Name: taste preference assay for adult drosophila
Uploaded: 2016-09-08T12:30:00
Description: Watch this Scientific Journal Video about Taste Preference Assay for Adult Drosophila at JoVE.com

We would like to thank members of the Tessier lab for critical reading of this manuscript and helpful suggestions during the preparation of this protocol.

1. المجاعة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إعداد يطير قارورة المجاعة التي تشبع القطن الكرة مع 18.2 MΩ الماء في قاع القارورة الطيران القياسية. بدلا من ذلك، تشبع بالمثل شريط صغير من ورق الترشيح مع 18.2 MΩ المياه ومكان في زاوية داخل القارورة. </li><li style=";text-align:ri...

<ol>
	<li>Herrero, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fruit+fly+behavior+in+response+to+chemosensory+signals.">Fruit fly behavior in response to chemosensory signals.</a> Peptides. 38 (2), 228-237 (2012).</li><li>Vosshall, L. B., Stocker, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+architecture+of+smell+and+taste+in+Drosophila.">Molecular architecture of smell and taste in Drosophila.</a> Annu Rev Neurosci. 30, 505-533 (2007).</li><li>Harris, D. T., Kallman, B. R., Mullaney, B. C., Scott, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Representations+of+Taste+Modality+in+the+Drosophila+Brain.">Representations of Taste Modality in the Drosophila Brain.</a> Neuron. 86 (6), 1449-1460 (2015).</li><li>Hong, E. J., Wilson, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+encoding+of+odors+by+channels+with+diverse+sensitivity+to+inhibition.">Simultaneous encoding of odors by channels with diverse sensitivity to inhibition.</a> Neuron. 85 (3), 573-589 (2015).</li><li>Kain, P., Dahanukar, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secondary+taste+neurons+that+convey+sweet+taste+and+starvation+in+the+Drosophila+brain.">Secondary taste neurons that convey sweet taste and starvation in the Drosophila brain.</a> Neuron. 85 (4), 819-832 (2015).</li><li>Masek, P., Worden, K., Aso, Y., Rubin, G. M., Keene, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dopamine-modulated+neural+circuit+regulating+aversive+taste+memory+in+Drosophila.">A dopamine-modulated neural circuit regulating aversive taste memory in Drosophila.</a> Curr Biol. 25 (11), 1535-1541 (2015).</li><li>Charlu, S., Wisotsky, Z., Medina, A., Dahanukar, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acid+sensing+by+sweet+and+bitter+taste+neurons+in+Drosophila+melanogaster.">Acid sensing by sweet and bitter taste neurons in Drosophila melanogaster.</a> Nat Commun. 4, 2042 (2013).</li><li>Clyne, P. J., Warr, C. 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=Candidate+taste+receptors+in+Drosophila.">Candidate taste receptors in Drosophila.</a> Science. 287 (5459), 1830-1834 (2000).</li><li>Scott, K., 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=A+chemosensory+gene+family+encoding+candidate+gustatory+and+olfactory+receptors+in+Drosophila.">A chemosensory gene family encoding candidate gustatory and olfactory receptors in Drosophila.</a> Cell. 104 (5), 661-673 (2001).</li><li>Kim, S. H., 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=Drosophila+TRPA1+channel+mediates+chemical+avoidance+in+gustatory+receptor+neurons.">Drosophila TRPA1 channel mediates chemical avoidance in gustatory receptor neurons.</a> Proc Natl Acad Sci U S A. 107 (18), 8440-8445 (2010).</li><li>Koh, T. W., 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=The+Drosophila+IR20a+clade+of+ionotropic+receptors+are+candidate+taste+and+pheromone+receptors.">The Drosophila IR20a clade of ionotropic receptors are candidate taste and pheromone receptors.</a> Neuron. 83 (4), 850-865 (2014).</li><li>Zhang, Y. V., Ni, J., Montell, C. <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+for+attractive+salt-taste+coding+in+Drosophila.">The molecular basis for attractive salt-taste coding in Drosophila.</a> Science. 340 (6138), 1334-1338 (2013).</li><li>Zhang, Y. V., Raghuwanshi, R. P., Shen, W. L., Montell, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Food+experience-induced+taste+desensitization+modulated+by+the+Drosophila+TRPL+channel.">Food experience-induced taste desensitization modulated by the Drosophila TRPL channel.</a> Nat Neurosci. 16 (10), 1468-1476 (2013).</li><li>Liman, E. R., Zhang, Y. V., Montell, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+coding+of+taste.">Peripheral coding of taste.</a> Neuron. 81 (5), 984-1000 (2014).</li><li>Rodrigues, V., Cheah, P. Y., Ray, K., Chia, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=malvolio,+the+Drosophila+homologue+of+mouse+NRAMP-1+(Bcg),+is+expressed+in+macrophages+and+in+the+nervous+system+and+is+required+for+normal+taste+behaviour.">malvolio, the Drosophila homologue of mouse NRAMP-1 (Bcg), is expressed in macrophages and in the nervous system and is required for normal taste behaviour.</a> EMBO J. 14 (13), 3007-3020 (1995).</li><li>Tanimura, T., Isono, K., Yamamoto, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Taste+sensitivity+to+trehalose+and+its+alteration+by+gene+dosage+in+Drosophila+melanogaster.">Taste sensitivity to trehalose and its alteration by gene dosage in Drosophila melanogaster.</a> Genetics. 119 (2), 399-406 (1988).</li><li>Weiss, L. A., Dahanukar, A., Kwon, J. Y., Banerjee, D., 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+and+cellular+basis+of+bitter+taste+in+Drosophila.">The molecular and cellular basis of bitter taste in Drosophila.</a> Neuron. 69 (2), 258-272 (2011).</li><li>French, A. S., 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=Dual+mechanism+for+bitter+avoidance+in+Drosophila.">Dual mechanism for bitter avoidance in Drosophila.</a> J Neurosci. 35 (9), 3990-4004 (2015).</li><li>Deshpande, S. A., 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=Quantifying+Drosophila+food+intake:+comparative+analysis+of+current+methodology.">Quantifying Drosophila food intake: comparative analysis of current methodology.</a> Nat Methods. 11 (5), 535-540 (2014).</li></ol>

