Name: techniques for investigating the anatomy of the ant visual system
Uploaded: 2017-11-27T15:00:00.000+00:00
Duration: 8 min 56 s
Description: Watch this Scientific Journal Video about Techniques for Investigating the Anatomy of the Ant Visual System at JoVE.com

우리는 곤충 해부학에 그들의 지식을 공유 하기 위한 감사 Jochen 제일, 폴 쿠퍼와 Birgit 발전 하 고 여러 기술을 위한 우리는 여기에 설명 된. 우리는 ANU에서 고급 현미경 및 현미경 검사 법 단위 MQU에 센터에서 재능과 지원 직원에 게 감사입니다. 이 작품은 호주 연구 협의회 (DE120100019, FT140100221, DP150101172)에서 보조금을 FRE 대학원 장학금에 의해 지원 되었다.

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저자 아무 경쟁 관심사를 선언합니다.

위에서 설명한 방법 중 스위트 개미와 다른 곤충의 광학 시스템에 대 한 효과적인 조사에 대 한 수 있습니다. 이러한 기술을 샘플링 해상도, 광학 감도 및 공부 되 고 눈의 잠재적인 분극 감도 대 한 우리의 이해를 게 알립니다. 이 지식은 그들의 시각적인 기능에 생리 및 행동 조사에 대 한 중요 한 기초를 제공합니다. 또한, 여기서 설명 하는 방법 개미 영상 시스템에 초점을 맞춘, 하는 동안 이러?...

비전 대부분의 동물에 대 한 중요 한 감각 양식 적임 이다. 시력은 정확히 파악 목표의 수립 및 노선, 준수 및 나침반 정보1,2를 얻기 위한 탐색의 맥락에서 특히 중요 합니다. 곤충 겹 눈의 쌍을 사용 하 여 시각적 정보를 감지 하 고, 경우에 따라 1 ~ 3 dorsally 배치 간단한 눈 이라고 ocelli3,,45.
개미 눈은 특정 관심의 개미 멋지고 다양 한 동안, 그들은 종에 걸쳐 몇 가지 중요 한 특성을 보존 하기 때문에. 해부학, 크기, 및 생태학에 있는 극적인 변화에도 불구 하 고 대부분의 eusocial 고 식민지;에 살고 그 결과, 다른 종 중앙 장소 및 리소스 사이 앞뒤로 탐색 하는 점에서 비슷한 시각 도전 얼굴. 개미에 걸쳐 동일한 기본적인 눈 bauplan 엄격히 야행성 종, 독점적으로 일주와 느린 시각적 포식 자6,7, 뛰어 드는 지 하 도보에서 몸 길이 0.5-26 m m에서 배열 하는 동물에서 관찰 될 수 있다 8,,910. 이러한 엄청난 차이 생태와 행동의 모든 동일한 기본 눈 구조에 맞게 다양 한 환경, 라이프 스타일, 바디 크기11,12의 무수 한 순열에 일으키. 결과적으로 개미의 시각적인 생태 공부 전함 매장 물 결정된 탐정에 게 가능성을 제공 합니다.
곤충의 시각 시스템을 이해 그들의 행동 능력에 대 한 통찰력을 확보에 필수적 이다. 이것은 명백한 멋지게 생태와 동작 몇 가지 곤충 그룹 (예를 들어, 참조13,,1415,16, 에서 큰 성공을 해부학을 결합 하는 통합 연구에서 17). 개미 탐색 및 개미 행동 분야 일반적으로 꽤 성공 했습니다, 비록 매우 작은 강조 몇 가지 선택한 종족 이외의 개미 비전에 배치 되었습니다. 여기, 우리는 개미의 눈 디자인 조사에 관련 된 기술에 정교한 것 이다. 우리는 개미에 집중할 것 이다, 하는 동안 이러한 기술은 적용할 수 있습니다, 다른 곤충에 약간의 수정도.

