Immunohistological analysis of transglutaminase factor XIIIA expression in mouse embryonic growth plate

Abstract

Previously we demonstrated the expression of Factor XIIIA (FXIIIA), a coagulatioli traasglutaminase, in avian embryonic growth plate. To explore whether FXIIIA is also expressed by chondrocytes of the iiiamliialian cartilage anlagen of bones, we analyzed tlie iiiouse embryonic growth plate by iiiiinuiiostainilig using anti-FXIIIA antibodies developed against human zind chicken proteins. We revealed the expression of FXIIIA in the epipliyseal growth plate, where FXIIIA appears first intracellularly in the zone of proliferation/iiiaturatioii, and remains intra-and extracellularly throughout the hypertrophic zone. Externalizatioii of FXIIIA O C C L I~S before mineralization. Transglutaminase activity was assayed in organ cultures using rliodamine-labeled synthetic substrate Pro-Val-Lys-Gly. Enzyiiiatic activity shows a restricted distribution in cartilage and correlates with FXIIIA expression pattern, suggesting that cartilagenous transglutaminase activity is due, at least partially, to the FXIIIA isoform. We conclude, that coagulation factor FXIIIA is expressed by chondrocytes of embryonic mouse long bone cartilages in a strictly regulated pattern.which correlates with chondrocyte differentiation and matrix mineralization.

Fetal Mesenchymal Stromal Cells Differentiating towards Chondrocytes Acquire a Gene Expression Profile Resembling Human Growth Plate Cartilage

“…Coagulation factor XIII ( F13A1 ) expression was up regulated, while other genes like complement component I ( C1R ), urokinase-type plasminogen activator ( PLAU ) and bradykinin receptor B2 ( BDKRB2 ) were downregulated during differentiation in our model. These proteins are associated with matrix mineralization [18]–[20] or matrix degradation in the growth plate [21]–[23], indicating early hypertrophic differentiation in the model, but not late hypertrophy. Proto-oncogene KIT (cluster F, hypertrophy related genes), transcription factor JUN and FOS are all associated with the regulation of cell proliferation [24], [25].…”

Comprehensive Profiling of Cartilage Extracellular Matrix Formation and Maturation Using Sequential Extraction and Label-free Quantitative Proteomics

“…Immunohistochemical staining was detected using ImmPact 3,3Ј-diaminobenzidine substrate (Vector Laboratories) until color was detected. Reactions were terminated by rinsing in tap water followed by dehydration of tissue sections using a graded ethanol series (30,50,70,90, and 100%). Sections were cleared in xylene and mounted using Pertex mounting medium.…”

FXIIIA and TGF-β over-expression produces normal musculo-skeletal phenotype in TG2-/- mice

“…The observation that TG2-/-mice have no overt skeletal phenotype, suggest that at least a second TG enzyme must influence matrix assembly and mineralisation. Previously, it has been described that, in addition to TG2, cultured osteoblasts also express the plasma transglutaminase Factor XIIIA (FXIIIA; Nurminskaya and Linsenmayer 2002;De Laurenzi and Melino 2001;Nurminskaya and Kaartinen 2006). To verify whether Factor XIIIA is involved, we analysed by western-blot its expression on protein extracts obtained from wild type and TG2-/-femurs (Fig.…”

“…These locations led to speculation that transglutaminases regulate endochondral ossification in particular both terminal cell differentiation and matrix calcification (Johnson 2007;Nurminskaya and Linsenmayer 2002;Johnson et al 2001) also FXIIIA-null mice show a dramatic decrease in COL I deposition in osteoblast cultures (Al-Jallad et al 2006). Indeed, many investigators confirmed this hypothesis in vitro, but to our knowledge no studies on the endochondral ossification of TG2-/-mice during are available.…”

“…Among the TGs family members, in the context of connective tissue formation, TG2 and Factor XIII are expressed in cartilage by hypertrophic chondrocytes (Aeschlimann et al 1996;Aeschlimann and Thomazy 2000;Nurminskaya and Linsenmayer 2002;De Laurenzi and Melino 2001;Nurminskaya et al 1998). In addition, several reports correlate in vitro TG2 expression, chondrocyte differentiation and matrix mineralization (Aeschlimann et al 1996;Aeschlimann and Thomazy 2000;Nurminskaya et al 1998;Nurminskaya and Linsenmayer 2002;Johnson and Terkeltaub 2005). Also, TGF-ß family is involved during endochondral ossification and bone remodelling (Janssens et al 2005).…”