We are grateful to anonymous reviewers for comments and suggestions that greatly improved this manuscript. We thank R. Bowes, R. Carreno, and T. Panhuis for assistance in collecting the fish. We also thank M. Lee for assistance with male preference trials. We are grateful to the Ohio Wesleyan University Department of Zoology for helpful advice and suggestions throughout this study and Arizona State University (McGraw Lab) for software advice.

모든 실험은 오하이오 웨슬리 언 대학의 기관 동물 관리 및 사용위원회의 승인을 실시 하였다. 분광 광도계를 사용하여 물고기의 1. 녹화 UV 반사율 <ol><li> 파장의 범위 알려진 흰색 표준 분광 광도계와 광원을 보정하는 장비 또는 소프트웨어의 권고에 따라 측정한다. </li><li> 에틸 3- 아미노 벤조 에이트, 메탄 설폰 산염의 0.5 % 수용액에 넣어 물고기 마취 (MS-222)가 물고기가 응답 될 때까지 중탄산 나트륨을 동일한 양의 버퍼. 응답하면, 즉시 블랙, 비 반사 배경에 물고기를 배치합니다. 참고 : MS-222는 잠재적으로 독성, 장갑은 항상 착용해야한다. </li><li> 분광계 광원의 이상을 초과 빛을 최소화하기 위해 어두운 방에있는 모든 측정을 수행. </li><li> 유기체의 몸에 대해 유지 광섬유 프로브를 사용하여 측정 반사율. 하다일부 프로브 설계에 내장 된 바와 같이, 광섬유와 본체 사이의 일관성 45 ° 각도를 형성해야. 주 : 측정 일관성을 위해, 다수의 스캔 (2 - 5)에 걸쳐 평균하는 것이 바람직 할 수있다 바디 영역마다. </li><li> 적절한 반사율 분석 소프트웨어와 함께 기록 반사율 데이터. 참고 : 통합 시간 = 2000 초, 평균 검사 = 2, 및 픽셀 다듬기 = 5. 데이터 복사 및 스프레드 시트에 붙여되었다 여기에 사용되는 소프트웨어로 설정했다. </li><li> 존재 또는 UV 반사율 및 생물에서 표현형 변이의 범위의 유무를 결정하기 위해 여러 신체 영역에 반복합니다. 주 : 예비 스캔이있는 경우, 본문 부분이 지속적으로 UV 마크를 표시하는 판별 할 필요가있다. </li><li> 완전히 회복 될 때까지 정상 아가미의 비트와 수영의 행동에 의해 입증, 잘 산소 유지 수족관에 물고기를 돌려줍니다. 참고 : 몸 피부 회복에 도움을 첨가 할 수있다에 상업 보호제는 점액 코팅을 복원 할 수 있습니다. </li><li> 대표원하는 경우 남녀 모두를 포함, 여러 개인에서 먹는다. </li><li> 모든 데이터가 X 축의 Y 축 및 파장에 대한 반사율, 산점도를 사용하여 플롯 데이터 복사 된 스프레드 시트. 자외선 영역에서의 피크 진폭 (300-400 나노 미터)가 물고기 UV 반사율의 존재를 나타낸다. </li></ol> 2. 이분법 적 결합 재판 주 : 초점 물고기, UV 필터, 불투명 필터를 대신하는 제어 관측을 수행. 불투명 필터가 제거 된 이들 실험에서, 초점 물고기 UV 광의 유무를 표시하지만, 잠재적 결합 파트너 결여 탱크의 일부를 노출한다. 