Effect of salivary gland removal on taste preference in mice.

To evaluate the effect of decreased salivary secretion on taste preference, we investigated taste preference for five basic tastes by a 48 h two-bottle preference test using a mouse model (desalivated mice) that underwent surgical removal of three major salivary glands: the parotid, submandibular, and sublingual glands. In the desalivated mice, the avoidance behaviors for bitter and salty tastes and the attractive behaviors for sweet and umami tastes were significantly decreased. We confirmed that saliva is necessary to maintain normal taste preference. To estimate the cause of the preference changes, we investigated the effects of salivary gland removal on the expression of taste-related molecules in the taste buds. No apparent changes were observed in the expression levels or patterns of taste-related molecules after salivary gland removal. When the protein concentration and composition in the saliva were compared between the control and desalivated mice, the protein concentration decreased and its composition changed after major salivary gland removal. These results suggest that changes in protein concentration and composition in the saliva may be one of the factors responsible for the changes in taste preferences observed in the desalivated mice.

Factors associated with the eruption of the impacted maxillary third molars after second molar extraction.

INTRODUCTION: In orthodontic treatment, the space left after extracting the maxillary second molar (MxM2) may be filled by the eruption of the impacted third molar (MxM3). However, little is known about the factors associated with the eruption of the impacted MxM3. We aimed to characterize the clinical factors associated with the time taken for MxM3 eruption after MxM2 extraction. METHODS: We analyzed factors associated with late MxM3 eruption (>500 days after MxM2 extraction) in 84 molars. Prespecified risk factors were entered into logistic regression models to estimate odds ratios (ORs). RESULTS: The median duration between MxM2 extraction and MxM3 eruption was 302 days (interquartile range, 140-424). Significant factors associated with late MxM3 eruption included the proximity of the MxM3 root to the maxillary sinus floor (OR, 51.72), the distance between the occlusal plane of the MxM3 and the apical third of the MxM2 roots (OR, 16.56), MxM3 angulation and depth of ≥20° (OR, 5.58), ANB angle of <2° (OR, 9.05), and ≥1.5 mm distal movement of the maxillary first molar (MxM1) from its original position at the time of MxM2 extraction and MxM3 eruption (OR, 12.9). The probability of late MxM3 eruption was 0% (0 out of 30) with no risk, 6.9% (2 out of 29) with 1 risk factor, and 52% (13 out of 25) with ≥2 risk factors. CONCLUSIONS: We identified 5 clinical factors associated with late MxM3 eruption after MxM2 extraction. The probability of late MxM3 eruption increased as the number of present risk factors increased. These findings can be used for risk stratification during orthodontic treatment.

Mechanoresponsive and lubricating changes of mandibular condylar cartilage associated with mandibular lateral shift and recovery in the growing rat.

OBJECTIVE: The in vivo mechanoresponsive and lubricating changes of the mandibular condylar cartilage (MCC) associated with mandibular lateral shift (MLS) and recovery are poorly understood. Using growing rats, we investigated whether the expression of mechanoresponsive factors, including proteoglycan-4 (PRG4), Indian hedgehog (Ihh) and transforming growth factor-ß1 (TGF-ß1), would be affected by MLS. We also investigated whether these changes could recover to the control level after a 2-week treatment reversal (TR). MATERIALS AND METHODS: The MLS appliances were placed for 2 or 4 weeks in 5-week-old rats and removed from 7-week-old rats in the TR group. The MCC was analysed histomorphometrically by toluidine blue staining. Reverse transcription-polymerase chain reaction and immunohistochemistry were performed to evaluate the expression of PRG4, Ihh, PTHrP (parathyroid hormone-related protein), TGF-ß1, Matrix metallopeptidase 13 (MMP-13) and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5). RESULTS: A thickened superficial layer and an enhanced expression of PRG4 were detected in MLS groups. PTHrP-Ihh expression correlated positively with the up-regulation of PRG4. TGF-ß1 expression decreased in the early stage of MLS but recovered to the control level in the TR group. A significantly enhanced expression of MMP-13 in MLS groups was detected. CONCLUSION: MLS treatment, which acted on the growth stage of rats, affected the morphology and expression of lubrication factor in the MCC. Elimination of this mechanical stimulus may help MCC recover to normal conditions. CLINICAL RELEVANCE: Our study supports that the adaptive changes of MCC, which are caused by mandibular functional deviation, could be largely recovered by early treatment.

Effect of functional lateral shift of the mandible on hyaluronic acid metabolism related to lubrication of temporomandibular joint in growing rats.

