ID 111877
Author
Kawahara, Kohichi Kumamoto University|Niigata University of Pharmacy and Applied Life Sciences
Hirata, Hiroshi Kumamoto University
Ohbuchi, Kengo Kumamoto University
Nishi, Kentaro Kumamoto University
Maeda, Akira Kumamoto University
Kuniyasu, Akihiko Sojo University
Yamada, Daisuke Niigata University of Pharmacy and Applied Life Sciences
Maeda, Takehiko Niigata University of Pharmacy and Applied Life Sciences
Sawada, Makoto Nagoya University
Nakayama, Hitoshi Kumamoto University
Keywords
development
GPNMB/osteoactivin
microglial heterogeneity
retinal pigment epithelium
Content Type
Journal Article
Description
To differentiate subtypes of microglia (MG), we developed a novel monoclonal antibody, 9F5, against one subtype (type 1) of rat primary MG. The 9F5 showed high selectivity for this cell type in Western blot and immunocytochemical analyses and no cross‐reaction with rat peritoneal macrophages (Mφ). We identified the antigen molecule for 9F5: the 50‐ to 70‐kDa fragments of rat glycoprotein nonmetastatic melanoma protein B (GPNMB)/osteoactivin, which started at Lys170. In addition, 9F5 immunoreactivity with GPNMB depended on the activity of furin‐like protease(s). More important, rat type 1 MG expressed the GPNMB fragments, but type 2 MG and Mφ did not, although all these cells expressed mRNA and the full‐length protein for GPNMB. These results suggest that 9F5 reactivity with MG depends greatly on cleavage of GPNMB and that type 1 MG, in contrast to type 2 MG and Mφ, may have furin‐like protease(s) for GPNMB cleavage. In neonatal rat brain, amoeboid 9F5+ MG were observed in specific brain areas including forebrain subventricular zone, corpus callosum, and retina. Double‐immunοstaining with 9F5 antibody and anti‐Iba1 antibody, which reacts with MG throughout the CNS, revealed that 9F5+ MG were a portion of Iba1+ MG, suggesting that MG subtype(s) exist in vivo. We propose that 9F5 is a useful tool to discriminate between rat type 1 MG and other subtypes of MG/Mφ and to reveal the role of the GPNMB fragments during developing brain.
Journal Title
GLIA
ISSN
08941491
10981136
NCID
AA10687234
AA11620396
Publisher
Wiley
Volume
64
Issue
11
Start Page
1938
End Page
1961
Published Date
2016-07-27
Rights
© 2016 The Authors. Glia Published by Wiley Periodicals, Inc.
This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
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DOI (Published Version)
URL ( Publisher's Version )
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language
eng
TextVersion
Publisher
departments
Bioscience and Bioindustry