Plectin stabilizes microtubules during osteoclastic bone resorption by acting as a scaffold for Src and Pyk2

Matsubara, Takuma; Yaginuma, Tatsuki; Addison, William N.; Fujita, Yūkō; Watanabe, Kouji; Yoshioka, Izumi; Hikiji, Hisako; Maki, Kenshi; Baron, Roland; Kokabu, Shoichiro

doi:10.1016/j.bone.2019.115209

Cited by 13 publications

(30 citation statements)

References 32 publications

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“…7a) [70]. Consistent with previous studies [71], RAW 264.7 cells were successfully induced into mature osteoclasts under conditioned culture medium, as reflected by the remarkable induction of representative osteoclastic markers including carbonic anhydrase II (CA-II) [72], nuclear factor of activated T-cells (NFATC) [73] and TRAP [74], as determined by RT-qPCR (Additional file 1: Figure S4, P < 0.001). Moreover, morphological changes also verified the maturation of RAW 264.7 cells into osteoclasts, such as multinucleated osteoclastlike cells with larger size, as reported [75], as shown in Fig.…”

Section: Nano-pfc Elicited No Effect On Osteoclastic Differentiation and Activitysupporting

confidence: 88%

Nanoscale perfluorocarbon expediates bone fracture healing through selectively activating osteoblastic differentiation and functions

et al. 2020

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Background and rationale: Fracture incidence increases with ageing and other contingencies. However, the strategy of accelerating fracture repair in clinical therapeutics remain a huge challenge due to its complexity and a longlasting period. The emergence of nano-based drug delivery systems provides a highly efficient, targeted and controllable drug release at the diseased site. Thus far, fairly limited studies have been carried out using nanomedicines for the bone repair applications. Perfluorocarbon (PFC), FDA-approved clinical drug, is received increasing attention in nanomedicine due to its favorable chemical and biologic inertness, great biocompatibility, high oxygen affinity and serum-resistant capability. In the premise, the purpose of the current study is to prepare nano-sized PFC materials and to evaluate their advisable effects on promoting bone fracture repair.Results: Our data unveiled that nano-PFC significantly enhanced the fracture repair in the rabbit model with radial fractures, as evidenced by increased soft callus formation, collagen synthesis and accumulation of beneficial cytokines (e.g., vascular endothelial growth factor (VEGF), matrix metalloprotein 9 (MMP-9) and osteocalcin). Mechanistic studies unraveled that nano-PFC functioned to target osteoblasts by stimulating their differentiation and activities in bone formation, leading to accelerated bone remodeling in the fractured zones. Otherwise, osteoclasts were not affected upon nano-PFC treatment, ruling out the potential target of nano-PFC on osteoclasts and their progenitors. Conclusions:These results suggest that nano-PFC provides a potential perspective for selectively targeting osteoblast cell and facilitating callus generation. This study opens up a new avenue for nano-PFC as a promising agent in therapeutics to shorten healing time in treating bone fracture.

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Section: Nano-pfc Elicited No Effect On Osteoclastic Differentiation and Activitysupporting

confidence: 88%

Nanoscale perfluorocarbon expediates bone fracture healing through selectively activating osteoblastic differentiation and functions

et al. 2020

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“…Differentiated cells were fixed with 10% formaldehyde for 10 min, ethanol-acetone (1:1) mixture for 1 min, and then washed with distilled water. Fixed cells were stained for 30 min with a TRAP solution containing Fast Red violet LB salt, naphthol-MX phosphate, and N , N -dimethylformamide in 50 mM acetate buffer as described before ( Matsubara et al, 2018a ; Matsubara et al, 2017 ; Matsubara et al, 2020 ). TRAP-positive multinuclear (> 3 nuclei) cells (TRAP (+) MNCs) were regarded as osteoclasts.…”

