Vitamin D-Binding Protein Influences Total Circulating Levels of 1,25-Dihydroxyvitamin D3 but Does Not Directly Modulate the Bioactive Levels of the Hormone in Vivo

• Published in: Endocrinology (Philadelphia) 2008-07, Vol.149 (7), p.3656-3667
• Main Authors: Zella, Lee A, Shevde, Nirupama K, Hollis, Bruce W, Cooke, Nancy E, Pike, J. Wesley
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Animals; Bioaccumulation; Biological activity; Biological and medical sciences; Blood levels; Blotting, Western; Calciferol; Calcitriol; Calcitriol - analogs & derivatives; Calcitriol - pharmacology; Calcium; Calcium - blood; Calcium homeostasis; Calcium metabolism; Cell culture; Chromatin Immunoprecipitation; Circulation; Dose-Response Relationship, Drug; Extracellular levels; Fundamental and applied biological sciences. Psychology; Homeostasis; In vivo methods and tests; Intestines - drug effects; Intestines - metabolism; Kidney - drug effects; Kidney - metabolism; Mice; Mice, Inbred C57BL; Mice, Knockout; Protein Binding - drug effects; Protein Binding - genetics; Proteins; Vertebrates: endocrinology; Vitamin D; Vitamin D - analogs & derivatives; Vitamin D - blood; Vitamin D - metabolism; Vitamin D - pharmacology; Vitamin D-binding protein; Vitamin D-Binding Protein - genetics; Vitamin D-Binding Protein - metabolism; Vitamin D-Binding Protein - physiology; Vitamin D3; Vitamin deficiency
• ISSN: 0013-7227; 1945-7170
• DOI: 10.1210/en.2008-0042

Mice deficient in the expression of vitamin D-binding protein (DBP) are normocalcemic despite undetectable levels of circulating 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. We used this in vivo mouse model together with cells in culture to explore the impact of DBP on the biological activity of 1,25(OH)2D3. Modest changes in the basal expression of genes involved in 1,25(OH)2D3 metabolism and calcium homeostasis were observed in vivo; however, these changes seemed unlikely to explain the normal calcium balance seen in DBP-null mice. Further investigation revealed that despite the reduced blood levels of 1,25(OH)2D3 in these mice, tissue concentrations were equivalent to those measured in wild-type counterparts. Thus, the presence of DBP has limited impact on the extracellular pool of 1,25(OH)2D3 that is biologically active and that accumulates within target tissues. In cell culture, in contrast, the biological activity of 1,25(OH)2D3 is significantly impacted by DBP. Here, although DBP deficiency had no effect on the activation profile itself, the absence of DBP strongly reduced the concentration of exogenous 1,25(OH)2D3 necessary for transactivation. Surprisingly, analogous studies in wild-type and DBP-null mice, wherein we explored the activity of exogenous 1,25(OH)2D3, produced strikingly different results as compared with those in vitro. Here, the carrier protein had virtually no impact on the distribution, uptake, activation profile, or biological potency of the hormone. Collectively, these experiments suggest that whereas DBP is important to total circulating 1,25(OH)2D3 and sequesters extracellular levels of this hormone both in vivo and in vitro, the binding protein does not influence the hormone’s biologically active pool.