<table><tbody><tr><td>Ant</td><td>Myrmecia midas</td><td></td><td></td></tr><tr><td>Stereomicroscope</td><td>Leica M205 FA</td><td></td><td></td></tr><tr><td>Sputter coater</td><td>Pro Sci Tech</td><td></td><td></td></tr><tr><td>Ethanol</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Petri dish</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Dissecting microscope</td><td>Leica MZ6</td><td></td><td></td></tr><tr><td>Insect Pin</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Colourless nail polish</td><td>Non branded: from any cosmetic store</td><td></td><td></td></tr><tr><td>Glass slide</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Razor blade</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Foreceps</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Cover slip</td><td>ProSciTech</td><td></td><td></td></tr><tr><td>Compound microscope</td><td>Leica DM5000 B</td><td></td><td></td></tr><tr><td>Glutaraldehyde</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Paraformalydehyde</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Potassium Chloride (KCl)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>di-Sodium Hydrogen phosphate (Na2HPO4)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Potassium di-Hydrogen Phosphate (KH2PO4)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Sodium Chloride (NaCl)</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Osmium tetroxide</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Acetone</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Araldite Epoxy Resin</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Pasteur pipette</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Toluidie Blue</td><td>Sigma Aldrich</td><td></td><td></td></tr><tr><td>Hotplate</td><td>Riechert HK120</td><td></td><td></td></tr></tbody></table>

techniques for investigating the anatomy of the ant visual system

이 문서는 가벼운 현미경 검사 법 (LM)에서 기술의 제품군 설명 및 전자 현미경 (EM) 곤충의 내부 및 외부 눈 해부학을 공부 하는 데 사용할 수 있습니다. 전통적인 조직학 기술 작품 개미 눈에 대 한 최적화 및 전송 전자 현미경 (TEM)과 주사 전자 현미경 (SEM)과 같은 다른 기술 함께에서 작동 하도록 적응 포함 됩니다. 이러한 기술을 훨씬 유용 하지만 어려울 수 있습니다 초보자 현미경에 대 한 큰 강조 다른 표본에 대 한 최적화 및 문제 해결이 문서에서 배치 되었습니다. 우리는 (사진-현미경과 sem의) 전체 표본에 대 한 이미징 기술에 대 한 정보를 제공 하 고 그들의 장점과 단점을 토론. 우리 전체 눈 렌즈 직경 결정에 사용 되는 기술 하 고 개선에 대 한 새로운 기술에 이야기. 마지막으로, 우리는 LM 및 가장, 샘플 준비에 관련 된 기술 토론 단면, 얼룩, 및 이러한 샘플 영상. 우리는 한 때 샘플 준비와 그들을 주위에 이동 하는 최선의 방법을 모른다는 장애물 논의.

여기에 설명 된 방법을 사용 간단한의 상세한 연구와 개미의 겹 눈. Z-스택 photomicrography 기술을 사용 하 여 머리의 등 쪽 보기 이미징 비주얼 시스템 (그림 1)의 레이아웃에 대 한 개요를 얻을 수 있습니다. 이것은 좋은 준비 해 필요한 단면 각도 결정 하 고입니다. 이 기술은 머리 폭, 눈 길이, ocellar 렌즈 직경 등 측정에 유용 합니다. SEM 이미지 또한 상?...

Watch this Scientific Journal Video about Techniques for Investigating the Anatomy of the Ant Visual System at JoVE.com

Techniques for Investigating the Anatomy of the Ant Visual System

이 문서는 빛과 곤충의 내부 및 외부 눈 해부학 공부에 전자 현미경 검사 법에 기술 제품군을 설명. 개미 눈, 상세한 문제 해결, 그리고 다른 표본 및 관심 영역에 대 한 최적화에 대 한 제안에 대 한 작업에 대 한 최적화의 전통적인 기법 여러 개 포함 됩니다.

개미 비주얼 시스템의 조사 기법

This article outlines a suite of techniques in light microscopy (LM) and electron microscopy (EM) which can be used to study the internal and external eye ...

techniques-for-investigating-the-anatomy-of-the-ant-visual-system

Universidad San Sebastian

Research

JoVE Journal

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13.2K 조회수.  Australian National University. 이 문서는 빛과 곤충의 내부 및 외부 눈 해부학 공부에 전자 현미경 검사 법에 기술 제품군을 설명. 개미 눈, 상세한 문제 해결, 그리고 다른 표본 및 관심 영역에 대 한 최적화에 대 한 제안에 대 한 작업에 대 한 최적화의 전통적인 기법 여러 개 포함 됩니다.

