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  1. Article ; Online: Follow Me! A Tale of Avian Heart Development with Comparisons to Mammal Heart Development.

    Lansford, Rusty / Rugonyi, Sandra

    Journal of cardiovascular development and disease

    2020  Volume 7, Issue 1

    Abstract: Avian embryos have been used for centuries to study development due to the ease of access. Because the embryos are sheltered inside the eggshell, a small window in the shell is ideal for visualizing the embryos and performing different interventions. The ...

    Abstract Avian embryos have been used for centuries to study development due to the ease of access. Because the embryos are sheltered inside the eggshell, a small window in the shell is ideal for visualizing the embryos and performing different interventions. The window can then be covered, and the embryo returned to the incubator for the desired amount of time, and observed during further development. Up to about 4 days of chicken development (out of 21 days of incubation), when the egg is opened the embryo is on top of the yolk, and its heart is on top of its body. This allows easy imaging of heart formation and heart development using non-invasive techniques, including regular optical microscopy. After day 4, the embryo starts sinking into the yolk, but still imaging technologies, such as ultrasound, can tomographically image the embryo and its heart in vivo. Importantly, because like the human heart the avian heart develops into a four-chambered heart with valves, heart malformations and pathologies that human babies suffer can be replicated in avian embryos, allowing a unique developmental window into human congenital heart disease. Here, we review avian heart formation and provide comparisons to the mammalian heart.
    Language English
    Publishing date 2020-03-07
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2777082-5
    ISSN 2308-3425 ; 2308-3425
    ISSN (online) 2308-3425
    ISSN 2308-3425
    DOI 10.3390/jcdd7010008
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Book: Molecular neuroscience

    Lansford, Rusty

    a laboratory manual

    2014  

    Author's details edited by Rusty Lansford
    MeSH term(s) Genetic Phenomena ; Genetic Techniques ; Molecular Imaging ; Neurons/physiology
    Language English
    Size xiv, 632 pages :, illustrations.
    Document type Book
    ISBN 9781621820130 ; 9781621820147 ; 1621820130 ; 1621820149
    Database Catalogue of the US National Library of Medicine (NLM)

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  3. Article ; Online: Using Avian Skin Explants to Study Tissue Patterning and Organogenesis.

    Jiang, Tingxin / Secor, Maeve / Lansford, Rusty / Widelitz, Randall B / Chuong, Cheng Ming

    Journal of visualized experiments : JoVE

    2023  , Issue 199

    Abstract: The developing avian skin during embryogenesis is a unique model that can provide valuable insights into tissue patterning. Here three variations on skin explant cultures to examine different aspects of skin development are described. First, ex vivo ... ...

    Abstract The developing avian skin during embryogenesis is a unique model that can provide valuable insights into tissue patterning. Here three variations on skin explant cultures to examine different aspects of skin development are described. First, ex vivo organ cultures and manipulations offer researchers opportunities to observe and study the development of feather buds directly. Skin explant culture can grow for 7 days enabling direct analysis of cellular behavior and 4D imaging at intervals during this growth period. This also allows for physical and molecular manipulations of culture conditions to visualize tissue response. For example, growth factor-coated beads can be applied locally to induce changes in feather patterning in a limited area. Alternatively, viral transduction can be delivered globally in the culture media to up or downregulate gene expression. Second, the skin recombination protocol allows researchers to investigate tissue interactions between the epidermis and mesenchyme that are derived from different skin regions, different life stages, or different species. This affords an opportunity to test the time window in which the epithelium is competent to respond to signals and its ability to form different skin appendages in response to signals from different mesenchymal sources. Third, skin reconstitution using dissociated dermal cells overlaid with intact epithelium resets skin development and enables the study of the initial processes of periodic patterning. This approach also enhances our ability to manipulate gene expression among the dissociated cells before creating the reconstituted skin explant. This paper provides the three culture protocols and exemplary experiments to demonstrate their utility.
    MeSH term(s) Animals ; Skin ; Epithelium/metabolism ; Feathers ; Organogenesis
    Language English
    Publishing date 2023-09-15
    Publishing country United States
    Document type Journal Article ; Video-Audio Media ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/65580
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Fluorescent Quail: A Transgenic Model System for the Dynamic Study of Avian Development.

