저자는 기술 지원을 크리스타 하세, 도리스 웨버와 브리짓 Korte은 감사합니다. WEIDINGER 실험실에서 작업은 도이치 Forschungsgemeinschaft 우리 4223 / 3-1의 보조금 지원, 4223 / 4-1 도이치 먼 대 Kardiologie 오스카 - 라플란드-Stipendium를 통해와 클라우스 - 게오르그 - 싶게 - Sigrid-에 의해 우리 Hengstberger-Forschungsstipendium.

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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+2A+peptides+allow+expression+of+multiple+proteins+from+a+single+ORF+in+transgenic+zebrafish+embryos.">Viral 2A peptides allow expression of multiple proteins from a single ORF in transgenic zebrafish embryos.</a> Genesis. 45 (10), 625-629 (2007).</li><li>Suster, M. L., Abe, G., Schouw, A., Kawakami, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transposon-mediated+BAC+transgenesis+in+zebrafish.">Transposon-mediated BAC transgenesis in zebrafish.</a> Nat Protoc. 6 (12), 1998-2021 (2011).</li><li>Bussmann, J., Schulte-Merker, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+BAC+selection+for+tol2-mediated+transgenesis+in+zebrafish.">Rapid BAC selection for tol2-mediated transgenesis in zebrafish.</a> Development. 138 (19), 4327-4332 (2011).</li><li>Yuan, S., Sun, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microinjection+of+mRNA+and+morpholino+antisense+oligonucleotides+in+zebrafish+embryos.">Microinjection of mRNA and morpholino antisense oligonucleotides in zebrafish embryos.</a> J Vis Exp. 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저자가 공개하는 게 없다.

성인 제브라 피쉬가 성공적으로 많은 내부 장기와 부속을 재생하는 놀라운 능력을 가지고있다. 관련된 분자 및 세포 메커니즘에 대한 철저한 이해는 유전자의 기능과 신호 전달 경로의 조직 특이 적 분석이 필요합니다. 이 무렵, 코스닥 시스템은 배아와 성인 제브라 피쉬의 시공간으로 제어 유전자 발현을위한 효율적인 도구를 제공합니다. 이 논문에 기술 된 TETON 시스템 구축 및 방법론이 성공적...