비디오: Techniques for Investigating the Anatomy of the Ant Visual System

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

나무 개미에 비주얼 고전 컨디셔닝

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

연구

행동분석학

The Gateway to the Brain: Dissecting the Primate Eye

The visual system in humans is considered the gateway to the world and plays a principal role in the plethora of sensory, perceptual and cognitive processes. It is therefore not surprising that quality of vision is tied to quality of life . Despite widespread clinical and basic research surrounding the causes of visual disorders, many forms of visual impairments, such as retinitis pigmentosa and macular degeneration, lack effective treatments. Non-human primates have the closest general features of eye development to that of humans. Not only do they have a similar vascular anatomy, but amongst other mammals, primates have the unique characteristic of having a region in the temporal retina specialized for high visual acuity, the fovea1. Here we describe a general technique for dissecting the primate retina to provide tissue for retinal histology, immunohistochemistry, laser capture microdissection, as well as light and electron microscopy. With the extended use of the non-human primate as a translational model, our hope is that improved understanding of the retina will provide insights into effective approaches towards attenuating or reversing the negative impact of visual disorders on the quality of life of affected individuals.

The non-human primate is an important translational species for our understanding of development and aging. The anatomical organization of the primate retina may provide important insights into normal and pathological conditions in humans.

두뇌에 게이트웨이 : 프리 메이트 &#39;눈&#39;을 분석 해 봅시다

The visual system in humans is considered the  gateway  to the world and plays a principal role in the plethora of sensory, perceptual and cognitive ...

생물학

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees (Apis mellifera L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker bees spend the first half of their life inside the dark hive, where they are nursing the larvae or building the regular hexagonal combs for food (e.g. pollen or nectar) and brood3. The antennae are extraordinary multisensory feelers and play a pivotal role in various tactile mediated tasks4, including hive building5 and pattern recognition6. Later in life, each single bee leaves the hive to forage for food. Then a bee has to learn to discriminate profitable food sources, memorize their location, and communicate it to its nest mates7. Bees use different floral signals like colors or odors7, 8, but also tactile cues from the petal surface9 to form multisensory memories of the food source. Under laboratory conditions, bees can be trained in an appetitive learning paradigm to discriminate tactile object features, such as edges or grooves with their antennae10, 11, 12, 13. This learning paradigm is closely related to the classical olfactory conditioning of the proboscis extension response (PER) in harnessed bees14. The advantage of the tactile learning paradigm in the laboratory is the possibility of combining behavioral experiments on learning with various physiological measurements, including the analysis of the antennal movement pattern.

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L.

In this protocol we show how to condition harnessed honey bees to tactile stimuli and introduce a 2D motion capture technique for analyzing the kinematics of fine-scale antennal sampling pattern.

촉각 시설과 꿀벌에 Antennal 샘플링 전략의 움직임 분석 (<em&gt; API를 mellifera</em&gt; L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker ...

신경과학

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging

The Drosophila brain and visual system are widely utilized model systems to study neuronal development, function and degeneration. Here we show three preparations of the brain and visual system that cover the range from the developing eye disc-brain complex in the developing pupae to individual eye and brain dissection from adult flies. All protocols are optimized for the live culture of the preparations. However, we also present the conditions for fixed tissue immunohistochemistry where applicable. Finally, we show live imaging conditions for these preparations using conventional and resonant 4D confocal live imaging in a perfusion chamber. Together, these protocols provide a basis for live imaging on different time scales ranging from functional intracellular assays on the scale of minutes to developmental or degenerative processes on the scale of many hours.

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging

This protocol describes three Drosophila preparations: 1) adult brain dissection, 2) adult retina dissection and 3) developing eye disc- brain complexes dissection. Emphasis is laid on special preparation techniques and conditions for live imaging, although all preparations can be used for fixed tissue immunohistochemistry.