이 실험에서 이분법 적 선택 시험의 유효성에 대한 자세한 내용은 참조하십시오 이러한 테스트도 가지고 있습니다하지만 파머와 Hankison (2015) 10, 11 단점. <ol><li> 초점 생물 등의 성적으로 받아들이는 개인을 사용합니다. 여성 P.를 사용하여 latipinna &lt;48 시간 산후 5차 남성은 이전에 성적 동기 부여의 가능성을 개선하기 위해 테스트에 최소 24 시간 동안입니다. </li><li> 이 선택 구획에 인접 중성 영역으로 하나의 초점 개인을 놓습니다. P.하십시오 두 개의 동일한 크기의 선택 구획에 의해 측면에 초점 개인에 대한 중앙 영역을 형성, 투명한 플렉시 유리 파티션에 의해 세 부분으로 나누어, latipinna는 75.7 L 테스트 수족관 (그림 1 30.5 cm X 30.5 cm X 75cm)를 사용합니다. 물이나 후각 단서 개별 탱크 섹션간에 공유되지 않았는지 확인합니다. 각 개인이 관찰 될 때까지 반복합니다. </li></ol><img alt="그림 1" src="/files/ftp_upload/54453/54453fig1.jpg" /> 그림 1. 실험 수족관 설치 세 부분으로 나누어 직사각형 수족관 :. 초점 개인과 개체 쌍을 개최 두 끝 구획을 유지 중앙 영역 (그림은 끝에 객체 남성과 중간에 여성을 보여줍니다). 교환 FIL어느 차단 또는 허용 자외선 반사율이 수족관의 남성과 여성의 섹션을 나누어 유리 파티션 위에 배치 할 수 TERS. 이 그림은 파머와 Hankison (2015) 10에서 수정됩니다. <a href="http://ecsource.jove.com/files/ftp_upload/54453/54453fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <ol start="3"><li> 풀 스펙트럼 조명 조건 (그 스펙트럼 UV-A 범위로 확장 조명)에서 테스트 수족관의 각 끝 구획 (선택 함) 하나의 잠재적 인 짝짓기 파트너를 찾아냅니다. 두 개의 격리 된 결합 파트너 사이의 주요 차이점은 그들이 자외선 광을 투과 블록 또는 필터 뒤에 표시할지 여부되도록 잠재적 인 짝짓기 파트너, 크기, 착색 및 디스플레이 특성에 최대한 가깝게 서로를 닮아 있는지 확인합니다. 주 :이 최상의 컬러 매칭 알 수 있도록 가시 광선 영역의 반사율을 기록하는 것이 바람직 할 수있다여기에 비록 일치가 눈에 의해 수행되었다. </li><li> 15 분 동안 시각 격리에서 개인 (불투명 필터) 적응. 그 자외선 +를 확인하고 - 필터가이 시간에 장소에 있습니다. 순응 후, 불투명 한 필터를 제거합니다. 그 측면 바이어스가 (에 관계없이 객체의 탱크의 한면에 대한 고유의 환경 설정) 존재하지 않습니다 보장하기 위해 무작위로 UV +와 UV를 할당 - 탱크의 측면을 각 각 재판. </li><li> 스톱워치를 이용하여 초점 개인 (약 2 표준 본체 길이,이 경우, 선택 구획 근방) 특정 환경 영역에서 소비되는 시간을 기록한다. 여성 물고기의 경우,이 남성 초점 물고기에 대한이 두 남성의 각 근처에 소비 시간의 양, 그리고 바이스​​ 마찬가지입니다. 주 : 이벤트 레코더 소프트웨어는 기록에 사용될 수있다. 15 분의 시험은 응답하지 않는 것으로 한 번 이상 결과에서 제외 된 각각의 환경 설정 영역을 방문하지 않은 실험과 생선에 사용되었다. </li><li> 기록 물고기 비어 UV + 및 UV 부근 보내는 시간 분석 - 물고기의 상대 선택에 영향을 줄 수있는 특정 조명 환경쪽으로 고유 바이어스가되지 않도록 구획. 개인이 특정 UV 조건을 선호하는 경우 결정하기 위해 쌍 t-테스트를​​ 사용합니다. </li></ol>

