우리는 원고를 개선하기 위해 자신의 작품에 대해 4 익명의 검토를 부탁드립니다. 우리는 MBG과 YG에 재정적 지원 센터 국립 드 라 공들인 Scientifique, 감사 및 MMS에 Université 드 부르고뉴 및 연구의 프랑스 교육부. YG의 실험실에서 연구는 유럽 연구위원회 (ERC 그랜트, GliSFCo-311403 시작), 직원은 국립 드 라 공들인 (ANR-JCJC, GGCB-2010), 헌법위원회 지역 드 부르고뉴 (페이버) 및 CNRS에 의해 투자된다.

1. 시작하기 전에 <ol><li> 안정적이고 강력한 행동 표현형 베어링 isogenized 기준 재고를 사용합니다. 모든 잠재적 인 컨트롤 이기종 배경 대립 유전자를 전달할 수 있기 때문에,이 주식을 선택에 대한 일반적인 규칙은 없다. </li><li> 이후 단계에 필요한 다른 모든 주식을 역 교배이 컨트롤 변형을 사용합니다. 이 교잡 단계는 일반적으로 2 ~ 3 동질 기준 남성 5에 (가능한 횡단 오버 상동 염색체 사이에 허용하는) 하나의 처녀 여성의 적어도 5 연속 십자가로 표시됩니다. 이 단계는 다른 비행 주식 사이의 유전 적 배경을 균질화하는 것이 중요합니다. </li><li> 25 ℃에서 12 시간 빛 / 어둠주기 항생제 (0.4 % 메틸 파라 하이드 록시 - 벤조 에이트) 보완 표준 옥수수 가루 (9 %), 효모 (10 %), 한천 배지 (1.5 %)에 초파리의 주식을 유지 ° C. </li><li> 온도 제어 방까지 붉은 빛 아래 (25 ℃)에서 화학 감각 실험 달성(시각적 단서의 기여를 제거하고, 화학 감각 신호에 집중). 정기적으로 각 실험 사이의 영역을 환기하는 방의 공기를 갱신. </li></ol> Y-미로 분석을 사용하여 2. 후각 응답 <ol><li> 테스트 전에 젖은 종이 타월을 포함하는 유리 튜브에 25 ° C에서 16 ~ 18 시간 동안 파리를 굶어. </li><li> 두 개의 유리 병에 더 작은 플라스틱 병 (로드 유리 병)에 Y 형 커넥터에 가입하세요. 세 개의 작은 유리 병에 커넥터를 연결하고 밀봉 Y-미로를 얻기 위해 거품 마개를 통과 1 ㎖ 피펫 팁을 사용합니다. 두 개의 &quot;함정&quot;튜브를 형성하기 위해, 하나의 피펫 팁의 넓은 끝이 &quot;로드&quot;를 형성하기 위해 (그것의 결정을 일단 비행의 반환을 방지하기 위해 1 ~ 2 mm 직경) 두 피펫 팁의 좁은 끝을 잘라 관 (그림 1A). </li><li> 다만 &quot;함정&quot;병 (그림 1B)를 연결하기 전에, 한 ~ 6mm 직경의 필터 종이를 배치각각의 병. 한 여과지에 부 취제 용액 40 μL 및 제 2 필터 종이에 대응하는 용매의 40 μL를 추가. </li><li> &quot;로드&quot;유리 병에 열 4~9일 된 파리를 소개합니다. 이 동작을 10에 강한 효과를 가지고 있기 때문에,이 전송 중에 CO 2 마취를 사용하지 마십시오. 오히려 얼음에 간단한 냉각을 사용합니다. 마취 파리의 적절한 조작 최대한 피사체에 스트레스를 제한하는 것이 중요하다. <ol><li> 시각적 자극에게 최대한 방지하기 위해 (가능한 열원을 제한하는 LED 전구를 사용하여) 원적외선 빛 아래에서 25 ° C에서 Y-미로 일련의 테스트를 수행합니다. (; 그림 1C 왼쪽, 또는 오른쪽에 악취 물질 함유 튜브 및로드 관 앞이나 뒤에있는) Y-미로의 방향을 교대로주의하십시오. </li><li> 파리에 대한 몇 시간이 악취 물질 또는 용매를 포함하는 트랩 유리 병에 입력 할 수 있습니다. participati을 높이기 위해 24 시간 후 파리를 계산에 80 % 이상까지 최대 후각 인덱스 값 (Simonnet, 개인 통신)을 제공한다. </li><li> - /로드 파리의 총 수 (용매 튜브 수 냄새 튜브 번호) : 다음과 같은 공식을 사용하여 그 결과 후각 인덱스를 계산합니다. </li></ol></li><li> 세척 Y-미로 셋업 다음과 같이 하룻밤 RBS 35 MD의 해체 셋업을 흡수. 철저하게 수돗물로 헹구어. 마지막으로 탈 이온수로 세척하고 건조 할 수 있습니다. </li></ol> 데이터의 3. 통계 분석 <ol><li> 데이터 및 변수들에 따라 t-테스트, 일방향 ANOVA 또는 양방향 ANOVA를 수행한다. </li></ol>

