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 reserpine- and 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 A1and tetanus toxin, was significantly suppressed byVPATgene 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 1 mm spermine. These results strongly suggest that VPAT is 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.
authors.affiliation: From the Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8530.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Takeuchi
authors.affiliation: From the Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8530.
authors.forename: Yuika
authors.initials: Y
authors.lastname: Harada
authors.affiliation: From the Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8530.
authors.forename: Satomi
authors.initials: S
authors.lastname: Moriyama
authors.affiliation: the Department of Immunobiology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8530, and.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Furuta
authors.affiliation: the Department of Immunobiology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8530, and.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tanaka
authors.affiliation: the Advanced Science Research Center, Okayama University, Okayama 700-8530, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Miyaji
authors.affiliation: From the Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8530.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Omote
authors.affiliation: From the Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8530, moriyama@pharm.okayama-u.ac.jp.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Moriyama
authors.affiliation: From the Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8530, hiasa@okayama-u.ac.jp.
authors.forename: Miki
authors.initials: M
authors.lastname: Hiasa
mesh_terms: D000818:Animals; D000001:Calcimycin / Q000737:chemistry; D002118:Calcium / Q000737:chemistry; D002402:Cathepsin D / Q000737:chemistry; D027682:Cation Transport Proteins / Q000378:metabolism*; D005089:Exocytosis; D006632:Histamine / Q000737:chemistry; D006636:Histamine Release; D007150:Immunohistochemistry; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D011073:Polyamines / Q000378:metabolism*; D050683:R-SNARE Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D022142:Secretory Vesicles / Q000378:metabolism; D012701:Serotonin / Q000737:chemistry; D013095:Spermidine / Q000378:metabolism; D013096:Spermine / Q000378:metabolism; D050988:Vesicle-Associated Membrane Protein 2 / Q000378:metabolism; D051040:Vesicle-Associated Membrane Protein 3 / Q000378:metabolism; D050493:Vesicular Monoamine Transport Proteins / Q000378:metabolism*
chemical_list: D027682:Cation Transport Proteins; D011073:Polyamines; D050683:R-SNARE Proteins; C493450:Slc18a1 protein, mouse; C493451:Slc18a1 protein, rat; C496927:Sybl1 protein, mouse; C494706:Vamp2 protein, rat; C496926:Vamp7 protein, rat; D050988:Vesicle-Associated Membrane Protein 2; D051040:Vesicle-Associated Membrane Protein 3; D050493:Vesicular Monoamine Transport Proteins; C494808:vesicle-associated membrane protein 2, mouse; C494953:vesicle-associated membrane protein 3, mouse; C494951:vesicle-associated membrane protein 3, rat; C000620983:vesicular polyamine transporter, mouse; C000620984:vesicular polyamine transporter, rat; D013096:Spermine; D012701:Serotonin; D000001:Calcimycin; D006632:Histamine; D002402:Cathepsin D; D002118:Calcium; D013095:Spermidine
keywords: histamine; mast cell; polyamine; secretory granules; spermine; transporter; vesicles; vesicular polyamine transporter