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The authors have nothing to disclose.

측정법은 P.에 UV 표시를 식별 성공 latipinna. P.의 두 남녀 latipinna는 측면을 따라 UV 표시를 가지고있다. 또한, 일부 남성은 자신의 등 지느러미에 UV 표시를했다, 특성은 이전에 여성 짝짓기 환경 설정 7에 중요한 것으로 나타났습니다. 우리는 UV 마크의 존재를 검출하기위한 메커니즘으로서 UV 분광 광도계의 사용을 권장. 또한 테스트는 짝짓기...

단서과 동물의 사회적 상호 작용에 사용되는 신호를 이해하는 것은 우리가 내 종 중 모두 표현형의 변화를 이해 할 수 있습니다. 이 변형은 개체군 발산 성 선택 및 분화와 같은 진화 과정에서 중요한 역할을한다. 종종, 그러나, 연구진은 인간의 감각 기관, 시각 또는 청각 영역 내에서 특히 가장 확실한 단서를 탐험 제한됩니다. 분광 광도법의 사용은, 그러나, 인간의 가시 스펙트럼을 넘어 다른 종의 사회적 상호 작용에 중요 할 수있다 파장에 우리의 조사를 확장하는 우리를 할 수 있습니다. 특히, 자외선에 의해 제공되는 단거리 통신 (UV; 300-400 ㎚) 감도 메이트 선택 일 동안 매우 유리할 가능성이있다. 조류 및 어류의 대부분 시각적 사냥 육식 동물, 예를 들어 UV 방사선을 검출 할 수 없다. 남성 정교하게 표시 시스템에서자외선 스펙트럼을 이용하기보다는 가시 스펙트럼 2,3-에서 검출 신호를 개발하여 메이트를 유치 할 수있는 능력을 유지하면서 하나의 유기체들 각각과 통신하는 가능성을 고려하지 못하면 암컷이 수컷 .. 포식의 위험을 감소시킬 다른 사용하여 이러한 &quot;개인 통신 채널&quot;, 행동과 진화의 중요한 드라이버를 놓칠 수있다. 이 프로토콜은 SailFin을 몰리, Poecilia latipinna, 기존에 UV를 감지하거나 UV 마크를 활용하는 능력을 알려져 있지 않은 일부 다처제 물고기 메이트 선택을위한 UV 큐의 사용에 대한 조사를 간략하게 설명합니다. 이 물고기 종은 다른 자외선에 민감한 라이브 베어링 물고기 (4)에 가까운 계통 근접을 가지고 있으며, microspectroscopic 증거가있다 P. 이러한 P. 다른 몰리 종과 함께 latipinna, 멕시 카나와 P. 포모사 공동을 담당 시각 세포 (콘 클래스를 가지고UV 파장 5에 가장 민감 LOR 비전). 이 성적으로 동종 이형 종에서는, 여성의 선택은 남성 &#39;밝은 색상과 확대 핀 6-9의 발전에 강한 역할을하고있다. 이 방법은 우리가 UV는 여성이 남성의 품질을 평가하는 추가 매체인지 여부를 탐색 할 수 있습니다. 검출 및 P.에 UV 마크의 측정 광섬유 프로브를 여기에 자세히 설명되어 latipinna 분광 광도계. 또한, 수용 여성 mollies가 차등 UV-A를 포함한 전체 스펙트럼 빛을 전송하는 광학 필터를 통해 본 남성과 연관 여부 ([UV +] 320-700 ㎚) 및 자외선 차단 필터를 통해 본 남성 ([자외선] (400) - 700 nm의)이 논의된다. 이 방법은 UV와 행동에서의 역할과 관련된 다양한 질문에 대한 연구를, 물고기와 다른 생물에 UV 감도와 컬러 패턴을 발견 허용하는 다양한 응용 프로그램이 있습니다.

<table><tbody><tr><td>Spectrophotometer, P1000</td><td>Ocean Optics</td><td></td><td>newer models are availabe</td></tr><tr><td>DT 1000 xenon UV light source</td><td>Ocean Optics</td><td></td><td>newer models are availabe</td></tr><tr><td>Ocean Optics Overture Software</td><td>Ocean Optics</td><td></td><td>newer software is available</td></tr><tr><td>R200-Angle-UV bifurcated fiber-optic probe (Guided Wave)</td><td>Ocean Optics</td><td></td><td>newer models are availabe</td></tr><tr><td>Certified reflectance standard, white</td><td>labsphere</td><td></td><td></td></tr><tr><td>75.7 L Aquarium, divided</td><td>Experimental Builder</td><td></td><td></td></tr><tr><td>Full Spectrum Bulb</td><td>Nature&#039;s Sunlight</td><td></td><td></td></tr><tr><td>UV blocking sheet</td><td>GAM UV Sheet</td><td></td><td></td></tr></tbody></table>

assays to detect uv-reflecting structures and determine their importance in mate preference using the sailfin molly poecilia latipinna

