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Visualization of microglia in living tissues using Iba1-EGFP transgenic mice.

Abstract

Microglia are thought to play important roles not only in repairing injured tissue but in regulating neuronal activity, and visualizing the cells is very useful as a means of further investigating the function of microglia in vivo. We previously cloned the ionized calcium-binding adaptor molecule 1 (Iba1) gene, which is expressed selectively in microglia/microphages. To generate new transgenic mice to visualize microglia with enhanced green fluorescent protein (EGFP), we here constructed a plasmid carrying EGFP cDNA under control of the Iba1 promoter. This construct was injected into C57B/6 mouse zygotes, and the Iba1-EGFP transgenic line was developed. Fluorescent in-situ hybridization analysis revealed that the Iba1-EGFP transgene was located on chromosome 11D. No obvious defects were observed during development or in adulthood, and the EGFP fluorescence remained invariant over the course of at least four generations. Judging from the immunoreactivity with anti-Iba1 antibody, all EGFP-positive cells in the adult brain were ramified microglia. In the developing transgenic embryos, EGFP signals were detected as early as embryonic Day 10.5. The most prominent EGFP signals were found in forebrain, spinal cord, eye, foreleg, yolk sac, liver, and vessel walls. At postnatal Day 6, clear EGFP signals were observed in the supraventricular corpus callosum, known as "fountain of microglia", where ameboid microglia migrate into the brain parenchyma and mature into ramified microglia. Iba1-EGFP transgenic mice thus permit observation of living microglia under a fluorescence microscope and provide a useful tool for studying the function of microglia in vivo.

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  • Authors+Show Affiliations

    ,

    Department of Neurochemistry, National Institute of Neuroscience, Kodaira, Tokyo, Japan.

    , , , , ,

    Source

    Journal of neuroscience research 81:3 2005 Aug 01 pg 357-62

    MeSH

    Animals
    Brain
    Calcium-Binding Proteins
    Cell Line
    Cercopithecus aethiops
    Chromosomes, Human, Pair 11
    Embryo, Mammalian
    Gene Expression Regulation, Developmental
    Green Fluorescent Proteins
    Humans
    Immunohistochemistry
    In Situ Hybridization, Fluorescence
    In Vitro Techniques
    Mice
    Mice, Inbred C57BL
    Mice, Transgenic
    Microfilament Proteins
    Microglia
    Transfection

    Pub Type(s)

    Comparative Study
    Journal Article
    Research Support, Non-U.S. Gov't

    Language

    eng

    PubMed ID

    15948177

    Citation

    Hirasawa, T, et al. "Visualization of Microglia in Living Tissues Using Iba1-EGFP Transgenic Mice." Journal of Neuroscience Research, vol. 81, no. 3, 2005, pp. 357-62.
    Hirasawa T, Ohsawa K, Imai Y, et al. Visualization of microglia in living tissues using Iba1-EGFP transgenic mice. J Neurosci Res. 2005;81(3):357-62.
    Hirasawa, T., Ohsawa, K., Imai, Y., Ondo, Y., Akazawa, C., Uchino, S., & Kohsaka, S. (2005). Visualization of microglia in living tissues using Iba1-EGFP transgenic mice. Journal of Neuroscience Research, 81(3), pp. 357-62.
    Hirasawa T, et al. Visualization of Microglia in Living Tissues Using Iba1-EGFP Transgenic Mice. J Neurosci Res. 2005 Aug 1;81(3):357-62. PubMed PMID: 15948177.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Visualization of microglia in living tissues using Iba1-EGFP transgenic mice. AU - Hirasawa,T, AU - Ohsawa,K, AU - Imai,Y, AU - Ondo,Y, AU - Akazawa,C, AU - Uchino,S, AU - Kohsaka,S, PY - 2005/6/11/pubmed PY - 2005/11/8/medline PY - 2005/6/11/entrez SP - 357 EP - 62 JF - Journal of neuroscience research JO - J. Neurosci. Res. VL - 81 IS - 3 N2 - Microglia are thought to play important roles not only in repairing injured tissue but in regulating neuronal activity, and visualizing the cells is very useful as a means of further investigating the function of microglia in vivo. We previously cloned the ionized calcium-binding adaptor molecule 1 (Iba1) gene, which is expressed selectively in microglia/microphages. To generate new transgenic mice to visualize microglia with enhanced green fluorescent protein (EGFP), we here constructed a plasmid carrying EGFP cDNA under control of the Iba1 promoter. This construct was injected into C57B/6 mouse zygotes, and the Iba1-EGFP transgenic line was developed. Fluorescent in-situ hybridization analysis revealed that the Iba1-EGFP transgene was located on chromosome 11D. No obvious defects were observed during development or in adulthood, and the EGFP fluorescence remained invariant over the course of at least four generations. Judging from the immunoreactivity with anti-Iba1 antibody, all EGFP-positive cells in the adult brain were ramified microglia. In the developing transgenic embryos, EGFP signals were detected as early as embryonic Day 10.5. The most prominent EGFP signals were found in forebrain, spinal cord, eye, foreleg, yolk sac, liver, and vessel walls. At postnatal Day 6, clear EGFP signals were observed in the supraventricular corpus callosum, known as "fountain of microglia", where ameboid microglia migrate into the brain parenchyma and mature into ramified microglia. Iba1-EGFP transgenic mice thus permit observation of living microglia under a fluorescence microscope and provide a useful tool for studying the function of microglia in vivo. SN - 0360-4012 UR - https://www.unboundmedicine.com/medline/citation/15948177/Visualization_of_microglia_in_living_tissues_using_Iba1_EGFP_transgenic_mice_ L2 - https://doi.org/10.1002/jnr.20480 DB - PRIME DP - Unbound Medicine ER -