ATP and UTP stimulate bone morphogenetic protein-2,-4 and -5 gene expression and mineralization by rat primary osteoblasts involving PI3K/AKT pathway

Ayala-Peña, Victoria B.; Scolaro, Luis Alberto; Santillán, Graciela

doi:10.1016/j.yexcr.2013.05.006

Cited by 45 publications

(50 citation statements)

References 59 publications

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“…In turn, ATP activates P2Y purinergic signaling pathways (Ayala-Peña et al, 2013;Rodrigues-Ribeiro et al, 2015). We also used external ATP stimulation to replace oscillatory fluid flow, and we obtained similar results for all our experiments; the results of ERK activation by fluid flow or ATP confirm the important role of ATP release in bone cell mechanotransduction.…”

Section: Discussionsupporting

confidence: 69%

GRK2 desensitizes flow-induced responses in osteoblasts

Xing

Gu²,

Shan

et al. 2017

Genet. Mol. Res.

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ABSTRACT. Bone desensitization after mechanical loading is essential for bone to adapt to its mechanical environment. However, the desensitization mechanism is unknown. Previous studies suggest that G protein-coupled receptors (GPCRs), including P2Y and parathyroid hormone receptors, play important roles in osteoblast mechanobiology. Thus, for the present research, we examined the role of G protein-coupled receptor kinase 2 (GRK2) in osteoblast desensitization after exposure to mechanical stimulation. We first showed the existence of osteoblast desensitization after mechanical stimulation based on cytosol Ca 2+ and phosphorylated ERK1/2 activities, detected using a fluorescent Ca 2+ -sensitive dye and western blotting, respectively. We then demonstrated that GRK2 overexpression in MC3T3-E1 cells inhibits flow-induced ERK1/2 phosphorylation, while siRNA knockdown of GRK2 enhances ERK1/2 phosphorylation. Additionally, we found that GRK2 overexpression in MC3T3-E1 cells inhibits cyclooxygenase-2 mRNA expression in the short term and alkaline phosphatase activity in the long term. More importantly, we discovered that GRK2 translocated to the cell membrane shortly after flow stimulation -a step necessary for GPCR desensitization. Previously, we have demonstrated that P2Y2 purinergic receptors, one type of GPCRs, are involved in various flowinduced osteoblastic responses. In this research, we also showed that GRK2 overexpression does not affect ATP release. Accordingly, GRK2 is able to inhibit flow-induced osteoblast responses possibly through desensitizing P2Y2 receptors.

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Section: Discussionsupporting

confidence: 69%

GRK2 desensitizes flow-induced responses in osteoblasts

Xing

Gu²,

Shan

et al. 2017

Genet. Mol. Res.

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“…BMP 2, BMP 5, and BMP 6 are among the most potent inducers and stimulators of osteoblast differentiation [65][66][67][68]. For normal osteogenesis to proceed, the expression of BMPs needs to be suppressed, as observed in human ESCs during osteogenic differentiation [69].…”

Section: Figmentioning

confidence: 99%

Comparative Analysis of Mouse-Induced Pluripotent Stem Cells and Mesenchymal Stem Cells During Osteogenic Differentiation In Vitro