대부분의 생물은 사회적 상호 작용하는 동안 인간의 감성을 넘어 단서 및 신호를 사용합니다. 그들의 행동과 생태를 이해하려고 할 때 유기체가 자신의 세계를 인식하는 방법을 고려하는 것이 중요하다. 자외선 스펙트럼 감도 (UV; 300-400 nm의)는 새, 물고기, 파충류, 양서류, 심지어 포유류의 여러 속에서 발견된다. 이 프로토콜은 UV 반사 구조의 존재와 이들 큐 메이트 원하는 맥락에서 사회적 신호로서 이용되는지 여부를 테스트하기위한 방법에 대해 유기체를 검사하는 기술을 설명한다. 분광 광도계는 개인 및 양성 사이의 반사 강도 UV 반사율 변동의 존재를 검출하는데 사용된다. 이 기술의 예는 이분법 메이트 선택 테스트 외관 전체 스펙트럼 또는 블록 UV 파장을 전송하거나 필터에 의해 조작 할 수있는 이성의 개인 성적으로 받아들이는 개인을 노출하는 표시됩니다. 이시스템은, 여성,하지만 남자하지 SailFin을의 mollies (Poecilia latipinna)이 자신의 짝짓기 결정의 일부로서 UV 표시를 사용했던 결정에 대해 허용. 연구의 이러한 종류의 UV를 활용 생물의 범위에 대한 우리의 지식을 확장하여 UV 자신의 삶에 중요한 역할을하는 방법에 대한 통찰력을 제공하는 역할을한다.

<html> 1 및도 2는 실험에 대한 설정 짝짓기 환경 수족관 및 UV 측정 사이트를 보여줍니다. UV 반사율 판정이 허용 측정 P. latipinna 이러한 특성의 개인차에 더하여, 특히 몸 (도 3)의 측면을 따라, UV 특성을 보유 할. UV 특성이 발견되면, 테스트는 여성이 아닌 남성, P. 밝혀 그들의 ?...

Watch this Scientific Journal Video about Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna at JoVE.com

Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

이 프로토콜은 생물에 자외선 반사 구조를 감지하는 분광 광도법의 사용을 설명합니다 (이 예에서 SailFin을 몰리 Poecilia latipinna) 및 추론 메이트 선택시 자외선 단서의 역할을 할 수 있도록 물고기에 대한 이분법 적 선택 테스트를 설명합니다.

분석은 UV 반사 구조를 검색 및 SailFin을 몰리를 사용하여 메이트 환경 설정에서 자신의 중요성을 확인하는 방법<em&gt; Poecilia latipinna</em

Many organisms use cues and signals beyond human sensitivity during social interactions. It is important to take into account how organisms perceive their ...

assays-to-detect-uv-reflecting-structures-determine-their-importance

Research

JoVE Journal

Neuroscience

Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

6.5K 조회수.  Ohio Wesleyan University.  이 프로토콜은 생물에 자외선 반사 구조를 감지하는 분광 광도법의 사용을 설명합니다 (이 예에서 SailFin을 몰리 Poecilia latipinna) 및 추론 메이트 선택시 자외선 단서의 역할을 할 수 있도록 물고기에 대한 이분법 적 선택 테스트를 설명합니다. 

비디오: Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American lobsters. Recent work in exploited crustacean populations (crabs and lobsters) suggests that there are circumstances where mature females are unable to achieve their full reproductive potential due to sperm limitation. To examine this possibility in different regions of the American lobster fishery, a reliable and noninvasive method was developed for sampling large numbers of female lobsters at sea. This method involves inserting a blunt-tipped needle into the female&#39;s seminal receptacle to determine the presence or absence of a sperm plug and to withdraw a sample that can be examined for the presence of sperm. A series of control studies were conducted at the dock and in the laboratory to test the reliability of this technique. These efforts entailed sampling 294 female lobsters to confirm that the presence of a sperm plug was a reliable indicator of sperm within the receptacle and thus, mating. This paper details the methodology and the results obtained from a subset of the total females sampled. Of the 230 female lobsters sampled from George&#39;s Bank and Cape Ann, MA (size range = 71-145 mm in carapace length), 90.3% were positive for sperm. Potential explanations for the absence of sperm in some females include: immaturity (lack of physiological maturity), breakdown of the sperm plug after being used to fertilize a clutch of eggs, and lack of mating activity. The surveys indicate that this technique for examining the mating success of female lobsters is a reliable proxy that can be used in the field to document reproductive activity in natural populations.

