Vesicular polyamine transporter mediates vesicular storage and release of polyamine from mast cells

Tomoya Takeuchi, Yuika Harada, Satomi Moriyama, Kazuyuki Furuta, Satoshi Tanaka, Takaaki Miyaji, Hiroshi Omote, Yoshinori Moriyama, Miki Hiasa

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Mast cells are secretory cells that play an important role in host defense by discharging various intragranular contents, such as histamine and serotonin, upon stimulation of Fc receptors. The granules also contain spermine and spermidine, which can act as modulators of mast cell function, although the mechanism underlying vesicular storage remains unknown. Vesicular polyamine transporter (VPAT), the fourth member of the SLC18 transporter family, is an active transporter responsible for vesicular storage of spermine and spermidine in neurons. In the present study, we investigated whether VPAT functions in mast cells. RT-PCR and Western blotting indicated VPAT expression in murine bone marrow-derived mast cells (BMMCs). Immunohistochemical analysis indicated that VPAT is colocalized with VAMP3 but not with histamine, serotonin, cathepsin D, VAMP2, or VAMP7. Membrane vesicles from BMMCs accumulated spermidine upon the addition of ATP in a reserpineand bafilomycin A1-sensitive manner. BMMCs secreted spermine and spermidine upon the addition of either antigen or A23187 in the presence of Ca2, and the antigen-mediated release, which was shown to be temperature-dependent and sensitive to bafilomycin A1 and tetanus toxin, was significantly suppressed by VPAT gene RNA interference. Under these conditions, expression of vesicular monoamine transporter 2 was unaffected, but antigen-dependent histamine release was significantly suppressed, which was recovered by the addition of 1mM spermine. These results strongly suggest thatVPATis expressed and is responsible for vesicular storage of spermine and spermidine in novel secretory granules that differ from histamine- and serotonin-containing granules and is involved in vesicular release of these polyamines from mast cells.

Original languageEnglish
Pages (from-to)3909-3918
Number of pages10
JournalJournal of Biological Chemistry
Volume292
Issue number9
DOIs
Publication statusPublished - Mar 3 2017

Fingerprint

Polyamines
Spermidine
Spermine
Mast Cells
Histamine
Serotonin
Bone
Antigens
Bone Marrow
Vesicle-Associated Membrane Protein 3
Vesicle-Associated Membrane Protein 2
Vesicular Monoamine Transport Proteins
Tetanus Toxin
Cathepsin D
Fc Receptors
Histamine Release
Calcimycin
Secretory Vesicles
Modulators
Neurons

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Vesicular polyamine transporter mediates vesicular storage and release of polyamine from mast cells. / Takeuchi, Tomoya; Harada, Yuika; Moriyama, Satomi; Furuta, Kazuyuki; Tanaka, Satoshi; Miyaji, Takaaki; Omote, Hiroshi; Moriyama, Yoshinori; Hiasa, Miki.

In: Journal of Biological Chemistry, Vol. 292, No. 9, 03.03.2017, p. 3909-3918.

Research output: Contribution to journalArticle

@article{c9c00aa3406647a1bf9e7108dfdbf8e6,
title = "Vesicular polyamine transporter mediates vesicular storage and release of polyamine from mast cells",
abstract = "Mast cells are secretory cells that play an important role in host defense by discharging various intragranular contents, such as histamine and serotonin, upon stimulation of Fc receptors. The granules also contain spermine and spermidine, which can act as modulators of mast cell function, although the mechanism underlying vesicular storage remains unknown. Vesicular polyamine transporter (VPAT), the fourth member of the SLC18 transporter family, is an active transporter responsible for vesicular storage of spermine and spermidine in neurons. In the present study, we investigated whether VPAT functions in mast cells. RT-PCR and Western blotting indicated VPAT expression in murine bone marrow-derived mast cells (BMMCs). Immunohistochemical analysis indicated that VPAT is colocalized with VAMP3 but not with histamine, serotonin, cathepsin D, VAMP2, or VAMP7. Membrane vesicles from BMMCs accumulated spermidine upon the addition of ATP in a reserpineand bafilomycin A1-sensitive manner. BMMCs secreted spermine and spermidine upon the addition of either antigen or A23187 in the presence of Ca2, and the antigen-mediated release, which was shown to be temperature-dependent and sensitive to bafilomycin A1 and tetanus toxin, was significantly suppressed by VPAT gene RNA interference. Under these conditions, expression of vesicular monoamine transporter 2 was unaffected, but antigen-dependent histamine release was significantly suppressed, which was recovered by the addition of 1mM spermine. These results strongly suggest thatVPATis expressed and is responsible for vesicular storage of spermine and spermidine in novel secretory granules that differ from histamine- and serotonin-containing granules and is involved in vesicular release of these polyamines from mast cells.",
author = "Tomoya Takeuchi and Yuika Harada and Satomi Moriyama and Kazuyuki Furuta and Satoshi Tanaka and Takaaki Miyaji and Hiroshi Omote and Yoshinori Moriyama and Miki Hiasa",
year = "2017",
month = "3",
day = "3",
doi = "10.1074/jbc.M116.756197",
language = "English",
volume = "292",
pages = "3909--3918",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "9",