BACKGROUND: Hyaluronic acid (HA) is a major molecular component of the articular cartilage of the temporomandibular joint (TMJ) influencing joint lubrication. Functional lateral shift of the mandible (FLSM) can lead to malocclusion. This study investigated the effects of FLSM on HA metabolism and lubrication of the TMJ in growing rats. METHODS: Thirty 5-week-old male Wistar rats were divided into shift, recovery, and control groups. Rats in the shift and recovery groups were fitted with guiding plates to produce a 2-mm FLSM which were removed from the rats in the recovery group 14 days later. Animals were sacrificed at 14 and 28 days after the appliances were attached. Immunohistochemistry of HA-binding protein (HABP), hyaluronan synthase (HAS), and hyaluronoglucosaminidases (HYALs) was examined. RESULTS: The thickness of HABP-positively stained areas in the lateral regions in the bilateral condyle was reduced during the experimental period in the shift group compared with that in the control group. The proportion of HAS2-stained areas was bilaterally decreased in different regions of condylar cartilage during the experimental period in the shift group. The reduction of the HYAL2-stained area proportion in the condylar cartilage was more significant than that of HYAL1 at 14 days after appliance attachment in the shift group. HAS2 staining was not recovered in the recovery group. LIMITATIONS: This research was based on animal experiments with a limited experimental period. CONCLUSION: FLSM altered lubrication related HA metabolism in the articular cartilage of the TMJ in growing rats.

Unilateral nasal obstruction induces degeneration of fungiform and circumvallate papillae in rats.

BACKGROUND: In clinical orthodontic treatment, chronic respiratory disturbance or mouth breathing has been concerned symptoms and screening criteria. In this study, to analyze the relation between nasal obstruction and taste sensing, a unilateral nasal obstruction model was performed to investigate the taste papillae and taste buds in rats. METHODS: Fourteen 6-day-old male Wistar rats were randomly divided into control and experimental groups (n = 7 each). The experimental group underwent unilateral nasal obstruction at 8 days of age. The rats were euthanized at 9-week-old. The morphology of the circumvallate papillae and taste buds were identified by immunohistochemical methods. The fungiform papillae were visualized with 1% methylene blue and sectioned for taste bud observation. RESULTS: Some defects in the gustatory epithelium were observed after unilateral nasal obstruction. Rats in the experimental group had significantly fewer fungiform papillae and smaller volumes of taste bud. In circumvallate papillae, smaller total taste bud area was found in experiment group. CONCLUSION: Findings in the present study suggest that nasal obstruction might have significant influences on the gustatory function via morphologic change in the taste papillae and taste buds in tongue area.

Degeneration of fungiform and circumvallate papillae following molar extraction in rats.

OBJECTIVE: Proper occlusion facilitates food intake and gustatory function is indispensable for the enjoyment of food. Although an interaction between dentoalveolar and gustatory afferent neurons has been suggested by previous studies, the relationship between occlusion and gustation remains unclear. This study investigated the effect of upper molar extraction which diminished occlusal support on peripheral gustatory receptors in rats. MATERIALS AND METHODS: Thirty-six 7-week-old male Wistar rats were randomly assigned to either the experimental or the control group. All maxillary molars were extracted from rats in the experimental group under anesthesia, while a sham operation was conducted in the control group. The rats were euthanized 7, 14 or 28 days after the procedure. The morphology of the circumvallate papillae and taste buds using immunohistochemical methods and the fungiform papillae were visualized with 1% methylene blue. RESULTS: Defects in the gustatory epithelium were observed after maxillary molar extraction. Rats in the experimental group had significantly fewer fungiform papillae, narrower circumvallate papillae, shallower trench depth, smaller trench area, smaller taste bud area, lower ratios of taste bud area to trench area and fewer taste buds than those in the control group. CONCLUSIONS: The findings indicate that molar extraction would affect peripheral gustatory receptors. This is the first study to characterize changes in rat fungiform and circumvallate papillae after maxillary molar extraction. This study suggests a possible synergic relationship between dentoalveolar perception and gustatory function, which has clinical implications that occlusion is closely correlated with gustatory perception.

P2X7R and P2X4R expression of mice submandibular gland in high-fat diet/streptozotocin-induced type 2 diabetes.

