Involvement of Collagen Synthesis in Tissue Reconstitution by Dissociated Sponge Cells

Cells dissociated from the sponge Haliclona permollis reconstituted a new body by the following three consecutive processes: (1) Reaggregation, cells aggregate as a spheroidal mass. (2) Spreading, aggregates flatten and spread. (3) Reconstruction, aggregates coalesce into a reconstituted body. Cells in the process of reconstitution incorporated ¹⁴C-proline into proteins, converting 17% of it to hydroxyproline and synthesize collagen molecules during this process. Inhibitors of collagen biosynthesis did not affect the reaggregation, but caused incomplete morphogenesis in the processes of spreading and reconstruction: cycloheximide and 2,2'-dipyridyl inhibited spreading while 3-aminopropionitrile induced incomplete reconstruction. These findings suggested that spreading and reconstruction, but not reaggregation require both synthesis and cross-linking of collagens. Three polypeptides with molecular weights of 58 K, 160 K and 180 K were identified in sponge cells as collagens by immunoblot analysis with antibodies against sea urchin collagen and studies on susceptibilities to collagenase and pepsin. The 58 KDa polypeptide appeared in reconstituted bodies but not in dissociated cells, suggesting its importance in tissue reconstitution by dissociated cells.