وصفناها بروتوكول بسيطة لكنها فعالة لتحديد طعم الأفضلية في ذبابة الفاكهة. وتستخدم إصدارات هذا الفحص بشكل روتيني في التجارب لتحديد مساهمات من المستقبلات الذوقية (الموارد الوراثية) إلى إدراك نوعيات مختلفة (المر والحلو والحامض والمالح، وأومامي) من المركبات الذوق. ...

الحيوانات تستخدم chemosensation للتمييز الشروط المواتية بصرف النظر عن ظروف الحرمان. هذا التصور يمكن أن تكون حاسمة لأشياء مثل تحديد أفضل المصادر الغذائية، وتجنب المواد السامة أو تحديد أفضل التزاوج شريك 1. وغالبا ما تنقسم Chemosensation إلى عنصرين الحسية: حواس الشم والحواس الذوقية. وهناك سمة مميزة الرئيسية لهذه الحواس هي أن الشم (الرائحة) يستخدم لأخذ عينات من البيئة الكيميائية الغازية المحيطة بينما ذوق حاسة (طعم) يتطلب اتصال جسدي مع الركيزة غير قلق. كلا طرائق الحسية تحفز الاستجابات العصبية التي يتم تجهيزها وفك الشفرة في الدماغ لإنتاج جذابة أو مثير للاشمئزاز السلوك المناسب 2. هذه الحواس هي بالتالي حاسمة لبقاء الحيوان. ذبابة الفاكهة ذبابة الفاكهة هو كائن النموذج الذي لا تزال تنمو في شعبية للاستخدام في فهمجي كيف تنظر الحشرات الشم والذوق. ذبابة الفاكهة توفر مزايا هائلة على النظم نموذج الأخرى نتيجة للثروة من الأدوات الجينية المتاحة للتشريح المسارات الجزيئية الخلوية، والسلوكية. وقد تم العمل على مدى السنوات ال 15 الماضية بشكل خاص دور فعال في تشخيص هويات الخلوية محددة، مستقبلات الخلايا العصبية، ويشير الآليات التي تشارك في كل من الشم والذوق. الآن، يتم استخدام قوة الوراثة ذبابة الفاكهة لزيادة توضيح كيف يتم ترميز هذه العمليات على مستوى الخلايا العصبية واحدة ودائرة واحدة 3-6. وبالتالي، المقايسات التي توفر سجل بسهولة قراءات من التعديلات على مسارات الحسية هي حيوية للتقدم المستمر في هذه المجالات. في حين أن الكثير معروف حول كيفية إشارات الشم يتم ترميز ومعالجتها في الدماغ، ومن المفهوم أقل بكثير عن آليات مماثلة في مسار الذوقية. نحن هنا وصف بروتوكول والتي يمكن استخدامها للتأكد preferen الذوقم في ذبابة الفاكهة. ذبابة الفاكهة، مثل الثدييات، يفضلون عموما المركبات تذوق الحلو بدلا من المركبات تذوق مرارة. ويمكن استخدام أي مزيج من هذه المصادر الغذائية في هذا التصميم التجريبي لتحديد كيفية تأثير تغيرات جينية معروفة الاختيار الذوق. وبالإضافة إلى ذلك، يمكن بالمثل استراتيجيات التدخل الدوائي يتم تقييم لآثارها على تفضيل طعم الحيوانات. سهولة ومرونة هذا الاختبار يجعلها نموذجا مفيدا لفهم طبيعة الإدراك الذوقي في ذبابة الفاكهة.

