Acid polysaccharides in the skeletal matrix and calicoblastic epithelium of the stony coral Mycetophyllia reesi

Like many corals the skeletal organic matrix and associated epithelium of Mycetophyllia reesi is physico-chemically unstable to preparative procedures for electron microscopy. Ethanol cryofracture of mineralized and demineralized material is accompanied by delamination of tissue and skeleton. Filamentous algae occur in the interface and account for some but not all of the separation artifact. Transmission microscopy accompanied by decalcification requires embedment in glycerol jelly to preserve the skeletal organic matrix. Even then, the matrix is not fixed and is not retained within the gel using standard double fixation with or without tannic acid as an additive. Ruthenium red, in combination with osmium, prevents the matrix from physical disruption, although positional artifacts relative to the calicoblastic epithelium are still evident. Inclusion of other glycan precipitating agents in the fixative sequence (Alcian blue, iron diamine or the detergent cetylpyridinium chloride) are more useful in preserving an acid polysaccharide-rich, fibrillar, extracellular matrix after demineralization. This material is not observed in SEM preparations. The calicoblast cells appear to be the source of this extracellular material that also appears to contribute to the composition of the mineralizing matrix. Moreover, a hyaluronan-like substance appears to play a significant role in matrix structure as suggested by its degradation by hyaluronidase. (C) 2001 Harcourt Publishers Ltd.