Type 2 diabetes mellitus (T2DM) is a chronic inflammatory disease that can compromise the functioning of various organs, including the salivary glands (SG). The purinergic system is one of the most important inflammatory pathways in T2DM condition, and P2X7R and P2X4R are the primary purinergic receptors in SG that regulate inflammatory homeostasis. This study aimed to evaluate P2X7R and P2X4R expression, and morphological changes in the submandibular gland (SMG) in T2DM. Twenty-four 5-week-old mice were randomly assigned to control (CON) and diabetes mellitus (DM) groups (n = 12 each). Body weight, diet, and blood glucose levels were monitored weekly. The histomorphology of the SMG and the expression of the P2X7R, and P2X7R was evaluated by immunohistochemistry (IHC) staining and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) at 11 and 13 weeks of age. Our findings indicate a significant increase in food consumption, body weight, and blood glucose levels in the DM group. Although a significant increase in P2X7R and P2X4R expression was observed in the DM groups, the receptor location remained unchanged. We also observed a significant increase in the acinar area in the DM13w group, and a significant decrease in the ductal area in the DM11w and DM13w groups. Targeting purinergic receptors may offer novel therapeutic methods for diabetic complications.

Expression of glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide in the rat submandibular gland is influenced by pre- and post-natal high-fat diet exposure.

Background: Incretins, i.e., glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) promote insulin secretion to reduce postprandial blood sugar. Previous studies found incretins in the salivary glands. However, the role of GLP-1 and GIP in the submandibular gland (SMG) is unclear. This study investigates the effects of a high-fat diet (HFD) on the expression of GLP-1 and GIP throughout the development of rat SMG. Methods: Pregnant 11-week-old Wistar rats were divided into two groups: those fed on a standard diet (n = 5) and those fed on a HFD (n = 5). From day 7 of pregnancy and throughout the lactation period, all the rats were fed on either a chow diet or HFD. The newborns were divided into four subgroups (n = 6): standard diet males (SM), HFD males (HM), standard diet females (SF), and HFD females (HF). The SMGs of 3- and 10-week-old rats from each subgroup were collected under general anesthesia. Moreover, body weight, food intake, and fasting blood sugar were measured. The mRNA expression of GLP-1 and GIP was quantified, and the localization was observed using immunohistochemistry (p < 0.05). Results: GLP-1 mRNA expression was statistically significantly more upregulated in HM than in HF at 3 weeks. Moreover, GLP-1 mRNA expression was significantly higher in HM than in both SM and HF at 10 weeks. Although a decreasing trend was observed in GIP mRNA expression in both 3- and 10-week-old rats fed on a HFD, a significant difference between HM and SM only occurred at 3 weeks. Furthermore, the GIP mRNA expression of HM was lower than that of HF at 10 weeks. Immunohistochemical staining revealed GLP-1 and GIP expression mainly in the SMG duct system. Moreover, vacuolated cytoplasm in the duct was observed in rats fed on a HFD. Conclusion: Exposure to HFD during pre- and post-natal periods increased GLP-1 mRNA expression in the SMGs of male rats. However, GIP expression decreased following the HFD in male newborns. Furthermore, a decreasing trend of GIP mRNA expression was observed in male newborns after HFD feeding. Sex influenced incretin hormones secretion and obesity-related conditions. HFD during pre- and post-natal periods reprograms the epigenome, contributing to subsequent disease development.

The Effect of Two-Generation Exposure to a High-Fat Diet on Craniofacial Morphology in Rats.

This study aimed to examine the sexual dimorphism effect of two-generation exposure to a high-fat diet (HFD) on the craniofacial growth of rat offspring. Ten eleven-week-old pregnant Wistar rats were fed either a control or HFD from day 7 of pregnancy until the end of lactation. Twelve male and female offspring from the control-diet-fed mothers were assigned to the CM (control male, n = 6) and CF (control female, n = 6) groups. The other twelve from the HFD-fed mothers were assigned to the HFD male (HFDM, n = 6) and HFD female (HFDF, n = 6) groups. HFDM and HFDF rats continued with an HFD. The offspring's weight and fasting blood sugar levels were measured every two weeks. The craniofacial and dental morphologies were studied from lateral X-rays of the head at ten weeks old. The HFDM rats showed an increased body weight and larger neurocranial parameters compared with the CM group. Furthermore, there were slightly significant differences in body weight and viscerocranial parameters between the rats in the HFDF and CF groups. In conclusion, two-generational exposure to an HFD had a greater effect on the male offspring's body weight and craniofacial morphology.

Two-generation exposure to a high-fat diet induces the change of salty taste preference in rats.

