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Modulation of 1α,25‐dihydroxyvitamin D 3 ‐membrane associated, rapid response steroid binding protein expression in mouse odontoblasts by 1α,25‐(OH) 2 D 3
Author(s) -
Teillaud Christophe,
Nemere Ilka,
Boukhobza Florine,
Mathiot Claire,
Conan Nicole,
Oboeuf Martine,
Hotton Dominique,
MacDougall Mary,
Berdal Ariane
Publication year - 2004
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.20275
Subject(s) - immunogold labelling , odontoblast , biology , microbiology and biotechnology , intracellular , blot , polyclonal antibodies , extracellular , membrane protein , cell culture , endocrinology , medicine , biochemistry , membrane , antibody , immunology , pathology , gene , genetics , dentin
The rapid, nongenomic effects of 1α,25‐dihydroxyvitamin D 3 (1α,25‐(OH) 2 D 3 ) have been related to a 1,25D 3 ‐membrane associated, rapid response steroid binding protein or 1,25D 3 ‐[MARRS]bp, with a molecular weight of 65 kDa, in several tissues and species. Currently, no information is available concerning the nongenomic responses to 1α,25‐(OH) 2 D 3 in dental tissues. In order to investigate the expression of 1,25D 3 ‐[MARRS]bp in dental cells, in the presence or absence of 1α,25‐(OH) 2 D 3 , we have used rabbit polyclonal antibodies directed against the N‐terminus of the 1,25D 3 ‐[MARRS]bp (Ab099) that recognizes the 1α,25‐(OH) 2 D 3 binding protein in chick intestinal basolateral membranes and a mouse odontoblast‐like cell line (MO6‐G3). Western blotting and flow cytometric analyses with Ab099 specifically detected 1,25D 3 ‐[MARRS]bp in MO6‐G3 cells. Moreover, 1,25D 3 ‐[MARRS]bp was up‐regulated, in vivo, in differentiated dental cells. Electron microscopic analysis confirmed the plasma membrane localization of this binding protein and also showed its intracellular presence. Incubation of MO6‐G3 cells with different doses of 1α,25‐(OH) 2 D 3 for 36 h resulted in an inhibition of 1,25D 3 ‐[MARRS]bp expression with a maximal effect at 50 nM steroid. In addition, the culture media of MO6‐G3 cells contains immunoreactive 1,25D 3 ‐[MARRS]bp. Immunogold positive membrane vesicle‐like structures are present in the extracellular matrix of MO6‐G3 cells. Altogether, these results indicate that the 1,25D 3 ‐[MARRS]bp expression in MO6‐G3 cells is modulated by 1α,25‐(OH) 2 D 3 . In conclusion, this 1α,25‐(OH) 2 D 3 binding protein could play an important role in the rapid, nongenomic responses to 1α,25‐(OH) 2 D 3 in dental cells. © 2004 Wiley‐Liss, Inc.