기금은 국립 과학 재단 (MF)에 의해 제공됩니다.

소개 OMT는 &quot;가상 플륨 시뮬레이터&quot;를 통합할 수 있도록 설계된 자기 테더 시스템 [1]의 적응이다. 다음 프로토콜은 설명 방법을 제대로 테더 빠르다 (1 부)과 자석 (2 부) 및 질량 흐름 규제 악취 전달 시스템 (Part3)을 설정하는 전략을 제공합니다. 여기에서 설명한 내용이 특정 시스템에 최적화된이며 다른 기술 사양과 구성 요소에 따라 다를 수 있습니다. 1 부 : Tethering의 파리 여기에서 설명한 tethering에 대한 단계는 많은 이전의 연구와 유사합니다 [2]하지만, OMT에 사용하기 위해 적응 [3]. 적절한 tethering는 강력하고 반복적인 실험 실험을 보장하기 위해 중요합니다. 조심 tethering의 중요성은 overstated 수 없습니다. <ol><li> 4-6일 게시물 eclosion을 날아 수집 4~6시간 그들을 굶어. kimwipe은 탈수를 방지하기 위해 foodless 유리병에 물을 moistened 놓습니다. </li><li> 작은 유리병에 병에서 파리의 배치를 전송하고 섭씨 약 4도 정도에서 설정 펠티어 냉각 단계에서 황동 블록에 삽입. 적정 온도는 약 30 초 내에 파리를 anesthetizes. </li><li> 냉각 단계의 차가운 표면에 파리를 정렬하십시오. 젖은 Kimwipe은 표피에서 지속적인 냉각과 수분 위킹을 보장합니다. tethering에 대한 가장 큰 여자를 선택합니다. </li><li> 좋은 브러시를 사용하여 하나의 야생 타입 여성 과일 비행을 수용하기 위해 모양 사용자 지정 - 내장 관의 위치에 하나의 파리 밀어. <img src="/files/ftp_upload/old/1063_2008_11_26_2/1063_Figure-01_thumb.png" alt="" /> 그림 1. <a href="https://jove.unicartagenaproxy.elogim.com/files/1063_2008_10_28_1/1063_Figure-01.png">그림의 더 큰 버전을 보시려면 여기를 클릭하십시오.</a> </li><li> 우리는 micromanipulator에 장착된 자석 막대와 minutien 핀 (벌금 과학 도구)를 위치. 핀은 무딘 끝이 단지 두 날개 사이에 머리 뒤에 지느러미 근육을 건드리면 이러한 정렬하는 것입니다. 머리에 눈을 향해 볼 때 핀 완벽하게 수직으로해야합니다. 파리와 핀 사이의 &#39;롤&#39;의 비록 작은 금액은 묵인할 수 없습니다. 핀과 신체의 긴 축 사이의 사이드 뷰 각도가 수평선 (그림 1)에 상대적인 30 도의 코를 접속 각도로해야합니다. 참고 : 자석 밧줄에 최적의 작업은 핀을 짧게 길이로 줄어들 수 할 수 있습니다. 이것은 서스펜션 자석 사이의 거리에 전적으로 달려 있습니다. </li><li> minutien 핀이 정렬되면, minutien 핀의 뭉툭한 끝에 UV - 활성 접착제 (ElectroLite 코프)의 작은 방울을 적용하는 얇은 강철 와이어를 사용합니다. </li><li> 접착제의 비말 그냥 머리 위의 파리의 흉부를 만지는 있도록 핀을 조작. 적절한 드롭 크기, 접착제 어닐링해야하며 흉부에 핀의 흐름. 너무 많은 접착제를 사용하지 않도록주의하십시오. </li><li> 자외선의 두 20초 파열로 접착제 치료. </li><li> 옆으로 고정 파리를 놓고 반복합니다. Kimwipe 작은 광장을 얇게하고 집게로 비행을 억제하기 위해 각 곁에 비행을 제공합니다. </li><li> 복구 한 시간 후, 닿는 파리 실험을 준비하고 있습니다. </li><li> 부적 절한 gluing는 자석 분야에서 수행하는 비행의 능력을 방해 수 있습니다. 모든 비행은 비행이 제대로 접착되도록하기 위해 모든 실험의 시작 원활한 회전을위한 확인하여야한다. 이것은 강한 optomotor 응답을 이끌어하기 위해 충분히 큰 공간 기간 줄무늬 패턴을 회전하여 수행할 수 있습니다 [4]. 비행이 잘못된 피치 각도로 접착하거나 롤 축에 대하여 핀에 졌다면, 즉시 수 있습니다 (I)하지 전혀 스핀하고, (ii) 운동의 제한 범위를 가지고, 또는 (iii) 일관성에 회전 속도. 위의 경우에는 즉시 폐기되어야하고 새로 교체. 이 문제를 해결하는 경우가 대부분 부적 절한 gluing의 경우되었습니다. (파리를 교체해도 문제가 해결되지 않으면, 부 2.4 참조) </li></ol> 2 부 : 자석 정렬 자석 (희토류 - magnets.com) 정렬하면 제대로 동작의 부드러운 360도 범위를 달성하기 위해 비행을 위해 필요합니다. 제대로 자석을 정렬하려면 다음 단계를 수행하십시오. <ol><li> 낮은 링 자석은 플라스틱 슬리브에 의해 수평으로 개최하고 맑은 진공 챔버 위에 직접 배치됩니다 (그림 1, 제 3의 자세한 설명). 위쪽 막대 자석은 낮은 자석 위에 수직으로 약 3 / 4 &quot;고정됩니다. </li><li> 대략 직접 낮은 고리 자석의 중앙 지점 위의 상단 막대 자석을 배치하여 시각 위턱과 아래턱에 자석을 맞춥니다. 