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

Fishery-induced changes to exploited crustacean fisheries, such as the American lobster fishery, could potentially influence their reproductive dynamics, leading to a reduction in mating success. This study&#39;s goal was to develop a noninvasive method for ascertaining the mating success of female lobsters that may be physiologically or functionally mature.

에 대한 비 침습적 방법<em&gt; 현장에서</em&gt; 여성 미국의 바닷가에 연결 성공 (의 결정<em&gt; Homarus의 americanus</em&gt;)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

연구

환경과학

Multimodal Optical Microscopy Methods Reveal Polyp Tissue Morphology and Structure in Caribbean Reef Building Corals

An integrated suite of imaging techniques has been applied to determine the three-dimensional (3D) morphology and cellular structure of polyp tissues comprising the Caribbean reef building corals Montastraeaannularis and M. faveolata. These approaches include fluorescence microscopy (FM), serial block face imaging (SBFI), and two-photon confocal laser scanning microscopy (TPLSM). SBFI provides deep tissue imaging after physical sectioning; it details the tissue surface texture and 3D visualization to tissue depths of more than 2 mm. Complementary FM and TPLSM yield ultra-high resolution images of tissue cellular structure. Results have: (1) identified previously unreported lobate tissue morphologies on the outer wall of individual coral polyps&#160;and (2) created the first surface maps of the 3D distribution and tissue density of chromatophores and algae-like dinoflagellate zooxanthellae endosymbionts. Spectral absorption peaks of 500 nm and 675 nm, respectively, suggest that M. annularis and M. faveolata contain similar types of chlorophyll and chromatophores. However, M. annularis and M. faveolata exhibit significant differences in the tissue density and 3D distribution of these key cellular components. This study focusing on imaging methods indicates that SBFI is extremely useful for analysis of large mm-scale samples of decalcified coral tissues. Complimentary FM and TPLSM reveal subtle submillimeter&#160;scale changes in cellular distribution and density in nondecalcified coral tissue samples. The TPLSM technique affords: (1) minimally invasive sample preparation,&#160;(2) superior optical sectioning ability,&#160;and (3) minimal light absorption and scattering, while still permitting deep tissue imaging.

An integrated suite of imaging techniques has been applied to determine polyp morphology and tissue structure in the Caribbean corals Montastraeaannularis and M. faveolata. Fluorescence, serial block face,&#160;and two-photon confocal laser scanning microscopy have identified lobate structure, polyp walls, and estimated chromatophore and zooxanthellae densities and distributions.

복합 광학 현미경 방법 카리브 암초 건물 산호의 폴립 조직 형태와 구조를 밝히기

An integrated suite of imaging techniques has been applied to determine the three-dimensional (3D) morphology and cellular structure of polyp tissues ...

The Identification of Sea Lamprey Pheromones Using Bioassay-Guided Fractionation

Bioassay-guided fractionation is an iterative approach that uses the results of physiological and behavioral bioassays to guide the isolation and identification of an active pheromone compound. This method has resulted in the successful characterization of the chemical signals that function as pheromones in a wide range of animal species. Sea lampreys rely on olfaction to detect pheromones that mediate behavioral or physiological responses. We use this knowledge of fish biology to posit functions of putative pheromones and to guide the isolation and identification of active pheromone components. Chromatography is used to extract, concentrate, and separate compounds from the conditioned water. Electro-olfactogram (EOG) recordings are conducted to determine which fractions elicit olfactory responses. Two-choice maze behavioral assays are then used to determine if any of the odorous fractions are also behaviorally active and induce a preference. Spectrometric and spectroscopic methods provide the molecular weight and structural information to assist with the structure elucidation. The bioactivity of the pure compounds is confirmed with EOG and behavioral assays. The behavioral responses observed in the maze should ultimately be validated in a field setting to confirm their function in a natural stream setting. These bioassays play a dual role to 1) guide the fractionation process and 2) confirm and further define the bioactivity of isolated components. Here, we report the representative results of a sea lamprey pheromone identification that exemplify the utility of the bioassay-guided fractionation approach. The identification of sea lamprey pheromones is particularly important because a modulation of its pheromone communication system is among the options considered to control the invasive sea lamprey in the Laurentian Great Lakes. This method can be readily adapted to characterize the chemical communication in a broad array of taxa and shed light on waterborne chemical ecology.

Here, we present a protocol to isolate and characterize the structure, olfactory potency, and behavioral response of putative pheromone compounds of sea lampreys.

