The Skeletal site‐specific role of connective tissue growth factor in prenatal osteogenesis

Lambi, Alex G.; Pankratz, Talia L.; Mundy, Christina; Gannon, Maureen; Barbe, Mary F.; Richtsmeier, Joan T.; Popoff, Steven N.

doi:10.1002/dvdy.23888

Cited by 27 publications

(38 citation statements)

References 55 publications

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“…Additionally, microCT analysis revealed a serpentine morphology of the nasal septum perhaps as a result of the shortened rostro-caudal length of the skull previously reported in the CCN2 knockout phenotype (Fig. 1e and f) (Lambi et al 2012). Coronal sections of WT and KO P0 heads revealed absence of the secondary palate in KO mice ( Fig.…”

Section: Resultssupporting

confidence: 62%

The pivotal role of CCN2 in mammalian palatogenesis

Tarr

Visser

Moon

et al. 2016

J. Cell Commun. Signal.

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Mammalian palatogenesis is a complex process involving a temporally and spatially regulated myriad of factors. Together these factors control the 3 vital processes of proliferation, elevation and fusion of the developing palate. In this study, we show for the first time the unequivocally vital role of CCN2 in development of the mammalian palate. We utilized CCN2 knockout (KO) mice and cranial neural crest derived mesenchymal cells from these CCN2 KO mice to investigate the 3 processes crucial to normal palatogenesis. Similar to previously published reports, the absence of CCN2 inhibits proliferation of cells in the palate specifically at the G1/S transition. Absence of CCN2 also inhibited palatal shelf elevation from the vertical to horizontal position. CCN2 KO mesenchymal cells demonstrated deficiencies in adhesion and spreading owing to an inability to activate Rac1 and RhoA. On the contrary, CCN2 KO mesenchymal cells exhibited increased rates of migration compared to WT cells. The addition of exogenous CCN2 to KO mesenchymal cells restored their ability to spread normally on fibronectin. Finally, utilizing an organ culture model we show that the palatal shelves of the CCN2 KO mice demonstrate an inability to fuse when apposed. Together, these data signify that CCN2 plays an indispensible role in normal development of the mammalian palate and warrants additional studies to determine the precise mechanism(s) responsible for these effects.

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Section: Resultssupporting

confidence: 62%

The pivotal role of CCN2 in mammalian palatogenesis

Tarr

Visser

Moon

et al. 2016

J. Cell Commun. Signal.

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“…Similarly, Ctgf overexpression in chondrocytes causes bone loss [22]. Targeted disruption of Ctgf in mice leads to severe skeletal developmental abnormalities, as a result of impaired cartilage/bone development [21, 23]. We demonstrated that the conditional inactivation of Ctgf in the limb bud or in differentiated osteoblasts results in osteopenia, confirming its direct role in skeletal development, and demonstrating that Ctgf is necessary for adult skeletal homeostasis [20].…”

Section: Introductionmentioning

confidence: 66%

Connective Tissue Growth Factor is a Target of Notch Signaling in Cells of the Osteoblastic Lineage

Canalis

Zanotti

Smerdel-Ramoya

2014

Bone

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Connective tissue growth factor (Ctgf) or CCN2 is a protein synthesized by osteoblasts necessary for skeletal homeostasis, although its overexpression inhibits osteogenic signals and bone formation. Ctgf is induced by bone morphogenetic proteins, transforming growth factor β and Wnt; and in the present studies, we explored whether Notch regulated Ctgf expression in osteoblasts. We employed RosaNotch mice, where the Notch intracellular domain (NICD) is expressed following the excision of a STOP cassette, placed between the Rosa26 promoter and NICD. Notch was activated by transduction of adenoviral vectors expressing Cre recombinase (Ad-CMV-Cre). Notch induced Ctgf mRNA levels in a time dependent manner and increased Ctgf heterogeneous nuclear RNA. Notch also destabilized Ctgf mRNA shortening its half-life from 13 h to 3 h. The effect of Notch on Ctgf expression was lost following Rbpjκ downregulation, demonstrating that it was mediated by Notch canonical signaling. However, downregulation of the classic Notch target genes Hes1, Hey1 and Hey2 did not modify the effect of Notch on Ctgf expression. Wild type osteoblasts exposed to immobilized Delta-like 1 displayed enhanced Notch signaling and increased Ctgf expression. In addition to the effects of Notch in vitro, Notch induced Ctgf in vivo, and calvariae and femurs from RosaNotch mice mated with transgenics expressing the Cre recombinase in cells of the osteoblastic lineage exhibited increased expression of Ctgf. In conclusion, Ctgf is a target of Notch canonical signaling in osteoblasts, and may act in concert with Notch to regulate skeletal homeostasis.

