Dexamethasone suppresses in vivo levels of bone collagen synthesis in neonatal mice.
The objective of this study was to determine the acute effects of glucocorticoids on in vivo levels of bone collagen synthesis in neonatal mice. Mice were injected with vehicle or dexamethasone at the start of the experiment. At 22 h, mice were given a 10 microCi injection of [3H]proline. At 24 h, the mice were sacrificed and the incorporation of [3H]proline into collagenase-digestible CDP labeling) and noncollagen (NCP labeling) protein in calvariae were determined by digestion with bacterial collagenase. Calvarial RNA was analyzed for COL 1A1 and osteocalcin mRNA levels by Northern blotting. After 24 h, vehicle-treated mice showed a 9.8 +/- 1.0% weight gain while dexamethasone-treated mice (1 mg/kg) had a 7.4 +/- 0.8% weight loss. Dexamethasone (1 mg/kg) decreased CDP and NCP labeling in calvariae by 51 +/- 4% and 17 +/- 4%, respectively (13 experiments). The inhibitory effect on protein labeling was selective for collagen since dexamethasone decreased the percent collagen synthesis from 25.4 +/- 1.6% to 16.6 +/- 1.0% (13 experiments). Dexamethasone at 3 mg/kg also decreased CDP labeling and the percent collagen synthesis in calvariae. There was a 30% reduction in COL1A1 mRNA levels and a 67% decrease in osteocalcin mRNA levels. To determine the reversibility of the inhibition of collagen synthesis, mice were given a single injection of dexamethasone (1 mg/kg) and then injected with [3H]proline 2 h prior to sacrifice at 24, 48, or 72 h. The reduction in CDP labeling observed at 24 h was fully reversed by 48-72 h. Moreover, by 72 h, the-rate of weight gain by dexamethasone-treated mice was similar to vehicle-treated controls. These data show that administration of dexamethasone to neonatal mice leads to a selective decrease in bone collagen synthesis within 24 h that is accompanied by down-regulation of osteocalcin and COL1A1 mRNA levels. This model will be useful in determining mechanisms by which high dose glucocorticoids inhibit bone formation in vivo.
Advani
S
S
Department of Medicine, University of Connecticut Health Center, Farmington 06030, USA.
LaFrancis
D
D
Bogdanovic
E
E
Taxel
P
P
Raisz
L G
LG
Kream
B E
BE
D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D015152:Blotting, Northern; D001846:Bone Development / Q000187:drug effects; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics; D003907:Dexamethasone / Q000008:administration & dosage / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005938:Glucocorticoids / Q000633:toxicity*; D007553:Isotope Labeling; D051379:Mice; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D011392:Proline / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012886:Skull / Q000166:cytology / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014316:Tritium / Q000378:metabolism
D005938:Glucocorticoids; D012333:RNA, Messenger; D014316:Tritium; D015675:Osteocalcin; D003907:Dexamethasone; D003094:Collagen; D011392:Proline