Effects of dexamethasone on tooth eruption in rats: differences in incisor and molar eruption.

A requirement for tooth eruption is the resorption of alveolar bone. Because bone resorption is stimulated by dexamethasone both in vivo and in vitro, dexamethasone 21-phosphate, a soluble form of dexamethasone, was injected into rats to determine its effect on tooth eruption. Such dexamethasone injections accelerate the time of intra-osseous eruption in rat incisors but do not accelerate the eruption time of rat molars when injected into rats. The injections of dexamethasone 21-phosphate also accelerate the time of eyelid opening in the postnatal rats, as well as retarding growth, as measured by body weight. These effects of dexamethasone 21-phosphate parallel the effects of epidermal growth factor injections, including the absence of an effect on molar eruption. This suggests that the molecular signals for the initiation of tooth eruption (i.e., onset of bone resorption) differ between rat incisors and molars. Given that rat incisors are teeth of continuous eruption whereas rat molars are teeth of limited eruption, as are human teeth, care must be taken in extrapolating results derived from rat incisors to human dentition. In vitro, dexamethasone has no effect on the gene expression of either osteoprotegerin or epidermal growth factor in dental follicle cells derived from molars. Because osteoprotegerin expression during normal tooth eruption is transitorily inhibited early postnatally in the molar dental follicle to allow osteoclast formation, the absence of inhibition of its expression by dexamethasone could explain why dexamethasone does not accelerate eruption in molars.

Enhancement of gene expression in rat dental follicle cells by parathyroid hormone-related protein.

Recent studies indicate that parathyroid hormone-related protein (PTHrP) is required for tooth eruption in mice. Localized in the stellate reticulum, PTHrP might exert a paracrine effect on cells of the adjacent dental follicle to initiate eruption. The presence of a follicle is needed for eruption and, at the cellular level, there is an influx of mononuclear cells in the follicle early postnatally in the first mandibular molar of the rat. In turn, these mononuclear cells fuse to form osteoclasts, which erode the alveolar bone to form an eruption pathway. At the molecular level, the dental follicle cells of the rat molar maximally express the genes for monocyte chemotactic protein-1 (MCP-1) and colony-stimulating factor-1 (CSF-1) at day 3 postnatally. Because day 3 also is the time of maximal influx of the mononuclear cells into the follicle, MCP-1 and CSF-1 could be involved in the recruitment/maturation of these cells. To determine if PTHrP can modulate gene expression in the dental follicle, cultured follicle cells were immunostained to show the receptor for PTHrP. The gene expression of this receptor was enhanced by incubating the cells with interleukin-1alpha (IL-1alpha). Next, the ability of PTHrP itself to enhance gene expression of either MCP-1 or CSF-1 in the dental follicle cells was determined by incubating the cells with PTHrP in either a time- or concentration-course manner (1-15 h or 1-100 ng/ml). By reverse transcription-polymerase chain reaction, it was demonstrated that PTHrP enhanced MCP-1 expression in a concentration-dependent fashion, with 50 ng PTHrP/ml inducing maximal expression of either MCP-1 or CSF-1. In the time-dependent studies, PTHrP caused maximal expression within 30 min for either MCP-1 or CSF-1. Immunoblotting revealed that PTHrP also enhanced secretion of MCP-1 by the follicle cells. Thus, one of the actions of PTHrP in tooth eruption may be that it enhances MCP-1 and CSF-1 gene expression and secretion in the dental follicle. Moreover, IL-1alpha may accentuate its action by enhancing the expression for the PTHrP receptor in the follicle cells.

Osteoprotegerin and osteoclast differentiation factor in tooth eruption.