괜찮아, 규모 조정 micromanipulator (시스키유 주)에 상단 자석을 첨부하는 데 유용합니다. </li><li> 에폭시는 상단 자석의 아래쪽 표면에 V - 보석 베어링 (소형 부품)는 회전 마찰을 최소화하고 비행 위치를 표준화합니다. </li><li> 경기장에서 비행기를 놓고 수평 및 V를 조정날 때까지 상단에 자석의 ertical 위치는 원활하고 꾸준히 편주 축에서 360도 회전시킬 수 있습니다. 모든 비행은 자석이 올바르게 정렬되고 개별 minutien의 핀이 (부 1.11 참조) 손상되지 않도록 모든 실험의 시작 원활한 회전을위한 확인하여야한다. 자석이 제대로 날아 있습니다 (I)하지 전혀 회전 정렬되지 않은 경우 (2) 운동의 제한 범위 (III)이 일치하지 판매율에서 회전합니다. 문제가 새로 비행을 대체하여 해결하는 경우, 그것은 (1 부 참조) 대부분 gluing 문제로 인해되었습니다. 이러한 문제는 모든 비행을 위해 존재한다면, 그것은 가능성 자석 추가적인 조정이 필요합니다의 정렬에 문제가 있습니다. </li><li> 경고 : 재미있을 수있는 희귀한 지구 자석과 함께 즐기면서, 그들은 매우 위험합니다. 손상된 장비 및 / 또는 가벼운 신체적 상해의 결과를 오용. 이러한 유혹 아직 쉽게 과소 평가 자석과 극단주의 운동. </li></ol> 파트 3 : 냄새 배달 <ol><li> 악취 전달 시스템이 이전에 설명한 하나를 기반으로 [5]하고 최적화하는 가장 힘든 부분은 (그림 1)입니다. 그것은 인내와 재판과 - 오류가 많이 필요합니다. 안정적인 냄새 추적을 달성하려면 다음 단계를 수행하십시오. </li><li> 악취 시스템의 초기 셋업은 사용자 정의 지어진 물 / 냄새 유리병에 가스 멀티플렉서 (사블 시스템 국제)에서 협궤 테플론 튜브 (소형 부품)를 실행해야합니다. 이러한 튜브의 콘센트는 동일한 테플론 튜브를 사용하여 피하 포트 튜브에 직접 연결됩니다. </li><li> 명확한 아크릴 진공 챔버는 자석을 지원하기 위해 무대 아래에 위치합니다. 우리는 비행을 가까이 개방 진공을 가지고 있으며 개방의 직경을 줄이기 위해 진공 챔버에 연결된 비행 아래 장착된 4mm 유리관을 사용합니다. </li><li> 포셉를 사용하여 무대에 닿는 파리를 놓습니다. </li><li> 매력적인 냄새 (애플 시더 식초가 잘 작동), 그리고 시각 비행은 그 냄새를 추적하는지 확인을 켭니다. 파리는 냄새에 대한 선호를 사용하지 않으려면 나타나는 경우, 다음 세 단계를 따르십시오. 이것은 몇 파리, 여러 시련을 요구할 수 있습니다. </li><li> 파리는 냄새를 추적하기 위해 나타날 때까지 악취 포트의 위치를​​ 조정합니다. 우리는 파리의 머리에 냄새 micromanipulators에 포트 (시스키유 주)와 위치 4mm 지느러미와 3mm 앞쪽에를 탑재. </li><li> 파리는 냄새를 추적하기 위해 나타날 때까지 질량 유량을 조정하십시오. 우리의 설정은 7ml/min (사블 시스템 국제 MFC - 4). </li><li> 파리는 냄새를 추적하기 위해 나타날 때까지 진공의 흐름 속도를 조정합니다. 우리는 흐름 레귤레이터 (콜 파머)를 사용 13l/min에서 진공 흐름 속도를 설정할 수 있습니다. 참고 : 빌딩 진공 공급 라인이 플륨 구조의 변화를 일으키는 시간이 지남에 따라 변동될 수 있습니다. 그것은 일정한 진공 흐름 조건에 대한 유량 조절기에 외부 진공 펌프를 연결하는 것이 유용할 수 있습니다. </li><li> 항구와 흐름 속도의 위치가 설정되면, 정기적으로 가스 멀티플렉서를 사용하여 전환을 사용하여 구현 물 증기 및 냄새의 증기 사이에 스위치와 시각 냄새가 때 비행은 그 냄새를 추적하는지 확인합니다. 때때로 그 냄새 포트 내부의 잔류 냄새 그 냄새가 꺼져 냄새 추적을 이끌어내는 수 있습니다. 이것은 에탄올과 피하 튜브를 작성하고 Kimwipe와 함께 배달 끝에 수집하여 제대로 위치 악취 포트를 제거하지 않고 제거하실 수 있습니다. 압축 공기를 사용하여 남아있는 에탄올을 날려와 실험을 재개. </li></ol> 대표 결과 : 다음 단계가 제대로 준수하면 두 가지가 관찰해야합니다. 첫째, 지속적인 회전 시각적 자극이 아래 OMT에 배치 닿는 파리가 원활하게 편주 축에서 360도 회전한다. &quot;손으로&quot;전환에 (saccades라고도 함) 지속적인 시각적 자극의 부재에서 자연 터닝 동작의 특징입니다. 파리는 분명히 원활하게 회전 수없는 경우, 부적 절한 gluing (1 부 참조)하고 고르지 자석이 (2 부 참조) 비난 수 있습니다. 둘째, 그 냄새가 켜져있을 때, 즉시 그 냄새의 방향으로 머리를해야합니다. 냄새가 해제되면 파리가 무대에 하나의 위치에 대한 설정을 자발적 saccade해야합니다. 파리 적극적으로 악취 플륨, 악취 포트의 위치, 공기 흐름 속도 및 / 또는 진공 흐름 속도를 추적하지 않는 경우 (파트 3 참조) 조정이 필요할 수 있습니다.

