An in situ hybridization study of decorin and biglycan mRNA in mouse osteoblasts in vivo
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ORIGINAL ARTICLE
An in situ hybridization study of decorin and biglycan mRNA in mouse osteoblasts in vivo Angammana Randilini1 · Kaoru Fujikawa2 · Shunichi Shibata1 Received: 26 August 2020 / Accepted: 4 November 2020 © Japanese Association of Anatomists 2020
Abstract In situ hybridization of decorin and biglycan mRNA, principal members of small leucine-rich proteoglycan, was performed using [35S]-labeled RNA probes, in the context of the hypothesis that they show different expression patterns associated with osteoblast differentiation in mice. We adopted two ossifying sites that can clearly follow the developmental process of bone formation: ossifying tympanic ring and developing bone collar of mandibular condylar cartilage. Decorin mRNA was expressed in osteoblasts of developing tympanic ring at E14.0, as well as of developing bone collar at E15.0, but biglycan mRNA was not, indicating decorin mRNA was expressed earlier in newly differentiating osteoblasts than biglycan. With maturation of osteoblasts, biglycan mRNA became expressed and maintained its expression both in the outer region (periosteum) and in the interior region (endosteum) of bone. By contrast, decorin mRNA expression was maintained in the outer region but diminished in the interior region. These results indicate that decorin and biglycan show differential expression patterns in differentiating osteoblasts and play specific roles in bone formation. Keywords Biglycan · Bone formation · Decorin · In situ hybridization · Osteoblasts
Introduction Small leucine-rich proteoglycans (SLRPs) have been identified as important components in the extracellular matrix and are involved in several biological and pathological processes in various tissues. They are proteoglycans with small core proteins of tandem leucine-rich repeats that are usually attached to one or more glycosaminoglycan chains, such as chondroitin sulfate (CS), dermatan sulfate (DS), or keratan sulfate. Eighteen SLRP genes have been identified to date, which are classified into five classes based on their characteristics at both genomic and protein levels (Schaefer and Iozzo 2008; Schaefer and Schaefer 2010; Nikitovic et al. 2012; Kram et al. 2020). Class I SLRPs are most commonly identified and include decorin and biglycan, which are the * Shunichi Shibata [email protected] 1
Department of Maxillofacial Anatomy, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1‑5‑45, Bunkyo‑ku, Yushima, Tokyo 113‑8549, Japan
Department of Oral Anatomy and Developmental Biology, Showa University School of Dentistry, Tokyo, Japan
2
most studied and relatively abundant. These SLRPs are found in various tissues, such as skin, tendon, bone, cartilage, muscle, and teeth (Neame and Kay 2000). They contain CS/DS chains: one in decorin and two in biglycan. Decorin plays a significant role in collagen fibrillogenesis and prevention of premature mineralization (Mochida et al. 2009), whereas biglycan is more significant in osteoblast differentiation and matrix mine
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