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    FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
    Authors:  Nadezhda S. Sabeva and Maria Bykhovskaia, date: 09/05/2017, view: 272, Q&A: 0
    [Abstract] We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are ...
    Microinjection of Virus into Lumbar Enlargement of Spinal Dorsal Horn in Mice
    Authors:  Zhi-Jun Zhang, Peng-Bo Jing and Yong-Jing Gao, date: 11/20/2016, view: 2003, Q&A: 0
    [Abstract] In order to explore the role of a specific gene/protein in the specific segment of spinal cord, the technique of intraspinal injection is particularly used to deliver viral vectors targeting the specific gene/protein. These viral vectors can knockdown or overexpress the specific gene/protein in specific cells (glial cells or neurons). In this ...
    In utero Electroporation of Mouse Cerebellar Purkinje Cells
    Author:  Yukari H. Takeo, date: 06/05/2016, view: 2751, Q&A: 0
    [Abstract] In utero electroporation (IUE) of mouse cerebellar Purkinje cells allows high expression levels of transgenes without toxicity (Nishiyama et al., 2012). This technique is suitable for co-transfection of multiple plasmid genes. Therefore, it is useful to express various sets of genes such as drug-inducible Cre/loxP constructs and ...
    Dissection and Staining of Mouse Brain Ventricular Wall for the Analysis of Ependymal Cell Cilia Organization
    [Abstract] In the developing and mature central nervous system (CNS) the ventricular lumen is lined by the neuroepithelium and ependymal, respectively. These ventricular epithelia perform important functions related to the development, morphogenesis and physiology of the brain. In the mature CNS, ependyma constitute a barrier between brain parenchyma and ...
    Fluoro-Jade B Staining for Neuronal Cell Death
    Authors:  Nathalie Laflamme, Paul Préfontaine and Serge Rivest, date: 01/05/2016, view: 3604, Q&A: 0
    [Abstract] Fluoro-Jade is a fluorescent derivative used for histological staining of degenerating neurons. This technique is simple and sensitive enough to label distal dendrites, axons, axon terminals as well as neuronal bodies. Fluoro-Jade has excitation and emission peak of 480 and 525 nanometer respectively. It can be visualized using a fluorescein/FITC ...
    Chick Neural Tube Explant Culture
    Authors:  Zhanna Alekseenko, Elisabet Andersson and José M. Dias, date: 10/05/2015, view: 2675, Q&A: 0
    [Abstract] The neural tube explant culture technique allows in vitro culturing of small pieces of neural tissue isolated from e.g., chick or mouse embryonic tissue in a matrix of collagen for defined periods of time. This method can be used to study the effects of defined molecules on developmental processes such as neural progenitor ...
    Electroporation of Embryonic Chick Eyes
    Authors:  Agustín Luz-Madrigal, Erika Grajales-Esquivel and Katia Del Rio-Tsonis, date: 06/20/2015, view: 3544, Q&A: 0
    [Abstract] The chick embryo has prevailed as one of the major models to study developmental biology, cell biology and regeneration. From all the anatomical features of the chick embryo, the eye is one of the most studied. In the chick embryo, the eye develops between 26 and 33 h after incubation (Stages 8-9, Hamburger and Hamilton, 1951). It originates from ...
    In utero Electroporation of the Embryonic Mouse Retina
    Authors:  Francisco Nieto-Lopez and Luisa Sanchez-Arrones, date: 10/05/2014, view: 5295, Q&A: 0
    [Abstract] This protocol is useful to manipulate gene expression in the embryonic retina and compare the result with the contralateral non electroporated retina. In addition, the electroporation of a membrane or cytoplasmic tagged GFP allows to determine the effects of gene manipulation on the outgrowth of retinal ganglion cell axons (Garcia-Frigola et al. ...
    Combined in situ Hybridization/Immunohistochemistry (ISH/IH) on Free-floating Vibratome Tissue Sections
    Author:  Manuel E. Lopez, date: 09/20/2014, view: 5655, Q&A: 0
    [Abstract] In situ hybridization and immunostaining are common techniques for localizing gene expression, the mRNA and protein respectively, within tissues. Both techniques can be applied to tissue sections to achieve similar goals, but in some cases, it is necessary to use them together. For example, complement C1q is a secreted protein complex ...
    Open-book Preparations from Chick Embryos and DiI Labeling of Commissural Axons
    Authors:  Nicole H. Wilson and Esther T. Stoeckli, date: 07/05/2014, view: 4325, Q&A: 0
    [Abstract] Successful neural circuit formation relies on the accurate navigation of axons towards their targets during development. Axons are guided by a combination of short-range and long-range, attractive and repulsive cues. The commissural axons of the developing spinal cord have provided an informative in vivo model for the identification of ...