Immunocytochemical assessment of cell differentiation of podoplanin-positive osteoblasts into osteocytes in murine bone

Nagai, Tomoya; Hasegawa, Tomoka; Yimin,; Yamamoto, Takanobu; Hongo, Hiromi; Abe, Miki; Yoshida, Taiji; Yokoyama, Akihisa; Freitas, Paulo Henrique Luiz de; Li, Minqi; Yokoyama, Atsuro; Amizuka, Norio

doi:10.1007/s00418-020-01937-y

Cited by 8 publications

(4 citation statements)

References 45 publications

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“…The lymph node expansion causes by the reduction of podoplanin-mediated contraction in inflamed lymph nodes where mature dendritic cells interact with fibroblastic reticular cells via podoplanin-CLEC-2 binding [21][22][23][24] . There is a detailed report on the expression of podoplanin in bone cells 25) . It shows that the CD44-podoplanin binding contributes to the osteoblast differentiation in the remodeling sites but not in modeling sites of bone.…”

Section: Introductionmentioning

confidence: 99%

Morphological Study for the Osteocytes in Podoplanin-Conditional Knockout Mice

Osawa,

Kanai,

Ushijima

et al. 2023

J. Hard Tissue Biology.

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We generated podoplanin-conditional knockout mice where the floxed podoplanin exon3 was deleted by the Dmp-1-driven Cre (Dmp1-Cre;Pdpn Δ/Δ ) and investigated the cell process elongation of podoplanin-deficient mouse osteocyte in vitro and in vivo. The expression of podoplanin is found in odontoblasts while not observed in odontoblasts of Dmp1-Cre; Pdpn Δ/Δ mice, indicating that the conditional knockout of podoplanin in Dmp1-expressing cells in Dmp1-Cre;Pdpn Δ/Δ mice is successful. There were no differences in the growth of wild-type and Dmp1-Cre;Pdpn Δ/Δ mice, and no differences in calcification and alkaline phosphatase activity in cultured calvarial osteoblasts of the wild-type and Dmp1-Cre;Pdpn Δ/Δ mice, in total this suggests that the podoplanin-cKO has no effect on generation of the bone. The cell process elongation was suppressed in cultured calvarial osteoblasts of Dmp1-Cre;Pdpn Δ/Δ mice compared with wild-type mice. In the electron microscopic study, there were no morphological differences in bone matrix formation and osteocyte distribution in Dmp1-Cre; Pdpn Δ/Δ and wild-type mice, whereas the cell process formation was sparser and the network with neighboring cells was more deficient in Dmp1-Cre;Pdpn Δ/Δ mice than in wild-type mice. In the quantitative analysis, the number and thickness of the cell processes were significantly smaller and thinner in Dmp1-Cre;Pdpn Δ/Δ mice than in wild-type mice. This could suggest that podoplanin plays a role in the formation of the osteocyte network created by the cell process elongation.

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Section: Introductionmentioning

confidence: 99%

Morphological Study for the Osteocytes in Podoplanin-Conditional Knockout Mice

Osawa,

Kanai,

Ushijima

et al. 2023

J. Hard Tissue Biology.

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“…Osteoblasts secrete bone matrix proteins and can become embedded in the bone matrix, where they differentiate into osteocytes. Immediately before becoming embedded into the bone matrix, osteoblasts rearrange the actin filament assembly along the cell membranes and the cytoplasmic processes, which resemble that of embedded osteocytes [ 24 ]. This implies that the osteoblasts approaching osteocytic differentiation and the newly-differentiated osteocytes decide the geometrical structure of the cellular network of their cytoplasmic processes.…”

Section: Osteocyte Network and The Biological Function Of Regulating ...mentioning

confidence: 99%

“…After primary mineralization, the bone mineral density becomes slowly and chronologically elevated in a phenomenon called secondary mineralization, which is independent of osteoblastic bone formation [ 2 ]. It is hypothesized that secondary mineralization is achieved by the physicochemical processes of mineral transport in the osteocytic network extended throughout the bone [ 24 ]. Therefore, the osteocytes and the meshwork of their cytoplasmic processes appear essential for secondary mineralization involving bone mineral transportation [ 25 , 26 , 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

Matrix Vesicle-Mediated Mineralization and Osteocytic Regulation of Bone Mineralization

Hasegawa

Hongo

Yamamoto

et al. 2022

IJMS

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Bone mineralization entails two mineralization phases: primary and secondary mineralization. Primary mineralization is achieved when matrix vesicles are secreted by osteoblasts, and thereafter, bone mineral density gradually increases during secondary mineralization. Nearby extracellular phosphate ions (PO43−) flow into the vesicles via membrane transporters and enzymes located on the vesicles’ membranes, while calcium ions (Ca2+), abundant in the tissue fluid, are also transported into the vesicles. The accumulation of Ca2+ and PO43− in the matrix vesicles induces crystal nucleation and growth. The calcium phosphate crystals grow radially within the vesicle, penetrate the vesicle’s membrane, and continue to grow outside the vesicle, ultimately forming mineralized nodules. The mineralized nodules then attach to collagen fibrils, mineralizing them from the contact sites (i.e., collagen mineralization). Afterward, the bone mineral density gradually increases during the secondary mineralization process. The mechanisms of this phenomenon remain unclear, but osteocytes may play a key role; it is assumed that osteocytes enable the transport of Ca2+ and PO43− through the canaliculi of the osteocyte network, as well as regulate the mineralization of the surrounding bone matrix via the Phex/SIBLINGs axis. Thus, bone mineralization is biologically regulated by osteoblasts and osteocytes.

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“…Moreover, osteoblasts interacting through the OLCS can undergo further differentiation into osteocytes, comprising approximately 90-95% of all bone cells (Florencio-Silva et al 2015). This has prompted Nagai et al (2021) to perform a detailed multi-immunohistochemical analysis, combined with pre-embedding peroxidase immunoelectron microscopy to investigate the process of late-stage osteoblast differentiation into osteocytes. They chose to combine immunostaining for podoplanin, a transmembrane glycoprotein expressed in late-stage osteoblasts and pre-osteocytes, together with some of its purported binding partners.…”

Section: A Bone To Pick…mentioning

confidence: 99%