<ol>
	<li>Mombaerts, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genes+and+ligands+for+odorant,+vomeronasal+and+taste+receptors.">Genes and ligands for odorant, vomeronasal and taste receptors.</a> Nat. Rev. Neurosci. 5, 263-278 (2004).</li><li>Silbering, A. F., Benton, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ionotropic+and+metabotropic+mechanisms+in+chemoreception:+'chance+or+design.">Ionotropic and metabotropic mechanisms in chemoreception: 'chance or design.</a> EMBO Rep. 11, 173-179 (2010).</li><li>Ziegler, A. B., Berthelot-Grosjean, M., Grosjean, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+smell+of+love+in+Drosophila.">The smell of love in Drosophila.</a> Front. Physiol. 4, (2013).</li><li>Silbering, A. F., 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=Complementary+function+and+integrated+wiring+of+the+evolutionarily+distinct+Drosophila+olfactory+subsystems.">Complementary function and integrated wiring of the evolutionarily distinct Drosophila olfactory subsystems.</a> J. Neurosci. 31, 13357-13375 (2011).</li><li>Grosjean, Y., 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=An+olfactory+receptor+for+food-derived+odours+promotes+male+courtship+in+Drosophila.">An olfactory receptor for food-derived odours promotes male courtship in Drosophila.</a> Nature. 478, 236-240 (2011).</li><li>Woodard, C., Huang, T., Sun, H., Helfand, S. L., Carlson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+analysis+of+olfactory+behavior+in+Drosophila:+a+new+screen+yields+the+ota+mutants.">Genetic analysis of olfactory behavior in Drosophila: a new screen yields the ota mutants.</a> Genetics. 123, 315-326 (1989).</li><li>Semmelhack, J. L., Wang, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Select+Drosophila+glomeruli+mediate+innate+olfactory+attraction+and+aversion.">Select Drosophila glomeruli mediate innate olfactory attraction and aversion.</a> Nature. 459, 218-223 (2009).</li><li>Budick, S. A., Dickinson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free-flight+responses+of+Drosophila+melanogaster+to+attractive+odors.">Free-flight responses of Drosophila melanogaster to attractive odors.</a> The Journal of Experimental Biology. 209, 3001-3017 (2006).</li><li>Martin, F., Charro, M. J., Alcorta, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+affecting+the+cAMP+transduction+pathway+modify+olfaction+in+Drosophila.">Mutations affecting the cAMP transduction pathway modify olfaction in Drosophila.</a> J. Comp. Physiol. A. 187, 359-370 (2001).</li><li>Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anaesthetising+Drosophila+for+behavioural+studies.">Anaesthetising Drosophila for behavioural studies.</a> Journal of Insect Physiology. 46, 439-442 (2000).</li><li>Ai, M., 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=Acid+sensing+by+the+Drosophila+olfactory+system.">Acid sensing by the Drosophila olfactory system.</a> Nature. 468, 691-695 (2010).</li><li>Becher, P. G., Bengtsson, M., Hansson, B. S., Witzgall, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flying+the+fly:+long-range+flight+behavior+of+Drosophila+melanogaster+to+attractive+odors.">Flying the fly: long-range flight behavior of Drosophila melanogaster to attractive odors.</a> J. Chem. Ecol. 36, 599-607 (2010).</li></ol>

저자가 공개하는 게 없다.

우리의 Y-미로 프로토콜은 이전에 설명한 프로토콜 9를 기반으로합니다. 그러나, 우리는 두 가지 주요 차이점을 소개합니다. 첫째, 우리는 그들이 용매 또는 용매 플러스 악취 물질을 포함하는 유리 병에 입력하는 것을 결정하면 돌아가는 비행을 방지하기 위해 좁은 피펫 팁을 사용합니다. 이 좁은 팁은 Y-미로의 악취 물질의 확산을 제한하는 데 유용합니다. 둘째, 우리는 Y-미로에 입력하?...