Section: Methodsmentioning

confidence: 99%

“…All reactions were performed in triplicate and analyzed. Expression levels were normalized by Gapdh expression ( Matsubara et al, 2020 ; Matsubara et al, 2018b ; Matsubara et al, 2018c ; Inoue et al, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Protein phosphatase 1 regulatory subunit 18 suppresses the transcriptional activity of NFATc1 via regulation of c-fos

et al. 2021

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The transcription factor NFATc1 and its binding partner AP-1 (a complex containing c-fos and c-Jun) play a central role in osteoclast differentiation. NFATc1 and AP-1 promote the expression of target genes such as Acp5, Ctsk and also auto-regulate NFATc1 expression as well. We previously reported that protein phosphatase 1 regulatory subunit 18 (PPP1r18) is a negative regulator of osteoclast bone resorption by inhibiting cell attachment to bone matrix. We also reported that PPP1r18 potentially regulates NFATc1 expression during osteoclast differentiation. To further explore this, in this study we have examined the effect of PPP1r18 on NFATc1 expression and activity by overexpressing PPP1r18 during the early stage of osteoclast differentiation. We found that PPP1r18 suppressed NFATc1 expression through inhibition of the transcriptional activity of NFATc1. Since PPP1r18 does not regulate NFATc1 directly, we next explored the involvement of AP-1. Our data showed that c-fos phosphorylation and nuclear localization were reduced by PPP1r18 overexpression. Further experiments showed that overexpression of c-fos together with PPP1r18 rescued NFATc1 expression and transcriptional activity. Moreover, c-fos activity inhibition by PPP1r18 was canceled by mutation of the phosphatase binding site of PPP1r18. Taken together, PPP1r18-regulated phosphatase activity targets c-fos phosphorylation and suppresses subsequent NFATc1 expression and activity.

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“…In addition, plectin, which connects vimentin to actin filaments and to microtubules (Leduc et al, 2015), is a podosomal protein (Gad et al, 2008). Plectin is required for actin organization and resorption activity in osteoclasts derived from RAW264.7 cells (Box 3), where it appears to control microtubule acetylation as well as Src and Pyk2 activities (Matsubara et al, 2017;Matsubara et al, 2020).…”

Section: Intermediate Filaments and Septinsmentioning

confidence: 99%

The osteoclast cytoskeleton – current understanding and therapeutic perspectives for osteoporosis

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Bompard

Guérit

et al. 2020

Journal of Cell Science

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Osteoclasts are giant multinucleated myeloid cells specialized for bone resorption, which is essential to preserve bone health throughout life. The activity of osteoclasts relies on the typical organization of osteoclast cytoskeleton components into a highly complex structure comprising actin, microtubules and other cytoskeletal proteins that constitutes the backbone of the bone resorption apparatus. The development of methods to obtain osteoclasts in culture and manipulate them genetically, as well as improvements in cell-imaging technologies, allowed shedding light onto the molecular mechanisms that control the structure and dynamics of the osteoclast cytoskeleton, and thus the mechanism of bone resorption. Although essential for normal bone physiology, abnormal osteoclast activity can cause bone defects, in particular their hyper-activation commonly associated with many pathologies, hormonal imbalance and medical treatments. Increased bone degradation by osteoclasts provokes progressive bone loss, leading to osteoporosis, with the resulting bone frailty resulting in fractures, loss of autonomy and premature death. In this context, the osteoclast cytoskeleton has recently proven to be a relevant therapeutic target for controlling pathological bone resorption levels. Here, we review our present knowledge about the regulatory mechanisms of the osteoclast cytoskeleton that control their bone resorption activity in normal and pathological conditions.

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Plectin stabilizes microtubules during osteoclastic bone resorption by acting as a scaffold for Src and Pyk2

Cited by 13 publications

References 32 publications

Nanoscale perfluorocarbon expediates bone fracture healing through selectively activating osteoblastic differentiation and functions

Nanoscale perfluorocarbon expediates bone fracture healing through selectively activating osteoblastic differentiation and functions

Protein phosphatase 1 regulatory subunit 18 suppresses the transcriptional activity of NFATc1 via regulation of c-fos

The osteoclast cytoskeleton – current understanding and therapeutic perspectives for osteoporosis

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