<ol>
	<li>Bender, J. A., Dickinson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+stimulation+of+saccades+in+magnetically+tethered+Drosophila.">Visual stimulation of saccades in magnetically tethered Drosophila.</a> J. Exp. Biol. 209, 3170-3182 (2006).</li><li>Tammero, L. F., 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=Collision-avoidance+and+landing+responses+are+mediated+by+separate+pathways+in+the+fruit+fly,+Drosophila+melanogaster.">Collision-avoidance and landing responses are mediated by separate pathways in the fruit fly, Drosophila melanogaster.</a> J. Exp. Biol. 205, 2785-2798 (2002).</li><li>Duistermars, B. J., Frye, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crossmodal+visual+input+for+odor+tracking+during+fly+flight.">Crossmodal visual input for odor tracking during fly flight.</a> Curr. Biol. 18, 270-275 (2008).</li><li>Reiser, M. B., Dickinson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+modular+display+system+for+insect+behavioral+neuroscience.">A modular display system for insect behavioral neuroscience.</a> J. Neurosci. Methods. 167, 127-139 (2008).</li><li>Frye, M. 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=Motor+output+reflects+the+linear+superposition+of+visual+and+olfactory+inputs+in+Drosophila.">Motor output reflects the linear superposition of visual and olfactory inputs in Drosophila.</a> J. Exp. Biol. 207, 123-131 (2004).</li></ol>