맛이나 후각을 통해 Chemoreception 동물의 생존을위한 중요한 감각 양상이다. 그것은 중요한 위험 또는 음식 소스의 검출에 필요한 신호뿐만 아니라 사회적 상호 작용을 제공한다. 또한 자신의 재생에 필요한 섹스 파트너를 찾기 위해 동물을하는 데 도움이됩니다. 20 년 이상, 노벨상은 &quot;후각 수용체와 후각 시스템의 조직의 그들의 발견에 대해&quot;2004 년 리처드 악셀과 린다 벅에 의해 작업 경력을 포함하여 집중적 인 연구는 분자 및 세포 기지를 나타 내기 위해 수행 된 후각, 2. 후각 인식을 해부하는 과학자의 마음에 드는 동물 모델 중 하나는 D.입니다 의 melanogaster. 이 곤충은 포유 동물과 유사한 세포 및 분자 냄새 코딩 전략을 공유한다. 과학계는이 과일 파리에있는 방향제의 역할을 연구하기 위해 다양한 행동 패러다임을 사용합니다. 이 테스트는 courtshi으로 복합 분석을 포함후각 등 다양한 감각 양식이, 남성의 구애 3을 유도하는 것이 중요합니다 P 테스트합니다. 다른 분석은 더 구체적 방향제의 역할을 해결하기 위해 개발되었다; 이들은 T-미로, Y-미로 트랩 분석, 4 개의 필드로 경기장과 바람 터널 4,5,6,7,8 있습니다. 이 문서에서 우리는 D를 사용하여 강력한 후각 응답을 제공하는 간단한 수정 된 Y-미로 분석을 제시 의 melanogaster. 우리의 셋업은 이전에 설명한 방법 9에 반대의 최종 팁을 사용합니다. 따라서, 우리의 Y-미로는 두 가지 장점이 있습니다. 파리가 그녀의 선택을 한 후에 첫째, 시스템의 반환을 방지 할 수 있습니다. 둘째, Y-미로의 모든 영역에서의 냄새 물질의 교환을 제한한다. 초파리는 종종 취제 포화를 방지하는 데 사용되는 공기 흐름에 매우 민감하기 때문에이 마지막 이점은 중요하다. 공기의 흐름과 실험 장치를 조정하려면 시간과 비용이 소모 될 것이다. 따라서, 우리의 Y-미로 분석은 EF를 나타냅니다ficient 초파리의 후각 성능을 테스트하는 빠른 방법.

<table border="0" cellpadding="0" cellspacing="0" width="835">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr>
			<td>Drosophila Polystyrene tube</td>
			<td>VWR europe</td>
			<td>734-2255</td>
			<td>30 x 25 mm&#160; 
			Y-maze</td>
		</tr>
		<tr>
			<td>Drosophila Borosilicate tube</td>
			<td>Dijon verre</td>
			<td></td>
			<td>95 X 25 mm 
			Y-maze</td>
		</tr>
		<tr>
			<td>Foam stopper</td>
			<td>Dutscher</td>
			<td>999038</td>
			<td> 
			Y-maze</td>
		</tr>
		<tr>
			<td>Y-shaped connector</td>
			<td>Europrix</td>
			<td>11020605</td>
			<td> 
			Y-maze</td>
		</tr>
		<tr>
			<td>100-1000&#181;l pipet tips</td>
			<td>Corning</td>
			<td>4868</td>
			<td>Join the following pipet tips to the Y-shaped connector. Cut 2 pipet tips at 65 mm from the wide end, and connect the narrow end (with a &#8764;2 mm opening) to 2 test vials. These openings will limit the U-turns once the flies enter the tubes containing the odors. Cut 1 pipet tip at 35 mm from the wide end, and connect it to the loading vial. 
			Y-maze</td>
		</tr>
		<tr>
			<td>Far-Red LED Bulb</td>
			<td>Rubin-Lacaque</td>
			<td>0RB180238</td>
			<td>625-630 nm</td>
		</tr>
		<tr>
			<td>Acetic Acid</td>
			<td>Sigma-Aldrich</td>
			<td>45725</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenylacetic Acid</td>
			<td>Sigma-Aldrich</td>
			<td>P16621</td>
			<td></td>
		</tr>
		<tr>
			<td>Yeast</td>
			<td>Sensient Flavors Strasbourg</td>
			<td>1018880464</td>
			<td></td>
		</tr>
		<tr>
			<td>Cornmeal</td>
			<td>eurogerm</td>
			<td>Farine de ma&#239;s</td>
			<td></td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>Kalys</td>
			<td>HP-697-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Methyl hydroxy 4 benzoate</td>
			<td>VWR international</td>
			<td>25605293</td>
			<td></td>
		</tr>
	</tbody>
</table>

