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Dependency of Estrogen’s Stimulatory Action on Cancellous Bone Formation in Male Mice
Academic Rheumatology (K.E.M., M.J.P., R.L.G., S.M.C., J.H.T.), University of Bristol, Bristol BS2 8HW, United Kingdom; and National Institute of Environmental Health Sciences (K.S.K.), National Institutes of Health, Research Triangle Park, North Carolina 27709
Address all correspondence and requests for reprints to: Dr. J. H. Tobias, Rheumatology Unit, Bristol Royal Infirmary, Bristol BS2 8HW, United Kingdom. E-mail: Jon.Tobias{at}bristol.ac.uk.
We examined whether estrogen receptor (ER)
is required for estrogen to stimulate cancellous bone formation in long bones of male mice. 17ß-Estradiol (E2) was administered to ER-/- male mice or wild-type (WT) littermate controls at 40, 400, or 4000 µg/kg by daily sc injection for 28 d and histomorphometric analysis performed at the distal femoral metaphysis. In WT mice, treatment with E2 (40 µg/kg·d) increased the proportion of cancellous bone surfaces undergoing mineralization and stimulated mineral apposition rate. In addition, higher doses of E2 induced the formation of new cancellous bone formation surfaces in WT mice. In contrast, E2 had little effect on any of these parameters in ER-/- mice. Immunohistochemistry was subsequently performed using an ER-specific C-terminal polyclonal antibody. In WT mice, ER was expressed both by cancellous osteoblasts and a significant proportion of mononuclear bone marrow cells. Immunoreactivity was also observed in cancellous osteoblasts of ER-/- mice, resulting from expression of the activation function-1-deficient 46-kDa ER isoform previously reported to be expressed in normal osteoblasts and bones of ER-/- mice. Taken together, our results suggest that estrogen stimulates bone formation in mouse long bones via a mechanism that requires the presence of full-length ER possessing activation function-1.
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