    Huss, David / Lansford, Rusty

    Methods in molecular biology (Clifton, N.J.)

    2017  Volume 1650, Page(s) 125–147

    Abstract: Real-time four-dimensional (4D, xyzt) imaging of cultured avian embryos is an ideal method for investigating the complex movements of cells and tissues during early morphogenesis. While methods that transiently label cells, such as electroporation, are ... ...

    Abstract Real-time four-dimensional (4D, xyzt) imaging of cultured avian embryos is an ideal method for investigating the complex movements of cells and tissues during early morphogenesis. While methods that transiently label cells, such as electroporation, are highly useful for dynamic imaging, they can also be limiting due to the number and type of cells that can be effectively targeted. In contrast, the heritable, stable, and long-term expression of a fluorescent protein driven by the exogenous promoter of a transgene overcomes these challenges. We have used lentiviral vectors to produce several novel transgenic quail lines that express fluorescent proteins either ubiquitously or in a cell-specific manner. These lines have proven to be useful models for dynamic imaging and analysis. Here, we provide detailed protocols for generating transgenic quail with the emphasis on producing high titer lentivirus , effectively introducing it into the early embryo and efficiently screening for G1 founder birds .
    MeSH term(s) Animals ; Animals, Genetically Modified ; Birds/growth & development ; Birds/metabolism ; Genetic Vectors ; Green Fluorescent Proteins/metabolism ; Lentivirus/genetics ; Models, Animal ; Morphogenesis ; Quail/growth & development ; Quail/metabolism ; Transgenes
    Chemical Substances Green Fluorescent Proteins (147336-22-9)
    Language English
    Publishing date 2017
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-7216-6_8
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Fast and efficient expression of multiple proteins in avian embryos using mrna electroporation

    Tran, Martin / Dave, Mohit / Lansford, Rusty

    Journal of visualized experiments. 2019 June 07, , no. 148

    2019  

    Abstract: We report that mRNA electroporation permits fluorescent proteins to label cells in living quail embryos more quickly and broadly than DNA electroporation. The high transfection efficiency permits at least 4 distinct mRNAs to be co-transfected with ~87% ... ...

    Abstract We report that mRNA electroporation permits fluorescent proteins to label cells in living quail embryos more quickly and broadly than DNA electroporation. The high transfection efficiency permits at least 4 distinct mRNAs to be co-transfected with ~87% efficiency. Most of the electroporated mRNAs are degraded during the first 2 h post-electroporation, permitting time-sensitive experiments to be carried out in the developing embryo. Finally, we describe how to dynamically image live embryos electroporated with mRNAs that encode various subcellular targeted fluorescent proteins.
    Keywords DNA ; electroporation ; embryogenesis ; fluorescent proteins ; image analysis ; messenger RNA ; quails ; transfection
    Language English
    Dates of publication 2019-0607
    Size p. e59664.
    Publishing place Journal of Visualized Experiments
    Document type Article
    ISSN 1940-087X
    DOI 10.3791/59664
    Database NAL-Catalogue (AGRICOLA)

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  6. Article ; Online: Fast and Efficient Expression of Multiple Proteins in Avian Embryos Using mRNA Electroporation.

    Tran, Martin / Dave, Mohit / Lansford, Rusty

    Journal of visualized experiments : JoVE

    2019  , Issue 148

    Abstract: We report that mRNA electroporation permits fluorescent proteins to label cells in living quail embryos more quickly and broadly than DNA electroporation. The high transfection efficiency permits at least 4 distinct mRNAs to be co-transfected with ~87% ... ...

