The Transient Receptor Potential Ion Channel TRPV6 Is Expressed at Low Levels in Osteoblasts and Has Little Role in Osteoblast Calcium Uptake

被引:14
作者
Little, Robert [1 ]
Muimo, Richmond [2 ]
Robson, Louise [3 ]
Harris, Kate [3 ]
Grabowski, Peter S. [1 ]
机构
[1] Univ Sheffield, Dept Human Metab, Mellanby Ctr Bone Res, Sheffield, S Yorkshire, England
[2] Univ Sheffield, Dept Infect & Immun, Sheffield, S Yorkshire, England
[3] Univ Sheffield, Dept Biomed Sci, Sheffield S10 2TN, S Yorkshire, England
关键词
PLASMA-MEMBRANE CA2+-ATPASE; CELL-LINE SAOS-2; CA2+ CHANNEL; BONE-CELLS; ABSORPTION; EPITHELIA; GENE; TRANSPORT; KIDNEY; ECAC2;
D O I
10.1371/journal.pone.0028166
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: TRPV6 ion channels are key mediators of regulated transepithelial absorption of Ca(2+) within the small intestine. Trpv6(-/-) mice were reported to have lower bone density than wild-type littermates and significant disturbances in calcium homeostasis that suggested a role for TRPV6 in osteoblasts during bone formation and mineralization. TRPV6 and molecules related to transepithelial Ca2+ transport have been reported to be expressed at high levels in human and mouse osteoblasts. Results: Transmembrane ion currents in whole cell patch clamped SaOS-2 osteoblasts did not show sensitivity to ruthenium red, an inhibitor of TRPV5/6 ion channels, and (45)Ca uptake was not significantly affected by ruthenium red in either SaOS-2 (P = 0.77) or TE-85 (P = 0.69) osteoblastic cells. In contrast, ion currents and (45)Ca uptake were both significantly affected in a human bronchial epithelial cell line known to express TRPV6. TRPV6 was expressed at lower levels in osteoblastic cells than has been reported in some literature. In SaOS-2 TRPV6 mRNA was below the assay detection limit; in TE-85 TRPV6 mRNA was detected at 6.90 +/- 1.9 x 10(-5) relative to B2M. In contrast, TRPV6 was detected at 7.7 +/- 3.0 x 10(-2) and 2.38 +/- 0.28 x 10(-4) the level of B2M in human carcinoma-derived cell lines LNCaP and CaCO-2 respectively. In murine primary calvarial osteoblasts TRPV6 was detected at 3.80 +/- 0.24 x 10(-5) relative to GAPDH, in contrast with 4.3 +/- 1.5 x 10(-2) relative to GAPDH in murine duodenum. By immunohistochemistry, TRPV6 was expressed mainly in myleocytic cells of the murine bone marrow and was observed only at low levels in murine osteoblasts, osteocytes or growth plate cartilage. Conclusions: TRPV6 is expressed only at low levels in osteoblasts and plays little functional role in osteoblastic calcium uptake.
引用
收藏
页数:8
相关论文
共 36 条
[1]   Expression of transient receptor potential (TRP) channels in human and murine osteoblast-like cells [J].
Abed, Elie ;
Labelle, Dominique ;
Martineau, Corine ;
Loghin, Andrew ;
Moreau, Robert .
MOLECULAR MEMBRANE BIOLOGY, 2009, 26 (03) :146-158
[2]   CALBINDIN-D9K IMMUNOLOCALIZATION AND VITAMIN-D-DEPENDENCE IN THE BONE OF GROWING AND ADULT-RATS [J].
BALMAIN, N ;
BERDAL, A ;
HOTTON, D ;
CUISINIERGLEIZES, P ;
MATHIEU, H .
HISTOCHEMISTRY, 1989, 92 (05) :359-365
[3]  
Berdal A, 1996, J BONE MINER RES, V11, P768
[4]   Marked disturbance of calcium homeostasis in mice with targeted disruption of the Trpv6 calcium channel gene [J].
Bianco, Suzy D. C. ;
Peng, Ji-Bin ;
Takanaga, Hitomi ;
Suzuki, Yoshiro ;
Crescenzi, Alessandra ;
Kos, Claudine H. ;
Zhuang, Liyan ;
Freeman, Michael R. ;
Gouveia, Cecilia H. A. ;
Wu, Jiangping ;
Luo, Hongyu ;
Mauro, Theodora ;
Brown, Edward M. ;
Hediger, Matthias A. .
JOURNAL OF BONE AND MINERAL RESEARCH, 2007, 22 (02) :274-285
[5]   ROLE OF NA+/CA2+ EXCHANGE IN TRANSCELLULAR CA2+ TRANSPORT ACROSS PRIMARY CULTURES OF RABBIT KIDNEY COLLECTING SYSTEM [J].
BINDELS, RJM ;
RAMAKERS, PLM ;
DEMPSTER, JA ;
HARTOG, A ;
VANOS, CH .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1992, 420 (5-6) :566-572
[6]   The annexin 2-S100A10 complex and its association with TRPV6 is regulated by cAMP/PKA/CnA in airway and gut epithelia [J].
Borthwick, Lee A. ;
Neal, Andy ;
Hobson, Lynsey ;
Gerke, Volker ;
Robson, Louise ;
Muimo, Richmond .
CELL CALCIUM, 2008, 44 (02) :147-157
[7]   Synthesis of calbindin-D28K during mineralization in human bone marrow stromal cells [J].
Faucheux, C ;
Bareille, R ;
Amedee, J .
BIOCHEMICAL JOURNAL, 1998, 333 :817-823
[8]   ROLE OF FACILITATED DIFFUSION OF CALCIUM BY CALBINDIN IN INTESTINAL CALCIUM-ABSORPTION [J].
FEHER, JJ ;
FULLMER, CS ;
WASSERMAN, RH .
AMERICAN JOURNAL OF PHYSIOLOGY, 1992, 262 (02) :C517-C526
[9]   IMPROVED PATCH-CLAMP TECHNIQUES FOR HIGH-RESOLUTION CURRENT RECORDING FROM CELLS AND CELL-FREE MEMBRANE PATCHES [J].
HAMILL, OP ;
MARTY, A ;
NEHER, E ;
SAKMANN, B ;
SIGWORTH, FJ .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1981, 391 (02) :85-100
[10]   Calcium absorption across epithelia [J].
Hoenderop, JGJ ;
Nilius, B ;
Bindels, RJM .
PHYSIOLOGICAL REVIEWS, 2005, 85 (01) :373-422