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Dentin matrix protein 1 gene cis-regulation: use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo.
J Biol Chem. 2005 May 27; 280(21):20680-90.JB

Abstract

Dentin matrix protein 1 (DMP1) is highly expressed in osteocytes and is mechanically responsive. To study osteocyte-specific and mechanically regulated DMP1 gene expression, the transcriptional activity of three cis-regulatory regions was first examined in an osteoblast differentiation model in vitro using a green fluorescent protein (GFP) reporter. Expression of the -9624 to +1996 bp (10 kb) and -7892 to +4439 bp (8 kb) DMP1 cis-regulatory regions dramatically increased in areas of mineralized matrix, in dendritic, osteocyte-like cells. Mineralizing cultures expressing the 8-kb construct show dramatic GFP increases in response to loading in cells with a dendritic morphology. Transgenic mice expressing the 8-kb DMP1-GFP and -2433 to +4439 bp (2.5 kb) DMP1-LacZ were generated. Osteocyte-specific expression was found with the 8 kb but not with the 2.5 kb in postnatal animals. However, the 2.5 kb could support expression in rapidly forming osteoblasts and pre-osteocytes in the embryo. Primary calvarial osteoblast cultures demonstrated that the 2.5 kb supports weak expression in a subset of osteoblasts and pre-osteocytes, but not in mature osteocytes. However, the 8 kb supports robust expression in primary bone marrow cultures. Therefore the region -7892 to -2433 bp, termed a 5.5-kb "Osteocyte Enhancer Module," appears to be required for osteocyte specificity. Ulnae of mice with the 8-kb DMP1-GFP were subjected to mechanical loading where GFP expression increased selectively and locally in osteocytes, distal to the mid-shaft and near the surface of the bone. Thus, the 8-kb region of the DMP1 gene is a target for mechanotransduction in osteocytes, and its cis-regulatory activity may be correlated to local strain in bone.

Authors+Show Affiliations

Department of Periodontics and Cellular and Structural Biology, University of Texas, San Antonio, Texas 78229, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15728181

Citation

Yang, Wuchen, et al. "Dentin Matrix Protein 1 Gene Cis-regulation: Use in Osteocytes to Characterize Local Responses to Mechanical Loading in Vitro and in Vivo." The Journal of Biological Chemistry, vol. 280, no. 21, 2005, pp. 20680-90.
Yang W, Lu Y, Kalajzic I, et al. Dentin matrix protein 1 gene cis-regulation: use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo. J Biol Chem. 2005;280(21):20680-90.
Yang, W., Lu, Y., Kalajzic, I., Guo, D., Harris, M. A., Gluhak-Heinrich, J., Kotha, S., Bonewald, L. F., Feng, J. Q., Rowe, D. W., Turner, C. H., Robling, A. G., & Harris, S. E. (2005). Dentin matrix protein 1 gene cis-regulation: use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo. The Journal of Biological Chemistry, 280(21), 20680-90.
Yang W, et al. Dentin Matrix Protein 1 Gene Cis-regulation: Use in Osteocytes to Characterize Local Responses to Mechanical Loading in Vitro and in Vivo. J Biol Chem. 2005 May 27;280(21):20680-90. PubMed PMID: 15728181.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Dentin matrix protein 1 gene cis-regulation: use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo. AU - Yang,Wuchen, AU - Lu,Yongbo, AU - Kalajzic,Ivo, AU - Guo,Dayong, AU - Harris,Marie A, AU - Gluhak-Heinrich,Jelica, AU - Kotha,Shiva, AU - Bonewald,Lynda F, AU - Feng,Jian Q, AU - Rowe,David W, AU - Turner,Charles H, AU - Robling,Alexander G, AU - Harris,Stephen E, Y1 - 2005/02/22/ PY - 2005/2/25/pubmed PY - 2005/8/16/medline PY - 2005/2/25/entrez KW - Non-programmatic SP - 20680 EP - 90 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 280 IS - 21 N2 - Dentin matrix protein 1 (DMP1) is highly expressed in osteocytes and is mechanically responsive. To study osteocyte-specific and mechanically regulated DMP1 gene expression, the transcriptional activity of three cis-regulatory regions was first examined in an osteoblast differentiation model in vitro using a green fluorescent protein (GFP) reporter. Expression of the -9624 to +1996 bp (10 kb) and -7892 to +4439 bp (8 kb) DMP1 cis-regulatory regions dramatically increased in areas of mineralized matrix, in dendritic, osteocyte-like cells. Mineralizing cultures expressing the 8-kb construct show dramatic GFP increases in response to loading in cells with a dendritic morphology. Transgenic mice expressing the 8-kb DMP1-GFP and -2433 to +4439 bp (2.5 kb) DMP1-LacZ were generated. Osteocyte-specific expression was found with the 8 kb but not with the 2.5 kb in postnatal animals. However, the 2.5 kb could support expression in rapidly forming osteoblasts and pre-osteocytes in the embryo. Primary calvarial osteoblast cultures demonstrated that the 2.5 kb supports weak expression in a subset of osteoblasts and pre-osteocytes, but not in mature osteocytes. However, the 8 kb supports robust expression in primary bone marrow cultures. Therefore the region -7892 to -2433 bp, termed a 5.5-kb "Osteocyte Enhancer Module," appears to be required for osteocyte specificity. Ulnae of mice with the 8-kb DMP1-GFP were subjected to mechanical loading where GFP expression increased selectively and locally in osteocytes, distal to the mid-shaft and near the surface of the bone. Thus, the 8-kb region of the DMP1 gene is a target for mechanotransduction in osteocytes, and its cis-regulatory activity may be correlated to local strain in bone. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/15728181/Dentin_matrix_protein_1_gene_cis_regulation:_use_in_osteocytes_to_characterize_local_responses_to_mechanical_loading_in_vitro_and_in_vivo_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=15728181 DB - PRIME DP - Unbound Medicine ER -