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. 기아 <ol><li> 표준 플라이 유리 병의 하단에 18.2 MΩ 물 목화 공을 포화에 의해 기아 튜브를 비행 준비합니다. 다르게는, 유사 바이알 내의 각도 18.2 MΩ 물 장소 여과지의 작은 조각을 포화. </li><li> 준비된 유리 병에 파리를 추가 한 후 CO 2 패드 ~ 100 동물의 세트로 파리를 수집합니다. 참고 : 최상의 결과 미만 5 일 이전입니다 동물 얻을 수있다. 그러나, 동물의 실제 나이는 시간에 기?...

<ol>
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우리는 초파리 취향 우선 순위를 결정하기위한 단순하지만 효율적인 프로토콜을 설명 하였다. 이 분석의 버전은 정기적으로 맛 화합물 (쓴맛, 단맛, 신맛, 짠맛,​​ 감칠맛과) 다른 특성을 지각에 미각 수용체 (GRS)의 기여를 결정하기 위해 실험에 사용된다. 초파리 게놈 대체 접합 8,9에 의해 68 식별 미각 수용체를 인코딩 약 60 유전자가 포함되어 있습니다. 그러나, 이러한 이...

동물 불리한 조건에서 떨어져 유리한 조건을 구별하는 chemosensation를 사용합니다. 이 인식은, 최고의 음식 소스를 결정하는 독성 물질을 피하거나 가장 짝짓기 파트너 일을 결정하는 등의 것들에 대한 중요 할 수 있습니다. 후각의 감각과 미각 감각 : Chemosensation은 종종 두 개의 감각 구성 요소로 나누어 져 있습니다. 이러한 감각의 주요 구별되는 특징은 후각 (냄새) 맛보기 (맛) 비 휘발성 기판과 물리적 접촉을 필요로하는 동안 주위의 가스 화학 환경을 샘플링하는 데 사용되는 것입니다. 두 감각 양식을 처리하고 적절한 매력적인 또는 척력이 동작을 생성하기 위해 뇌에서 디코딩되는 신경 반응을 자극한다. 이 감각은​​ 동물의 생존을위한 따라서 중요합니다. 초파리 비행 열매는 이해에 사용하기 위해 인기가 성장을 계속하고 모델 생물이다곤충은 냄새와 맛을 감지하는 방법을 보내고. 초파리 때문에, 분자 세포, 행동 경로의 해부에 사용할 유전 도구의 재산에 다른 모델 시스템에 비해 엄청난 장점을 제공한다. 지난 15 년 동안 작업은 특정 세포 정체성, 신경 수용체를 특성화하고, 냄새와 맛 모두에 관여하는 메커니즘을 신호에 특히 쓸모있다. 지금, 초파리 유전학의 힘은 더욱 이러한 프로세스는 하나의 신경 세포 및 단일 회로 레벨 3-6에서 코딩하는 방법을 명료하게 사용되고있다. 따라서, 제공하는 분석은 쉽게 감각 경로에 변화의 판독이 분야의 지속적인 사전에 매우 중요 얻었습니다. 많은 코딩 뇌에서 처리하는 방법 후각 신호에 대해 알려져 있지만, 훨씬 덜은 미각 경로와 유사한 메커니즘에 대한 이해된다. 우리는 여기서 맛 preferen을 확인하는 데 사용될 수있는 프로토콜을 설명쓴 맛 화합물 반대로 초파리. 초파리의 CE 마크는 포유 동물처럼, 일반적으로 달콤한 맛 화합물을 선호합니다. 이러한 음식 소스의 조합은 알려진 유전 적 변화가 맛 선택에 영향을 미치는 방법을 결정하기 위해이 실험 설계에 이용 될 수있다. 또한, 약리학 적 개입 전략은 유사 동물의 맛이 환경에 미치는 영향에 대해 평가 될 수있다. 이 분석의 용이성과 유연성 초파리 미각 지각의 특성을 이해하는 것이 유용에게 패러다임을 만든다.

<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.

맛을 선호 분석에서 몇 가지 전형적인 결과는 다음과 같습니다. 대부분의 실험에서 복부 색상의 강도에 약간의 변화는 (그림 1) 볼 수 있습니다. 강한 또는 약한 여부 복부의 모든 색상이 양의 섭취로 간주됩니다. 잠재적 인 편견을 제한하기 위해 연구자들은 실험 조건에있는 동안 블라인드 동물을 득점하는 것은 그러므로 것이 좋습니다. <p class="jove_content" ...

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.

성인을위한 맛의 기본 설정 분석<em&gt; 초파리</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

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

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

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

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