Name: a novel method to model chronic traumatic encephalopathy in drosophila
Uploaded: 2017-07-04T13:00:00.000+00:00
Duration: 10 min 15 s
Description: Watch this Scientific Journal Video about A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila at JoVE.com

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1. 스트라이크 장치의 조립 ( 그림 1 ) <ol><li> 1mL 투베르쿨린 주사기에서 플런저를 제거합니다. 배럴을 1 mL 표시로 자릅니다. </li><li> 200 μL 피펫 팁에서 에어로졸 장벽 (높이 3 mm x 4 mm)을 제거하고 임팩터로 사용하십시오. 충격 장치를 시린지 배럴 안에 놓으십시오. 천천히 배럴을 탭하여 임팩터를 팁 끝으로 움직이십시오. 평평한면이 노즐 입구를 덮습니다. </li><li> Drosophila 마취 스테이션의 이산화탄소 (CO2) 유량 조절기에 연결된 플라스틱 튜빙에 배럴 팁 끝을 연결하십시오. </li><li> 배럴을 수직으로 잡고 표준 클램프 홀더 스탠드에 끼워서 충격 장치가 통의 바닥에 머 무르도록하십시오. </li><li> 플라이 홀더를 만들기 위해 200 μL 피펫 팁을 수정하십시오. <ol><li> 끝에서 4mm를 잘라 0.8mm 지름의 구멍을 만들어 플라이 헤드 만 노출되게하십시오. 참고 : tho뗏목 및 파리 몸의 다른 모든 부분은 피펫 팁 안에 머물 것입니다. </li></ol></li><li> 커넥터를 만들기 위해 1,000 μL 피펫 팁과 1 mL 주사기 바늘 캡을 수정하십시오. <ol><li> 팁의 입구에서 44mm 떨어져 자릅니다. 1 ML 주사기 바늘 캡의 6mm 길이를 가지고 1,000 μL 피펫 팁의 나머지 세그먼트에 단단히 밀어. </li></ol></li></ol> 2. 스트라이크 장치의 작동 <ol><li> 플라이 패드 (fly pad)에서 CO 2 를 사용하여 2 일 된 성인 여성 플라이에게 마취를하십시오. </li><li> 미세한 브러시로 플라이 홀더로 부드럽게 옮깁니다. 플라이 헤드가 팁 끝 외부에 보이도록 홀더를 부드럽게 두드립니다. 가요 성 코의 가위가 팁 외부에 노출 된 경우, 천천히 1 mL 주사기 바늘로 팁 안쪽으로 다시 넣습니다. 참고 : Fly proboscope를 홀더 안에 넣어야합니다. 그렇지 않으면, 파리는 빠는 코의 부상으로 사망 할 수 있습니다. </li><li> 플라이 호를 조이십시오.플라이 헤드가 아래쪽을 향하도록 커넥터가있는 주사기 배럴에 연결하십시오. </li><li> 가스 압력을 100 kPa로 설정하십시오. 실험 설계에 따라 유속을 조정하십시오. </li><li> 임팩트 터가 플라이 헤드를 한 번 치기 위해 유량 조절기 토글 스위치를 빠르게 켜고 끕니다. </li><li> 플라이 홀더를 들어 올려 플라이 패드 위로 옮깁니다. 플라이 홀더를 뒤집고 가볍게 측면을 가볍게 두 드리십시오. 빈 유리 병에 파리를 넣어 복구하십시오. </li></ol> 3. 비디오 지원 이동 추적 <ol><li> 추적 경기장을 만들기 위해 투명 실리콘 엘라스토머가있는 6cm 직경의 페트리 접시를 채 웁니다. 파리와 파리 사이에 3mm의 간격을 두어 파리가 자유롭게 걷고 날 수 없게하십시오. </li><li> 매번 샴 또는 치료 그룹에서 4 개의 파리를 마취하여 경기장에 배치하십시오. 파리를 22 ° C에서 1 시간 동안 방치하십시오. </li><li> 경기장 위에 CCD (Charge Coupled Device) 카메라를 배치하고 5 분 동안 기록합니다.</li><li> Ctrax 소프트웨어 (Caltech에서 무료로 구할 수 있음)를 사용하여 기록 된 운동 궤도를 분석하십시오. 추적 된 데이터를 프로그래밍 언어 (예 : Matlab) 호환 형식으로 내보내고 프레임 당 이동 한 거리를 기반으로 데이터를 분석합니다. 각 비행에 대한 평균 도보 거리를 계산하고 다른 모든 기록 된 파리 / 그룹과 결합하여 같은 그룹의 파일 집단이 이동 한 평균 누적 거리를 얻습니다. </li></ol>

