Continuous infusion of PTH in vivo results in active
bone resorption. To investigate the molecular basis of the catabolic effect of PTH in vivo, we evaluated the role of OPG and RANKL, which are known to influence osteoclast formation and function. Weanling rats fed a
calcium-free diet were parathyroidectomized and infused with PTH via an Alzet pump to examine: 1) the changes of serum-ionized
calcium and osteoclast number, 2) the expression of OPG/RANKL
mRNA and
protein, and 3) the expression of osteoblast phenotype bone formation-associated genes such as osteoblast specific
transcription factor,
osteocalcin,
bone sialoprotein, and
type I collagen.
PTH (1--38) (0.01--20 microg/100 g) continuous infusion for 1--24 h resulted in a dose-dependent increase in serum-ionized
calcium in parathyroidectomized rats and a corresponding dose-dependent increase in osteoclast number, indicating an increased
bone resorption. At 20 microg/100 g PTH dose level, serum-ionized
calcium was 2.1-fold of the vehicle control and not different from the
Sham-parathyroidectomized rats, and osteoclast number was 3-fold of the vehicle control and 1.7-fold of the
Sham-parathyroidectomized rats. In the distal femur, RANKL
mRNA expression was increased (27-fold) and OPG
mRNA expression was decreased (4.6-fold). The changes in RANKL and OPG
mRNA levels were rapid (as early as 1 h), dose dependent, and sustained over a 24-h period that was examined. Immunohistochemical evaluation of bone sections confirmed that OPG level was reduced in proximal tibial metaphysis upon PTH infusion. Circulating OPG
protein level was also decreased by 32% when compared with the parathyroidectomized control. The expression of genes that mark the osteoblast phenotype was significantly decreased [osteoblast specific
transcription factor (2.3-fold),
osteocalcin (3-fold),
bone sialoprotein (2.8-fold), and
type I collagen (5-fold)]. These results suggest that the catabolic effect of PTH infusion in vivo in this well-established resorption model is associated with a reciprocal expression of OPG/RANKL and a co-ordinate decrease in the expression of bone formation-related genes. We propose that the rapid and sustained increase in RANKL and decrease in OPG initiate maintain and favor the cascade of events in the differentiation/recruitment and activation of osteoclasts.