title: Growth hormone binds to a single high affinity receptor site on mouse osteoblasts: modulation by retinoic acid and cell differentiation.
abstract: Growth hormone (GH) exerts direct differentiative and proliferative effects on osteoblasts. We studied 125I-labeled human (h) GH binding to primary mouse osteoblasts derived from collagenase-treated 18-day fetal mouse calvaria. Scatchard analysis of the data revealed a single class of high affinity GH receptors (apparent Ka = 5.74 x 10(9) M-1) with 2200 sites per cell. Affinity crosslinking and SDS-PAGE electrophoresis showed two bands with apparent molecular masses of 120 and 70 kDa. Mouse osteoblasts express GH receptor mRNA with gene transcripts of 4.2 and 1.2 kb, at levels which reach approximately 1/6 of those in mouse liver and 1/3 of those in mouse muscle. Two populations of undifferentiated and diffentiated osteoblasts, obtained by sequential collagenase digestion of mouse calvaria, were used to study the relationship between osteoblastic phenotype and GH receptor expression. Although the affinity of the receptors in undifferentiated and differentiated cells was the same, the capacity was significantly higher (1.45 +/- 1.0% vs 2.39 +/- 0.9%, P = 0.03) in differentiated cells. This stresses the specific importance of the osteoblast as a target cell for GH. The differentiating potential of the vitamin A derivative retinoic acid was subsequently used experimentally to induce differentiation in the cells. Retinoic acid increased 125I-hGH binding to preosteoblasts (153%, P = 0.02). Together, these data demonstrate the presence of a high affinity GH receptor in mouse osteoblasts which is related to differentiation.
authors.affiliation: Department of Endocrinology, Free University Hospital, Amsterdam, Netherlands.
authors.forename: M C
authors.initials: MC
authors.lastname: Slootweg
authors.forename: J P
authors.initials: JP
authors.lastname: Salles
authors.forename: C
authors.initials: C
authors.lastname: Ohlsson
authors.forename: C P
authors.initials: CP
authors.lastname: de Vries
authors.forename: M J
authors.initials: MJ
authors.lastname: Engelbregt
authors.forename: J C
authors.initials: JC
authors.lastname: Netelenbos
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D013006:Growth Hormone / Q000378:metabolism*; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D011986:Receptors, Somatotropin / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D011986:Receptors, Somatotropin; D014212:Tretinoin; D013006:Growth Hormone