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Abstract
Bone sialoprotein (BSP) expression is restricted to cells associated with the mineralization of bones and teeth. We previously identified a homeodomain binding element in a 2.5 kb fragment of the murine Bsp promoter that is required for osteoblast-selective expression in cell culture. To examine the role of this element (called OSHE1; osteoblast-specific homeodomain element 1) in the tissue-specific expression of Bsp in vivo, we generated transgenic mice using the wild-type 2472 bp promoter or the same promoter containing a 2 bp mutation in OSHE1. Promoter constructs driving both luciferase and lacZ reporter genes were microinjected into fertilized eggs from (C57BL/6 X SJL)F1 mice. Four lines containing the wild-type promoter and 5 lines containing the mutated promoter were established, and the tissue specificity of g -galactosidase staining and luciferase expression was examined. g -gal staining was observed in osteoblasts of calvaria and trabecular regions of tibia and femur in 12-day-old mice while chondrocytes, kidney, heart, muscle, spleen, liver, skin, stomach, and lung were negative. Whole tissue luciferase activity was also much higher in mineralized tissues although some soft tissue expression was detected. In contrast, analysis of OSHE1 mutant lines revealed expression of luciferase and g -gal in kidney, skin, liver, and lung. g -gal expression in these tissues was restricted to specific cell populations. Trabecular regions were devoid of g -gal staining in the tibia and femur of the mutant mice, while staining was seen in the chondrocytes. We therefore hypothesize that the OSHE1 site is involved in both the expression of Bsp in mineralizing tissues and the suppression of transcription in nonmineralizing tissues.