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신경 생리 학적 변화, 신경 병리학 적 특징, 신경 행동 적 결함을 포함하여 CTE 특성을 충실하게 모델링 한 동물 모델은 질병 기전을 밝히고 진단 및 치료 목표를 개발하는 데 필수적입니다. 인간 질병의 동물 모델이 임상 적으로 관련된 모든 종말점을 모방 할 때 완벽한 것은 아니라는 것을 이해할 수 있습니다. 그러나 우리는 견고한 CTE 모델이 다음 세 가지 요구 사항을 충족해야한다고 생각합니?...

만성 외상성 뇌병증 (CTE)은 최근 알츠하이머 병과 같은 다른 타우 병증과 별개로 별개의 신경 퇴행성 장애로 인정 받고 있습니다. 가장 중요한 위험 요소가 나이와 치매의 가족력이있는 알츠하이머 병 및 다른 일반적인 타우 병증과는 달리 CTE는 그 이름에서 알 수 있듯이 접촉 스포츠 선수들에게서 볼 수있는 뇌 외상의 병력과 밀접한 관련이 있으며, 사각 팬티, 축구 선수, 군대 참전 용사 2 , 3 , 4 , 5 등이 있습니다. 그것은 반복적으로 뇌에 뇌졸중과 뇌졸중을 가하는 것으로 시작된다고 생각됩니다. 환자는 알츠하이머 병, 전두 측두엽과 유의 한 중복되는인지 장애, 기분 및 행동 변화, 운동 장애와 같은 증상 및 징후를 나타낼 수 있습니다치매, 레위 몸 치매, 파킨슨 병 6 . 대조적으로, 뇌 조직의 사후 검사는 다른 퇴행성 질환에서 관찰되지 않는 병리학 적 특징 인 피질 상피의 깊이에서 작은 혈관을 둘러싼과 인산화 된 타우 축적의 뚜렷한 패턴을 나타낸다. 그러나 지금까지 질병 발현으로 이어지는 병인에 대해서는 거의 알려지지 않았다. 이것은 충실한 동물 모델이 부족하여 많은 부분을 차지합니다 - 단지 최근에 설치류 모델이 생성되었습니다 ( 5 , 8) . 이러한 모델 생물은 비용 집중 치료와 상대적으로 긴 수명의 단점을 가지고 있으며 신경 퇴행성 질환 연구에 적합하지 않습니다. 포유류 대조 물과 비교하여, Drosophila 와 같은 무척추 동물은 비용 효율적인 유지 관리와 함께 훌륭한 대안이며,유전 적 결정 요인 해부를위한 광범위한 도구 및 상대적으로 짧은 수명 9 . 놀랍게도, 파리와 인간의 두뇌는 진화 적으로 보존 된 분자 및 세포 경로뿐만 아니라 해부학 적 유사성을 공유한다 10 , 11 , 12 . 외상성 뇌 손상을 연구하기위한 두 개의 독창적 인 초파리 모델이 이전에 13 , 14 번 보고되었습니다. Katzenberger와 동료가 고안 한 최초의 &quot;고 충격 손상&quot;(HIT) 장치는 금속 스프링 13 , 15 의 자유 단에 연결된 플라스틱 바이알에 자유롭게 움직이는 파리가 들어있었습니다. 플라스틱 병이 수직으로 기울어 졌을 때 폴리 우레탄 패드를 치고 파리 벽에 부딪쳐 튀어 나와 반발했다. 대조적으로, Barekat과 동료들은 다른 전달 방법을 설계했습니다.Omni Bead Ruptor-24 호 모지 나이저 플랫폼 14 . 파리는 이산화탄소로 무력화되어 호 모지 나이저에 고정되고 사전 프로그램 된 진탕 조건에 적용되는 2 mL 스크류 캡 튜브에 두었다. 조직 균질 기 시스템을 사용하는 한 가지 이점은 실험자가 상해의 강도, 상해의 지속 시간 및 부상의 횟수를 조절할 수 있다는 것입니다. 그러나 두 정권 모두 동일한 단점을 갖고있다 : 머리에 대한 일차적 인 상해는 충격 위치와 힘의 측면에서 무작위로 가해진 다. 또한, 두 가지 방법 모두 신체 및 내부 기관의 다른 부분에 불가피한 부수적 인 손상으로 인해 상당한 사망률을 초래했습니다. 여기, 우리는 과일 파리에서 mTBI를 유도하는 새로운 방법을 설명합니다. 우리의 장치는 가스 추진 탄도 충격기로 구성되어 있습니다. 기존의 Drosophila 모델 14 , 15에 비해 우리의 방법은 meas자유롭게 움직일 수있는 플라이 헤드에서만 지시되는 유랑 충격 (lrable impact)은 충격 강도, 충격 사이의 시간 간격 및 지속 된 충격의 총 횟수와 같은 다양한 요인을 정확하게 제어 할 수 있습니다.

