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  JoVE Developmental Biology


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October, 2006
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Gastrula: The developmental stage that follows Blastula. It is characterized by the morphogenetic cell movements including invagination, ingression, and involution. Gastrulation begins with the formation of the Primitive streak, and ends with the formation of three Germ layers, the body plan of the mature organism.
 JoVE Biology

Tissue Targeted Embryonic Chimeras: Zebrafish Gastrula Cell Transplantation

1Department of Biological Sciences, Smith College

JoVE 1422

Zebrafish cell transplantation enables the combination of genetics and embryology to generate tissue specific chimeras. This video demonstrates gastrula staged cell transplantations that have allowed our lab to investigate the roles of astroglial populations and specific guidance cues during commissure formation in the forebrain.

 JoVE Biology

Primary Cell Cultures from Drosophila Gastrula Embryos

1Department of Genetics, Harvard Medical School, 2Howard Hughes Medical Institute

JoVE 2215

We provide a detailed protocol for preparing primary cells dissociated from Drosophila embryos. The ability to carry out the effective RNAi perturbation, together with other molecular, biochemical and cell imaging methods will allow a variety of questions to be addressed in Drosophila primary cells.

 JoVE Biology

Generating Chimeric Zebrafish Embryos by Transplantation

1HHMI and Division of Basic Sciences, Fred Hutchinson Cancer Research Center - FHCRC

JoVE 1394

A step-by-step guide to generating targeted chimeric zebrafish embryos by transplantation at the blastula or gastrula stage.

 JoVE Developmental Biology

Manipulation and In Vitro Maturation of Xenopus laevis Oocytes, Followed by Intracytoplasmic Sperm Injection, to Study Embryonic Development

1Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, 2Department of Zoology, University of Cambridge

JoVE 52496

We describe methods of manipulating Xenopus laevis immature oocytes, in vitro maturation of oocytes to eggs, and intracytoplasmic sperm injection. This protocol allows degradation of some maternal proteins and overexpression of genes of interest at fertilization, and hence is valuable to study roles of specific factors in early embryonic development.

 JoVE Developmental Biology

Genome-wide Snapshot of Chromatin Regulators and States in Xenopus Embryos by ChIP-Seq

1Division of Systems Biology, MRC National Institute for Medical Research

JoVE 52535

The question of how chromatin regulators and chromatin states affect the genome in vivo is key to our understanding of how early cell fate decisions are made in the developing embryo. ChIP-Seq—the most popular approach to investigate chromatin features at a global level—is outlined here for Xenopus embryos.

 JoVE Biology

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

1The Healing Foundation Centre, Faculty of Life Sciences, University of Manchester, 2Department of Developmental Biology, Washington University School of Medicine

JoVE 2010

This video protocol demonstrates a method for generating transgenic Xenopus laevis by introduction of transgenes into sperm nuclei followed by nuclear transplantation into unfertilized eggs.

 JoVE Developmental Biology

Expression of Fluorescent Proteins in Branchiostoma lanceolatum by mRNA Injection into Unfertilized Oocytes

1Département de Biologie du Développement et Cellules Souches, Institut Pasteur, 2Laboratoire de Biologie du Développement de Villefranche-sur-Mer (UMR7009 CNRS/UPMC Univ Paris 06), Sorbonne Universités, 3Equipe Epigenetic Control of Normal and Pathological Hematopoiesis, Centre de Recherche en Cancérologie de Marseille, 4Unité de Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR5235/DAA/cc107/Université Montpellier II, 5Plateforme BioEmergences IBiSA FBI, CNRS-NED, Institut de Neurobiologie Alfred Fessard

JoVE 52042

We report here the robust and efficient expression of fluorescent proteins after mRNA injection into unfertilized oocytes of Branchiostoma lanceolatum. The development of the microinjection technique in this basal chordate will pave the way for far-reaching technical innovations in this emerging model system, including in vivo imaging and gene-specific manipulations.

 Science Education: Essentials of Biology 2

Zebrafish Reproduction and Development

JoVE Science Education

The zebrafish (Danio rerio) has become a popular model for studying genetics and developmental biology. The transparency of these animals at early developmental stages permits the direct visualization of tissue morphogenesis at the cellular level. Furthermore, zebrafish are amenable to genetic manipulation, allowing researchers to determine the effect of gene expression on the development of a vertebrate with a high degree of genetic similarity to humans. This video provides a brief overview of the major phases of zebrafish development, with particular focus on the first 24 hours post fertilization (hpf). The discussion begins with a zygote consisting of a single cell, or blastomere, atop a large ball of yolk. Cleavage of the blastomere is then shown to produce an embryo containing thousands of cells within a matter of hours. Next, the dramatic cellular movements known as epiboly and gastrulation are explained, revealing how they contribute to reshaping a mass of cells into a moving embryo with a beating heart in just 1 day. The presentation follows embryo development through the hatching phase, when they become swimming, feeding larvae. Important considerations for caring for larvae are incorporated, including a brief review of how fish are raised to adulthood in a dedicated facility known as the nursery. Finally, the video concludes with some commo

 JoVE Biology

Blastomere Explants to Test for Cell Fate Commitment During Embryonic Development

1Department of Biological Sciences, The George Washington University, 2Department of Anatomy and Regenerative Biology, The George Washington University

JoVE 4458

The fate of an individual embryonic cell can be influenced by inherited molecules and/or by signals from neighboring cells. Utilizing fate maps of the cleavage stage Xenopus embryo, single blastomeres can be identified for culture in isolation to assess the contributions of inherited molecules versus cell-cell interactions.

