>&nbsp;</td>
<tr>
<td>Electric cable</td>
<td>Electrophysiology Equipment</td>
<td>Pomona</td>
<td>B-36-0</td>
<td>Test Cable Assembly</td>
</tr>
<tr>
<td>2 electrode holders</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>N/A</td>
<td>Electrode holders (set of 2) for tungsten wire electrode</td>
</tr>
<tr>
<td>AC probe</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>N/A</td>
<td>Universal single ended probe (10xAC)</td>
</tr>
<tr>
<td>Tungsten electrodes</td>
<td>Electrophysiology Equipment</td>
<td>Microprobes</td>
<td>M210</td>
<td>straight tungsten rods, 0.005&#x201C;x3&#x201C;</td>
</tr>
<tr>
<td>Potassium hydroxide</td>
<td>Electrophysiology Equipment</td>
<td>Sigma-Aldrich</td>
<td>221473</td>
<td>&nbsp;</td>
<tr>
<td>Syringe</td>
<td>Electrophysiology Equipment</td>
<td>BD</td>
<td>301625</td>
<td>20 mL</td>
</tr>
<tr>
<td>Power supply</td>
<td>Electrophysiology Equipment</td>
<td>WILD HEERBRUGG 6V 40W</td>
<td>e.g MTR32</td>
<td>&nbsp;</td>
<tr>
<td>Vertical puller</td>
<td>Insect prep</td>
<td>Narishige</td>
<td>PB-7</td>
<td>&nbsp;</td>
<tr>
<td>Razor blade</td>
<td>Insect prep</td>
<td>VWR</td>
<td>55411-050</td>
<td>&nbsp;</td>
<tr>
<td>Dental wax</td>
<td>Insect prep</td>
<td>Patterson</td>
<td>091-1503</td>
<td>&nbsp;</td>
<tr>
<td>Microscope slide</td>
<td>Insect prep</td>
<td>FisherBrand</td>
<td>12-550A</td>
<td>&nbsp;</td>
<tr>
<td>Cover glass</td>
<td>Insect prep</td>
<td>FisherBrand</td>
<td>12-541A</td>
<td>18X18 #1.5</td>
</tr>
<tr>
<td>Polypropylene mesh</td>
<td>Insect prep</td>
<td>Small Parts inc.</td>
<td>CMP-0500-B</td>
<td>&nbsp;</td>
<tr>
<td>Glass electrode</td>
<td>Insect prep</td>
<td>Frederick Haer & Co.</td>
<td>27-32-0-075</td>
<td>Capillary tubing borosilicate 1.5mm OD x 1.12mm ID x 75 mm</td>
</tr>
<tr>
<td>Double-sided tape (3M)</td>
<td>Insect prep</td>
<td>3M</td>
<td>MMM6652P3436</td>
<td>Double-sided tape (3M)</td>
</tr>
<tr>
<td>Forceps</td>
<td>Insect prep</td>
<td>Fine Science Tools</td>
<td>021x0053</td>
<td>Dumont #5 Mirror Finish Forceps</td>
</tr>
<tr>
<td>Small plastic cup</td>
<td>Insect prep</td>
<td>VWR</td>
<td>89009-662</td>
<td>7 x 5.7 (23/4 x 21/4)</td>
</tr>
<tr>
<td>Electric aspirator</td>
<td>Insect prep</td>
<td>Gempler&#x2019;s</td>
<td>RHM200</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

single sensillum recordings in the insects drosophila melanogaster and anopheles gambiae

حاسة الشم أمر أساسي للحشرات للعثور على الأطعمة ، والاصحاب ، الحيوانات المفترسة ، ومواقع oviposition 3. يتم تضمين الخلايا العصبية الحسية الشمية الحشرات (OSNs) في الشعر الحسية ودعا sensilla ، التي تغطي سطح أجهزة الشمية. وتغطي سطح كل sensillum مع مسام صغيرة ، يمكن من خلالها تمرير odorants وتذوب في السائل دعا الليمفاوية sensillum الذي يغمر التشعبات الحسية من OSNs يضم في sensillum معين. وأعرب عن التشعبات OSN مستقبلات الرائحة (OR) والبروتينات ، والتي تؤدي وظيفة والحشرات في قنوات ايون رائحة بوابات 4 و 5. تفاعل مع نسب الأرجحية odorants إما زيادة أو نقصان معدل اطلاق القاعدية من OSN. هذا نشاط الخلايا العصبية في شكل امكانات العمل يجسد التمثيل الأولى من الجودة ، والكثافة ، والخصائص الزمنية للالرائحة 6 ، 7. نظرا لسهولة الوصول إلى هذه الشعرات الحسية ، فمن الممكن أن تؤدي التسجيلات من خارج الخلية OSNs واحد عن طريق إدخال القطب تسجيل في sensillum الليمفاوية ، في حين يوضع القطب المرجع في الليمفاوية من العين أو جسم الحشرة. في ذبابة الفاكهة ، sensilla منزل ما بين واحد وأربعة OSNs ، ولكن كل OSN يعرض عادة على ارتفاع السعة مميزة. تقنيات الفرز ارتفاع تجعل من الممكن لتعيين الاستجابات التشويك لOSNs الفردية. هذا التسجيل sensillum واحد (SSR) تقنية مراقبة الفرق في محتملة بين الليمفاوية sensillum والمرجعية باعتبارها القطب التموج الكهربائية التي تتولد عن هذا النشاط على مستقبلات OSNs 1 ، 2 ، 8. تغييرات في عدد من المسامير ردا على الرائحة تمثل الأساس الخلوي للرائحة الترميز في الحشرات. هنا ، نحن تصف طريقة إعداد المستخدمة حاليا في المختبر لإجراء إصلاح قطاع الأمن في melanogaster ذبابة الفاكهة والأنوفيلة الغامبية ، وتظهر آثار الناجمة عن ممثل odorants بطريقة sensillum محددة.

