Functional alpha1- and beta2-adrenergic receptors in human osteoblasts.
Central (hypothalamic) control of bone mass is proposed to be mediated through beta2-adrenergic receptors (beta2-ARs). While investigations in mouse bone cells suggest that epinephrine enhances both RANKL and OPG mRNA via both beta-ARs and alpha-ARs, whether alpha-ARs are expressed in human bone cells is controversial. The current study investigated the expression of alpha1-AR and beta2-AR mRNA and protein and the functional role of adrenergic stimulation in human osteoblasts (HOBs). Expression of alpha1B- and beta2-ARs was examined by RT-PCR, immunofluorescence microscopy and Western blot (for alpha1B-ARs). Proliferation in HOBs was assessed by (3)H-thymidine incorporation and expression of RANKL and OPG was determined by quantitative RT-PCR. RNA message for alpha1B- and beta2-ARs was expressed in HOBs and MG63 human osteosarcoma cells. alpha1B- and beta2-AR immunofluorescent localization in HOBs was shown for the first time by deconvolution microscopy. alpha1B-AR protein was identified in HOBs by Western blot. Both alpha1-agonists and propranolol (beta-blocker) increased HOB replication but fenoterol, a beta2-agonist, inhibited it. Fenoterol nearly doubled RANKL mRNA and this was inhibited by propranolol. The alpha1-agonist cirazoline increased OPG mRNA and this increase was abolished by siRNA knockdown of alpha1B-ARs in HOBs. These data indicate that both alpha1-ARs and beta2-ARs are present and functional in HOBs. In addition to beta2-ARs, alpha1-ARs in human bone cells may play a role in modulation of bone turnover by the sympathetic nervous system.
Huang
H H
HH
Department of Physiology, School of Medical Sciences, University of Sydney, Sydney, NSW, Australia.
Brennan
T C
TC
Muir
M M
MM
Mason
R S
RS