NanoSIMS: insights into the organization of the proteinaceous scaffold within Hexactinellid sponge spicules

Abstract

The giant basal spicules (GBS) from Monorhaphis chuni (Porifera [sponges], Hexactinellida) represent the largest biosilica structures on Earth and can reach lengths of 300 cm (diameter of 1.1 cm). The amorphous silica of the inorganic matrix is formed enzymatically by silicatein. During this process, the enzyme remains trapped inside the lamellar-organized spicules. In order to localize the organic silicatein scaffold, the inside of a lamella has been analyzed by nano-secondary ion mass spectrometry (NanoSIMS). It is shown that the GBSs are composed of around 245 concentrically arranged individual siliceous lamellae. These surround an internal siliceous axial cylinder. The lamellae adjacent to the cylinder are thicker (10-30 mum) than the more peripheral lamellae (2-10 microm). One lamella of a thickness of 18 mum has been selected for further analysis. This lamella itself is composed of three sublamellae with an individual thickness of 2-6 microm each, which are then further organized into three cylindrical slats (thickness: 1.6-1.8 microm). Other than the main lamellae, the sublamellae are not separated from each other by gaps. The element analysis of the sublamellae by NanoSIMS revealed that the siliceous matrix is embedded in an organic matrix that consists of up to 6-10 wt/% of C. The pattern of C distribution reflects a distinct zonation of the organic material within the solid intralamellar biosiliceous material. A growth model for the lamella starting from nanosized silica particles is proposed: During formation of a lamella nanosized silica particles fuse, through biosintering processes, to slats that build the individual sublamellae, which then finally form the lamellae. In turn, those lamellae may form the higher structural entity, the axial cylinder.