testing drosophila olfaction with a y-maze assay

동물은 자신의 생존을 보장하는 환경에서 신호를 검출하는 것은 필수적이다. 이 목표로, 그들은 직접 접촉을 통해 이상 범위에서 동작하는 휘발성 분자에 대한 응답을 나타냅니다 후각을 통해, 같은 시각, mechanoreception, 청각, 맛을 통해 chemoperception 등의 환경 신호를 사용합니다. 휘발성 화학 분자 위험, 음식의 소스, 또는 개인 간의 통신의 검출에 대부분의 동물에 매우 중요한 신호입니다. 초파리는 후각의 세포 및 분자 기초를 탐구하는 과학자에 대한 가장 일반적인 생물학적 모델 중 하나입니다. 이 작은 곤충의 후각 능력을 강조하기 위해, 우리는 고전적인 마우스와 함께 사용되는 Y-미로 시험에 기초하여 수정 된 선택의 프로토콜을 설명합니다. Y-미로로 얻은 데이터는 더 나은 동물들은 끊임없이 변화하는 환경에 대처하는 방법을 이해하는 데 유용한 정보를 제공합니다. 우리는 공부를 단계별 프로토콜을 소개합니다이 Y-미로 분석을 사용하여 비행 탐색 응답에 냄새 물질의 영향.

그림 1D이 Y-미로 분석을 사용하여 두 개의 대표적인 응답을 보여줍니다. 그들은 크게 10 % 페닐 아세트산을 피하지 않는 반면, 주-S의 남성은 강하게, 증류수에 희석하여 10 % 초산을 피하십시오. 이러한 분석은로드 유리 병에 함께 배치 복제 당 10 명의 남성을 기준으로합니다. 이 프로토콜은 때때로 평균의 큰 표준 오차가 발생할 수 있습니다. 필요하다면, 대신 단지 10. 수학적으로, 한 ?...

Watch this Scientific Journal Video about Testing Drosophila Olfaction with a Y-maze Assay at JoVE.com

Testing Drosophila Olfaction with a Y-maze Assay

우리는 Y-미로 분석을 사용하여 초파리의 행동에 냄새 물질의 영향을 나타 내기 위해 선택 테스트를 제시한다.

테스트<em&gt; 초파리</emY-미로 분석과&gt; 후각

Detecting signals from the environment is essential for animals to ensure their survival. To this aim, they use environmental cues such as vision, ...

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Neuroscience

Testing Drosophila Olfaction with a Y-maze Assay

14.8K 조회수.  UMR-6265 CNRS.  우리는 Y-미로 분석을 사용하여 초파리의 행동에 냄새 물질의 영향을 나타 내기 위해 선택 테스트를 제시한다. 

비디오: Testing Drosophila Olfaction with a Y-maze Assay

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We developed a novel nanoinjection/electrophysiology assay that allows the detection of bioactive modulatory effects of compounds on the function of a neuronal circuit that mediates the escape response in Drosophila melanogaster3,4. Our in vivo assay, which uses the Drosophila Giant Fiber System (GFS, Figure 1) allows screening of different types of compounds, such as small molecules or peptides, and requires only minimal quantities to elicit an effect. In addition, the Drosophila GFS offers a large variety of potential molecular targets on neurons or muscles. The Giant Fibers (GFs) synapse electrically (Gap Junctions) as well as chemically (cholinergic) onto a Peripheral Synapsing Interneuron (PSI) and the Tergo Trochanteral Muscle neuron (TTMn)5. The PSI to DLMn (Dorsal Longitudinal Muscle neuron) connection is dependent on D&alpha;7 nicotinic acetylcholine receptors (nAChRs)6. Finally, the neuromuscular junctions (NMJ) of the TTMn and the DLMn with the jump (TTM) and flight muscles (DLM) are glutamatergic7-12. Here, we demonstrate how to inject nanoliter quantities of a compound, while obtaining electrophysiological intracellular recordings from the Giant Fiber System13 and how to monitor the effects of the compound on the function of this circuit. We show specificity of the assay with methyllycaconitine citrate (MLA), a nAChR antagonist, which disrupts the PSI to DLMn connection but not the GF to TTMn connection or the function of the NMJ at the jump or flight muscles.
Before beginning this video it is critical that you carefully watch and become familiar with the JoVE video titled "Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster " from Augustin et al7, as the video presented here is intended as an expansion to this existing technique. Here we use the electrophysiological recordings method and focus in detail only on the addition of the paired nanoinjections and monitoring technique.

