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. भुखमरी <ol><li> एक मानक मक्खी शीशी के नीचे 18.2 MΩ पानी के साथ एक कपास की गेंद को संतृप्त द्वारा भुखमरी शीशियों के लिए उड़ान भरने को तैयार है। वैकल्पिक रूप से, इसी तरह शीशी के भीतर एक कोण पर 18.2 MΩ पानी और जगह के साथ ...

<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>

हम ड्रोसोफिला में स्वाद वरीयता निर्धारित करने के लिए एक सरल लेकिन प्रभावी प्रोटोकॉल का वर्णन किया है। इस परख के संस्करण नियमित तौर पर अलग अलग गुणों (कड़वा, मीठा, खट्टा, नमकीन, और umami) स्वाद यौगिकों की म...

पशु chemosensation का उपयोग हानिकर की स्थिति से अलग लाभप्रद शर्तों भेद करने के लिए। यह धारणा है, सबसे अच्छा भोजन स्रोत का पता लगाने जहरीले पदार्थ से बचने या सबसे अच्छा संभोग साथी 1 निर्धारण के रूप में ऐसी बातों के लिए महत्वपूर्ण हो सकता है। घ्राण होश और स्वाद इंद्रियों: Chemosensation अक्सर दो संवेदी घटकों में विभाजित है। इन इंद्रियों की एक मुख्य विशिष्ठ विशेषता है कि महक (गंध) के आसपास के गैसीय रासायनिक वातावरण का नमूना है, जबकि स्वाद (स्वाद) एक nonvolatile सब्सट्रेट के साथ शारीरिक संपर्क की आवश्यकता है प्रयोग किया जाता है। दोनों संवेदी तौर तरीकों न्यूरोलॉजिकल प्रतिक्रियाएं जो संसाधित और मस्तिष्क उचित आकर्षक या प्रतिकारक व्यवहार 2 का उत्पादन करने में डीकोड कर रहे हैं प्रोत्साहित। इन इंद्रियों इसलिए पशु अस्तित्व के लिए महत्वपूर्ण हैं। फल मक्खी ड्रोसोफिला मेलानोगास्टर एक मॉडल जीव जो समझ में उपयोग के लिए लोकप्रियता में वृद्धि जारी हैआईएनजी कैसे कीड़े गंध और स्वाद मानता है। फल मक्खियों आनुवंशिक आणविक, सेलुलर, और व्यवहार रास्ते के विच्छेदन के लिए उपलब्ध उपकरणों के धन के कारण अन्य मॉडल प्रणाली पर जबरदस्त लाभ प्रदान करते हैं। पिछले 15 वर्षों से काम विशिष्ट सेलुलर पहचान, न्यूरोनल रिसेप्टर्स निस्र्पक, और दोनों गंध और स्वाद में शामिल तंत्र संकेत में विशेष रूप से महत्वपूर्ण भूमिका निभाई है। अब, ड्रोसोफिला आनुवंशिकी की शक्ति आगे स्पष्ट करने के लिए कैसे इन प्रक्रियाओं एक न्यूरॉन और सिंगल सर्किट स्तर 3-6 से कोडित रहे हैं इस्तेमाल किया जा रहा है। इसलिए, assays जो प्रदान आसानी से रन बनाए संवेदी रास्ते के लिए परिवर्तन की readouts इन क्षेत्रों के सतत अग्रिम के लिए महत्वपूर्ण हैं। एक बड़ा सौदा कैसे घ्राण संकेतों कोडित और मस्तिष्क में कार्रवाई कर रहे हैं के बारे में जाना जाता है, बहुत कम स्वाद मार्ग में इसी प्रकार के तंत्र के बारे में समझा जाता है। हम यहाँ एक प्रोटोकॉल जो स्वाद preferen पता लगाने के लिए इस्तेमाल किया जा सकता का वर्णनड्रोसोफिला। ड्रोसोफिला में सीई, स्तनधारियों की तरह, आम तौर पर मीठा स्वाद यौगिकों के रूप में पसंद करते हैं कड़वा स्वाद यौगिकों करने का विरोध किया। इन खाद्य स्रोतों का कोई भी संयोजन निर्धारित करने के लिए कैसे जाना जाता आनुवंशिक परिवर्तन स्वाद पसंद को प्रभावित इस प्रयोगात्मक डिजाइन में उपयोग किया जा सकता है। इसके अलावा, औषधीय हस्तक्षेप रणनीतियों इसी तरह &#39;जानवरों के स्वाद पसंद पर उनके प्रभाव के लिए मूल्यांकन किया जा सकता है। आसानी और इस परख का लचीलापन यह ड्रोसोफिला में स्वाद धारणा की प्रकृति को समझने के लिए एक उपयोगी प्रतिमान बना देता है।

<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.

स्वाद पसंद assays से कुछ विशिष्ट परिणाम नीचे दिखाई जाती हैं। सबसे प्रयोगों में पेट रंग की तीव्रता में कुछ बदलाव (चित्रा 1) में देखा जाएगा। चाहे वह तीव्र या कमजोर पेट में किसी भी रंग एक सकार...

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.

एडल्ट के लिए स्वाद पसंद परख<em&gt; ड्रोसोफिला</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

परख के लिए तरीके ड्रोसोफिला व्यवहार

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

Drosophila Adult Olfactory Shock Learning

Drosophila Adult Olfactory Shock Learning

ड्रोसोफिला वयस्क घ्राण शॉक प्रशिक्षण

Drosophila have been used in classical conditioning experiments for over 40 years, thus greatly facilitating our understanding of memory, including the ...

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

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

में सहज और Circadian नियमित Photobehavior अध्ययन करने के लिए लाइट पसंद परख<em&gt; ड्रोसोफिला</em&gt; लार्वा

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

A Single-fly Assay for Foraging Behavior in Drosophila

Foraging व्यवहार के लिए एक एकल मक्खी परख में<em&gt; ड्रोसोफिला</em

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video ...

Membrane Potential Dye Imaging of Ventromedial Hypothalamus Neurons From Adult Mice to Study Glucose Sensing

झिल्ली वयस्क चूहों से ventromedial Hypothalamus न्यूरॉन्स की संभावित डाई इमेजिंग ग्लूकोज सेंसिंग अध्ययन करने के लिए

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

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

से electrophysiological रिकॉर्डिंग<em&gt; ड्रोसोफिला</em&gt; Labellar Sensilla स्वाद

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

Novel Assay for Cold Nociception in Drosophila Larvae

में शीत Nociception के लिए उपन्यास परख<em&gt; ड्रोसोफिला</em&gt; लार्वा

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

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

Drosophila द्वीप परख में हरकत व्यवहार के उच्च प्रवाह विश्लेषण

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

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

Drosophila melanogaster लार्वा और वयस्क दिमाग का चयापचय विश्लेषण

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

व्यवहार और उत्तेजनाओं प्रतिक्रियाओं के दौरान पूरे मस्तिष्क इमेजिंग के लिए वयस्क ड्रोसोफिला मेलानोगेस्टर तैयार करना

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