<table><tbody><tr><td>Aerosol Barrier</td><td>USA Scientific</td><td>1120-8810</td><td>Used as an impactor</td></tr><tr><td>200 &amp;#956;L Pipette Tip</td><td>USA Scientific</td><td>1111-0706</td><td>Used as a fly head holder</td></tr><tr><td>1000 &amp;#956;L Pipette Tip</td><td>USA Scientific</td><td>1122-1830</td><td>Used as a connector</td></tr><tr><td>1 mL Tuberculin Syringe</td><td>Becton Dickinson</td><td>309625</td><td></td></tr><tr><td>60 mm Petri Dishes</td><td>Fisher Scientific</td><td>FB0875713A</td><td>Used as a tracking arenas</td></tr><tr><td>Flow Regulator</td><td>Genesee Scientific</td><td>59-122WC</td><td></td></tr><tr><td>Standard Clamp Holder/stand</td><td>EISCO Scientific</td><td>CH0688</td><td></td></tr><tr><td>Fine Brush</td><td>Genesee Scientific</td><td>59-204</td><td></td></tr><tr><td>Flypad</td><td>Genesee Scientific</td><td>59-114</td><td></td></tr><tr><td>Sylgard Silicone Elastomer</td><td>Dow Corning</td><td>4019862</td><td></td></tr><tr><td>CCD Camera</td><td>Microsoft&amp;nbsp;</td><td>HD-5000</td><td></td></tr><tr><td>Ctrax Walking Fly Tracker</td><td>Caltech</td><td>Ctrax 0.2.11</td><td></td></tr><tr><td>MATLAB Image Processing Toolbox</td><td>MATLAB</td><td>R2015b</td><td></td></tr></tbody></table>

a novel method to model chronic traumatic encephalopathy in drosophila

만성 외상성 뇌병증 (CTE)은 반복적 인 경미한 외상성 뇌 손상 (mTBI)에 대한 노출과 밀접하게 관련된 확립 된 신경 퇴행성 질환입니다. 복잡한 병리학 적 변화를 초래하는 기전은 신경 병리학 적 기준을 정의하는 최근의 합의에도 불구하고 크게 애매하다. 여기에서 우리는 Drosophila melanogaster ( Drosophila )에서 CTE 모델을 개발하여 뇌에서 특유의 과산화 된 타우 축적 및 신경 세포 죽음을 유도하는 핵심 유전자와 경로를 확인하려는 새로운 방법을 설명합니다. 경미한 닫힌 상처를 입히기 위해 조정 가능한 강도의 충격이 헤드를 급 가속 및 감속하도록하여 플라이 헤드에 직접 전달됩니다. 우리의 방법은 다른 Drosophila mTBI 모델에 내재 된 잠재적 문제를 제거합니다 ( 예 : 동물의 죽음은신체의 다른 부위 또는 내부 장기). 적은 노동력과 비용 집약적 인 동물 관리, 짧은 수명 및 방대한 유전 도구로 인해 CTE 병인을 연구하고 게놈 전반에 걸친 전방 유전자 및 약리학 적 스크리닝을 수행하는 것이 이상적입니다. 우리는 모델의 지속적인 특성화가 질병 예방 및 치료 방법에 대한 중요한 기계 론적 통찰력을 창출 할 것으로 기대한다.