개발 및 성인의 준비 Drosophila 두뇌와 라이브 영상을위한 Retinae

The Drosophila brain and visual system are widely utilized model systems to study neuronal development, function and degeneration. Here we show three ...

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization

The Drosophila optic lobe, comprised of four neuropils: the lamina, medulla, lobula and lobula plate, is an excellent model system for exploring the developmental mechanisms that generate neural diversity and drive circuit assembly. Given its complex three-dimensional organization, analysis of the optic lobe requires that one understand how its adult neuropils and larval progenitors are positioned relative to each other and the central brain. Here, we describe a protocol for the dissection, immunostaining and mounting of larval and adult brains for optic lobe imaging. Special emphasis is placed on the relationship between mounting orientation and the spatial organization of the optic lobe. We describe three mounting strategies in the larva (anterior, posterior and lateral) and three in the adult (anterior, posterior and horizontal), each of which provide an ideal imaging angle for a distinct optic lobe structure.

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization

This protocol describes three steps to prepare larval and adult Drosophila optic lobes for imaging: 1) brain dissections, 2) immunohistochemistry and 3) mounting. Emphasis is placed on step 3, as distinct mounting orientations are required to visualize specific optic lobe structures.&#160;

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization(시신경엽 시각화를 위한 유충 및 성체 초파리 뇌의 해부, 면역조직화학 및 마운팅)

The Drosophila optic lobe, comprised of four neuropils: the lamina, medulla, lobula and lobula plate, is an excellent model system for exploring the ...

발생학

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila

~Neurons are precisely interconnected to form circuits essential for the proper function of the brain. The Drosophila olfactory system provides an excellent model to investigate this process since 50 types of olfactory receptor neurons (ORNs) from the antennae and maxillary palps project their axons to 50 identifiable glomeruli in the antennal lobe and form synaptic connections with dendrites from 50 types of second-order projection neurons (PNs). Previous studies mainly focused on identifying important molecules that regulate the precise targeting in the olfactory circuit using fixed tissues. Here, an antennae-brain explant system that recapitulates key developmental milestones of olfactory circuit assembly in culture is described. Through dissecting the external cuticle and cleaning opaque fat bodies covering the developing pupal brain, high quality images of single neurons from live brains can be collected using two-photon microscopy. This allows time-lapse imaging of single ORN axon targeting from live tissue. This approach will help reveal important cell biological contexts and functions of previously identified important genes and identify mechanisms underpinning the dynamic process of circuit assembly.

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila

This protocol describes the dissection procedure, culture condition, and live imaging of an antennae-brain explant system for the study of the olfactory circuit assembly.

초파리의 후각 회로 어셈블리의 타임랩스 이미징 연구를위한 Explant 시스템

~Neurons are precisely interconnected to form circuits essential for the proper function of the brain. The Drosophila olfactory system provides an ...

Analyzing Dendritic Morphology in Columns and Layers

In many regions of the central nervous systems, such as the fly optic lobes and the vertebrate cortex, synaptic circuits are organized in layers and columns to facilitate brain wiring during development and information processing in developed animals. Postsynaptic neurons elaborate dendrites in type-specific patterns in specific layers to synapse with appropriate presynaptic terminals. The fly medulla neuropil is composed of 10 layers and about 750 columns; each column is innervated by dendrites of over 38 types of medulla neurons, which match with the axonal terminals of some 7 types of afferents in a type-specific fashion. This report details the procedures to image and analyze dendrites of medulla neurons. The workflow includes three sections: (i) the dual-view imaging section combines two confocal image stacks collected at orthogonal orientations into a high-resolution 3D image of dendrites; (ii) the dendrite tracing and registration section traces dendritic arbors in 3D and registers dendritic traces to the reference column array; (iii) the dendritic analysis section analyzes dendritic patterns with respect to columns and layers, including layer-specific termination and planar projection direction of dendritic arbors, and derives estimates of dendritic branching and termination frequencies. The protocols utilize custom plugins built on the open-source MIPAV (Medical Imaging Processing, Analysis, and Visualization) platform and custom toolboxes in the matrix laboratory language. Together, these protocols provide a complete workflow to analyze the dendritic routing of Drosophila medulla neurons in layers and columns, to identify cell types, and to determine defects in mutants.