<table border="0" cellpadding="0" cellspacing="0" width="1031">
	<tbody>
		<tr height="17">
			<td height="17" style="height:17px;width:543px;">Blue Food Coloring (Water, Propylene Glycol, FD&#38;C Blue 1 and Red 40, Propylparaben)</td>
			<td style="width:128px;">McCormick</td>
			<td style="width:111px;">N/A</td>
			<td style="width:251px;"></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Cryo/Freezer Boxes w/o Dividers</td>
			<td>Fisher</td>
			<td>03-395-455</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Dumont #5 Forceps</td>
			<td>Fine Science Tools</td>
			<td>11251-20</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Glacial Acetic Acid</td>
			<td>Fisher</td>
			<td>BP2401-500</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Leica S6 E Stereozoom 0.63x-4.0x microscope</td>
			<td>W. Nuhsbaum, Inc.</td>
			<td>10446294</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Petri Dish (100 x 15 mm)</td>
			<td>BD Falcon</td>
			<td>351029</td>
			<td>Reuseable if thoroughly washed and dried</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Quick-Snap Microtubes</td>
			<td>Alkali Scientific Inc.</td>
			<td>C3017</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Red Food Coloring (Water, Propylene Glycol, FD&#38;C Reds 40 and 3, Propylparaben)</td>
			<td>McCormick</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sucrose</td>
			<td>IBI Scientific</td>
			<td>IB37160</td>
			<td></td>
		</tr>
	</tbody>
</table>

taste preference assay for adult drosophila

Olfactory and gustatory perception of the environment is vital for animal survival. The most obvious application of these chemosenses is to be able to distinguish good food sources from potentially dangerous food sources. Gustation requires physical contact with a chemical compound which is able to signal through taste receptors that are expressed on the surface of neurons. In insects, these gustatory neurons can be located across the animal's body allowing taste to play an important role in many different behaviors. Insects typically prefer compounds containing sugars, while compounds that are considered bitter tasting are avoided. Given the basic biological importance of taste, there is intense interest in understanding the molecular mechanisms underlying this sensory modality. We describe an adult Drosophila taste assay which reflects the preference of the animals for a given tastant compound. This assay may be applied to animals of any genetic background to examine the taste preference for a desired soluble compound.

وأظهرت بعض النتائج نموذجية من المقايسات تفضيل طعم أدناه. في معظم التجارب سوف ينظر بعض الاختلاف في شدة التلوين البطن (الشكل 1). يعتبر أي التلوين في البطن سواء مكثفة أو ضعيفة على ابتلاع إيجابي. ولذلك فمن المستحسن للباحثين ليسجل الحيوانات في حي...