 JoVE Developmental Biology

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

1Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory

JoVE 52654

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.

 JoVE Biology

Analysis of Gene Function and Visualization of Cilia-Generated Fluid Flow in Kupffer's Vesicle

1Department of Cell and Developmental Biology, State University of New York, Upstate Medical University, 2Department of Neurobiology and Anatomy, Eccles Institute of Human Genetics, University of Utah

JoVE 50038

Cilia-generated fluid flow in Kupffer’s Vesicle (KV) controls left-right patterning of the zebrafish embryo. Here, we describe a technique to modulate gene function specifically in KV cells. In addition, we show how to deliver fluorescent beads into KV to visualize fluid flow.

 JoVE Developmental Biology

Microbead Implantation in the Zebrafish Embryo

1Department of Biological Sciences, University of Notre Dame

JoVE 52943

The zebrafish is an excellent model system for genetic and developmental studies. Bead implantation is a valuable tissue manipulation technique that can be used to interrogate developmental mechanisms by introducing alterations in local cellular environments. This protocol describes how to perform microbead implantation in the zebrafish embryo.

 JoVE Biology

Single Cell Fate Mapping in Zebrafish

1Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, 2Division of Hematology/Oncology, Cincinnati Children's Hospital Medical Center

JoVE 3172

A method is described to photoactivate single cells containing a caged fluorescent protein using two-photon absorption from a Ti:Sapphire femtosecond laser oscillator. To fate map the photoactivated cell, immunohistochemistry is used. This technique can be applied to any cell type.

 JoVE Neuroscience

Dissection and Lateral Mounting of Zebrafish Embryos: Analysis of Spinal Cord Development

1Biology Department, Skidmore College

JoVE 50703

Developmental processes such as proliferation, patterning, differentiation, and axon guidance can be readily modeled in the zebrafish spinal cord. In this article, we describe a mounting procedure for zebrafish embryos, which optimizes visualization of these events.

 JoVE Biology

Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae

1Department of Development and Cell Biology, Department of Anatomy and Neurobiology, University of California, Irvine (UCI)

JoVE 200

This video demonstrates the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. Views of live cultures show neurite outgrowth and imaging of calcium levels using Fura-2.

 JoVE Biology

Imaging Glycans in Zebrafish Embryos by Metabolic Labeling and Bioorthogonal Click Chemistry

1Department of Biochemistry, Albert Einstein College of Medicine, Yeshiva University, 2Macromolecular Therapeutics Development Facility, Albert Einstein College of Medicine, Yeshiva University, 3Developmental and Molecular Biology, Albert Einstein College of Medicine, Yeshiva University

JoVE 2686

A click-chemistry based method that allows for the rapid, noninvasive, and robust labeling of alkyne-tagged glycans in zebrafish embryos is described. Fucosylated glycans in the enveloping layer of zebrafish embryos in the late gastrulation stage were imaged in this study.

 JoVE Biology

Microinjection of mRNA and Morpholino Antisense Oligonucleotides in Zebrafish Embryos.

1Department of Genetics, Yale University School of Medicine

JoVE 1113

Microinjection is a well-established and effective method for introducing foreign substances into fertilized zebrafish embryos. Here, we demonstrate a robust microinjection technique for performing mRNA overexpression, and morpholino oligonucleotide gene knockdown studies in zebrafish.

 Science Education: Essentials of Developmental Biology

Fate Mapping

JoVE Science Education

Fate mapping is a technique used to understand how embryonic cells divide, differentiate, and migrate during development. In classic fate mapping experiments, cells in different areas of an embryo are labeled with a chemical dye and then tracked to determine which tissues or structures they form. Technological improvements now allow for individual cells to be marked and traced throughout embryonic development and adulthood. This video reviews the concepts behind fate mapping, and then details a fate mapping protocol in zebrafish using photoactivatable fluorescent proteins. Finally, specific applications and modifications of this unique technique are discussed.

 JoVE Biology

Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos

1Department of Development and Cell Biology, Department of Anatomy and Neurobiology, University of California, Irvine (UCI)

JoVE 226

This video demonstrates the preparation of primary neuronal cultures from midgastrula stage Drosophila embryos. Views of live cultures show cells 1 hour after plating and differentiated neurons after 2 days of growth in a bicarbonate-based defined medium. The neurons are electrically excitable and form synaptic connections.

 JoVE Biology

Polarized Translocation of Fluorescent Proteins in Xenopus Ectoderm in Response to Wnt Signaling

1Department of Developmental and Regenerative Biology, Mount Sinai School of Medicine

JoVE 2700

Xenopus embryonic ectoderm has become an attractive model for studies of cell polarity. An assay is described, in which subcellular distribution of fluorescent proteins is assessed in ectoderm cells. This protocol will help address questions related to spatial control of signaling.

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