생물 검정 기반 분류를 사용 하 여 바다 심고 페로몬의 식별

Bioassay-guided fractionation is an iterative approach that uses the results of physiological and behavioral bioassays to guide the isolation and ...

생화학

Individual Culturing of Tigriopus Copepods and Quantitative Analysis of Their Mate-guarding Behavior

Copepods of the genus Tigriopus, which are common zooplankton in rocky tide pools, show precopulatory mate-guarding behavior where a male clasps a potential mate to form a pair. While this phenomenon has attracted interest of researchers, methods for its analysis have not been well described. Here we describe procedures for: 1) individual culturing and staging of Tigriopus juveniles and adults, and 2) video-based analysis of their mate-guarding behavior. The culturing method enables experimental control of paring experience of animals as well as the ability to track their development before behavioral tests. The analysis method allows quantitative evaluation of several aspects of the mate-guarding behavior, including capturing attempts by males and swimming trajectory of mate-guarding pairs. Although these methods were originally established for ethological studies on Tigriopus, with proper modifications they can also be applied to studies of other zooplankton in different research fields, such as physiology, toxicology, and ecological genetics.

Individual Culturing of Tigriopus Copepods and Quantitative Analysis of Their Mate-guarding Behavior

Mate-guarding behavior plays an important role in reproduction of intertidal copepods of the genus Tigriopus. However, methods for studying this behavior have not been well described. Here we describe methods for: 1) individual culture of virgin Tigriopus animals, and 2) quantitative analysis of their mate-guarding behavior.

Tigriopus Copepods 및 그들의 동료를 지키는 행동의 정량 분석의 개별 경작

Copepods of the genus Tigriopus, which are common zooplankton in rocky tide pools, show precopulatory mate-guarding behavior where a male clasps a ...

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the appearance and behavior of visual stimuli, compared to manipulating live animals. Here, we present the FishSim Animation Toolchain, a software framework developed to provide researchers with an easy-to-use method for implementing 3D computer animations in behavioral experiments with fish. The toolchain offers templates to create virtual 3D stimuli of five different fish species. Stimuli are customizable in both appearance and size, based on photographs taken of live fish. Multiple stimuli can be animated by recording swimming paths in a virtual environment using a video game controller. To increase standardization of the simulated behavior, the prerecorded swimming path may be replayed with different stimuli. Multiple animations can later be organized into playlists and presented on monitors during experiments with live fish.
In a case study with sailfin mollies (Poecilia latipinna), we provide a protocol on how to conduct a mate-choice copying experiment with FishSim. We utilized this method to create and animate virtual males and virtual model females, and then presented these to live focal females in a binary choice experiment. Our results demonstrate that computer animation may be used to simulate virtual fish in a mate-choice copying experiment to investigate the role of female gravid spots as an indication of quality for a model female in mate-choice copying.
Applying this method is not limited to mate-choice copying experiments but can be used in various experimental designs. Still, its usability depends on the visual capabilities of the study species and first needs validation. Overall, computer animations offer a high degree of control and standardization in experiments and bear the potential to &#39;reduce&#39; and &#39;replace&#39; live stimulus animals as well as to &#39;refine&#39; experimental procedures.

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Using the novel FishSim Animation Toolchain, we present a protocol for non-invasive visual manipulation of public information in the context of mate-choice copying in sailfin mollies. FishSim Animation Toolchain provides an easy-to-use framework for the design, animation and presentation of computer-animated fish stimuli for behavioral experiments with live test fish.

FishSim 애니메이션 툴체인을 사용 하 여 물고기 동작 조사: 메이 트 선택 복사 Sailfin Mollies에 대 한 사례 연구

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the ...

행동분석학

Behavioral Tracking and Neuromast Imaging of Mexican Cavefish

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among these traits, foraging behavior is one of the useful windows into functional advantages of behavioral trait evolution. Presented herein are updated methods for analyzing vibration attraction behavior (VAB: an adaptive foraging behavior) and imaging of associated mechanosensors of cave-adapted tetra, Astyanax mexicanus. In addition, methods are presented for high-throughput tracking of a series of additional cavefish behaviors including hyperactivity and sleep-loss. Cavefish also show asociality, repetitive behavior and higher anxiety. Therefore, cavefish serve as an animal model for evolved behaviors. These methods use free-software and custom-made scripts that can be applied to other types of behavior. These methods provide practical and cost-effective alternatives to commercially available tracking software.

