Targeted expression of a dominant-negative N-cadherin in vivo delays peak bone mass and increases adipogenesis

Castro, Charlles Heldan de Moura; Shin, Chan Soo; Stains, Joseph P.; Cheng, Su‐Li; Sheikh, Sharmin; Mbalaviele, Gabriel; Szejnfeld, Vera Lúcia; Civitelli, Roberto

doi:10.1242/jcs.01133

Cited by 97 publications

(128 citation statements)

References 39 publications

(27 reference statements)

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“…(33) Cells were seeded in 96-well plate at a density of 100,000 cells/well and left undisturbed for 7 days. Cells were fed with ␣-MEM medium containing 10% FBS, ascorbic acid (50 g/ml), and ␤-glycerophosphate (10 mM) for 3 weeks.…”

Section: Osteogenesis Of Bone Marrow Cellsmentioning

confidence: 99%

Four and Half Lim Protein 2 (FHL2) Stimulates Osteoblast Differentiation

Lai

Bai

Uthgenannt

et al. 2006

Journal of Bone and Mineral Research

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FHL2, a molecule that interacts with many integrins and transcription factors, was found to play an important role in osteoblast differentiation. Overexpression of FHL2 increases the accumulation of osteoblast differentiation markers and matrix mineralization, whereas FHL2 deficiency results in inhibition of osteoblast differentiation and decreased bone formation.Introduction: Integrin-matrix interaction plays a critical role in osteoblast function. It has been shown that the cytoplasmic domains of integrin ␤ subunits mediate signal transduction induced by integrin-matrix interaction. We reasoned that the identification of proteins interacting with ␤-cytoplasmic tails followed by analysis of the function of these proteins would enhance our understanding on integrin signaling and the roles of these proteins in osteoblast activities. Materials and Methods: Yeast two hybrid assay was used to identify proteins interacting with the cytoplasmic domain of integrin ␤5 subunit. The association of these proteins with integrin ␣v␤5 was confirmed by confocal analysis and co-immunoprecipitation. A stable MC3T3-E1 cells line overexpressing Four and Half Lim Protein 2 (FHL2) and mouse osteoblasts deficient in FHL2 were used to study the roles of FHL2 in osteoblast differentiation and bone formation. Matrix protein expression was determined by mRNA analysis and Western blotting. Matrix mineralization was detected by Alizarin red staining. Alkaline phosphatase activity was also measured. CT was used to determine bone histomorphometry. Results and Conclusions: FHL2 and actin-binding proteins, palladin and filamin A, were identified as proteins interacting with ␤5 cytoplasmic domain. FHL2 co-localized with ␣v␤5 at the focal adhesion sites in association with palladin and filamin A. FHL2 was also present in nuclei. Osteoblasts overexpressing FHL2 exhibited increased adhesion to and migration on matrix proteins. Conversely, FHL2 stimulation of CREB activity was dependent on integrin function because it was inhibited by Gly-Arg-Gly-Asp-Ser (GRGDS) peptide. The expression of osteoblast differentiation markers and Msx2 was upregulated, and bone matrix mineralization was increased in FHL2 overexpressing cells. In contrast, FHL2-deficient bone marrow cells and osteoblasts displayed decreased osteoblast colony formation and differentiation, respectively, compared with wildtype cells. Moreover, FHL2-deficient female mice exhibited greater bone loss than the wildtype littermates after ovariectomy. Thus, FHL2 plays an important role in osteoblast differentiation and bone formation.

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Section: Osteogenesis Of Bone Marrow Cellsmentioning

confidence: 99%

Four and Half Lim Protein 2 (FHL2) Stimulates Osteoblast Differentiation

Lai

Bai

Uthgenannt

et al. 2006

Journal of Bone and Mineral Research

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“…Consistent with an early role for cell adhesion molecules in lineage commitment, mice expressing the mutant N-cadherin also displayed an osteogenic to adipogenic shift, with 27% increase in the percent of body fat relative to controls. Bone marrow mesenchymal cells from these animals were skewed towards adipogenic commitment rather than osteogenic (Castro et al, 2004). Osteoblast differentiation was delayed in calvaria isolated from these transgenic mice (Castro et al, 2004).…”

Section: Regulation Of Osteogenic Differentiation By Cell-to-cell Adhmentioning

confidence: 95%

“…This defect is cell autonomous since osteoblast function is impaired in vitro. The difficulty of the embryonic lethality of N-cadherin deficiency has been overcome by using a conditional genetic knock-out approach (Castro et al, 2004). Mice expressing a conditional dominant negative N-cadherin mutant showed a delay in reaching peak bone mass as a result of impaired osteogenic differentiation.…”