High-fat diet (HFD) leads to multiple complications, including taste alteration. This study observed the effect of a two-generation exposure to an HFD on the peripheral taste system in offspring. Ten pregnant Wistar rats were assigned a standard diet (SD) (n = 5) or HFD (n = 5) from day 7 of pregnancy through the lactation. Thirty-six male and female 3-week-old offspring were measured for body weight and blood glucose level, and the circumvallate papillae were collected. The other twenty-four 3-week-old offspring were weaned on the same diet as their mothers and raised individually. The taste preference behaviors were studied using the two-bottle taste preference test and analyzed five basic tastes (sweet, bitter, umami, sour, and salty). The expressions of epithelial sodium channel alpha subunit (ENaCα) and angiotensin II receptor type 1 (AT1) in the circumvallate papilla were analyzed by immunohistochemical staining and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). We found increased body weight and salty taste preference of offspring from the HFD group in both sexes. Correspondingly, the AT1 level of the taste bud cells significantly increased in 3-week-old female offspring from the HFD group. An increase in AT1 levels may be a risk factor for changes in salty taste preference.

Does Orthodontic Treatment Change the Preferred Chewing Side of Patients with Malocclusion?

Whether orthodontic treatment can change the preferred chewing side (PCS) is unknown. This study examined (1) if the PCS changes after orthodontic treatment and (2) which factors contribute to this change. Two hundred fifty patients who visited the orthodontic clinic at Tokyo Medical and Dental University Hospital between 2017 and 2020 were included in the study. Mandibular kinesiograph (MKG) was taken at pre- and post-treatment, and PCS was determined. Patients who showed a change in PCS to the opposite side and those who showed no change in PCS at post-treatment were pooled into the PCS-changed and PCS-unchanged groups, respectively. The demographic, clinical, and cephalometric parameters were compared between the groups. Significant factors associated with changes in were of age < 20 years at the beginning of orthodontic treatment (odds ratio (OR), 2.00), maximum lateral mandibular movement to PCS ≥ 10.0 mm at pre-treatment (OR, 6.51), and change in occlusal canting of ≥1.0° (OR, 2.72). The predicted probability of change in PCS was 13.2%, 36.0%, and 67.5% for no factor, one factor, and two factors associated with PCS change, respectively. Orthodontic treatment may change PCS due to patient age, maximum lateral mandibular movement to PCS, and change in occlusal canting.

The effect of diabetes mellitus on rat mandibular bone formation and microarchitecture.

The aim of this study was to assess the effect of type 1 diabetes mellitus (DM) on the structure of mandibular bone and on the changes of alveolar/jaw bone formation. Experimental DM was induced in 3-wk-old male Wistar rats by a single dose of 60 mg/kg body weight of streptozotocin. All rats were injected with calcein on days 21 and 28. The rats were killed when 8 wk of age. Bone structure was analyzed by bone histomorphometry, microcomputed tomography (micro-CT), and histological section. Histomorphometric analysis showed that the mineral apposition and the bone formation rates in most of the mandibular regions were significantly decreased in the DM group compared with the control group. Micro-CT analysis showed significant deterioration of the bone quality in rats with DM. For a histometric measure of bone resorption, the number of osteoclasts along the distal surface of the alveolar wall was counted. The number of osteoclasts was significantly lower in the rats with DM than in the controls. These findings suggest that uncontrolled DM decreases mandibular bone formation, reduces the rate of bone turnover in the alveolar wall surrounding the root, and affects the quality of bone structure resulting in retardation of its skeletal development.

Occlusional Modifications Reversibly Alter Aquaporin 5 Expression and Localization in Rat Salivary Glands.

BACKGROUND: Aquaporin 5 (AQP5) is a water channel-forming protein that plays a key role in saliva secretion. A decrease in masticatory function associated with the molar extraction adversely affects the submandibular salivary gland (SMG) in rats, inducing hypertrophic changes in the acinar cells and the expression of AQP5 in acinar cells or intercalated duct of the SMG. However, changes in AQP5 expression and localization in the SMG in association with occlusal modification have not been fully characterized. METHODS: We examined the influence of the decline and recovery of masticatory function on expression and localization of AQP5 in the rat SMG by inserting and removing an incisor bite plate (IBP). Thirty 5-week-old male Wistar rats were randomly divided into IBP (n = 12), recovery (REC) (n = 6), and control (CON) (n = 12) groups. Each rat in both the IBP and REC groups was fitted with the IBP on its maxillary incisors. Rats without the IBPs served as controls. All rats were fed powder diet and water ad libitum. Rats in the IBP and CON groups were sacrificed after 14 (n = 6) and 28 (n = 6) days after the IBP attachment. In the REC group, the IBP was detached on the 14th day and sacrificed on 28th day after the IBP attachment. AQP5 mRNA expression was quantified by reverse transcription-polymerase chain reaction. Changes in the localization of AQP5 were tracked by immunohistochemical staining. RESULTS: Attachment of IBP resulted in a decrease in the expression of AQP5 in the IBP group. Changes in the localization of AQP5 were observed between 14 and 28 days in the IBP group. In contrast, changes in the expression and localization of AQP5 were not observed in the REC group. CONCLUSION: Findings suggested that a loss of molar occlusion, due to the IBP attachment, altered AQP5 expression and localization in the rat SMG. However, removal of the bite plate allowed the recovery of both AQP5 expression and its normal localization in the SMGs.