성인 Drosophila를 사용하여 CTE 모델을 구축하기 위해 단일 폐쇄 머리 손상을 가할 때 우리 장치의 효율성을 결정했습니다. 유전자형, 성별 또는 나이와 관련된 변이를 없애기 위해 우리는 2 일 된 Canton-S WT 암컷 파리를 실험에 사용했습니다. 우리는 100 kPa의 일정한 가스 압력에서 CO 2 의 유량을 조절함으로써 임팩터의 강도를 쉽게 제어 할 수 있습니다. 가장 높...

Watch this Scientific Journal Video about A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila at JoVE.com

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

여기, 우리는 Drosophila melanogaster 에서 닫힌 머리 외상성 뇌 손상을 일으키는 새로운 접근법을 설명합니다. 우리의 방법은 조정 가능한 강도로 반복적 인 충격을 머리에 직접 전달하는 장점이 있습니다. 무척추 동물 시스템의 추가 탐구는 만성 외상성 뇌증의 병인을 밝히는 데 도움이 될 것입니다.

만성 외상성 뇌증 모델링을위한 새로운 방법<em&gt; Drosophila</em

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic ...

a-novel-method-to-model-chronic-traumatic-encephalopathy-in-drosophila

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Neuroscience

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

8.2K 조회수.  Johns Hopkins University School of Medicine.  여기, 우리는 Drosophila melanogaster 에서 닫힌 머리 외상성 뇌 손상을 일으키는 새로운 접근법을 설명합니다. 우리의 방법은 조정 가능한 강도로 반복적 인 충격을 머리에 직접 전달하는 장점이 있습니다. 무척추 동물 시스템의 추가 탐구는 만성 외상성 뇌증의 병인을 밝히는 데 도움이 될 것입니다. 

비디오: A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of these diseases, including Alzheimer's, Parkinson's and Huntington's disease are characterized by age-dependent deterioration in learning and memory functions and movement coordination2. Here we use behavioral assays, including the negative geotaxis assay3 and the aversive phototaxic suppression assay (APS assay)4,5, to show that some of the behavior characteristics associated with human neurodegeneration can be recapitulated in flies. In the negative geotaxis assay, the natural tendency of flies to move against gravity when agitated is utilized to study genes or conditions that may hinder locomotor capacities. In the APS assay, the learning and memory functions are tested in positively-phototactic flies trained to associate light with aversive bitter taste and hence avoid this otherwise natural tendency to move toward light. Testing these trained flies 6 hours post-training is used to assess memory functions. Using these assays, the contribution of any genetic or environmental factors toward developing neurodegeneration can be easily studied in flies.

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Behavior assays for measuring locomotor functions, learning, and memory abilities in Drosophila.

에 시금 운동력이있는 사람, 학습, 및 메모리 적자 Drosophila</emNeurodegeneration의> 모델

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of ...

연구

신경과학

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the proteins involved in these pathways are evolutionarily conserved between mammals and other eukaryotes, such as the fruit fly Drosophila melanogaster, suggesting that similar organizing principles exist during the development of these organisms. Importantly, Drosophila has been used extensively to identify cellular and molecular mechanisms regulating processes that are required in mammals including neurogenesis, differentiation, axonal guidance, and synaptogenesis. Flies have also been used successfully to model a variety of human neurodevelopmental diseases. Here we describe a protocol for the step-by-step microdissection, fixation, and immunofluorescent localization of proteins within the adult Drosophila brain. This protocol focuses on two example neuronal populations, mushroom body neurons and retinal photoreceptors, and includes optional steps to trace individual mushroom body neurons using Mosaic Analysis with a Repressible Cell Marker (MARCM) technique. Example data from both wild-type and mutant brains are shown along with a brief description of a scoring criteria for axonal guidance defects. While this protocol highlights two well-established antibodies for investigating the morphology of mushroom body and photoreceptor neurons, other Drosophila brain regions and the localization of proteins within other brain regions can also be investigated using this protocol.

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

This protocol describes the dissection and immunostaining of adult Drosophila melanogaster brain tissues. Specifically, this protocol highlights the use of Drosophila mushroom body and photoreceptor neurons as example neuronal subsets that can be accurately used to uncover general principles underlying many aspects of neuronal development.

해 부 및 Immunofluorescent 얼룩의 버섯 몸과 성인 초파리 melanogaster 두뇌 신경 세포 포토 리셉터

Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the ...