파리 Tethering이 분야에서 안정적인 냄새 추적을 도출하는 것은 비교적 간단합니다. 그것은 연습, 인내와 시행 착오가 필요합니다. 이 분야의 많은 부분이 더욱 최적화된 또는 실험적인 잠재력에 변화를 얻을 수로 대체 수 있습니다. 예를 들어, 진공 챔버의 최신 버전이 훨씬 작은하며 무대 아래에 더 많은 공간을 허용합니다. 또한, 강도와 희토류 자석의 종류가 약간 변경될 수 있으며, 추가로 악취 포트는 냄새 유...

<table><tbody><tr><td>Cooling Stage</td><td>Teca</td><td>LHP-300CP</td><td></td></tr><tr><td>Brass Cooling Block</td><td>Custom Made</td><td></td><td>~4 x 8 x 2cm</td></tr><tr><td>Sarcophagus</td><td>Custom Made</td><td></td><td>Contact us for details</td></tr><tr><td>UV-Activated Glue</td><td>Kemxert corp</td><td>KOA300</td><td>1oz bottle lasts years</td></tr><tr><td>Panel System</td><td>Caltech</td><td>http://www.dickinson.caltech.edu/panels</td><td>[4]</td></tr><tr><td>Minutien Pins</td><td>Fine Science Tools</td><td>26002-20</td><td></td></tr><tr><td>Lower Ring Magnets</td><td>Rare-earth-magnets.com</td><td>NSN0615</td><td>5 - 0.75&quot;o.d. x 0.375&quot; i.d. x 0.125&quot;</td></tr><tr><td>Upper Rod Magnet</td><td>Rare-earth-magnets.com</td><td>NSN0750</td><td>0.125&quot; x 1&quot;</td></tr><tr><td>V-Jewel Bearing</td><td>Small Parts, Inc.</td><td>VJ-0469-01</td><td></td></tr><tr><td>MFC-4 Flow Regulators</td><td>Sable Systems International</td><td></td><td></td></tr><tr><td>Gas Multiplexer</td><td>Sable Systems International</td><td></td><td></td></tr><tr><td>Teflon Tubing</td><td>Small Parts, Inc.</td><td>STT-20-C</td><td></td></tr><tr><td>20 Gauge Hypodermic Tubes</td><td>Small Parts, Inc.</td><td>HTX-20R-06</td><td></td></tr><tr><td>Glass Tubing</td><td></td><td></td><td>4mm o.d.</td></tr><tr><td>Vacuum Flow Regulator</td><td>Cole-Parmer</td><td>EW-32464-52</td><td></td></tr></tbody></table>

a magnetic tether system to investigate visual and olfactory mediated flight control in drosophila

그것은 곤충들이 바람 스트림의 제목을 안정화시키기 위해 시각적 단서를 사용하는 것이 여러 해 동안 명확되었습니다. 많은 동물이 바람에 실시 냄새를 추적할 수 있습니다. 맞바람의 등, 영상 안정화는 냄새 추적에 직접 에이즈 추적. 그러나 후각 신호를 직접 바람의 단서에서 독립적으로 시각 추적 행동에 영향을합니까? 또한, neurophysiology 및 Drosophila에서 후각의 neurobehavioral 유전학 연구의 최근 홍수가 훨씬 더 기술적으로 세련되고 양적 행동 assays 동기가있다. 여기, 우리는 원래 좁은 판상 흐름 냄새 plumes과 무대를 갖춘하여 시력 실험 고안 자기 테더 시스템을 수정했습니다. 비행기는 작은 강철 핀 접착하고 자유롭게 편주 수 있도록 자기 분야에서 일시 중지되었습니다. 작은 직경의 음식 냄새 plumes은 배고픈 파리로 안정적인 추적을 도출, 비행의 머리 아래로 이동합니다. 여기서 우리는 자석을 정렬 악취 플륨을 고안하고, 안정적인 냄새 추적을 확인, tethering의 중요한 역학에 중점을두고 있습니다.

