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01.08.2003 | Laboratory Investigations | Ausgabe 2/2003

Calcified Tissue International 2/2003

Regulated Expression of G Protein-Coupled Receptor Kinases (GRK’s) and β-Arrestins in Osteoblasts

Zeitschrift:
Calcified Tissue International > Ausgabe 2/2003
Autor:
R. F. Spurney

Abstract

Desensitization of G-protein coupled receptors (GPCR’s) is largely mediated by a family of enzymes and protein co-factors termed GRKs and arrestins, respectively. In the present studies, we investigated expression of GRKs and arrestins in osteoblastic cell lines concentrating on the enzymes (GRK2 and GRK3) and protein co-factors (β-arrestint 1 and β-arrestin 2) that play dominant roles in regulating GPCR responsiveness in most tissues and cell types. We found that osteoblastic cells express similar amounts of GRK2 with either undetectable or lesser amounts of GRK3. In contrast, expression of β-arrestin 1 and β-arrestin 2 by osteoblastic cells varied between cell lines. To determine if GRK2 or β-arrestin expression is modulated during osteoblast development, we assessed expression of GRK2 and β-arrestin proteins during differentiation of the mouse osteoblastic cell line MC3T3-E1 cells over a 21-day period. We found that expression of GRK2 and β-arrestin 2 increased to maximal levels by day 7 and then decreased 4-fold by day 21. In contrast, expression of β-arrestin 1 increased to maximal levels by day 14 and then decreased 2-fold by day 21. Over this same time period (days 7–21), PTH/PTHrP receptor number decreased to a greater extent than the decrease in PTH(l-34)-induced cAMP generation, suggesting that responsiveness of individual PTH/PTHrP receptors was enhanced in differentiated cells. We conclude that (l) osteoblastic cell lines differentially express the enzymes and protein co-factors that modulate GPCR responsiveness and (2) expression of both GRK2 and β-arrestins is temporally regulated during osteoblast development. These data are consistent with the notion that GPCR responsiveness may be differentially regulated in osteoblastic cell lines and during osteoblast development.

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