발생학

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

Drosophila melanogaster is a valuable model organism to study aging and pathological degenerative processes in the nervous system. The advantages of the fly as an experimental system include its genetic tractability, short life span and the possibility to observe and quantitatively analyze complex behaviors. The expression of disease-linked genes in specific neuronal populations of the Drosophila brain, can be used to model human neurodegenerative diseases such as Parkinson's and Alzheimer's 5.
Dopaminergic (DA) neurons are among the most vulnerable neuronal populations in the aging human brain. In Parkinson's disease (PD), the most common neurodegenerative movement disorder, the accelerated loss of DA neurons leads to a progressive and irreversible decline in locomotor function. In addition to age and exposure to environmental toxins, loss of DA neurons is exacerbated by specific mutations in the coding or promoter regions of several genes. The identification of such PD-associated alleles provides the experimental basis for the use of Drosophila as a model to study neurodegeneration of DA neurons in vivo. For example, the expression of the PD-linked human &alpha;-synuclein gene in Drosophila DA neurons recapitulates some features of the human disease, e.g. progressive loss of DA neurons and declining locomotor function 2. Accordingly, this model has been successfully used to identify potential therapeutic targets in PD 8.
Here we describe two assays that have commonly been used to study age-dependent neurodegeneration of DA neurons in Drosophila: a climbing assay based on the startle-induced negative geotaxis response and tyrosine hydroxylase immunostaining of whole adult brain mounts to monitor the number of DA neurons at different ages. In both cases, in vivo expression of UAS transgenes specifically in DA neurons can be achieved by using a tyrosine hydroxylase (TH) promoter-Gal4 driver line 3, 10.

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

Here we describe two assays that have been established to study age-dependent neurodegeneration of dopaminergic (DA) neurons in Drosophila: the climbing/startle-induced negative geotaxis assay which allows to study the functional effects of DA neurons degeneration and the tyrosine hydroxylase immunostaining which is used to identify and count DA neurons in whole brain mounts.

에 신약 표현형을 평가<em&gt; 초파리</em&gt; 등산 분석 실험에 의한 도파민 뉴런과 뇌 전체의 면역 염색

Drosophila melanogaster is a valuable model organism to study aging and pathological degenerative processes in the nervous system. The advantages of the ...

Quantitative Analysis of Climbing Defects in a Drosophila Model of Neurodegenerative Disorders

Locomotive defects resulting from neurodegenerative disorders can be a late onset symptom of disease, following years of subclinical degeneration, and thus current therapeutic treatment strategies are not curative. Through the use of whole exome sequencing, an increasing number of genes have been identified to play a role in human locomotion. Despite identifying these genes, it is not known how these genes are crucial to normal locomotive functioning. Therefore, a reliable assay, which utilizes model organisms to elucidate the role of these genes in order to identify novel targets of therapeutic interest, is needed more than ever. We have designed a sensitized version of the negative geotaxis assay that allows for the detection of milder defects earlier and has the ability to evaluate these defects over time. The assay is performed in a glass graduated cylinder, which is sealed with a wax barrier film. By increasing the threshold distance to be climbed to 17.5 cm and increasing the experiment duration to 2 min&#160;we have observed a greater sensitivity in detecting mild mobility dysfunctions. The assay is cost effective and does not require extensive training to obtain highly reproducible results. This makes it an excellent technique for screening candidate drugs in Drosophila mutants with locomotion defects.

We present an optimized inexpensive and reliable negative geotaxis assay in Drosophila melanogaster as a model for neurodegenerative disorders. Being more sensitive to mild locomotor defects, this assay will help screen for potential genetic interactions and drug targets.

퇴행성 신경 질환의 초파리 모델에서 등반 결함의 정량 분석

Locomotive defects resulting from neurodegenerative disorders can be a late onset symptom of disease, following years of subclinical degeneration, and ...

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Traumatic brain injury (TBI) affects millions of people each year, causing impairment of physical, cognitive, and behavioral functions and death. Studies using Drosophila have contributed important breakthroughs in understanding neurological processes. Thus, with the goal of understanding the cellular and molecular basis of TBI pathologies in humans, we developed the High Impact Trauma (HIT) device to inflict closed head TBI in flies. Flies subjected to the HIT device display phenotypes consistent with human TBI such as temporary incapacitation and progressive neurodegeneration. The HIT device uses a spring-based mechanism to propel flies against the wall of a vial, causing mechanical damage to the fly brain. The device is inexpensive and easy to construct, its operation is simple and rapid, and it produces reproducible results. Consequently, the HIT device can be combined with existing experimental tools and techniques for flies to address fundamental questions about TBI that can lead to the development of diagnostics and treatments for TBI. In particular, the HIT device can be used to perform large-scale genetic screens to understand the genetic basis of TBI pathologies.