    Abstract We report that mRNA electroporation permits fluorescent proteins to label cells in living quail embryos more quickly and broadly than DNA electroporation. The high transfection efficiency permits at least 4 distinct mRNAs to be co-transfected with ~87% efficiency. Most of the electroporated mRNAs are degraded during the first 2 h post-electroporation, permitting time-sensitive experiments to be carried out in the developing embryo. Finally, we describe how to dynamically image live embryos electroporated with mRNAs that encode various subcellular targeted fluorescent proteins.
    MeSH term(s) Animals ; Electroporation/methods ; Embryo, Nonmammalian/metabolism ; Gene Expression ; Luminescent Proteins/genetics ; Quail/embryology ; RNA, Messenger/genetics ; Time Factors ; Transfection
    Chemical Substances Luminescent Proteins ; RNA, Messenger
    Language English
    Publishing date 2019-06-07
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/59664
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Adaptive patterning of vascular network during avian skin development: Mesenchymal plasticity and dermal vasculogenesis.

    Ou, Kuang-Ling / Chen, Chih-Kuan / Huang, Junxiang J / Chang, William Weijen / Hsieh Li, Shu-Man / Jiang, Ting-Xin / Widelitz, Randall B / Lansford, Rusty / Chuong, Cheng-Ming

    Cells & development

    2024  , Page(s) 203922

    Abstract: A vasculature network supplies blood to feather buds in the developing skin. Does the vasculature network during early skin development form by sequential sprouting from the central vasculature or does local vasculogenesis occur first that then connect ... ...

    Abstract A vasculature network supplies blood to feather buds in the developing skin. Does the vasculature network during early skin development form by sequential sprouting from the central vasculature or does local vasculogenesis occur first that then connect with the central vascular tree? Using transgenic Japanese quail Tg(TIE1p.H2B-eYFP), we observe that vascular progenitor cells appear after feather primordia formation. The vasculature then radiates out from each bud and connects with primordial vessels from neighboring buds. Later they connect with the central vasculature. Epithelial-mesenchymal recombination shows local vasculature is patterned by the epithelium, which expresses FGF2 and VEGF. Perturbing noggin expression leads to abnormal vascularization. To study endothelial origin, we compare transcriptomes of TIE1p.H2B-eYFP
    Language English
    Publishing date 2024-04-28
    Publishing country Netherlands
    Document type Journal Article
    ISSN 2667-2901
    ISSN (online) 2667-2901
    DOI 10.1016/j.cdev.2024.203922
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article: A Novel Egg-In-Cube System Enables Long-Term Culture and Dynamic Imaging of Early Embryonic Development.

    Dave, Mohit / Levin, Joshua / Ruffins, Seth Walter / Sato, Yuki / Fraser, Scott / Lansford, Rusty / Kawahara, Tomohiro

    Frontiers in physiology

    2022  Volume 13, Page(s) 893736

    Abstract: The avian egg is a closed system that protects the growing embryo from external factors but prevents direct observation of embryo development. Various culture systems exist in the literature to study the development of the embryo for short periods of ... ...

    Abstract The avian egg is a closed system that protects the growing embryo from external factors but prevents direct observation of embryo development. Various culture systems exist in the literature to study the development of the embryo for short periods of incubation (from 12 h up to a maximum of 60 h of egg incubation). A common flaw to these culture techniques is the inability to culture the unincubated avian blastoderm with intact tissue tensions on its native yolk. The goal of this work is to create a unique novel egg-in-cube system that can be used for long-term quail embryo culture initiated from its unincubated blastoderm stage. The egg-in-cube acts as an artificial transparent eggshell system that holds the growing embryo, making it amenable to microscopy. With the egg-in-cube system, quail embryos can be grown up to 9 days from the unincubated blastoderm (incubated in air, 20.9% O
    Language English
    Publishing date 2022-05-12
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2564217-0
    ISSN 1664-042X
    ISSN 1664-042X
    DOI 10.3389/fphys.2022.893736
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Generation and analysis of lentivirus expressing a 2A peptide-linked bicistronic fluorescent construct.

    Huss, David / Lansford, Rusty

    Cold Spring Harbor protocols

    2014  Volume 2014, Issue 12, Page(s) 1290–1311

    Abstract: This weeklong protocol for making and testing lentivirus has been used in the Advanced Topics in Molecular Neuroscience (ATMN) lecture and laboratory course at Cold Spring Harbor Laboratory (CSHL) for nearly a decade. Lentiviruses are derived from HIV-1 ... ...

