Common Genetic Denominators for Ca -Based Skeleton in Metazoa: Role of Osteoclast-Stimulating Factor and of Carbonic Anhydrase in a Calcareous SpongeReportar como inadecuado




Common Genetic Denominators for Ca -Based Skeleton in Metazoa: Role of Osteoclast-Stimulating Factor and of Carbonic Anhydrase in a Calcareous Sponge - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Calcium-based matrices serve predominantly as inorganic, hard skeletal systems in Metazoa from calcareous sponges phylum Porifera; class Calcarea to proto- and deuterostomian multicellular animals. The calcareous sponges form their skeletal elements, the spicules, from amorphous calcium carbonate ACC. Treatment of spicules from Sycon raphanus with sodium hypochlorite NaOCl results in the disintegration of the ACC in those skeletal elements. Until now a distinct protein-enzyme involved in ACC metabolism could not been identified in those animals. We applied the technique of phage display combinatorial libraries to identify oligopeptides that bind to NaOCl-treated spicules: those oligopeptides allowed us to detect proteins that bind to those spicules. Two molecules have been identified, the putative enzyme carbonic anhydrase and the putative osteoclast-stimulating factor OSTF, that are involved in the catabolism of ACC. The complete cDNAs were isolated and the recombinant proteins were prepared to raise antibodies. In turn, immunofluorescence staining of tissue slices and qPCR analyses have been performed. The data show that sponges, cultivated under standard condition 10 mM CaCl2 show low levels of transcripts-proteins for carbonic anhydrase or OSTF, compared to those animals that had been cultivated under Ca2+-depletion condition 1 mM CaCl2. Our data identify with the carbonic anhydrase and the OSTF the first two molecules which remain conserved in cells, potentially involved in Ca-based skeletal dissolution, from sponges sclerocytes to human osteoclast.



Autor: Werner E. G. Müller , Xiaohong Wang, Vlad A. Grebenjuk, Michael Korzhev, Matthias Wiens, Ute Schloßmacher, Heinz C. Schröder

Fuente: http://plos.srce.hr/



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