제브라 피쉬의 개발, 생리학, 질병, 및 재생의 여러 측면을 연구하는 기초가 튼튼한 척추 동물 모델 생물이다. 유전자 발현 또는 신호 전달 경로의 유도, 조직 특이 적 조작 후 배아 생물학적 과정, 실험 도구에 대한 모델로 제브라 피쉬의 성장 채택으로 점점 더 중요 해지고있다. 특히, 성인 제브라 피쉬의 기관 및 부속 재생에 대한 연구는 이러한 재생 과정에서 신호 전달 경로의 시공간 요구 사항의 해부 도구의 부족으로 고통을. 현재, 세가지 다른 시스템 조건, 조직 - 특이 성인 지브라 피쉬의 장기를 재생 유전자 발현 달성하는데 사용되어왔다 : Cre 호텔-LOX 시스템 트랜스포존 매개 체세포 형질 전환을 사용하여 열 - 충격 유도 유전자의 모자이크 식 및 코스닥 시스템 1 -3. 코스닥은 테트라 사이클린 계속의 변형을 말한다발현은 테트라 사이클린 항생 물질 또는 유도체, 예 독시사이클린의 존재 하에서 활성화 롤 전사 활성 시스템. 그것은 지금까지 성인 물고기에 사용 된 바와 같이 Cre 호텔-LOX 시스템은, 발현 공간적으로 조직 특이 적 조절 요소에 의해 제한되는 타목시펜 제어 Cre 호텔 재조합 효소 (CreERT2)에 의존한다. STOP 카세트의 CRE 구동 제거는 모든 세포 유형 1 활성화되어야 프로모터에 의해 구동되는 목적 유전자의 발현을 촉진한다. mosaically 표현 체세포 유전자의 트랜스포존 매개 창조 개별 세포 계통의 유도 유전자 발현 시스템을 제공한다. 전형적 재생 기관 (2)의 분리 된 세포 계통의 유전자를 운반하는 키메라 개인에 열 충격 프로모터 결과의 전사 제어하에 관심있는 유전자를 운반하는 트랜스포존과 TOL2 지브라 피쉬의 수정란 사출. 두 시스템은 조건부 조직 SPE를 허용하는 동안cific 유전자 발현은 LOX-Cre 호텔 시스템은 가역적없고, 트랜스포존 기반 클론 표지를 사용하는 전략은 스토캐스틱 자연에서 겪고있다. 따라서, 우리 등 최근 시간적 및 가산 및 가역 가변 3-5 인 공간적 조절 유전자 발현 촉진 제브라에 사용하기위한 유전자 코스닥 시스템을 채택 하였다. 여기서 사용 코스닥 시스템은 독시사이클린 (D​​OX) -inducible 전사 활성이 (역방향의 테트라 사이클린 transactivator, irtTA 짧은 TETA 개선) 조직 특이 적 유전자 조절 서열의 제어하에있는 트랜스 제닉 드라이버 라인 (TetActivator)을 포함한다. (; TetRE 구정 응답 요소) (그림 1A) 둘째, 유전자 변형 응답자 라인 테트라 사이클린 연산자의 전사 제어에 관심의 유전자를 품고 (TetResponder)가 필요합니다. 따라서 TetActivator 및 TetResponder 선의 특정 조합의 사용은 조건부 조직 정시 허용유전자 발현의 FIC 조작. 우리는 최근 제브라 피쉬의 꼬리 지느러미 3 재생 성인에서의 Wnt / 베타 카테닌 신호의 조직 특이 적 기능을 위해 프로브 TETON 시스템을 활용했다. 여기에 설명 된 프로토콜에서 우리는 지느러미 재생의 연구에 특히 제브라 피쉬에 Teton 시스템의 설정 및 사용에 대한 작업 흐름을 설명합니다. 이것은 상세한 안정적인 형질 전환 TetActivator과 TetResponder 라인을 생성하는 방법에 대한 지침 및 배아와 성인 제브라 피쉬의 형질 전환 유전자의 유도를위한 프로토콜을 포함한다. 더욱이, 우리는 성인 핀 저온부 지브라 피쉬의 제조를위한 프로토콜을 포함하여 핀 재생성에 조직 특이 적 유전자 발현의 확인을위한 기술을 설명한다. 또한, 우리는 TetResponder 식의 TetActivator 형질 전환 유전자의 설계, 형질 전환 방법의 선택, 및 검출을위한 고려 사항에 대해 설명합니다. 따라서,이 프로토콜의 전반적인 목표는 청사진 역할을하는 것입니다설정해서 기능성 코스닥 시스템 지브라 피쉬의 관심있는 조직에 적용 할 수있는 조건부 조직 특이 적 유전자 발현을 달성하기위한. 우리는 I-SceI 제한 효소 또는 짧은 유전자 조절 서열 사용하여 안정적인 TetActivator 라인의 TOL2 매개 세대를 허용 TetActivator 벡터 만든 (증강 조각, WEIDINGER 랩 플라스미드 데이터베이스없이 1247;. 그림 1B). 이 구조는 단순 포진 바이러스 VP16의 전이 활성화 도메인 파생 3F [irtTAM2 (3 층)]와 융합 역 구정 리프레 도메인의 M2 돌연변이 변종으로 구성된 TetActivator 카세트가 포함되어 있습니다. 이 공동 발현 동일한 오픈 리딩 프레임으로부터의 형광 단으로 AmCyan 때문에 TetActivator (TETA)의 발현을 용이하게 모니터링 할 수있다; 1 비율 5,6 : P2A 펩타이드는 1에서 분리 된 단백질과 같은 TETA과 AmCyan의 생산을 초래할한다 리보솜 건너 뛰기를, 중재. 구조는 또한 T의 &#39;poylinker 5 포함etActivator 카세트는 종래의 클로닝 방법을 사용하여 관심의 유전자 조절 서열의 삽입을 용이하게한다. (큰 게놈 영역을 포함하는 박테리아 인공 염색체로 재조합 될 수있다 (BAC), 또한, 우리는 전술 TetActivator 카세트 플러스 카나마이신 선택 카세트 (도 1C. WEIDINGER 랩 플라스미드 데이터베이스 없음 1180)로 이루어진 구조를 작성한 일반적으로 발현 패턴 트랜스 의해 모방 함) 유전자의 개시 코돈으로. 두 구조는 요청시 WEIDINGER 실험실에서 사용할 수 있습니다.