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Here we describe a method to inflict closed head traumatic brain injury (TBI) in Drosophila. This method provides a gateway to investigate the cellular and molecular mechanisms that underlie TBI pathologies using the vast array of experimental tools and techniques available for flies.

폐쇄 머리 외상성 뇌 손상을 입힐 수있는 방법<em&gt; 초파리</em

Traumatic brain injury (TBI) affects millions of people each year, causing impairment of physical, cognitive, and behavioral functions and death. Studies ...

Mass Histology to Quantify Neurodegeneration in Drosophila

Progressive neurodegenerative diseases like Alzheimer&#39;s disease (AD) or Parkinson&#39;s disease (PD) are an increasing threat to human health worldwide. Although mammalian models have provided important insights into the underlying mechanisms of pathogenicity, the complexity of mammalian systems together with their high costs are limiting their use. Therefore, the simple but well-established Drosophila model-system&#160;provides an alternative for investigating the molecular pathways that are affected in these diseases. Besides behavioral deficits, neurodegenerative diseases are characterized by histological phenotypes such as neuronal death and axonopathy. To quantify neuronal degeneration and to determine how it is affected by genetic and environmental factors, we use a histological approach that is based on measuring the vacuoles in adult fly brains. To minimize the effects of systematic error and to directly compare sections from control and experimental flies in one preparation, we use the &#39;collar&#39; method for paraffin sections. Neurodegeneration is then assessed by measuring the size and/or number of vacuoles that have developed in the fly brain. This can either be done by focusing on a specific region of interest or by analyzing the entire brain by obtaining serial sections that span the complete head. Therefore, this method allows one to measure not only severe degeneration but also relatively mild phenotypes that are only detectable in a few sections, as occurs during normal aging.

Mass Histology to Quantify Neurodegeneration in Drosophila

Drosophila is widely used as a model system to study neurodegeneration. This protocol describes a method by which degeneration, as determined by vacuole formation in the brain, can be quantified. It also minimizes effects due to the experimental procedure by processing and sectioning control and experimental flies as one sample.

질량 조직학은에 신경 퇴행을 정량화하기<em&gt; 초파리</em

Progressive neurodegenerative diseases like Alzheimer&#39;s disease (AD) or Parkinson&#39;s disease (PD) are an increasing threat to human health ...

Purification of Transcripts and Metabolites from Drosophila Heads

For the last decade, we have tried to understand the molecular and cellular mechanisms of neuronal degeneration using Drosophila as a model organism. Although fruit flies provide obvious experimental advantages, research on neurodegenerative diseases has mostly relied on traditional techniques, including genetic interaction, histology, immunofluorescence, and protein biochemistry. These techniques are effective for mechanistic, hypothesis-driven studies, which lead to a detailed understanding of the role of single genes in well-defined biological problems. However, neurodegenerative diseases are highly complex and affect multiple cellular organelles and processes over time. The advent of new technologies and the omics age provides a unique opportunity to understand the global cellular perturbations underlying complex diseases. Flexible model organisms such as Drosophila are ideal for adapting these new technologies because of their strong annotation and high tractability. One challenge with these small animals, though, is the purification of enough informational molecules (DNA, mRNA, protein, metabolites) from highly relevant tissues such as fly brains. Other challenges consist of collecting large numbers of flies for experimental replicates (critical for statistical robustness) and developing consistent procedures for the purification of high-quality biological material. Here, we describe the procedures for collecting thousands of fly heads and the extraction of transcripts and metabolites to understand how global changes in gene expression and metabolism contribute to neurodegenerative diseases. These procedures are easily scalable and can be applied to the study of proteomic and epigenomic contributions to disease.

Purification of Transcripts and Metabolites from Drosophila Heads

We describe here the procedures for the extraction and purification of mRNA and metabolites from Drosophila heads. We are applying these techniques to better understand the cellular perturbations underlying neuronal degeneration. These methodologies can be easily scaled and adapted for other "omic" projects.

성적 증명서와 Metabolites에서의 정화<em&gt; Drosophila</em&gt; 헤드

For the last decade, we have tried to understand the molecular and cellular mechanisms of neuronal degeneration using Drosophila as a model organism. ...