Here, we present methods for high-throughput study of a series of the Mexican cavefish behaviors and vital staining of a mechanosensory system. These methods use free-software and custom-made scripts, providing a practical and cost-effective method for the studies of behaviors.

행동 추적 및 멕시코 Cavefish의 Neuromast 이미징

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among ...

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 ...

신경과학

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

Zebrafish의 행동 증후군에 대한 대담, 침략, 그리고이 얕아 분석

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

의학

Analysis of Skeletal Muscle Defects in Larval Zebrafish by Birefringence and Touch-evoke Escape Response Assays

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular dystrophies1-3, congenital myopathies4,5, and disruptions in sarcomeric assembly6,7, due to high genomic and structural conservation with mammals8. Muscular disorganization and locomotive impairment can be quickly assessed in the zebrafish over the first few days post-fertilization. Two assays to help characterize skeletal muscle defects in zebrafish are birefringence (structural) and touch-evoked escape response (behavioral).
Birefringence is a physical property in which light is rotated as it passes through ordered matter, such as the pseudo-crystalline array of muscle sarcomeres9. It is a simple, noninvasive approach to assess muscle integrity in translucent zebrafish larvae early in development. Wild-type zebrafish with highly organized skeletal muscle appear very bright amidst a dark background when visualized between two polarized light filters, whereas muscle mutants have birefringence patterns specific to the primary muscular disorder they model. Zebrafish modeling muscular dystrophies, diseases characterized by myofiber degeneration followed by repeated rounds of regeneration, exhibit degenerative dark patches in skeletal muscle under polarized light. Nondystrophic myopathies are not associated with necrosis or regenerative changes, but result in disorganized myofibers and skeletal muscle weakness. Myopathic zebrafish typically show an overall reduction in birefringence, reflecting the disorganization of sarcomeres.
The touch-evoked escape assay involves observing an embryo&#39;s swimming behavior in response to tactile stimulation10-12. In comparison to wild-type larvae, mutant larvae frequently display a weak escape contraction, followed by slow swimming or other type of impaired motion that fails to propel the larvae more than a short distance12. The advantage of these assays is that disease progression in the same fish type can be monitored in vivo for several days, and that large numbers of fish can be analyzed in a short time relative to higher vertebrates.

The zebrafish is now an established and powerful tool for modeling muscular dystrophies, congenital myopathies, and related neuromuscular diseases. Birefringence and touch-evoked escape behavior are two common noninvasive assays used to determine the degree of muscular disorganization and locomotive impairment of zebrafish embryos during early development.

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular ...

생물학

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

Health monitoring systems are developed and used in zebrafish research facilities because pathogens of Danio rerio such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa have the potential to impair animal welfare and research. The fish are typically analyzed post mortem to detect microbes. The use of sentinels is a suggested way to improve the sensitivity of the surveillance and to reduce the number of animals to sample. The setting of a pre-filtration sentinel tank out of a recirculating system is described. The technique is developed to prevent water pollution and to represent the fish population by a careful selection of age, gender, and strains. In order to use the minimum number of animals, techniques to screen the environment are also detailed. Polymerase Chain Reaction (PCR) on surface sump swabs is used to significantly improve the detection of some prevalent and pathogenic mycobacterial species such as Mycobacterium fortuitum, Mycobacterium haemophilum, and Mycobacterium chelonae. Another environmental method consists of processing the sludge at the bottom of a holding tank or sump to look for P. tomentosa eggs. This is a cheap and fast technique that can be applied in quarantine where a breeding device is submerged into the holding tank of imported animals. Finally, PCR is applied to the sludge sample and A. hydrophila is detected at the sump&#39;s bottom and surface. Generally, these environmental screening techniques applied to these specific pathogens have led to an increased sensitivity compared to the testing of pre-filtration sentinels.

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

This protocol describes the use of sump swabs and sludge analysis of zebrafish systems, which leads to increased detection compared to the sole use of sentinels to detect pathogens such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa. A system to monitor P. tomentosa eggs in quarantine is also proposed.

분석은 UV 반사 구조를 검색 및 SailFin을 몰리를 사용하여 메이트 환경 설정에서 자신의 중요성을 확인하는 방법

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