<table><tbody><tr><td>Breeding boxes</td><td>Aqua Schwarz</td><td>AquaBox 1</td><td></td></tr><tr><td>Compound fluorescent microscope</td><td>e.g., Leica, Zeiss</td><td>varies with the manufacturer</td><td>to image fluorescent tissue sections</td></tr><tr><td>Confocal microscope</td><td>e.g., Leica, Zeiss</td><td>varies with the manufacturer</td><td>to image fluorescent tissue sections</td></tr><tr><td>Cryostat</td><td>e.g., Leica, Thermo-Scientific</td><td>varies with the manufacturer</td><td>for cryosectioning</td></tr><tr><td>4&amp;rsquo;, 6- diamidino-2-phenylinodole (Dapi)</td><td>Sigma-Aldrich</td><td>D9542</td><td>use 1/5,000 in PBS&lt;br/&gt; for visualization of nuclei</td></tr><tr><td>Doxycycline</td><td>Sigma-Aldrich</td><td>D9891</td><td>prepare stocks in 50% EtOH at 50 mg/ml (97 mM)&lt;br/&gt; for TetResponder induction</td></tr><tr><td>Paraformaldehyde (PFA)</td><td>Sigma-Aldrich</td><td>P6148</td><td>4% (w/v) paraformaldehyde in PBS, pH 7.5&lt;br/&gt; for fixation</td></tr><tr><td>1x Phosphat-buffer saline (PBS)</td><td></td><td></td><td>1.7 mM KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;, 5.2 mM Na&lt;sub&gt;2&lt;/sub&gt;HPO&lt;sub&gt;4&lt;/sub&gt;, 150 mM NaCl, pH 7.5</td></tr><tr><td>1x Phosphat-buffer saline + Tween 20 (PBT)</td><td></td><td></td><td>1x PBS with 0.1% Tween 20</td></tr><tr><td>Superfrost Ultra Plus adhesion microscope slides</td><td>Thermo Scientific&amp;nbsp;</td><td>1014356190</td><td>for collection of tissue sections</td></tr><tr><td>Stereo fluorescent microscope</td><td>e.g., Leica, Zeiss</td><td>varies with the manufacturer</td><td>for fluorescence-based genotyping</td></tr><tr><td>Thermocycler</td><td>e.g., Biorad, Applied Biosystems</td><td>varies with the manufacturer</td><td>for PCR-based genotyping</td></tr><tr><td>Tissue freezing medium (TFM)</td><td>Triangel Biomedical Sciences</td><td>TFM-C</td><td>for embedding of tissue samples</td></tr><tr><td>Tricaine (L-Ethyl-m-amino-benzoate-methane sulfonate/MS-222)</td><td>Sigma-Aldrich</td><td>E10521</td><td>for anesthesia&lt;br/&gt; use at 1 mg/ml in E3 embryo medium</td></tr></tbody></table>

use of the teton system to study molecular mechanisms of zebrafish regeneration

The zebrafish has become a very important model organism for studying vertebrate development, physiology, disease, and tissue regeneration. A thorough understanding of the molecular and cellular mechanisms involved requires experimental tools that allow for inducible, tissue-specific manipulation of gene expression or signaling pathways. Therefore, we and others have recently adapted the TetON system for use in zebrafish. The TetON system facilitates temporally and spatially-controlled gene expression and we have recently used this tool to probe for tissue-specific functions of Wnt/beta&#8211;catenin signaling during zebrafish tail fin regeneration. Here we describe the workflow for using the TetON system to achieve inducible, tissue-specific gene expression in the adult regenerating zebrafish tail fin. This includes the generation of stable transgenic TetActivator and TetResponder lines, transgene induction and techniques for verification of tissue-specific gene expression in the fin regenerate. Thus, this protocol serves as blueprint for setting up a functional TetON system in zebrafish and its subsequent use, in particular for studying fin regeneration.

조직 특이 유도 유전자 발현, 유전자 변형 TetActivator 및 TetResponder 라인을위한 기능 TETON 시스템을 구축하기 위해 생성 (그림 1A) 할 필요가있다. 초기 제브라 피쉬 배아 및 후속 세균 라인 통합으로 구축 또는 TetResponder (그림 1E) -이 TetActivator (C 그림 1B)를 마이크로 인젝션하여 수행됩니다. 기능성 TetActivator 구조물 중 하나, 또는 관심 유전자 (도 1C)의...

Watch this Scientific Journal Video about Use of the TetON System to Study Molecular Mechanisms of Zebrafish Regeneration at JoVE.com

Use of the TetON System to Study Molecular Mechanisms of Zebrafish Regeneration

Here we outline the workflow for using the TetON system to achieve tissue-specific gene expression in the adult regenerating zebrafish tail fin.