    Abstract This weeklong protocol for making and testing lentivirus has been used in the Advanced Topics in Molecular Neuroscience (ATMN) lecture and laboratory course at Cold Spring Harbor Laboratory (CSHL) for nearly a decade. Lentiviruses are derived from HIV-1 and are ideal vehicles for the delivery of multiple genes of interest into target cells. In this protocol, 2A peptide-linked sequences are used to create a bicistronic lentiviral construct containing a ubiquitous promoter (chick β actin with a cytomegalovirus [CMV] early enhancer) driving dual expression of two fluorescent proteins (FP): H2B-Cerulean (a nuclear-localized blue FP) and Dendra2 (a photoactivatable green FP that converts to red after exposure to UV light). Polymerase chain reaction (PCR) amplification of the bicistronic insert is followed by subcloning into a lentiviral vector and transfection into a packaging cell line. The resulting viral supernatants can be used to prepare concentrated stocks and infect cells for imaging via epifluorescent and confocal microscopy.
    MeSH term(s) Animals ; Base Sequence ; Chickens ; DNA/metabolism ; Electrophoresis, Agar Gel ; Escherichia coli/metabolism ; Genetic Techniques ; Genetic Vectors/metabolism ; Green Fluorescent Proteins/metabolism ; HEK293 Cells ; Humans ; Lentivirus/metabolism ; Mice ; Microscopy, Fluorescence ; Molecular Sequence Data ; NIH 3T3 Cells ; Oligonucleotides/genetics ; Peptides/metabolism ; Plasmids/metabolism ; Polymerase Chain Reaction ; Transfection ; Transformation, Genetic
    Chemical Substances Oligonucleotides ; Peptides ; Green Fluorescent Proteins (147336-22-9) ; DNA (9007-49-2)
    Language English
    Publishing date 2014-12-01
    Publishing country United States
    Document type Journal Article
    ISSN 1559-6095
    ISSN (online) 1559-6095
    DOI 10.1101/pdb.prot081422
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Transgenesis and imaging in birds, and available transgenic reporter lines.

    Sato, Yuki / Lansford, Rusty

    Development, growth & differentiation

    2013  Volume 55, Issue 4, Page(s) 406–421

    Abstract: Avian embryos are important model organism to study higher vertebrate development. Easy accessibility to developing avian embryos enables a variety of experimental applications to understand specific functions of molecules, tissue-tissue interactions, ... ...

    Abstract Avian embryos are important model organism to study higher vertebrate development. Easy accessibility to developing avian embryos enables a variety of experimental applications to understand specific functions of molecules, tissue-tissue interactions, and cell lineages. The whole-mount ex ovo culture technique for avian embryos permits time-lapse imaging analysis for a better understanding of cell behaviors underlying tissue morphogenesis in physiological conditions. To study mechanisms of blood vessel formation and remodeling in developing embryos by using a time-lapse imaging approach, a transgenic quail model, Tg(tie1:H2B-eYFP), was generated. From a cell behavior perspective, Tg(tie1:H2B-eYFP) quail embryos are a suitable model to shed light on how the structure and pattern of blood vessels are established in higher vertebrates. In this manuscript, we give an overview on the biological and technological background of the transgenic quail model and describe procedures for the ex ovo culture of quail embryos and time-lapse imaging analysis.
    MeSH term(s) Animals ; Animals, Genetically Modified/genetics ; Bacterial Proteins/metabolism ; Birds ; Culture Techniques ; Embryo Culture Techniques/methods ; Genes, Reporter ; Luminescent Proteins/metabolism ; Microscopy, Confocal/methods ; Quail/genetics ; Software ; Transgenes
    Chemical Substances Bacterial Proteins ; Luminescent Proteins ; yellow fluorescent protein, Bacteria
    Language English
    Publishing date 2013-05
    Publishing country Japan
    Document type Journal Article ; Review
    ZDB-ID 280433-5
    ISSN 1440-169X ; 0012-1592
    ISSN (online) 1440-169X
    ISSN 0012-1592
    DOI 10.1111/dgd.12058
    Database MEDical Literature Analysis and Retrieval System OnLINE

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