Watch this Scientific Journal Video about Taste Preference Assay for Adult Drosophila at JoVE.com

Taste Preference Assay for Adult Drosophila

Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

طعم الفحص تفضيل الكبار<em&gt; ذبابة الفاكهة</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

The overall goal of this procedure is to determine the relative preference of adult Drosophila from one chemical tastant over another. This method can help answer key questions in the Drosophila taste field. Such as which genes are necessary for selecting between sweet compounds or potentially toxic, bitter compounds.The main advantage of this technique, is that it is straightforward and does not require any specialized instrumentation. Demonstrating the procedure will be Andrew Bantel, a technician from my laboratory. To begin the experiment, saturate a cotton ball with water at the bottom of a standard fly vial.Next, collect flies in sets of 100 animals on a CO2 pad and then add the flies to a prepared vial. Use a cotton ball to secure the vials closed. Then, place the vials on their side in an environmentally controlled incubator.Prepare the controlled tastant of one millimolar sucrose, by combining 10 microliters of 100 millimolar sucrose solution, 13 microliters of red food coloring, and 977 microliters of water. Next, prepare the experimental tastant of five millimolar sucrose. Then, create the assay chambers, by obtaining a 100 millimeter by 15 millimeter plastic petri dish, and placing three microliter drops of controlled tastant as close to the edge of the plate as possible at the 12 o'clock position.At the three o'clock position, place three 10 microliter drops of experimental tastant as close to the edge of the plate as possible. Place another three drops of the controlled tastant at the six o'clock position. Then, place another three drops of the experimental tastant at the nine o'clock position.Next, empty one vial of 100 starved flies onto a CO2 pad, just long enough to anaesthetize all animals. Brush the animals into the middle of a prepared assay chamber, and cover it with the dish lid. Place the assay chamber into an opaque cardboard box.Then, label the outside of the box with the condition and genotype being tested. Move the entire setup into an incubator. After two hours, place the assay chambers still contained within cardboard boxes, directly into a negative 20 degree Celsius freezer until they are ready for quantitation.Allow a single assay chamber to warm up to room temperature. Under a dissection microscope, use a brush to group the animals based on the color of their abdomen. Red, blue, or purple.Record the number of animals in each grouping. This graph shows the effects of varying concentrations of blue dye, while maintaining a constant concentration of red dye, revealing an optimum combination of 1.3%red, to 1.0%blue. Drosophila are naturally attracted to high sucrose concentrations, as seen by a preference index value near one, when given the choice between five millimolar sucrose and one millimolar sucrose.This preference was reversed with acid, which dropped the preference index to near zero. While performing this procedure, it's important to remember to test the food coloring additives, were there affects on the preference assay? The concentrations of each dye should have no impact on the performance index.

Research

JoVE Journal

Neuroscience

Taste Preference Assay for Adult Drosophila

Methods to Assay Drosophila Behavior

Methods to Assay Drosophila Behavior

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Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

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ضوء الفحص ميول لدراسة الساعة البيولوجية الفطرية وPhotobehavior الخاضعة للرقابة في<em&gt; ذبابة الفاكهة</em&gt; يرقات

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer ...

A Single-fly Assay for Foraging Behavior in Drosophila

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Studies of neuronal activity are often performed using neurons from rodents less than 2 months of age due to the technical difficulties associated with ...

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

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تسجيل الكهربية من<em&gt; ذبابة الفاكهة</em&gt; Labellar طعم الشعيرات

The peripheral taste response of insects can be powerfully investigated with electrophysiological techniques. The method described here allows the ...

Novel Assay for Cold Nociception in Drosophila Larvae

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رواية الفحص لالباردة حس الألم في<em&gt; ذبابة الفاكهة</em&gt; يرقات

How organisms sense and respond to noxious temperatures is still poorly understood. Further, the mechanisms underlying sensitization of the sensory ...

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

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تحليل الفائق للسلوك الحركي في مقايسة الجزيرة المورفولوجية

Advances in next-generation sequencing technologies contribute to the identification of (candidate) disease genes for movement disorders and other ...

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التحليل الأيضي اليرقات melanogaster المورفولوجية و "العقول الكبار"

This protocol describes a method for measuring the metabolism in Drosophila melanogaster larval and adult brains. Quantifying metabolism in whole organs ...

Preparing Adult Drosophila melanogaster for Whole Brain Imaging during Behavior and Stimuli Responses

Preparing Adult Drosophila melanogaster for Whole Brain Imaging during Behavior and Stimuli Responses

إعداد الكبار Drosophila melanogaster لتصوير الدماغ كله أثناء السلوك والمحفزات الاستجابات

We present a method developed specifically to image the whole Drosophila brain during ongoing behavior such as walking. Head fixation and dissection are ...