Article

Visualizing levels of osteoblast differentiation by a two-color promoter-GFP strategy: Type I collagen-GFPcyan and osteocalcin-GFPtpz

  1. I. Bilic-Curcic1,
  2. M. Kronenberg1,
  3. X. Jiang1,
  4. J. Bellizzi1,
  5. M. Mina2,
  6. I. Marijanovic1,
  7. E.M. Gardiner3,
  8. D.W. Rowe1,*

Article first published online: 7 SEP 2005

DOI: 10.1002/gene.20156

Issue

genesis

genesis

Volume 43, Issue 2, pages 87–98, October 2005

Additional Information

How to Cite

Bilic-Curcic, I., Kronenberg, M., Jiang, X., Bellizzi, J., Mina, M., Marijanovic, I., Gardiner, E.M. and Rowe, D.W. (2005), Visualizing levels of osteoblast differentiation by a two-color promoter-GFP strategy: Type I collagen-GFPcyan and osteocalcin-GFPtpz. Genesis, 43: 87–98. doi: 10.1002/gene.20156

Author Information

  1. 1

    Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, Connecticut

  2. 2

    Department of Pediatric Dentistry, University of Connecticut Health Center, Farmington, Connecticut

  3. 3

    Bone and Mineral Research Program and Molecular Modeling Facility, Garvan Institute of Medical Research, Sydney NSW, Australia

*Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, CT 06030

Publication History

  1. Issue published online: 7 SEP 2005
  2. Article first published online: 7 SEP 2005
  3. Manuscript Accepted: 28 JUN 2005
  4. Manuscript Received: 15 DEC 2004

Funded by

  • Public Health Service (PHS). Grant Numbers: R01 AR43457, U01 DK63478

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Keywords:

  • GFP;
  • osteocalcin promoter;
  • osteoblast differentiation;
  • type I collagen promoter;
  • primary osteoblast culture;
  • bone histology

Abstract

A 3.9 kb DNA fragment of human osteocalcin promoter and 3.6 kb DNA fragment of the rat collagen type1a1 promoter linked with visually distinguishable GFP isomers, topaz and cyan, were used for multiplex analysis of osteoblast lineage progression. Three patterns of dual transgene expression can be appreciated in primary bone cell cultures derived from the transgenic mice and by histology of their corresponding bones. Our data support the interpretation that strong pOBCol3.6GFPcyan alone is found in newly formed osteoblasts, while strong pOBCol3.6GFPcyan and hOC-GFPtpz are present in osteoblasts actively making a new matrix. Osteoblasts expressing strong hOC-GFPtpz and weak pOBCol3.6GFPcyan are also present and may or may not be producing mineralized matrix. This multiplex approach reveals the heterogeneity within the mature osteoblast population that cannot be appreciated by current histological methods. It should be useful to identify and isolate populations of cells within an osteoblast lineage as they progress through stages of differentiation. genesis 43:87–98, 2005. © 2005 Wiley-Liss, Inc.

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