Section: Regulation Of Osteogenic Differentiation By Cell-to-cell Adhmentioning

confidence: 99%

“…Bone marrow mesenchymal cells from these animals were skewed towards adipogenic commitment rather than osteogenic (Castro et al, 2004). Osteoblast differentiation was delayed in calvaria isolated from these transgenic mice (Castro et al, 2004). Nevertheless, further investigations need to be conducted in order to dissect the specific contribution of each cadherin to osteogenic differentiation and osteoblast function, as well as to clarify the specific mechanism and signalling pathways by which they act during these processes.…”

Section: Regulation Of Osteogenic Differentiation By Cell-to-cell Adhmentioning

confidence: 99%

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The Retinoblastoma Protein in Osteogenesis and Osteosarcoma Formation

Santiago‐Cardona¹

2012

Osteogenesis

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“…23 Quantitative RT-PCR Expression levels of different liver mRNAs were determined by quantitative real-time PCR. Briefly, tissue explants from 12 h fasted mice were removed, snap frozen in liquid nitrogen and stored at À 80 1C until use.…”

Section: Body Compositionmentioning

confidence: 99%

Bradykinin inhibits hepatic gluconeogenesis in obese mice

Barros

Haro

Russo

et al. 2012

Laboratory Investigation

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The kallikrein-kinin system (KKS) has been previously linked to glucose homeostasis. In isolated muscle or fat cells, acute bradykinin (BK) stimulation was shown to improve insulin action and increase glucose uptake by promoting glucose transporter 4 translocation to plasma membrane. However, the role for BK in the pathophysiology of obesity and type 2 diabetes remains largely unknown. To address this, we generated genetically obese mice (ob/ob) lacking the BK B2 receptor (obB2KO). Despite similar body weight or fat accumulation, obB2KO mice showed increased fasting glycemia (162.3±28.2 mg/dl vs 85.3±13.3 mg/dl), hyperinsulinemia (7.71±1.75 ng/ml vs 4.09±0.51 ng/ml) and impaired glucose tolerance when compared with ob/ob control mice (obWT), indicating insulin resistance and impaired glucose homeostasis. This was corroborated by increased glucose production in response to a pyruvate challenge. Increased gluconeogenesis was accompanied by increased hepatic mRNA expression of forkhead box protein O1 (FoxO1, four-fold), peroxisome proliferator-activated receptor gamma co-activator 1-alpha (seven-fold), phosphoenolpyruvate carboxykinase (PEPCK, three-fold) and glucose-6-phosphatase (eight-fold). FoxO1 nuclear exclusion was also impaired, as the obB2KO mice showed increased levels of this transcription factor in the nucleus fraction of liver homogenates during random feeding. Intraportal injection of BK in lean mice was able to decrease the hepatic mRNA expression of FoxO1 and PEPCK. In conclusion, BK modulates glucose homeostasis by affecting hepatic glucose production in obWT. These results point to a protective role of the KKS in the pathophysiology of type 2 diabetes mellitus. KEYWORDS: bradykinin; gluconeogenesis; kinin receptor; leptin; ob/ob; obesity The pathophysiology of type 2 diabetes mellitus (T2DM) involves innumerous metabolic and endocrine alterations, including changes in the carbohydrate, lipid and protein metabolisms. T2DM is directly linked with insulin resistance and the inability of pancreatic beta cells to compensate for these changes. 1 There is also a strong association between T2DM and obesity 1,2 that persists independently of gender or ethnic group. In this interaction, the weight gain of obese subjects seems to precede the development of T2DM. 3 Kinins are vasoactive peptides produced in the circulatory system and other tissues by the action of serine proteases called kallikreins. The kallikrein-kinin system (KKS) modulates pain, vasodilatation, vascular permeability, inflammation and edema and has been linked to insulin resistance. [4][5][6][7] Bradykinin (BK), the agonist of the constitutively expressed BK B2 receptor (BKB2R), has been reported to increase glucose uptake in skeletal muscle and adipose tissue. 8 At the same time, the kinin B1 receptor and its agonist, the des-arg 9 -kinin, have been shown to control leptin sensitivity 7 and insulin secretion. 9 Despite evidence that acute BK stimulation improves glucose uptake, the role for BK and BKB2R in the pathophysiology of T2DM is s...

show abstract

Targeted expression of a dominant-negative N-cadherin in vivo delays peak bone mass and increases adipogenesis

Cited by 97 publications

References 39 publications

Four and Half Lim Protein 2 (FHL2) Stimulates Osteoblast Differentiation

Four and Half Lim Protein 2 (FHL2) Stimulates Osteoblast Differentiation

The Retinoblastoma Protein in Osteogenesis and Osteosarcoma Formation

Bradykinin inhibits hepatic gluconeogenesis in obese mice

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