Watch this Scientific Journal Video about A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila at JoVE.com

A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila

여기 우리는 어떻게 밧줄 후각 자기 - 밧줄 (OMT) 기기에서 플라이를 설명합니다. 우리는 희토류 자석 및 악취 포트 및 최적의 냄새 추적을 달성하기 위해 자극 전달 및 진공 흡입 모두 질량 흐름 속도를 설정할 수를 정렬하는 방법에 대해 설명합니다.

Drosophila에서 Visual과 강한 중재 플라이트 컨트롤을 맡은 이후로 자기 테더 시스템

It has been clear for many years that insects use visual cues to stabilize their heading in a wind stream. Many animals track odors carried in the wind. ...

a-magnetic-tether-system-to-investigate-visual-olfactory-mediated

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

11.2K 조회수.  University of California, Los Angeles. 여기 우리는 어떻게 밧줄 후각 자기 - 밧줄 (OMT) 기기에서 플라이를 설명합니다. 우리는 희토류 자석 및 악취 포트 및 최적의 냄새 추적을 달성하기 위해 자극 전달 및 진공 흡입 모두 질량 흐름 속도를 설정할 수를 정렬하는 방법에 대해 설명합니다.

비디오: A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila

Early Metamorphic Insertion Technology for Insect Flight Behavior Monitoring

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on insects during their metamorphic development. Here, the implants are fused within the structure and function of the neuromuscular system as a result of metamorphic tissue remaking. The implants emerge with the insect where the development of tissue around the electronics during pupal development results in a bioelectrically and biomechanically enhanced tissue interface. This relatively more reliable and stable interface would be beneficial for many researchers exploring the neural basis of the insect locomotion with alleviated traumatic effects caused during adult stage insertions. In this article, we implant our electrodes into the indirect flight muscles of Manduca sexta. Located in the dorsal-thorax, these main flight powering dorsoventral and dorsolongitudinal muscles actuate the wings and supply the mechanical power for up and down strokes. Relative contraction of these two muscle groups has been under investigation to explore how the yaw maneuver is neurophysiologically coordinated. To characterize the flight dynamics, insects are often tethered with wires and their flight is recorded with digital cameras. We also developed a novel way to tether Manduca sexta on a magnetically levitating frame where the insect is connected to a commercially available wireless neural amplifier. This set up can be used to limit the degree of freedom to yawing &#8220;only&#8221; while transmitting the related electromyography signals from dorsoventral and dorsolongitudinal muscle groups.

We present a novel surgical procedure to implant electrodes in Manduca sexta during its early metamorphic stages. This technique allows mechanically stable and electrically reliable coupling with the neuromuscular tissue to study flight neurophysiology dynamics. We also present a novel magnetic levitation platform for tethered studies of insect yaw.

곤충 비행 동작 모니터링에 대한 초기 변성 삽입 기술

Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on ...

연구

행동분석학

High-resolution Measurement of Odor-Driven Behavior in Drosophila Larvae

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral paradigm, the experimenter usually assumes that the rapid diffusion of odorant molecules from the source leads to the creation of a stable gradient in the dish. To establish a quantitative correlation between sensory inputs and behavioral responses, it is necessary to achieve a more thorough characterization of the odorant stimulus conditions. In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

Drosophila의 유충에서 냄새 기반 행동의 고해상도 측정

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral ...

생물학

Operant Learning of Drosophila at the Torque Meter

For experiments at the torque meter, flies are kept on standard fly medium at 25&deg;C and 60% humidity with a 12hr light/12hr dark regime. A standardized breeding regime assures proper larval density and age-matched cohorts. Cold-anesthetized flies are glued with head and thorax to a triangle-shaped hook the day before the experiment. Attached to the torque meter via a clamp, the fly's intended flight maneuvers are measured as the angular momentum around its vertical body axis. The fly is placed in the center of a cylindrical panorama to accomplish stationary flight. An analog to digital converter card feeds the yaw torque signal into a computer which stores the trace for later analysis. The computer also controls a variety of stimuli which can be brought under the fly's control by closing the feedback loop between these stimuli and the yaw torque trace. Punishment is achieved by applying heat from an adjustable infrared laser.

