Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells

Thomas Clemens, K. P. Garrett, X. Y. Zhou, J. W. Pike, M. R. Haussler, D. W. Dempster

Research output: Contribution to journalArticle

Abstract

We have used a monoclonal antibody (9A7) against the purified avian 1,25-dihydroxyvitamin D3 receptor to develop an immunocytochemical technique for visualization of the protein in fixed tissues and cultured cells. In Bouin's-fixed, chick intestine, 1,25-dihydroxyvitamin D3 receptor-like immunoreactivity was localized mainly in nuclei of epithelial cells and was more abundant in the crypt than in the villar cells. Receptor staining was low or undetectable in liver hepatocytes but was present in nuclei of cells lining the hepatic sinusoids. In rat brain, receptor-like immunoreactivity was abundant and widely distibuted, but did not always coincidence with the presence of vitamin D-dependent calcium-binding protein; 1,25-dihydroxyvitamin D3 receptor was absent from cerebellar Purkinje cells that contained abundant calcium-binding protein. In disaggregated rat bone cells, receptor immunoreactivity was present in mononuclear cells including osteoblasts and fibroblasts but was absent from osteoblasts-like, rat osteosarcoma cells, shown in previous studies to be either receptor positive (17/2.8) or negative (24.1), demonstrated nuclear immunoreactivity in exact concordance with receptor levels as determined by ligand binding. The phenomenon of hormone-induced up-regulation of receptor was visualized in receptor-positive 3T6 fibroblasts by demonstration of markedly enhanced nuclear reactivity in cells treated with 10-7 M 1,25-dihydroxyvitamin D3 for 48 h. Our studies demonstrate the feasibility of the immunocytochemical approach to visualize the 1,25-dihydroxyvitamin D3 receptor in target tissues and show that it is predominantly a nuclear protein in the relatively unoccupied and fully activated states. Moreover, the vitamin D-dependent calcium binding is not a universal marker for 1,25-dihydroxyvitamin D3 action. Rather, our observations suggest that the expression of the 1,25-dihydroxyvitamin D3 receptor may be connected with the state of cellular differentiation.

Original languageEnglish (US)
Pages (from-to)1224-1230
Number of pages7
JournalEndocrinology
Volume122
Issue number4
StatePublished - 1988
Externally publishedYes

Fingerprint

Calcitriol Receptors
Calcitriol
Osteoblasts
Hepatocytes
S100 Calcium Binding Protein G
Fibroblasts
Calcium-Binding Proteins
Purkinje Cells
Feasibility Studies
Osteosarcoma
Nuclear Proteins
Cell Nucleus
Vitamin D
Intestines
Cultured Cells
Up-Regulation
Epithelial Cells
Monoclonal Antibodies
Hormones
Staining and Labeling

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Clemens, T., Garrett, K. P., Zhou, X. Y., Pike, J. W., Haussler, M. R., & Dempster, D. W. (1988). Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells. Endocrinology, 122(4), 1224-1230.

Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells. / Clemens, Thomas; Garrett, K. P.; Zhou, X. Y.; Pike, J. W.; Haussler, M. R.; Dempster, D. W.

In: Endocrinology, Vol. 122, No. 4, 1988, p. 1224-1230.

Research output: Contribution to journalArticle

Clemens, T, Garrett, KP, Zhou, XY, Pike, JW, Haussler, MR & Dempster, DW 1988, 'Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells', Endocrinology, vol. 122, no. 4, pp. 1224-1230.
Clemens T, Garrett KP, Zhou XY, Pike JW, Haussler MR, Dempster DW. Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells. Endocrinology. 1988;122(4):1224-1230.
Clemens, Thomas ; Garrett, K. P. ; Zhou, X. Y. ; Pike, J. W. ; Haussler, M. R. ; Dempster, D. W. / Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells. In: Endocrinology. 1988 ; Vol. 122, No. 4. pp. 1224-1230.
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