{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T04:47:33Z","timestamp":1790484453203,"version":"4.1.0"},"reference-count":38,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2003,2,1]],"date-time":"2003-02-01T00:00:00Z","timestamp":1044057600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2003,2,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Matrix Gla protein (MGP) belongs to the family of vitamin K-dependent, Gla-containing proteins, and in mammals, birds, and Xenopus, its mRNA was previously detected in extracts of bone, cartilage, and soft tissues (mainly heart and kidney), whereas the protein was found to accumulate mainly in bone. However, at that time, it was not evaluated if this accumulation originated from protein synthesized in cartilage or in bone cells because both coexist in skeletal structures of higher vertebrates and Xenopus. Later reports showed that MGP also accumulated in costal calcified cartilage as well as at sites of heart valves and arterial calcification. Interestingly, MGP was also found to accumulate in vertebra of shark, a cartilaginous fish. However, to date, no information is available on sites of MGP expression or accumulation in teleost fishes, the ancestors of terrestrial vertebrates, who have in their skeleton mineralized structures with both bone and calcified cartilage. To analyze MGP structure and function in bony fish, MGP was acid-extracted from the mineralized matrix of either bone tissue (vertebra) or calcified cartilage (branchial arches) from the bony fish, Argyrosomus regius,1The A. regius MGP and BGP cDNA sequences were submitted to the GenBank database with accession numbers AF334473 and AF459030, respectively. separated from the mineral phase by dialysis, and purified by Sephacryl S-100 chromatography. No MGP was recovered from bone tissue, whereas a protein peak corresponding to the MGP position in this type of gel filtration was obtained from an extract of branchial arches, rich in calcified cartilage. MGP was identified by N-terminal amino acid sequence analysis, and the resulting protein sequence was used to design specific oligonucleotides suitable to amplify the corresponding DNA by a mixture of reverse transcription-polymerase chain reaction (RT-PCR) and 5\u2032rapid amplification of cDNA (RACE)-PCR. In parallel, ArBGP (bone Gla protein, osteocalcin) was also identified in the same fish, and its complementary DNA cloned by an identical procedure. Tissue distribution\/accumulation was analyzed by Northern blot, in situ hybridization, and immunohistochemistry. In mineralized tissues, the MGP gene was predominantly expressed in cartilage from branchial arches, with no expression detected in the different types of bone analyzed, whereas BGP mRNA was located in bone tissue as expected. Accordingly, the MGP protein was found to accumulate, by immunohistochemical analysis, mainly in the extracellular matrix of calcified cartilage. In soft tissues, MGP mRNA was mainly expressed in heart but in situ hybridization, indicated that cells expressing the MGP gene were located in the bulbus arteriosus and aortic wall, rich in smooth muscle and endothelial cells, whereas no expression was detected in the striated muscle myocardial fibers of the ventricle. These results show that in marine teleost fish, as in mammals, the MGP gene is expressed in cartilage, heart, and kidney tissues, but in contrast with results obtained in Xenopus and higher vertebrates, the protein does not accumulate in vertebra of non-osteocytic teleost fish, but only in calcified cartilage. In addition, our results also indicate that the presence of MGP mRNA in heart tissue is due, at least in fish, to the expression of the MGP gene in only two specific cell types, smooth muscle and endothelial cells, whereas no expression was found in the striated muscle fibers of the ventricle. In light of these results and recent information on expression of MGP gene in these same cell types in mammalian aorta, it is likely that the levels of MGP mRNA previously detected in Xenopus, birds, and mammalian heart tissue may be restricted toregions rich in smooth muscle and endothelial cells. Our results also emphasize the need to re-evaluate which cell types are involved in MGP gene expression in other soft tissues and bring further evidence that fish are a valuable model system to study MGP gene expression and regulation.<\/jats:p>","DOI":"10.1359\/jbmr.2003.18.2.244","type":"journal-article","created":{"date-parts":[[2006,4,27]],"date-time":"2006-04-27T05:39:46Z","timestamp":1146116386000},"page":"244-259","source":"Crossref","is-referenced-by-count":41,"title":["Purification of Matrix Gla Protein From a Marine Teleost Fish, <i>Argyrosomus regius<\/i>: Calcified Cartilage and Not Bone as the Primary Site of MGP Accumulation in Fish"],"prefix":"10.1093","volume":"18","author":[{"given":"DC","family":"Simes","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, CCMar University of Algarve, Faro, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"MK","family":"Williamson","sequence":"additional","affiliation":[{"name":"Division of Biology, University of California San Diego, La Jolla, California, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"JB","family":"Ortiz-Delgado","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, CCMar University of Algarve, Faro, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Csb","family":"Viegas","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, CCMar University of Algarve, Faro, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"PA","family":"Price","sequence":"additional","affiliation":[{"name":"Division of Biology, University of California San Diego, La Jolla, California, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"ML","family":"Cancela","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, CCMar University of Algarve, Faro, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2009,12,2]]},"reference":[{"key":"2024013019163423000_bib1","doi-asserted-by":"crossref","first-page":"14971","DOI":"10.1016\/S0021-9258(18)95688-7","article-title":"Primary structure of bovine matrix Gla protein, a new vitamin K-dependent bone protein","volume":"260","author":"Price","year":"1985","journal-title":"J Biol Chem"},{"key":"2024013019163423000_bib2","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1002\/pro.5560030511","article-title":"Conserved phosphorylation of serine in the Ser-X-glu\/Ser(P) sequences of the vitamin K-dependent matrix Gla protein from shark, lamb, rat, cow and human","volume":"3","author":"Price","year":"1994","journal-title":"Protein Sci"},{"key":"2024013019163423000_bib3","doi-asserted-by":"crossref","first-page":"1611","DOI":"10.1359\/jbmr.2001.16.9.1611","article-title":"Matrix Gla protein in Xenopus laevis: molecular cloning, tissue distribution and evolutionary considerations","volume":"16","author":"Cancela","year":"2001","journal-title":"J Bone Miner Res"},{"key":"2024013019163423000_bib4","doi-asserted-by":"crossref","first-page":"5820","DOI":"10.1016\/S0021-9258(18)60639-8","article-title":"The identification of matrix Gla protein in cartilage","volume":"263","author":"Hale","year":"1988","journal-title":"J Biol Chem"},{"key":"2024013019163423000_bib5","doi-asserted-by":"crossref","first-page":"1400","DOI":"10.1161\/01.ATV.18.9.1400","article-title":"Warfarin causes rapid calcification of the elastic lamellae in rat arteries and heart valves","volume":"18","author":"Price","year":"1998","journal-title":"Arterioscler Thromb Vasc Biol"},{"key":"2024013019163423000_bib6","doi-asserted-by":"crossref","first-page":"1764","DOI":"10.1055\/s-0037-1614911","article-title":"Matrix Gla protein synthesis and gamma-carboxylation in the aortic vessel wall and proliferating vascular smooth muscle cells","volume":"82","author":"Wallin","year":"1999","journal-title":"Thromb Haemost"},{"key":"2024013019163423000_bib7","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1161\/01.ATV.20.2.317","article-title":"Warfarin-induced artery calcification is accelerated by growth and vitamin D","volume":"20","author":"Price","year":"2000","journal-title":"Arterioscler Thromb Vasc Biol"},{"issue":"Suppl 2","key":"2024013019163423000_bib8","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1007\/s003920070101","article-title":"Expression of mineralization-regulating proteins in association with human vascular calcification","volume":"89","author":"Shanahan","year":"2000","journal-title":"Z. 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