Watch this Scientific Journal Video about Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae at JoVE.com

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

ويمكن قياس ردود الكهربية للخلايا العصبية الحسية الشمية إلى odorants في الحشرات باستخدام التسجيلات sensillum واحد. في هذه المقالة سوف نقوم الفيديو لشرح كيفية تنفيذ تسجيلات sensillum واحدة في هوائيات على ذبابة الخل (<em&gt; ذبابة الفاكهة السوداء البطن</em&gt;) والفك العلوي palps البعوض الذي ينقل الملاريا (<em&gt; الأنوفيلة الغامبية</em&gt;).

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 ...

single-sensillum-recordings-insects-drosophila-melanogaster-anopheles

Nanyang Technological University

Research

JoVE Journal

Neuroscience

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

25.1K Views.  Rockefeller University. ويمكن قياس ردود الكهربية للخلايا العصبية الحسية الشمية إلى odorants في الحشرات باستخدام التسجيلات sensillum واحد. في هذه المقالة سوف نقوم الفيديو لشرح كيفية تنفيذ تسجيلات sensillum واحدة في هوائيات على ذبابة الخل ( ذبابة الفاكهة السوداء البطن) والفك العلوي palps البعو...

Video: Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

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" ...

Visualization of Proprioceptors in Drosophila Larvae and Pupae

Proprioception is the ability to sense the motion, or position, of body parts by responding to stimuli arising within the body. In fruitflies and other insects proprioception is provided by specialized sensory organs termed chordotonal organs (ChOs) 2. Like many other organs in Drosophila, ChOs develop twice during the life cycle of the fly. First, the larval ChOs develop during embryogenesis. Then, the adult ChOs start to develop in the larval imaginal discs and continue to differentiate during metamorphosis.
The development of larval ChOs during embryogenesis has been studied extensively 10,11,13,15,16. The centerpiece of each ChO is a sensory unit composed of a neuron and a scolopale cell. The sensory unit is stretched between two types of accessory cells that attach to the cuticle via specialized epidermal attachment cells 1,9,14. When a fly larva moves, the relative displacement of the epidermal attachment cells leads to stretching of the sensory unit and consequent opening of specific transient receptor potential vanilloid (TRPV) channels at the outer segment of the dendrite 8,12. The elicited signal is then transferred to the locomotor central pattern generator circuit in the central nervous system.
Multiple ChOs have been described in the adult fly 7. These are located near the joints of the adult fly appendages (legs, wings and halters) and in the thorax and abdomen. In addition, several hundreds of ChOs collectively form the Johnston's organ in the adult antenna that transduce acoustic to mechanical energy 3,5,17,4.
In contrast to the extensive knowledge about the development of ChOs in embryonic stages, very little is known about the morphology of these organs during larval stages. Moreover, with the exception of femoral ChOs 18 and Johnston's organ, our knowledge about the development and structure of ChOs in the adult fly is very fragmentary.
Here we describe a method for staining and visualizing ChOs in third instar larvae and pupae. This method can be applied together with genetic tools to better characterize the morphology and understand the development of the various ChOs in the fly.

Visualization of Proprioceptors in Drosophila Larvae and Pupae

A method to immunostain and visualize chordotonal organs in larvae and pupae of Drosophila melanogaster is described.

تصور في Proprioceptors<em&gt; ذبابة الفاكهة</em&gt; يرقات وشرانق

Proprioception is the ability to sense the motion, or position, of body parts by responding to stimuli arising within the body. In fruitflies and other ...

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

In this article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the lipophilic fluorescent membrane marker DiI. This method allows the visualization of neuronal cell morphology in great detail. It is possible to label any cell in the CNS: cell bodies of target neurons are visualized under DIC optics or by expression of a fluorescent genetic marker such as GFP. After labeling, the DiI can be transformed into a permanent brown stain by photoconversion to allow visualization of cell morphology with transmitted light and DIC optics. Alternatively, the DiI-labeled cells can be observed directly with confocal microscopy, enabling genetically introduced fluorescent reporter proteins to be colocalised. The technique can be used in any animal, irrespective of genotype, making it possible to analyze mutant phenotypes at single cell resolution.

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

We present a technique for labeling single neurons in the central nervous system (CNS) of Drosophila embryos, which allows the analysis of neuronal morphology by either transmitted light or confocal microscopy.