Alterations in the gustatory papillae after anterior bite plate insertion in growing rats.

OBJECTIVES: To determine whether the modification of dental occlusion, without molar extraction, affected the gustatory papillae located in the tongue of growing rats. MATERIALS AND METHODS: Five-week-old male Wistar rats were randomly divided into an anterior bite plate (ABP) group and a control group. Under general anesthesia, ABPs were placed on the occlusal surfaces of the maxillary incisors, while metal caps covered the mandibular incisal edges of the rats in the ABP group. The control group rats underwent a sham operation. The rats in both groups were euthanized 14 days after the procedure. The circumvallate papillae and taste buds were analyzed by immunohistochemical methods, and the fungiform papillae were observed and counted after immersion of the tongue in 1% methylene blue. RESULTS: Two weeks after ABP insertion and mandibular incisal cap placement, the gustatory papillae exhibited morphological and structural changes. The rats in the ABP group had exhibited significantly fewer fungiform papillae, and narrower circumvallate papillae, with greater trench depths, larger trench profile areas, smaller taste bud profile areas, lower ratios of the taste bud profile area to the trench profile area, and more taste buds than those in the control group. CONCLUSIONS: Our findings support the association between occlusal and taste functions and provide a basis for further studies on the gustatory function. In conclusion, loss of molar occlusion, resulting from the ABP and metal cap insertion, altered the peripheral gustatory receptors in the growing rats.

Unilateral nasal obstruction alters sweet taste preference and sweet taste receptors in rat circumvallate papillae.

Nasal obstruction causes mouth breathing, and affects the growth and development of craniofacial structures, muscle function in the stomatognathic system, and the taste perceptive system. However, the detailed mechanism underlying the effects of nasal obstruction on taste perception has not been fully elucidated. In this study, we investigated this mechanism using the two-bottle taste preference test, immunohistological analysis, and quantification of the mRNA expression of taste-related molecules in the circumvallate papillae. Neonatal male Wistar rats were divided randomly into control and experimental groups. Rats in the experimental group underwent unilateral nasal obstruction by cauterization of the external nostril at the age of 8 days. Arterial oxygen saturation (SpO2) was recorded in awake rats using collar clip sensors. Taste preference for five basic taste solutions was evaluated. Immunohistochemical analysis and quantitative real-time polymerase chain reaction (RT-PCR) were conducted to evaluate the expressions of taste-related molecules in the taste cells of the circumvallate papillae. Body weights were similar between the two groups throughout the experimental period. The SpO2 in the 7- to 12-week-old rats in the experimental group was significantly lower than that in the age-matched rats in the control group. In the two-bottle taste preference test, the sensitivities to sweet taste decreased in the experimental group. The mRNA expression of T1R2, T1R3, α-gustducin, and PLCß2 was significantly lower in the experimental group than in the control group as determined by quantitative RT-PCR, and the immunohistochemical staining for α-gustducin and PLCß2 was less prominent. These findings suggest that nasal obstruction may affect sweet taste perception via the reduced expression of taste-related molecules in the taste cells in rat circumvallate papillae.

Effect of experimental diabetes on craniofacial growth in rats.

OBJECTIVES: The goal of this study was to assess the effect of type 1 diabetes mellitus (T1DM) on the craniofacial growth and skeletal maturation using the STZ-DM rat model. DESIGN: Experimental T1DM was induced in 3-week-old male Wistar rats by a single dose of 60 mg/kg body weight of streptozotocin (STZ). Lateral and dorsoventral X-rays of the head were taken at the age of 7 weeks. The X-rays were scanned, digitised and selected linear distances were measured and analysed statistically. RESULTS: In STZ-DM statistical analysis of results revealed a reduction in growth of most of the linear measurements in the neurocranium and mandible by X-ray analysis, and all measurements were significantly lower in viscerocranium. CONCLUSIONS: Uncontrolled T1DM reduces craniofacial growth, resulting in retardation of skeletal development.

