Dentin matrix protein 1 gene cis-regulation: Use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo

Wuchen Yang, Yongbo Lu, Ivo Kalajzic, Dayong Guo, Marie A. Harris, Jelica Gluhak-Heinrich, Shiva Kotha, Lynda Bonewald, Jian Q. Feng, David W. Rowe, Charles H. Turner, Alexander Robling, Stephen E. Harris

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)20680-20690
Number of pages11
JournalJournal of Biological Chemistry
Volume280
Issue number21
DOIs
StatePublished - May 27 2005

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Osteocytes
Dentin
Gene expression
Green Fluorescent Proteins
Osteoblasts
Proteins
Bone
Nucleic Acid Regulatory Sequences
In Vitro Techniques
Cell culture
Bone and Bones
Animals
Ulna
Genes
Transgenic Mice
Embryonic Structures
Bone Marrow

ASJC Scopus subject areas

  • Biochemistry

Cite this

Dentin matrix protein 1 gene cis-regulation : Use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo. / Yang, Wuchen; Lu, Yongbo; Kalajzic, Ivo; Guo, Dayong; Harris, Marie A.; Gluhak-Heinrich, Jelica; Kotha, Shiva; Bonewald, Lynda; Feng, Jian Q.; Rowe, David W.; Turner, Charles H.; Robling, Alexander; Harris, Stephen E.

In: Journal of Biological Chemistry, Vol. 280, No. 21, 27.05.2005, p. 20680-20690.

Research output: Contribution to journalArticle

Yang, W, Lu, Y, Kalajzic, I, Guo, D, Harris, MA, Gluhak-Heinrich, J, Kotha, S, Bonewald, L, Feng, JQ, Rowe, DW, Turner, CH, Robling, A & Harris, SE 2005, 'Dentin matrix protein 1 gene cis-regulation: Use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo', Journal of Biological Chemistry, vol. 280, no. 21, pp. 20680-20690. https://doi.org/10.1074/jbc.M500104200
Yang, Wuchen ; Lu, Yongbo ; Kalajzic, Ivo ; Guo, Dayong ; Harris, Marie A. ; Gluhak-Heinrich, Jelica ; Kotha, Shiva ; Bonewald, Lynda ; Feng, Jian Q. ; Rowe, David W. ; Turner, Charles H. ; Robling, Alexander ; Harris, Stephen E. / Dentin matrix protein 1 gene cis-regulation : Use in osteocytes to characterize local responses to mechanical loading in vitro and in vivo. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 21. pp. 20680-20690.
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AU - Harris, Marie A.

AU - Gluhak-Heinrich, Jelica

AU - Kotha, Shiva

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