Measuring the yaw torque of tethered Drosophila with the torque meter allows the neuroscientist exquisite control of the stimulus situation of the experimental animal. Together with the unique genetic tools available in the fruit fly, this paradigm is used for a wide variety of neurobiological research.

토크 미터에서 Drosophila의 Operant 학습

For experiments at the torque meter, flies are kept on standard fly medium at 25&deg;C and 60% humidity with a 12hr light/12hr dark regime. A standardized ...

Visually Mediated Odor Tracking During Flight in Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Additionally, recent advances in olfactory molecular genetics and neurophysiology have motivated novel quantitative behavioral analyses to assess the behavioral influence of (e.g.) genetically inactivating specific olfactory activation circuits. We modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. Here we focus on experiments that can be performed after a fly is tethered and is able to navigate in the magnetic arena. We show how to acquire video images optimized for measuring body angle, how to judge stable odor tracking, and we illustrate two experiments to examine the influence of visual cues on odor tracking.

Here we describe how to optimize the acquired video image for an olfactory magnetic-tether (OMT) apparatus. We also describe two sample experimental protocols for studying visuo-olfactory fusion.

Drosophila의 비행 중 시각 인한 악취 추적

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual ...

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.

의 이스케이프 문제의 Optogenetic의 자극<em&gt; Drosophila melanogaster</em

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

신경과학

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To complicate matters even further, the odors detected by the insect nervous system usually are not single compounds but mixtures whose composition and concentration ratios vary. This leads to an almost infinite amount of different olfactory stimuli which have to be evaluated by the nervous system.
To understand which aspects of an odor stimulus determine its evaluation by the fly, it is therefore desirable to efficiently examine odor-guided behavior towards many odorants and odor mixtures. To directly correlate behavior to neuronal activity, behavior should be quantified in a comparable time frame and under identical stimulus conditions as in neurophysiological experiments. However, many currently used olfactory bioassays in Drosophila neuroethology are rather specialized either towards efficiency or towards resolution.
Flywalk, an automated odor delivery and tracking system, bridges the gap between efficiency and resolution. It allows the determination of exactly when an odor packet stimulated a freely walking fly, and to determine the animal&#180;s dynamic behavioral reaction.

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

The automated tracking system Flywalk is used for high-resolution quantification of odor-guided behavior in Drosophila melanogaster.

에서 냄새 유도 행동의 고해상도 정량화<em&gt; 초파리</em사용&gt;<em&gt; Flywalk</em&gt; 패러다임

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

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 ...

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an excellent model system for such investigations due to its complex behaviors, powerful genetic techniques, and compact nervous system. Laboratory behavioral assays have long been used with Drosophila to simulate properties of the natural environment and study the neural mechanisms underlying the corresponding behaviors (e.g. phototaxis, chemotaxis, sensory learning and memory)1-3. With the recent availability of large collections of transgenic Drosophila lines that label specific neural subsets, behavioral assays have taken on a prominent role to link neurons with behaviors4-11. Versatile and reproducible paradigms, together with the underlying computational routines for data analysis, are indispensable for rapid tests of candidate fly lines with various genotypes. Particularly useful are setups that are flexible in the number of animals tested, duration of experiments and nature of presented stimuli. The assay of choice should also generate reproducible data that is easy to acquire and analyze. Here, we present a detailed description of a system and protocol for assaying behavioral responses of Drosophila flies in a large four-field arena. The setup is used here to assay responses of flies to a single olfactory stimulus; however, the same setup may be modified to test multiple olfactory, visual or optogenetic stimuli, or a combination of these. The olfactometer setup records the activity of fly populations responding to odors, and computational analytical methods are applied to quantify fly behaviors. The collected data are analyzed to get a quick read-out of an experimental run, which is essential for efficient data collection and the optimization of experimental conditions.