A critical cellular event in tooth eruption is the formation of osteoclasts that are needed for bone resorption to form an eruption pathway. To analyze molecular regulation of osteoclast formation and activation, we examined the expression of osteoprotegerin (OPG), an inhibitor of osteoclast formation. In vivo, the gene expression of OPG is reduced in the dental follicle of the first mandibular molar of the rat at day 3 post-natally and in the mouse at day 5. This correlates with the days of maximal mononuclear cell influx and osteoclast numbers in the rat and mouse. Thus, inhibition of OPG gene expression on these days might allow osteoclasts to be formed and/or activated. In vitro studies demonstrated that both colony-stimulating factor-1 and parathyroid hormone-related protein reduced OPG gene expression in follicle cells, suggesting that these are candidate molecules for the in vivo inhibition of OPG expression. Osteoclast differentiation factor (ODF) immunolocalizes to the alveolar bone stromal cells adjacent to the follicle, whereby it might act to stimulate fusion of the mononuclear cells in the follicle.

Implications for tooth eruption of the effect of interleukin-1alpha on nuclear factor-kappaB gene expression in the rat dental follicle.

Tooth eruption is a localized event and a cascade of molecular signals generated in the dental follicle and stellate reticulum appears to initiate its onset. Consequently, mononuclear cells are recruited into the follicle and, in turn, fuse to become osteoclasts needed to resorb the alveolar bone to form an eruption pathway. One of the transcription factors involved in the sequence of molecular signalling may be nuclear factor (NF)kappaB. This study shows that NFkappaB is expressed and synthesized by cultured dental follicle cells. Moreover, its transcription, activation and translocation were enhanced by interleukin (IL)-1alpha, a potential eruption molecule. The enhancement of transcription of the NFkappaB gene by IL-1alpha was blocked by a tyrosine-specific kinase inhibitor, suggesting that the enhancement may require the phosphorylation of the NFkappaB complex. In vivo, NFkappaB is maximally expressed in the dental follicle of the rat first mandibular molar at day 3 postnatally, the age at which there is a peak influx of mononuclear cells into the follicle. Thus, a transcription factor apparently required for eruption (NFkappaB) is present in the tissue required for eruption, the dental follicle, and its gene expression is maximal at a critical time in eruption.

Activation of signal transducers and activators of transcription 1 and 3 by leukemia inhibitory factor, oncostatin-M, and interferon-gamma in adipocytes.

We have recently demonstrated that signal transducers and activators of transcription (STATs) 1, 3, 5A, 5B, and 6 are expressed in both cultured and native adipocytes. Our current studies have focused on the activation of STATs 1 and 3 by leukemia inhibitory factor (LIF), oncostatin-M (OSM), and interferon-gamma (IFNgamma) in 3T3-L1 adipocytes. IFNgamma is shown to be a potent activator of STAT 1 as indicated by both tyrosine phosphorylation and nuclear translocation. However, LIF and OSM, which are potent inducers of STAT 3, are less potent activators of STAT 1 as measured by both tyrosine phosphorylation and nuclear translocation. Both STATs 1 and 3 were translocated to the nucleus in a time-dependent fashion following LIF treatment. In addition, IFNgamma resulted in a time- and dose-dependent effect on STATs 1 and 3 nuclear translocation. Growth hormone, a potent activator of STATs 5A and 5B, had a minimal effect on STAT 1 and STAT 3 tyrosine phosphorylation. Preincubation with either insulin or growth hormone had no detectable effects on the tyrosine phosphorylation or nuclear translocation of STATs 1 and 3 induced by LIF, OSM, or IFNgamma. The effects of LIF and IFNgamma on STAT 1 and 3 tyrosine phosphorylation and nuclear translocation were confirmed in native rat adipocytes. In 3T3-L1 adipocytes, a low level of serine phosphorylation of STAT 3 on residue 727 was observed and was markedly enhanced by insulin, LIF, or OSM. This increase in STAT 3 Ser727 phosphorylation was dependent upon the activation of MAPK, since the MAPK kinase inhibitor (PD98059) reduced STAT 3 Ser727 phosphorylation to basal levels. The inhibition of MAPK had no effect on the ability of STATs 1 and 3 to be tyrosine-phosphorylated or translocate to the nucleus. These studies demonstrate the highly specific and quantitative activation of STATs 1 and 3 by LIF, OSM, and IFNgamma in adipocytes and indicate that STAT 3 is a substrate for MAPK in adipocytes.