코스닥 시스템의 사용은 분자 메커니즘 Zebrafish의 재생의 연구를

The zebrafish has become a very important model organism for studying vertebrate development, physiology, disease, and tissue regeneration. A thorough ...

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10.6K 조회수.  Ulm University. Here we outline the workflow for using the TetON system to achieve tissue-specific gene expression in the adult regenerating zebrafish tail fin.

비디오: Use of the TetON System to Study Molecular Mechanisms of Zebrafish Regeneration

Kidney Regeneration in Adult Zebrafish by Gentamicin Induced Injury

The kidney is essential for fluid homeostasis, blood pressure regulation and filtration of waste from the body. The fundamental unit of kidney function is the nephron. Mammals are able to repair existing nephrons after injury, but lose the ability to form new nephrons soon after birth. In contrast to mammals, adult fish produce new nephrons (neonephrogenesis) throughout their lives in response to growth requirements or injury. Recently, lhx1a has been shown to mark nephron progenitor cells in the adult zebrafish kidney, however mechanisms controlling the formation of new nephrons after injury remain unknown. Here we show our method for robust and reproducible injury in the adult zebrafish kidney by intraperitoneal (i.p.) injection of gentamicin, which uses a noninvasive visual screening process to select for fish with strong but nonlethal injury. Using this method, we can determine optimal gentamicin dosages for injury and go on to demonstrate the effect of higher temperatures on kidney regeneration in zebrafish.

Here we present a reliable method to study adult kidney regeneration by inducing acute kidney injury by gentamicin injection. We show that injury is dependent on gentamicin dosage and environmental temperature using in situ hybridization to label lhx1a+ developing new nephrons.

겐타 마이신 유도 부상으로 성인 Zebrafish의에서 신장 재생

The kidney is essential for fluid homeostasis, blood pressure regulation and filtration of waste from the body. The fundamental unit of kidney function is ...

연구

발생학

Capturing Tissue Repair in Zebrafish Larvae with Time-lapse Brightfield Stereomicroscopy

The zebrafish larval tail fin is ideal for studying tissue regeneration due to the simple architecture of the larval fin-fold, which comprises of two layers of skin that enclose undifferentiated mesenchyme, and because the larval tail fin regenerates rapidly within 2-3 days. Using this system, we demonstrate a method for capturing the repair dynamics of the amputated tail fin with time-lapse video brightfield stereomicroscopy. We demonstrate that fin amputation triggers a contraction of the amputation wound and extrusion of cells around the wound margin, leading to their subsequent clearance. Fin regeneration proceeds from proximal to distal direction after a short delay. In addition, developmental growth of the larva can be observed during all stages. The presented method provides an opportunity for observing and analyzing whole tissue-scale behaviors such as fin development and growth in a simple microscope setting, which is easily adaptable to any stereomicroscope with time-lapse capabilities.

We present a protocol for capturing the dynamics of zebrafish larval tail fin regeneration on a whole-tissue scale using brightfield-based stereomicroscopy. This technique enables capturing the regeneration dynamics with single cell resolution. This methodology can be adapted to any stereomicroscope equipped with a CCD camera and time-lapse software.

시간 경과 브라이트 실체 현미경으로 Zebrafish의 애벌레에서 조직 복구를 캡처

The zebrafish larval tail fin is ideal for studying tissue regeneration due to the simple architecture of the larval fin-fold, which comprises of two ...

Nanoparticle-mediated siRNA Gene-silencing in Adult Zebrafish Heart

Mammals have a very limited capacity to regenerate the heart after myocardial infarction. On the other hand, the adult zebrafish regenerates its heart after apex resection or cryoinjury, making it an important model organism for heart regeneration study. However, the lack of loss-of-function methods for adult organs has restricted insights into the mechanisms underlying heart regeneration. RNA interference via different delivery systems is a powerful tool for silencing genes in mammalian cells and model organisms. We have previously reported that siRNA-encapsulated nanoparticles successfully enter cells and result in a remarkable gene-specific knockdown in the regenerating adult zebrafish heart. Here, we present a simple, rapid, and efficient protocol for the dendrimer-mediated siRNA delivery and gene-silencing in the regenerating adult zebrafish heart. This method provides an alternative approach for determining gene functions in adult organs in zebrafish and can be extended to other model organisms as well.

It remains a major challenge to develop conditional gene-knockout or effective gene-knockdown in adult zebrafish organs. Here we report a protocol for performing nanoparticle-mediated siRNA gene-silencing in adult zebrafish heart, thus providing a new loss-of-function method for studying adult organs in zebrafish and other model organisms.