생물학

Morphological and Functional Evaluation of Axons and their Synapses during Axon Death in Drosophila melanogaster

Axon degeneration is a shared feature in neurodegenerative disease and when nervous systems are challenged by mechanical or chemical forces. However, our understanding of the molecular mechanisms underlying axon degeneration remains limited. Injury-induced axon degeneration serves as a simple model to study how severed axons execute their own disassembly (axon death). Over recent years, an evolutionarily conserved axon death signaling cascade has been identified from flies to mammals, which is required for the separated axon to degenerate after injury. Conversely, attenuated axon death signaling results in morphological and functional preservation of severed axons and their synapses. Here, we present three simple and recently developed protocols that allow for the observation of axonal morphology, or axonal and synaptic function of severed axons that have been cut-off from the neuronal cell body, in the fruit fly Drosophila. Morphology can be observed in the wing, where a partial injury results in axon death side-by-side of uninjured control axons within the same nerve bundle. Alternatively, axonal morphology can also be observed in the brain, where the whole nerve bundle undergoes axon death triggered by antennal ablation. Functional preservation of severed axons and their synapses can be assessed by a simple optogenetic approach coupled with a post-synaptic grooming behavior. We present examples using a highwire loss-of-function mutation and by over-expressing dnmnat, both capable of delaying axon death for weeks to months. Importantly, these protocols can be used beyond injury; they facilitate the characterization of neuronal maintenance factors, axonal transport, and axonal mitochondria.

Morphological and Functional Evaluation of Axons and their Synapses during Axon Death in Drosophila melanogaster

Here, we provide protocols to perform three simple injury-induced axon degeneration (axon death) assays in Drosophila melanogaster to evaluate the morphological and functional preservation of severed axons and their synapses.

드로소필라 멜라노가스터에서 축사 사망 시축과 시냅스의 형태학적 및 기능적 평가

Axon degeneration is a shared feature in neurodegenerative disease and when nervous systems are challenged by mechanical or chemical forces. However, our ...

Tracking Freely Walking Flies: A Method to Assess Locomotion in Drosophila

Source: Sun, M. and Chen, L. L. <a href="https://www.jove.com/video/55602/" target="_blank">A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila</a>. J. Vis. Exp. (2017).
This video describes how to track freely walking flies to assess their general locomotion ability. The featured protocol demonstrates the assay&#39;s setup and data analysis using video tracking software.

자유롭게 걷는 파리 추적: 초파리의 이동을 평가하는 방법

Source: Sun, M. and Chen, L. L. A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila. J. Vis. Exp. (2017).
This video describes how to ...

JoVE Encyclopedia of Experiments

Drosophila melanogaster (fruit fly)

Adult

A Technique to Inflict Closed-Head Traumatic Brain Injury in Drosophila Flies

Source: Sun, M., et al. <a href="https://app.jove.com/v/55602">A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila.</a> J. Vis. Exp. (2017).
This video demonstrates a procedure to inflict a closed-head traumatic injury in anesthetized Drosophila flies using a strike device with a gas-propelled impactor. The injury affects neuronal function and impairs walking ability.

초파리 파리에서 폐쇄성 두부 외상성 뇌 손상을 입히는 기술

1. Assembly of the Strike Device (Figure 1)

	Remove the plunger from a 1 mL tuberculin syringe. Cut the barrel at the 1 mL mark.
	Remove an aerosol ...

만성 외상성 뇌증 모델링을위한 새로운 방법

요약

더 많은 비디오 탐색

이 비디오의 챕터

에 시금 운동력이있는 사람, 학습, 및 메모리 적자 Drosophila 모델

해 부 및 Immunofluorescent 얼룩의 버섯 몸과 성인 초파리 melanogaster 두뇌 신경 세포 포토 리셉터

에 신약 표현형을 평가

퇴행성 신경 질환의 초파리 모델에서 등반 결함의 정량 분석

폐쇄 머리 외상성 뇌 손상을 입힐 수있는 방법

질량 조직학은에 신경 퇴행을 정량화하기

성적 증명서와 Metabolites에서의 정화

드로소필라 멜라노가스터에서 축사 사망 시축과 시냅스의 형태학적 및 기능적 평가

자유롭게 걷는 파리 추적: 초파리의 이동을 평가하는 방법

초파리 파리에서 폐쇄성 두부 외상성 뇌 손상을 입히는 기술