Antigen-presenting cells expressing glutamate decarboxylase 67 were identified as epithelial cells in glutamate decarboxylase 67-GFP knock-in mouse thymus.
Glutamate decarboxylase (GAD), which has two isoforms, GAD65, and GAD67, is responsible for synthesis of the major inhibitory neurotransmitter, gamma-aminobutyric acid. GAD is expressed predominantly in the central nervous system; recent reports suggest that GAD is also expressed in non-neuronal organs including the pancreas. In the pancreatic islets, GAD serves as one of the autoantigens in type I diabetes mellitus. Recent flow cytometric analyses have shown that a variety of self-antigens, including GAD, are ectopically transcribed and expressed in particular cell populations of the thymus, although consensus concerning the cellular phenotype has not been obtained. The aim of this study was to clarify the localization and cellular phenotype of GAD67-expressing cells in the thymus at a cellular level with a novel approach using GAD67-green fluorescent protein (GFP) knock-in mice, in which GFP is expressed specifically in GAD67-positive cells. GFP-positive cells were detected in the thymic medulla and were identified as epithelial cells by immunohistochemistry. Almost all GFP-positive cells were positive for major histocompatibility complex (MHC) class II antigen staining and were positive for both cytokeratin and Ulex Europaeus Agglutinin I, markers of medullary thymic epithelial cells, but were negative for CD11c, Gr-1, and CD45, markers of dendritic cells, macrophages, and B-lymphocytes, respectively.
Maemura
K
K
Department of Anatomy, Osaka Medical College, Japan.
Yanagawa
Y
Y
Obata
K
K
Dohi
T
T
Egashira
Y
Y
Shibayama
Y
Y
Watanabe
M
M
D000818:Animals; D000938:Antigen-Presenting Cells / Q000201:enzymology*; D004847:Epithelial Cells / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D015870:Gene Expression; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D037121:Plant Lectins / Q000378:metabolism; D011384:Proinsulin / Q000378:metabolism; D011806:Quinolinium Compounds; D013950:Thymus Gland / Q000166:cytology / Q000201:enzymology*
C481052:Alexa fluor 546; D005456:Fluorescent Dyes; D007527:Isoenzymes; D037121:Plant Lectins; D011806:Quinolinium Compounds; C036850:Ulex europaeus lectins; D049452:Green Fluorescent Proteins; D007633:Keratins; D011384:Proinsulin; D005968:Glutamate Decarboxylase