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

A rapid in vivo assay to test for neuromodulatory compounds using the Giant Fiber System (GFS) of Drosophila melanogaster is described. Nanoinjections in the head of the animal along with electrophysiological recordings of the GFS can reveal bioactivity of compounds on neurons or muscles.

를 사용하여 화합물 Bioactivity위한 이점 Nanoinjection 및 화면에 대한 전기 생리학 분석 Drosophila melanogaster 거대 섬유 시스템

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We ...

연구

신경과학

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

분석의 방법 Drosophila 행동

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

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance imaging. However, use of such methods in animal models requires anesthesia and hence both alters the brain state and prevents behavioral measures. An alternative method is the use of in vivo microdialysis to take continuous measurement of brain extracellular fluid concentrations of glucose, lactate, and related metabolites in awake, unrestrained animals. This technique is especially useful when combined with tasks designed to rely on specific brain regions and/or acute pharmacological manipulation; for example, hippocampal measurements during a spatial working memory task (spontaneous alternation) show a dip in extracellular glucose and rise in lactate that are suggestive of enhanced glycolysis1-3,4-5, and intrahippocampal insulin administration both improves memory and increases hippocampal glycolysis6. Substances such as insulin can be delivered to the hippocampus via the same microdialysis probe used to measure metabolites. The use of spontaneous alternation as a measure of hippocampal function is designed to avoid any confound from stressful motivators (e.g. footshock), restraint, or rewards (e.g. food), all of which can alter both task performance and metabolism; this task also provides a measure of motor activity that permits control for nonspecific effects of treatment. Combined, these methods permit direct measurement of the neurochemical and metabolic variables regulating behavior.

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Modulation of hippocampally-dependent spatial working memory by direct intrahippocampal microinjection, accompanied and followed by in vivo microdialysis for metabolites in conscious, behaving animals.

Hippocampal 인슐린 Microinjection 및<em&gt; 생체에</em공간 메모리 테스트 기간 동안&gt; Microdialysis

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance ...

A Single-fly Assay for Foraging Behavior in Drosophila

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster. In a light-tight box illuminated by red light that is invisible to fruit flies, a camera linked to custom data acquisition software monitors the position of six flies simultaneously. Each fly is confined to walk in individual arenas containing a food odor at the center. The testing arenas rest on a porous floor that functions to prevent odor accumulation. Latency to locate the odor source, a metric that reflects olfactory sensitivity under different physiological states, is determined by software analysis. Here, we discuss the critical mechanics of running this behavioral paradigm and cover specific issues regarding fly loading, odor contamination, assay temperature, data quality, and statistical analysis.

A Single-fly Assay for Foraging Behavior in Drosophila

In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster.

먹이 찾아 돌아다 행동에 대한 단일 비행 분석에<em&gt; 초파리</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 ...

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response is a vision-based, reflexive behavior that has been observed in all vertebrates tested. Tadpole eyes are small so the tail flip response was used as alternative measure, which requires a trained technician to record the subtle response. We developed an alternative behavior assay based on the fact that tadpoles prefer to swim on the white side of a tank when placed in a tank with both black and white sides. The assay presented here is an inexpensive, simple alternative that creates a response that is easily measured. The setup consists of a tripod, webcam and nested testing tanks, readily available in most Xenopus laboratories. This article includes a movie showing the behavior of tadpoles, before and after severing the optic nerve. In order to test the function of one eye, we also include representative results of a tadpole in which each eye underwent retinal axotomy on consecutive days. Future studies could develop an automated version of this assay for testing the vision of many tadpoles at once.

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Xenopus laevis tadpoles prefer swimming on the white side of a black/white tank. This behavior is guided by their vision. Based on this behavior, we present a simple assay to test the visual function of tadpoles.