Calcitonin and vitamin D3 have high therapeutic potential for improving diabetic mandibular growth.

The goal of this study was to assess the effect of the intermittent combination of an antiresorptive agent (calcitonin) and an anabolic agent (vitamin D3) on treating the detrimental effects of Type 1 diabetes mellitus (DM) on mandibular bone formation and growth. Forty 3-week-old male Wistar rats were divided into four groups: the control group (normal rats), the control C+D group (normal rats injected with calcitonin and vitamin D3), the diabetic C+D group (diabetic rats injected with calcitonin and vitamin D3) and the diabetic group (uncontrolled diabetic rats). An experimental DM condition was induced in the male Wistar rats in the diabetic and diabetic C+D groups using a single dose of 60 mg·kg(-1) body weight of streptozotocin. Calcitonin and vitamin D3 were simultaneously injected in the rats of the control C+D and diabetic C+D groups. All rats were killed after 4 weeks, and the right mandibles were evaluated by micro-computed tomography and histomorphometric analysis. Diabetic rats showed a significant deterioration in bone quality and bone formation (diabetic group). By contrast, with the injection of calcitonin and vitamin D3, both bone parameters and bone formation significantly improved (diabetic C+D group) (P < 0.05). These findings suggest that these two hormones might potentially improve various bone properties.

Expression of glucose-dependent insulinotropic polypeptide and its receptor in the rat major salivary glands.

Glucose-dependent insulinotropic polypeptide receptors (GIPR) are expressed throughout the body. The expression of its ligand, glucose-dependent insulinotropic polypeptide (GIP) however, has only been reported in a limited numbers of organs. Although the rat submandibular salivary gland (SMG) has been found to express GIP, its biological role is still not understood. Moreover, nothing is known about the expression of GIP in other types of salivary glands, i.e. the parotid (PG) and sublingual (SLG) glands. We detected the expression of GIP mRNA in the rat PG, SMG and SLG. Immunohistochemical analyses revealed that GIP and GIPR were expressed only in the ductal area of all types of major salivary glands, and no immunostaining was found in the acini area. We also found GIP expression in the rat SMG to be age dependent, with 8-week-old rats showing 2-3-fold higher than those of 9- and 11-week-old rats, respectively. This is the first study to indicate both GIP and GIPR expression in the rat major salivary glands, as well as its variation in the rat SMG during the growth period. These findings are crucial for a better understanding of the physiological function of GIP in rat major salivary gland.

Expression of glucagon-like peptide-1 receptor and glucose­dependent insulinotropic polypeptide receptor is regulated by the glucose concentration in mouse osteoblastic MC3T3-E1 cells.

Glucose-dependent insulinotropic polypeptide receptor (GIPR) and glucagon-like peptide-1 receptor (GLP­1R) are incretin receptors that play important roles in regulating insulin secretion from pancreatic ß cells. Incretin receptors are also thought to play a potential role in bone metabolism. Osteoblasts in animals and humans express GIPR; however, the presence of GLP-1R in these cells has not been reported to date. Thus, the aim of this study was to determine whether GLP-1R and GIPR are expressed in osteoblastic cells, and whether their expression levels are regulated by the extracellular glucose concentration. Mouse osteoblastic MC3T3-E1 cells were cultured in medium containing normal (5.6 mM) or high (10, 20 or 30 mM) glucose concentrations, with or without bone morphogenetic protein-2 (BMP-2). RT-PCR, western blot analysis and immunofluorescence were carried out to determine GIPR and GLP-1R mRNA and protein expression levels. Cell proliferation was also assessed. The GLP-1R and GIPR mRNA expression levels were higher in the MC3T3-E1 cells cultured in medium containing high glucose concentrations with BMP-2 compared with the cells cultured in medium containing normal glucose concentrations with or without BMP-2. GLP-1R protein expression increased following culture in high-glucose medium with BMP-2 compared with culture under normal glucose conditions. However, the cellular localization of GLP-1R was not affected by either glucose or BMP-2. In conclusion, our data demonstrate that the expression of GLP-1R and GIPR is regulated by glucose concentrations in MC3T3-E1 cells undergoing differentiation induced by BMP-2. Our results reveal the potential role of incretins in bone metabolism.