Olfactory Behaviors Assayed by Computer Tracking Of Drosophila in a Four-quadrant Olfactometer

We describe here a behavioral setup and data analysis method for assaying olfactory responses of up to 100 vinegar flies (Drosophila melanogaster). This system may be used with single or multiple olfactory stimuli, and adaptable for optogenetic activation or silencing of neuronal subsets.

의 컴퓨터 추적하여있는 분석 후각 행동<em&gt; 초파리</em&gt; 네 사분면의 후각에

A key challenge in neurobiology is to understand how neural circuits function to guide appropriate animal behaviors. Drosophila melanogaster is an ...

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a considerable amount of information has been generated regarding ORN responses to odorants, the role of specific ORNs in driving behavioral responses remains poorly understood. Complications in behavior analyses arise due to different volatilities of odorants that activate individual ORNs, multiple ORNs activated by single odorants, and the difficulty in replicating naturally observed temporal variations in olfactory stimuli using conventional odor-delivery methods in the laboratory. Here, we describe a protocol that analyzes Drosophila larval behavior in response to simultaneous optogenetic stimulation of its ORNs. The optogenetic technology used here allows for specificity of ORN activation and precise control of temporal patterns of ORN activation. Corresponding larval movement is tracked, digitally recorded, and analyzed using custom written software. By replacing odor stimuli with light stimuli, this method allows for a more precise control of individual ORN activation in order to study its impact on larval behavior. Our method could be further extended to study the impact of second-order projection neurons (PNs) as well as local neurons (LNs) on larval behavior. This method will thus enable a comprehensive dissection of olfactory circuit function and complement studies on how olfactory neuron activities translate in to behavior responses.

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

This protocol analyzes navigational behavior of Drosophila larva in response to simultaneous optogenetic stimulation of its olfactory neurons. Light of 630 nm wavelength is used to activate individual olfactory neurons expressing a red-shifted channel rhodopsin. Larval movement is simultaneously tracked, digitally recorded, and analyzed using custom-written software.

초파리 애벌레 동작 Optogenetic 후 각 신경의 자극에 대 한 응답 추적

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a ...