테스트 시각 기능에 대한 간단한 행동 분석<em&gt;는 Xenopus laevis의</em

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response ...

Live Imaging of Drosophila Larval Neuroblasts

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given their relevance to development and disease, understanding the mechanisms that govern asymmetric stem cell division has been a robust area of study. Because they are genetically tractable and undergo successive rounds of cell division about once every hour, the stem cells of the Drosophila central nervous system, or neuroblasts, are indispensable models for the study of stem cell division. About 100 neural stem cells are located near the surface of each of the two larval brain lobes, making this model system particularly useful for live imaging microscopy studies. In this work, we review several approaches widely used to visualize stem cell divisions, and we address the relative advantages and disadvantages of those techniques that employ dissociated versus intact brain tissues. We also detail our simplified protocol used to explant whole brains from third instar larvae for live cell imaging and fixed analysis applications.

Live Imaging of Drosophila Larval Neuroblasts

This protocol details a streamlined method used to conduct live cell imaging in the context of an intact larval brain. Live cell imaging approaches are invaluable for the study of asymmetric neural stem cell divisions as well as other neurogenic and developmental processes, consistently uncovering mechanisms that were previously overlooked.

의 라이브 영상<em&gt; 초파리</em&gt; 애벌레 Neuroblasts

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by stressed conspecifics due to an odorant released during stress known as the Drosophila stress odorant (dSO). Through the use of the T-maze apparatus, one can quantify the avoidance of the dSO by responder flies in a very affordable and robust assay. Conditions necessary to obtain a strong performance are presented here. A stressful experience is necessary for the flies to emit dSO, as well as enough emitter flies to cause a robust avoidance response to the presence of dSO. Genetic background, but not their group size, strongly altered the avoidance of the dSO by the responder flies. Canton-S and Elwood display a higher performance in avoiding the dSO than Oregon and Samarkand strains. This behavioral assay will allow identification of mechanisms underlying this social behavior, and the assessment of the influence of genes and environmental conditions on both emission and avoidance of the dSO. Such an assay can be included in batteries of simple diagnostic tests used to identify social deficiencies of mutants or environmental conditions of interest.

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster.

사회 회피의 정량화에 대한 간단한 분석<em&gt; 초파리</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

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.

의 Mechanosensation에 대한 행동 분석 클론의 MARCM은 기반<em&gt; 초파리</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 ...

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.

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 ...

Dynamic Quantitative Sensory Testing to Characterize Central Pain Processing

Central facilitation and modulation of incoming nociceptive signals play an important role in the perception of pain. Disruption in central pain processing is present in many chronic pain conditions and can influence responses to specific therapies. Thus, the ability to precisely describe the state of central pain processing has profound clinical significance in both prognosis and prediction. Because it is not practical to record neuronal firings directly in the human spinal cord, surrogate behavior tests become an important tool to assess the state of central pain processing. Dynamic QST is one such test, and can probe both the ascending facilitation and descending modulation of incoming nociceptive signals via TS and CPM, respectively. Due to the large between-individual variability in the sensitivity to noxious signals, standardized TS and CPM tests may not yield any meaningful data in up to 50% of the population due to floor or ceiling effects. We present methodologies to individualize TS and CPM so we can capture these measures in a broader range of individuals than previously possible. We have used these methods successfully in several studies at the lab, and data from one ongoing study will be presented to demonstrate feasibility and potential applications of the methods.

By assessing pain in response to repetitive or different types of standardized stimuli, dynamic quantitative sensory testing (QST) can reveal changes in the central processing of pain. We present methods to optimize and individualize two dynamic QST measures: temporal summation (TS) and conditioned pain modulation (CPM).

테스트

요약

더 많은 비디오 탐색

이 비디오의 챕터

를 사용하여 화합물 Bioactivity위한 이점 Nanoinjection 및 화면에 대한 전기 생리학 분석 Drosophila melanogaster 거대 섬유 시스템

분석의 방법 Drosophila 행동

Hippocampal 인슐린 Microinjection 및

먹이 찾아 돌아다 행동에 대한 단일 비행 분석에

테스트 시각 기능에 대한 간단한 행동 분석

의 라이브 영상

사회 회피의 정량화에 대한 간단한 분석

의 Mechanosensation에 대한 행동 분석 클론의 MARCM은 기반

성인을위한 맛의 기본 설정 분석

동적 정량적 감각 테스트는 중앙 통증 처리의 특성을