وضع العلامات واحدة من الخلايا في الجهاز العصبي المركزي<em&gt; ذبابة الفاكهة الدروسوفيلا</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

A growing number of genetically encoded tools are becoming available that allow non-invasive manipulation of the neural activity of specific neurons in Drosophila melanogaster1. Chief among these are optogenetic tools, which enable the activation or silencing of specific neurons in the intact and freely moving animal using bright light. Channelrhodopsin (ChR2) is a light-activated cation channel that, when activated by blue light, causes depolarization of neurons that express it. ChR2 has been effective for identifying neurons critical for specific behaviors, such as CO2 avoidance, proboscis extension and giant-fiber mediated startle response2-4. However, as the intense light sources used to stimulate ChR2 also stimulate photoreceptors, these optogenetic techniques have not previously been used in the visual system. Here, we combine an optogenetic approach with a mutation that impairs phototransduction to demonstrate that activation of a cluster of loom-sensitive neurons in the fly's optic lobe, Foma-1 neurons, can drive an escape behavior used to avoid collision. We used a null allele of a critical component of the phototransduction cascade, phospholipase C-&beta;, encoded by the norpA gene, to render the flies blind and also use the Gal4-UAS transcriptional activator system to drive expression of ChR2 in the Foma-1 neurons. Individual flies are placed on a small platform surrounded by blue LEDs. When the LEDs are illuminated, the flies quickly take-off into flight, in a manner similar to visually driven loom-escape behavior. We believe that this technique can be easily adapted to examine other behaviors in freely moving flies.

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Genetically encoded optogenetic tools enable noninvasive manipulation of specific neurons in the Drosophila brain. Such tools can identify neurons whose activation is sufficient to elicit or suppress particular behaviors. Here we present a method for activating Channelrhodopsin2 that is expressed in targeted neurons in freely walking flies.

Optogenetic تحفيز السلوك في الهروب<em&gt; ذبابة الفاكهة الدروسوفيلا</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

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 ...

Conditions Affecting Social Space in Drosophila melanogaster

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a straightforward manner. As we previously demonstrated 1, in a two-dimensional chamber, we first force the flies to form a tight group, subsequently allowing them to take their preferred distance from each other. After the flies have settled, we measure the distance to the closest neighbor (or social space), processing a static picture with free online software (ImageJ). The analysis of the distance to the closest neighbor allows researchers to determine the effects of genetic and environmental factors on social interaction, while controlling for potential confounding factors. Diverse factors such as climbing ability, time of day, sex, and number of flies, can modify social spacing of flies. We thus propose a series of experimental controls to mitigate these confounding effects. This assay can be used for at least two purposes. First, researchers can determine how their favorite environmental shift (such as isolation, temperature, stress or toxins) will impact social spacing 1,2. Second, researchers can dissect the genetic and neural underpinnings of this basic form of social behavior 1,3. Specifically, we used it as a diagnostic tool to study the role of orthologous genes thought to be involved in social behavior in other organisms, such as candidate genes for autism in humans 4.

Conditions Affecting Social Space in Drosophila melanogaster

The effect of genes and environment on social space of Drosophila melanogaster can be quantified through a powerful but straightforward analytical paradigm. We show here different factors that can affect this social space, and thus need to be taken into consideration when designing experiments in this paradigm.

الظروف التي تؤثر في الفضاء الاجتماعي<em&gt; ذبابة الفاكهة السوداء البطن</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.

النسخ في والسلوكية الفحص لMechanosensation من MARCM المستندة<em&gt; ذبابة الفاكهة السوداء البطن</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

طريقة بسيطة لقياس التغيرات في السلوك المكافأة التماس طريق<em&gt; ذبابة الفاكهة</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of the right type and quantity of food therefore has a major influence on quality of life. Research on feeding behavior focuses on the underlying processes that ensure actual feeding and unravels the role of factors regulating internal energy homeostasis and the neuronal bases of decision-making. The model organism Drosophila melanogaster, with its great variety of genetically traceable tools for labeling and manipulating single neurons, allows mapping of neuronal networks and identification of molecular signaling cascades involved in the regulation of food intake. This report demonstrates the CApillary FEeder assay (CAFE) and shows how to measure food intake in a group of flies for time spans ranging from hours to days. This easy-to-use assay consists of glass capillaries filled with liquid food that flies can freely access and feed on. Food consumption in the assay is accurately determined using simple measurement tools. Herein we describe step-by-step the method from setup to successful execution of the CAFE assay, and provide practical examples to analyze the food intake of a group of flies under controlled conditions. The reader is guided through possible limitations of the assay, and advantages and disadvantages of the method compared to other feeding assays in D. melanogaster are evaluated.

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder (CAFE) assay is a simple, budget-friendly, highly reliable method for investigating mechanisms underlying food intake. Used with the highly versatile genetic model organism Drosophila melanogaster, it provides a powerful means of gaining new insights into regulatory mechanisms of food intake.

الشعيرات الدموية المغذية الفحص يقيس كمية الطعام في<em&gt; ذبابة الفاكهة</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...