Egusa

Kayashima

Miura

et al. 2014

Stem Cells and Development

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Induced pluripotent stem cells (iPSCs) can differentiate into mineralizing cells and are, therefore, expected to be useful for bone regenerative medicine; however, the characteristics of iPSC-derived osteogenic cells remain unclear. Here, we provide a direct in vitro comparison of the osteogenic differentiation process in mesenchymal stem cells (MSCs) and iPSCs from adult C57BL/6J mice. After 30 days of culture in osteogenic medium, both MSCs and iPSCs produced robustly mineralized bone nodules that contained abundant calcium phosphate with hydroxyapatite crystal formation. Mineral deposition was significantly higher in iPSC cultures than in MSC cultures. Scanning electron microscopy revealed budding matrix vesicles in early osteogenic iPSCs; subsequently, the vesicles propagated to exhibit robust mineralization without rich fibrous structures. Early osteogenic MSCs showed deposition of many matrix vesicles in abundant collagen fibrils that became solid mineralized structures. Both cell types demonstrated increased expression of osteogenic marker genes, such as runx2, osterix, dlx5, bone sialoprotein (BSP), and osteocalcin, during osteogenesis; however, real-time reverse transcription-polymerase chain reaction array analysis revealed that osteogenesis-related genes encoding mineralization-associated molecules, bone morphogenetic proteins, and extracellular matrix collagens were differentially expressed between iPSCs and MSCs. These data suggest that iPSCs are capable of differentiation into mature osteoblasts whose associated hydroxyapatite has a crystal structure similar to that of MSCassociated hydroxyapatite; however, the transcriptional differences between iPSCs and MSCs could result in differences in the mineral and matrix environments of the bone nodules. Determining the biological mechanisms underlying cell-specific differences in mineralization during in vitro iPSC osteogenesis may facilitate the development of clinically effective engineered bone.

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“…In a recent study, however, ATP and UTP were shown to increase mineralization and ALP activity in primary rat osteoblasts. Along with this, an increase in expression of BMP4 and BMP5, proteins involved in regulation of bone matrix formation, was seen upon stimulation with ATP and UTP [8]. Furthermore, Xing et al showed that primary osteoblasts from P2Y2R KO mice formed less mineralized nodules and had lower ALP activity compared to osteoblasts from Wt mice [14].…”

Section: Discussionmentioning

confidence: 91%

“…Stimulation of primary rat osteoblast cultures with ATP or UTP (1-100 μM) has been shown to inhibit mineralized bone nodule formation [7] by a mechanism not involving P2Y4R [6]. However, ATP and UTP (100 μM) have also been shown to increase mineralized bone nodule formation in primary rat osteoblasts, possibly through a mechanism involving upregulation of bone morphogenetic proteins (BMP) [8]. The expression of P2Y2R in osteoclasts is less clear.…”

Section: Introductionmentioning

confidence: 99%

Bone turnover is altered in transgenic rats overexpressing the P2Y2 purinergic receptor

Ellegaard

Ağca

Petersen

et al. 2017

Purinergic Signalling

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It is now widely recognized that purinergic signaling plays an important role in the regulation of bone remodeling. One receptor subtype, which has been suggested to be involved in this regulation, is the P2Y2 receptor (P2Y2R). In the present study, we investigated the effect of P2Y2R overexpression on bone status and bone cell function using a transgenic rat. Three-month-old female transgenic Sprague Dawley rats overexpressing P2Y2R (P2Y2R-Tg) showed higher bone strength of the femoral neck. Histomorphometry showed increase in resorptive surfaces and reduction in mineralizing surfaces. Both mineral apposition rate and thickness of the endocortical osteoid layer were higher in the P2Y2R-Tg rats. μCT analysis showed reduced trabecular thickness and structural model index in P2Y2R-Tg rats. Femoral length was increased in the P2Y2R-Tg rats compared to Wt rats. In vitro, there was an increased formation of osteoclasts, but no change in total resorption in cultures from P2Y2R-Tg rats. The formation of mineralized nodules was significantly reduced in the osteoblastic cultures from P2Y2R-Tg rats. In conclusion, our study suggests that P2Y2R is involved in regulation of bone turnover, due to the effects on both osteoblasts and osteoclasts and that these effects might be relevant in the regulation of bone growth.

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ATP and UTP stimulate bone morphogenetic protein-2,-4 and -5 gene expression and mineralization by rat primary osteoblasts involving PI3K/AKT pathway

Cited by 45 publications

References 59 publications

GRK2 desensitizes flow-induced responses in osteoblasts

GRK2 desensitizes flow-induced responses in osteoblasts

Comparative Analysis of Mouse-Induced Pluripotent Stem Cells and Mesenchymal Stem Cells During Osteogenic Differentiation In Vitro

Bone turnover is altered in transgenic rats overexpressing the P2Y2 purinergic receptor

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