나노-중재 siRNA 유전자 입을 성인 Zebrafish 마음에

Mammals have a very limited capacity to regenerate the heart after myocardial infarction. On the other hand, the adult zebrafish regenerates its heart ...

Spinal Cord Transection in the Larval Zebrafish

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability to reinitiate spinal neurogenesis. However, some anamniotes including the zebrafish Danio rerio exhibit both sensory and functional recovery even after complete transection of the spinal cord. The adult zebrafish is an established model organism for studying regeneration following spinal cord injury, with sensory and motor recovery by 6&#160;weeks post-injury. To take advantage of in vivo analysis of the regenerative process available in the transparent larval zebrafish as well as genetic tools not accessible in the adult, we use the larval zebrafish to study regeneration after spinal cord transection. Here we demonstrate a method for reproducibly and verifiably transecting the larval spinal cord. After transection, our data shows sensory recovery beginning at 2&#160;days post-injury (dpi), with the C-bend movement detectable by 3 dpi and resumption of free swimming by 5 dpi. Thus we propose the larval zebrafish as a companion tool to the adult zebrafish for the study of recovery after spinal cord injury.

After spinal transection, adult zebrafish have functional recovery by six weeks post-injury. To take advantage of larval transparency and faster recovery, we present a method for transecting the larval spinal cord. After transection, we observe sensory recovery beginning at 2&#160;days post-injury, and C-bend movement by 3 days post-injury.

애벌레 Zebrafish의 척수 절개

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability ...

신경과학

Stab Wound Injury of the Zebrafish Adult Telencephalon: A Method to Investigate Vertebrate Brain Neurogenesis and Regeneration

Adult zebrafish have an amazing capacity to regenerate their central nervous system after injury. To investigate the cellular response and the molecular mechanisms involved in zebrafish adult central nervous system (CNS) regeneration and repair, we developed a zebrafish model of adult telencephalic injury.
In this approach, we manually generate an injury by pushing an insulin syringe needle into the zebrafish adult telencephalon. At different post injury days, fish are sacrificed, their brains are dissected out and stained by immunohistochemistry and/or in situ hybridization (ISH) with appropriate markers to observe cell proliferation, gliogenesis, and neurogenesis. The contralateral unlesioned hemisphere serves as an internal control. This method combined for example with RNA deep sequencing can help to screen for new genes with a role in zebrafish adult telencephalon neurogenesis, regeneration, and repair.

To shed light on the cellular and molecular mechanisms of zebrafish adult neurogenesis and regeneration, we developed a protocol for invasive surgery causing mechanical injuries in the zebrafish adult telencephalon and subsequent monitoring of changes in the stabbed hemisphere by immunohistochemistry or in situ hybridization.

Zebrafish의 성인 Telencephalon의 자상 부상 : 척추 동물의 뇌 신경 발생과 재생을 조사하는 방법

Adult zebrafish have an amazing capacity to regenerate their central nervous system after injury. To investigate the cellular response and the molecular ...

Examining Muscle Regeneration in Zebrafish Models of Muscle Disease

Skeletal muscle has a remarkable ability to regenerate following injury, which is driven by obligate tissue resident muscle stem cells. Following injury, the muscle stem cell is activated and undergoes cell proliferation to generate a pool of myoblasts, which subsequently differentiate to form new muscle fibers. In many muscle wasting conditions, including muscular dystrophy and ageing, this process is impaired resulting in the inability of muscle to regenerate. The process of muscle regeneration in zebrafish is highly conserved with mammalian systems providing an excellent system to study muscle stem cell function and regeneration, in muscle wasting conditions such as muscular dystrophy. Here, we present a method to examine muscle regeneration in zebrafish models of muscle disease. The first step involves the use of a genotyping platform that allows the determination of the genotype of the larvae prior to eliciting an injury. Having determined the genotype, the muscle is injured using a needle stab, following which polarizing light microscopy is used to determine the extent of muscle regeneration. We therefore provide a high throughput pipeline which allows the examination of muscle regeneration in zebrafish models of muscle disease.

Skeletal muscle regeneration is driven by tissue resident muscle stem cells, which are impaired in many muscle diseases such as muscular dystrophy, and this results in the inability of muscle to regenerate. Here, we describe a protocol that allows the examination of muscle regeneration in zebrafish models of muscle disease.

근육 질환의 제브라피시 모델에서 근육 재생 검토

Skeletal muscle has a remarkable ability to regenerate following injury, which is driven by obligate tissue resident muscle stem cells. Following injury, ...

