TY - JOUR
T1 - Vesicular nucleotide transporter mediates ATP release and migration in neutrophils
AU - Harada, Yuika
AU - Kato, Yuri
AU - Miyaji, Takaaki
AU - Omote, Hiroshi
AU - Moriyama, Yoshinori
AU - Hiasa, Miki
N1 - Funding Information:
This work was supported in part by Japan Society for the Promotion of Sci-ence Fellows Grant-in Aid 15J03354 (to Y. H.); Japanese Ministry of Educa-tion, Culture, Sports, Science, and Technology of Japan Grants-in-Aid 26893154 (to Y. K.), 17K0733609 (to H. O.), 25253008 (to H. O. and Y. M.), and 16K08230 (to M. H.); the Japan Agency for Medical Research and Development (AMED) Grant JP17gm5910019 (to T. M.); and funds from the Narishige Neuroscience Research Foundation (to M. H.). The authors declare that they have no conflicts of interest with the contents of this article.
Funding Information:
This work was supported in part by Japan Society for the Promotion of Science Fellows Grant-in Aid 15J03354 (to Y. H.); Japanese Ministry of Education, Culture, Sports, Science, and Technology of Japan Grants-in-Aid 26893154 (to Y. K.), 17K0733609 (to H. O.), 25253008 (to H. O. and Y. M.), and 16K08230 (to M. H.); the Japan Agency for Medical Research and Development (AMED) Grant JP17gm5910019 (to T. M.); and funds from the Narishige Neuroscience Research Foundation (to M. H.). The authors declare that they have no conflicts of interest with the contents of this article.
Publisher Copyright:
© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2018/3/9
Y1 - 2018/3/9
N2 - Neutrophils migrate to sites infected by pathogenic microorganisms. This migration is regulated by neutrophil-secreted ATP, which stimulates neutrophils in an autocrine manner through purinergic receptors on the plasma membrane. Although previous studies have shown that ATP is released through channels at the plasma membrane of the neutrophil, it remains unknown whether it is also released through alternate secretory systems involving vesicular mechanisms. In this study, we investigated the possible involvement of vesicular nucleotide transporter (VNUT), a key molecule for vesicular storage and nucleotide release, in ATP secretion from neutrophils. RT-PCR and Western blotting analysis indicated that VNUT is expressed in mouse neutrophils. Immunohistochemical analysis indicated that VNUT mainly colocalized with matrix metalloproteinase-9 (MMP-9), a marker of tertiary granules, which are secretory organelles. In mouse neutrophils, ATP release was inhibited by clodronate, which is a potent VNUT inhibitor. Furthermore, neutrophils from VNUT/ mice did not release ATP and exhibited significantly reduced migration in vitro and in vivo. These findings suggest that tertiary granule-localized VNUT is responsible for vesicular ATP release and subsequent neutrophil migration. Thus, these findings suggest an additional mechanism through which ATP is released by neutrophils.
AB - Neutrophils migrate to sites infected by pathogenic microorganisms. This migration is regulated by neutrophil-secreted ATP, which stimulates neutrophils in an autocrine manner through purinergic receptors on the plasma membrane. Although previous studies have shown that ATP is released through channels at the plasma membrane of the neutrophil, it remains unknown whether it is also released through alternate secretory systems involving vesicular mechanisms. In this study, we investigated the possible involvement of vesicular nucleotide transporter (VNUT), a key molecule for vesicular storage and nucleotide release, in ATP secretion from neutrophils. RT-PCR and Western blotting analysis indicated that VNUT is expressed in mouse neutrophils. Immunohistochemical analysis indicated that VNUT mainly colocalized with matrix metalloproteinase-9 (MMP-9), a marker of tertiary granules, which are secretory organelles. In mouse neutrophils, ATP release was inhibited by clodronate, which is a potent VNUT inhibitor. Furthermore, neutrophils from VNUT/ mice did not release ATP and exhibited significantly reduced migration in vitro and in vivo. These findings suggest that tertiary granule-localized VNUT is responsible for vesicular ATP release and subsequent neutrophil migration. Thus, these findings suggest an additional mechanism through which ATP is released by neutrophils.
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U2 - 10.1074/jbc.M117.810168
DO - 10.1074/jbc.M117.810168
M3 - Article
C2 - 29363573
AN - SCOPUS:85043704896
VL - 293
SP - 3770
EP - 3779
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 10
ER -