Browsing by Author "Fonseca, Vera Alexandra Garcia"
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- Up-regulated genes during in vitro mineralization of Sparus aurata osteoblast-like cellsPublication . Fonseca, Vera Alexandra Garcia; Cancela, Leonor; Laizé, VicentVertebrate bone formation involves numerous and complex mechanisms and despite many studies, essentially performed in mammals, most of them remain largely unclear. Few in vitro studies, all in mammalian systems, have identified various genes differentially expressed during mineralization but no data is available yet from lower vertebrates. In the present study, it was used the recently developed Sparus aurata osteoblast-like cell line VSa16 to construct a cDNA subtractive library (control vs. mineralizing conditions) aimed at the identification of genes associated with fish in vitro mineralization. Suppression Subtractive Hybridization (SSH) combined with Mirror Otientation Selection (MOS) identified 194 cDNA clones representing 20 different up-regulated osteoblast-related genes. Among them, four were related to calcium mechanisms: osteopontin-like (SaOP-like), S100-like (SaS100-like), mucin-like (SaMUC) and transgelin2 (SaTGL2). The full length cDNA of SaOP-like (2138 bp) and SaS100-like (8648 bp) was obtained by RACE-PCR. SaOP-like and SaS100-like relative gene expression was analysed in 1) larvae and juvenile fish representing different developmental stages, 2) a broad selection of adult tissues, and 3) cell lines derived from bone and cartilage recently developed in our laboratory. Op-like Was shown to be expressed essentially in calcified tissues and to be important for late stages of mineralization. In contrary, SIOO-like was ubiquitously expressed and found to be important at early stage of larval development. The role of the proteins encoded by these two genes in the process of mineralization is likely to be related to their capacity of binding calcium and hydroxyapatite cxystals.