}

TY - JOUR

T1 - Vesicular polyamine transporter mediates vesicular storage and release of polyamine from mast cells

AU - Takeuchi, Tomoya

AU - Harada, Yuika

AU - Moriyama, Satomi

AU - Furuta, Kazuyuki

AU - Tanaka, Satoshi

AU - Miyaji, Takaaki

AU - Omote, Hiroshi

AU - Moriyama, Yoshinori

AU - Hiasa, Miki

PY - 2017/3/3

Y1 - 2017/3/3

N2 - Mast cells are secretory cells that play an important role in host defense by discharging various intragranular contents, such as histamine and serotonin, upon stimulation of Fc receptors. The granules also contain spermine and spermidine, which can act as modulators of mast cell function, although the mechanism underlying vesicular storage remains unknown. Vesicular polyamine transporter (VPAT), the fourth member of the SLC18 transporter family, is an active transporter responsible for vesicular storage of spermine and spermidine in neurons. In the present study, we investigated whether VPAT functions in mast cells. RT-PCR and Western blotting indicated VPAT expression in murine bone marrow-derived mast cells (BMMCs). Immunohistochemical analysis indicated that VPAT is colocalized with VAMP3 but not with histamine, serotonin, cathepsin D, VAMP2, or VAMP7. Membrane vesicles from BMMCs accumulated spermidine upon the addition of ATP in a reserpineand bafilomycin A1-sensitive manner. BMMCs secreted spermine and spermidine upon the addition of either antigen or A23187 in the presence of Ca2, and the antigen-mediated release, which was shown to be temperature-dependent and sensitive to bafilomycin A1 and tetanus toxin, was significantly suppressed by VPAT gene RNA interference. Under these conditions, expression of vesicular monoamine transporter 2 was unaffected, but antigen-dependent histamine release was significantly suppressed, which was recovered by the addition of 1mM spermine. These results strongly suggest thatVPATis expressed and is responsible for vesicular storage of spermine and spermidine in novel secretory granules that differ from histamine- and serotonin-containing granules and is involved in vesicular release of these polyamines from mast cells.

AB - Mast cells are secretory cells that play an important role in host defense by discharging various intragranular contents, such as histamine and serotonin, upon stimulation of Fc receptors. The granules also contain spermine and spermidine, which can act as modulators of mast cell function, although the mechanism underlying vesicular storage remains unknown. Vesicular polyamine transporter (VPAT), the fourth member of the SLC18 transporter family, is an active transporter responsible for vesicular storage of spermine and spermidine in neurons. In the present study, we investigated whether VPAT functions in mast cells. RT-PCR and Western blotting indicated VPAT expression in murine bone marrow-derived mast cells (BMMCs). Immunohistochemical analysis indicated that VPAT is colocalized with VAMP3 but not with histamine, serotonin, cathepsin D, VAMP2, or VAMP7. Membrane vesicles from BMMCs accumulated spermidine upon the addition of ATP in a reserpineand bafilomycin A1-sensitive manner. BMMCs secreted spermine and spermidine upon the addition of either antigen or A23187 in the presence of Ca2, and the antigen-mediated release, which was shown to be temperature-dependent and sensitive to bafilomycin A1 and tetanus toxin, was significantly suppressed by VPAT gene RNA interference. Under these conditions, expression of vesicular monoamine transporter 2 was unaffected, but antigen-dependent histamine release was significantly suppressed, which was recovered by the addition of 1mM spermine. These results strongly suggest thatVPATis expressed and is responsible for vesicular storage of spermine and spermidine in novel secretory granules that differ from histamine- and serotonin-containing granules and is involved in vesicular release of these polyamines from mast cells.

UR - http://www.scopus.com/inward/record.url?scp=85014686896&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85014686896&partnerID=8YFLogxK

U2 - 10.1074/jbc.M116.756197

DO - 10.1074/jbc.M116.756197

M3 - Article

C2 - 28082679

AN - SCOPUS:85014686896

VL - 292

SP - 3909

EP - 3918

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 9

ER -