Animal Behavior

<h4>Drosophila as a Model Organism</h4>
The common fruit fly, Drosophila melanogaster, is a widely used model organism in biology, though it may be more commonly recognized as a fruit-loving pest. There are multiple reasons that make D. melanogaster an excellent experimental organism: their roughly two-week generation time allows studying multiple generations, they are easily maintained in very small tubes, and their sexual dimorphism enables investigators to easily distinguish between males that have black coloration towards the posterior and females that have banded abdomens.
Thomas Hunt Morgan and his students were among the first to make use of Drosophila, in the early 20th century. In 1933, Morgan received a Nobel Prize for his discovery of the role of chromosomes in heredity, which was founded on his observations of mutant Drosophila. Morgan continued his studies by generating new mutants with radiation. The presence of multiple mutants with discrete effects on the appearance of the fly enabled him to look at patterns of multifactor inheritance. A key aspect of his early work was the discovery of apparent violations of Mendel&#8217;s second law, which states that genes segregate independently of one another. Morgan&#8217;s observations indicated that genes will not segregate independently if they are joined together on the same chromosome. Furthermore, it became possible to determine gene locations on a chromosome relative to each other through analyses of how often they recombined thus triggering gene mapping studies to identify the location of a gene and distances between genes. Consequently, associates of Morgan&#8217;s lab received two additional Nobel Prizes for other studies of Drosophila genetics. Morgan&#8217;s success and the accumulating information about Drosophila inspired others to adopt them as experimental organisms, which added to our knowledge of them and progressively increased their value as a model organism. Other scientists have developed many mutations. Over 27,000 unique Drosophila lines are now maintained as stock cultures at the Drosophila stock centers and are readily available to laboratories that use them as tools for studying diverse aspects of biology, including behavior1.
<h4>Drosophila Ethology</h4>
Animal behavior, or ethology, is the study of how animals interact with each other and their environments. Although it is a relatively young biological field, ethology has important implications since an organism&#8217;s survival depends on how it behaves with regard to possible mates, food sources, and natural obstacles. Studying these organisms helps scientists to understand the contexts of these behaviors and the underlying mechanisms as to how they have evolved. Thus, a goal of the study of animal behavior is to find a genetic basis for the behaviors exhibited by individuals or groups of individuals. For example, kin selection, first proposed by William Hamilton, suggests that individuals will behave altruistically when making a sacrifice for a family member rather than a stranger2. This is because related individuals share more genes than unrelated ones, and the future reproduction by their relatives contributes in a way to their own fitness, or reproductive success. Hence, observing how organisms like Drosophila interact with their environment can allow us to understand how these behaviors are hard-wired. Additionally, these behaviors provide insight into how these organisms have adapted to cope with their environment and survive.
Directional behaviors are commonly studied to understand the basis of how and why an organism moves in its environment. Kinesis and taxis are two forms of directional behaviors. Kinesis involves stimulus-induced movements in random directions, such as the random movement of flies when chased away. On the other hand, taxis involves stimulus-induced movement towards a specific direction, such as moths flying towards a lightbulb. Taxis behaviors are positive if the animal moves towards the stimulus and negative if the animal moves away from the stimulus. Furthermore, taxis behaviors are named based on the specific stimulus that induces them. For example, geotaxis is a response to gravity, where positive geotaxis means that the organism moves with gravity. Phototaxis is movement in response to light, such as positive phototaxis of moths towards a lightbulb. Similarly, chemotaxis is attraction to or avoidance of an airborne chemical cue, such as the negative chemotaxis of many predators, including humans, in response to the smell of skunk spray. Furthermore, there are many other types of taxes that are studied by scientists, including aerotaxis, barotaxis, hydrotaxis, and magnetotaxis, that are movements in response to oxygen, pressure, water, and magnetic fields, respectively.
<h4>Study of Drosophila taxis behavior</h4>
A common and simple method to study Drosophila taxis behavior is to use a choice chamber, which allows the observation of a directional response of the fly to a particular stimulus, such as gravity, light, or a chemical. Drosophila are inserted in the center of the choice chamber and then allowed to wander freely about the chamber, which is designed so that alternative stimuli can be presented at opposing sides. After a period of wandering, the number of flies is counted on either side of the choice chamber. The number of flies on each side of the chamber is then analyzed with the Chi Square test to gauge whether or not there is a preference for a stimulus.
Comparison of directional behavior of mutant and wild-type Drosophila allows researchers to determine the role of the mutated gene in that specific behavior. Moreover, scientists are also trying to understand how animals decide what to do in the presence of combinations of stimuli, including conflicting stimuli3. Together, these studies can help predict how organisms will react to certain stimuli or changes in environmental factors.
<h4>References</h4>
<ol class="references">
	<li>Cook KR, Parks AL, Jacobus LM, Kaufman TC, Matthews K. New research resources at the Bloomington Drosophila Stock Center. Fly. 2010, Vol. 4, 1: 88-91.</li>
	<li>WD, Hamilton. Altruism and related phenomena, mainly in social insects. Ann Rev Ecol Systematics. 1972, Vol. 3, 193-202.</li>
	<li>Gepner R, Skanata MM, Bernat NM, Kaplow M, Gershow M. Computations underlying Drosophila photo-taxis, odor-taxis, and multi-sensory integration. eLife. 2015, Vol. 4, e06229.</li>
</ol>

Animal Behavior: Studying Taxis Behavior of Drosophila - Concept

동물 행동

Drosophila as a Model Organism
The common fruit fly, Drosophila melanogaster, is a widely used model organism in biology, though it may be more commonly ...

Drosophila에서 Visual과 강한 중재 플라이트 컨트롤을 맡은 이후로 자기 테더 시스템

요약

더 많은 비디오 탐색

이 비디오의 챕터

곤충 비행 동작 모니터링에 대한 초기 변성 삽입 기술

Drosophila의 유충에서 냄새 기반 행동의 고해상도 측정

토크 미터에서 Drosophila의 Operant 학습

Drosophila의 비행 중 시각 인한 악취 추적

의 이스케이프 문제의 Optogenetic의 자극

에서 냄새 유도 행동의 고해상도 정량화

의 Mechanosensation에 대한 행동 분석 클론의 MARCM은 기반

의 컴퓨터 추적하여있는 분석 후각 행동

초파리 애벌레 동작 Optogenetic 후 각 신경의 자극에 대 한 응답 추적

동물 행동