Here, we show how to analyze dendritic routing of Drosophila medulla neurons in columns and layers. The workflow includes a dual-view imaging technique to improve the image quality and computational tools for tracing, registering dendritic arbors to the reference column array and for analyzing the dendritic structures in 3D space.

열 및 레이어에 수지상 형태학 분석

In many regions of the central nervous systems, such as the fly optic lobes and the vertebrate cortex, synaptic circuits are organized in layers and ...

<meta charset="utf8"></head><body>후각 경험 유도 시냅스 리모델링(Olfactory Experience-Induced Synaptic Remodeling in Drosophila's Juvenile Brain)

Experience-Dependent Remodeling of Juvenile Brain Olfactory Sensory Neuron Synaptic Connectivity in an Early-Life Critical Period

Early-life olfactory sensory experience induces dramatic synaptic glomeruli remodeling in the Drosophila juvenile brain, which is experientially dose-dependent, temporally restricted, and transiently reversible only in a short, well-defined critical period. The directionality of brain circuit synaptic connectivity remodeling is determined by the specific odorant acting on the respondent receptor class of olfactory sensory neurons. In general, each neuron class expresses only a single odorant receptor and innervates a single olfactory synaptic glomerulus. In the Drosophila genetic model, the full array of olfactory glomeruli has been precisely mapped by odorant responsiveness and behavioral output. Ethyl butyrate (EB) odorant activates Or42a receptor neurons innervating the VM7 glomerulus. During the early-life critical period, EB experience drives dose-dependent synapse elimination in the Or42a olfactory sensory neurons. Timed periods of dosed EB odorant exposure allow investigation of experience-dependent circuit connectivity pruning in juvenile brain. Confocal microscopy imaging of antennal lobe synaptic glomeruli is done with Or42a receptor-driven transgenic markers that provide quantification of synapse number and innervation volume. The sophisticated Drosophila genetic toolkit enables the systematic dissection of the cellular and molecular mechanisms mediating brain circuit remodeling.

<meta charset="utf8"></head><body>저자 스포트라이트: Exploring Glial Influence in Experience-Dependent Synaptic Pruning During Critical Periods

We describe here methods for inducing and analyzing olfactory experience-dependent remodeling of antennal lobe synaptic glomeruli in the Drosophila juvenile brain.

Experience-Dependent Remodeling of Juvenile Brain Olfactory Sensory Neuron Synaptic Connectivity in an Early-Life Critical Period(어린 뇌 후각 감각 뉴런 시냅스 연결성의 경험 의존적 리모델링)

개미 비주얼 시스템의 조사 기법

요약

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나무 개미에 비주얼 고전 컨디셔닝

두뇌에 게이트웨이 : 프리 메이트 '눈'을 분석 해 봅시다

촉각 시설과 꿀벌에 Antennal 샘플링 전략의 움직임 분석 (

개발 및 성인의 준비 Drosophila 두뇌와 라이브 영상을위한 Retinae

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization(시신경엽 시각화를 위한 유충 및 성체 초파리 뇌의 해부, 면역조직화학 및 마운팅)

초파리의 후각 회로 어셈블리의 타임랩스 이미징 연구를위한 Explant 시스템

열 및 레이어에 수지상 형태학 분석

Experience-Dependent Remodeling of Juvenile Brain Olfactory Sensory Neuron Synaptic Connectivity in an Early-Life Critical Period(어린 뇌 후각 감각 뉴런 시냅스 연결성의 경험 의존적 리모델링)

Experience-Dependent Remodeling of Juvenile Brain Olfactory Sensory Neuron Synaptic Connectivity in an Early-Life Critical Period(어린 뇌 후각 감각 뉴런 시냅스 연결성의 경험 의존적 리모델링)

Experience-Dependent Remodeling of Juvenile Brain Olfactory Sensory Neuron Synaptic Connectivity in an Early-Life Critical Period(어린 뇌 후각 감각 뉴런 시냅스 연결성의 경험 의존적 리모델링)