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“…For cDNA synthesis, 1 μg of cDNA was transcribed from total RNA using SuperScript® First-Strand Synthesis System (Invitrogen). Gene expression for connective tissue growth factor ( CTGF ), runt-related transcription factor 2 ( Runx2 ), alkaline phosphatase ( Alp ), collagen type I alpha 1 ( Col1a1 ), osteocalcin ( Oc ), bone morphogenetic protein receptor Ia ( BMPR-Ia ), and bone morphogenetic protein receptor Ib ( BMPR-Ib ) was determined by qPCR, as described previously (Lambi et al, 2012). The primer information for the abovementioned genes has been previously published (Lambi et al, 2012; Pache et al, 2006).…”

Section: Methodsmentioning

confidence: 99%

“…In a study in which osteoblasts derived from these KO mice exhibited delayed osteoblast maturation and mineralization in culture, suggesting that CTGF is essential for normal osteoblast differentiation (Kawaki et al, 2008). Our lab conducted an in-depth characterization of CTGF KO mice and found these mice to exhibit numerous site-specific defects in the axial, appendicular, and craniofacial skeleton (Lambi et al, 2012). Interestingly, we found that mRNA expression of Runx2, Alp, and Oc differed depending on the skeletal site (Lambi et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

“…Our lab conducted an in-depth characterization of CTGF KO mice and found these mice to exhibit numerous site-specific defects in the axial, appendicular, and craniofacial skeleton (Lambi et al, 2012). Interestingly, we found that mRNA expression of Runx2, Alp, and Oc differed depending on the skeletal site (Lambi et al, 2012). In addition to the CTGF KO model, others have generated transgenic models in which CTGF is over-expressed under the control of different promoters (Nakanishi et al, 2001; Smerdel-Ramoya et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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Connective Tissue Growth Factor (CTGF/CCN2) Negatively Regulates BMP‐2 Induced Osteoblast Differentiation and Signaling

Mundy

Gannon

Popoff

2014

Journal Cellular Physiology

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Connective tissue growth factor (CTGF/CCN2) and bone morphogenetic protein (BMP)-2 are both produced and secreted by osteoblasts. Both proteins have been shown to have independent effects in regulating osteoblast proliferation, maturation and mineralization. However, how these two proteins interact during osteoblast differentiation remains unknown. In this study, we utilized two cell culture model systems, osteoblasts derived from CTGF knockout (KO) mice and osteoblasts infected with an adenovirus which over-expresses CTGF (Ad-CTGF), to investigate the effects of CTGF and BMP-2 on osteoblast development and function in vitro. Contrary to a previously published report, osteoblast maturation and mineralization were similar in osteogenic cultures derived from KO and WT calvaria in the absence of BMP-2 stimulation. Interestingly, in KO and WT osteoblast cultures stimulated with BMP-2, the KO osteoblasts exhibited enhanced osteoblast differentiation. This increase in osteoblast differentiation was accompanied by increased protein levels of phosphorylated Smad 1/5/8 and mRNA expression levels of bone morphogenetic protein receptor Ib. We also examined osteoblast differentiation in cultures that were infected with an adenoviral-CTGF vector (Ad-CTGF) and in controls. Continuous over-expression of CTGF resulted in decreased osteoblast maturation and mineralization in both unstimulated and BMP-2 stimulated cultures. Impaired osteoblast differentiation in cultures over-expressing CTGF was accompanied by decreased protein levels of phosphorylated Smad 1/5/8. Collectively, the data from these studies demonstrate that CTGF acts to negatively regulate BMP-2 induced signaling and osteoblast differentiation, and warrant additional studies to determine the precise mechanism(s) responsible for this effect.

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The Skeletal site‐specific role of connective tissue growth factor in prenatal osteogenesis

Cited by 27 publications

References 55 publications

The pivotal role of CCN2 in mammalian palatogenesis

The pivotal role of CCN2 in mammalian palatogenesis

Connective Tissue Growth Factor is a Target of Notch Signaling in Cells of the Osteoblastic Lineage

Connective Tissue Growth Factor (CTGF/CCN2) Negatively Regulates BMP‐2 Induced Osteoblast Differentiation and Signaling

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