Stab Wound Injury Model of the Adult Optic Tectum Using Zebrafish and Medaka for the Comparative Analysis of Regenerative Capacity

While zebrafish have a superior capacity to regenerate their central nervous system (CNS), medaka has a lower CNS regenerative capacity. A brain injury model was developed in the adult optic tectum of zebrafish and medaka and comparative histological and molecular analyses were performed to elucidate the molecular mechanisms regulating the high regenerative capacity of this tissue across these fish species. Here a stab wound injury model is presented for the adult optic tectum using a needle and histological analyses for proliferation and differentiation of the neural stem cells (NSCs). A needle was manually inserted into the central region of the optic tectum, and then the fish were intracardially perfused, and their brains were dissected. These tissues were then cryosectioned and evaluated using immunostaining against the appropriate NSC proliferation and differentiation markers. This tectum injury model provides robust and reproducible results in both zebrafish and medaka, allowing for comparing NSC responses after injury. This method is available for small teleosts, including zebrafish, medaka, and African killifish, and enables us to compare their regenerative capacity and investigate unique molecular mechanisms.

A mechanical brain injury model in the adult zebrafish is described to investigate the molecular mechanisms regulating their high regenerative capacity. The method explains to create a stab wound injury in the optic tectum of multiple species of small fish to evaluate the regenerative responses using fluorescent immunostaining.

재생 능력의 비교 분석을 위해 Zebrafish와 Medaka를 사용한 성인 시신경의 찔린 상처 부상 모델

While zebrafish have a superior capacity to regenerate their central nervous system (CNS), medaka has a lower CNS regenerative capacity. A brain injury ...

<meta charset="utf8"></head><body>재생 및 개발 연구를 위한 강력한 제브라피시 세포 이식

High-resolution Cell Transplantation in Embryonic and Larval Zebrafish

Development and regeneration occur by a process of genetically encoded spatiotemporally dynamic cellular interactions. The use of cell transplantation between animals to track cell fate and to induce mismatches in the genetic, spatial, or temporal properties of donor and host cells is a powerful means of examining the nature of these interactions. Organisms such as chick and amphibians have made crucial contributions to our understanding of development and regeneration, respectively, in large part because of their amenability to transplantation. The power of these models, however, has been limited by low genetic tractability. Likewise, the major genetic model organisms have lower amenability to transplantation. 
The zebrafish is a major genetic model for development and regeneration, and while cell transplantation is common in zebrafish, it is generally limited to the transfer of undifferentiated cells at the early blastula and gastrula stages of development. In this article, we present a simple and robust method that extends the zebrafish transplantation window to any embryonic or larval stage between at least 1 and 7 days post fertilization. The precision of this approach allows for the transplantation of as little as one cell with near-perfect spatial and temporal resolution in both donor and host animals. While we highlight here the transplantation of embryonic and larval neurons for the study of nerve development and regeneration, respectively, this approach is applicable to a wide range of progenitor and differentiated cell types and research questions.

<meta charset="utf8"></head><body>저자 스포트라이트: 발달 및 재생 중 제브라피시 신경계의 세포 연결성 매핑

Here we present a protocol to transplant cells with high spatial and temporal resolution in zebrafish embryos and larvae at any stage between at least 1 and 7 days post fertilization.

배아 및 유충 제브라피시에서의 고분해능 세포 이식

Development and regeneration occur by a process of genetically encoded spatiotemporally dynamic cellular interactions. The use of cell transplantation ...

코스닥 시스템의 사용은 분자 메커니즘 Zebrafish의 재생의 연구를

요약

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이 비디오의 챕터

겐타 마이신 유도 부상으로 성인 Zebrafish의에서 신장 재생

시간 경과 브라이트 실체 현미경으로 Zebrafish의 애벌레에서 조직 복구를 캡처

나노-중재 siRNA 유전자 입을 성인 Zebrafish 마음에

애벌레 Zebrafish의 척수 절개

Zebrafish의 성인 Telencephalon의 자상 부상 : 척추 동물의 뇌 신경 발생과 재생을 조사하는 방법

근육 질환의 제브라피시 모델에서 근육 재생 검토

재생 능력의 비교 분석을 위해 Zebrafish와 Medaka를 사용한 성인 시신경의 찔린 상처 부상 모델

배아 및 유충 제브라피시에서의 고분해능 세포 이식

배아 및 유충 제브라피시에서의 고분해능 세포 이식

배아 및 유충 제브라피시에서의 고분해능 세포 이식