Biomedical subjects
María-J Uriz
Publications and source records attributed to María-J Uriz.
Siliceous spicules and skeleton frameworks in sponges: origin, diversity, ultrastructural patterns, and biological functions.
Silica deposition is a fundamental process in sponges. Most sponges in the Classes Demospongiae and Hexactinellida secrete siliceous elements, which can subsequently fuse, interlock with each other, or form three-dimensional structures connected by spongin. The resulting skeletal frameworks allow sponges to grow upwards and facilitate water exchange with minimal metabolic cost. Several studies on sponge skeletogenesis have been published. We are beginning to understand the mechanisms of spicule secretion and the role of spicules and skeletal frameworks in the biology, ecology, and evolution of sponges. Molecular techniques and ecological experiments have demonstrated the genetic control of the process and the contribution of environmental factors to the expression of a sponge spicule, respectively. However, other classic topics such as the role of membranes in silicon transport or whether spicules are formed in situ or secreted anywhere in the sponge mesohyl and then transported to the skeletal framework require further investigation. We review the process of silica deposition in sponges at the molecular and cellular levels, as well as the biological and ecological functions of spicules and skeletons. The genetic control of spicule shapes makes them useful in the reconstruction of sponge phylogeny, although recent experiments have demonstrated the influence of environmental factors in modulating spicule size, shape, and the presence or absence of one or more spicule types. The implications of such variations in sponge taxonomy may be important. Besides supporting sponge cells, spicules can help larvae stay buoyant while in the plankton or reach the bottom at settlement, enhance reproduction success, or catch prey. Conversely, the role of spicules and skeletons in deterring predation has not been demonstrated. Knowledge of several aspects is still based on a single or a few species and extrapolations should be made only with caution. With the advent of new molecular techniques, new lines of research are presently open and active in this field.
Long-term culture of sponge explants: conditions enhancing survival and growth, and assessment of bioactivity.
Sponges are an important source of secondary metabolites with pharmaceutical interest. This is the main reason for the increasing interest of sponge culture recent years. The optimal culture system depends on the species to be cultured: while some species easily produce sponge aggregates after dissociation (primmorphs), others show a great capacity to regenerate after fragmentation (explants). Corticium candelabrum is a Mediterranean bacteriosponge that can undergo asexual reproduction. We have taken advantage of this capability and cultured C. candelabrum explants under several experimental conditions. To find the best conditions for obtaining functional explants, we assayed a range of conditions, including seasons of collection, culture temperature, filtered versus filtered-sterile seawater, addition of antibiotics and proportion of ectosome. We monitored the changes in shape and ultrastructure during the formation of explants. After 24 h, TEM images showed the aquiferous system disarranged, in particular at the sponge periphery. From 2 to 4 weeks later, the aquiferous system regenerated, and fragments became functional sponges (explants). Explants were cultured under two regimes: in vitro and in a closed aquarium system. Antibiotics were only added to the in vitro culture to assess their effect on the symbiotic bacteria, which remained healthy despite the presence of antibiotics. Two food regimens (marine bacteria and green algae) were assayed for their ability to satisfy the metabolic requirements of explants. We monitored explant survival and growth. Explants showed a high long-term survival rate (close to 100%). Growth rates were higher in the closed aquarium system, without antibiotic addition, and fed with algae. Explants cultures were hardly contaminated because manipulation was reduced to a minimum and we used sterilized seawater. C. candelabrum produces bioactive molecules, which may play a defensive role in the sponge and may have pharmaceutical interest. The bioactivity of the explants was similar to that of wild sponges.
Quantitative assessment of natural toxicity in sponges: toxicity bioassay versus compound quantification.
Microtox assay was used to assess the natural toxicity of two sponges, Dysidea avara and Ircinia variabilis. The activity of crude extracts and major metabolites were compared. Methanol extract of D. avara was more toxic than that of acetone and was as toxic as pure avarol, thus suggesting that the toxicity of the sponge was mainly due to this metabolite. We also quantified palinurin, the major metabolite of I. variabilis, in specimens from several habitats. With the same methanol extracts used for palinurin quantification, we ran the Microtox assay and found a positive significant regression between toxicity and concentration of this metabolite. Pure palinurin was tested at the same concentration present in the extract, and the toxicity recorded was higher than that of the methanol extract. As with avarol from D. avara, palinurin is the main secondary metabolite that confers toxicity to I. variabilis. The results confirm that the standardized Microtox assay is an accurate and reproducible tool for assessing the toxicity of crude extracts and pure metabolites of marine organisms. These results also suggest that methanol is more suitable than acetone for the detection of species toxicity by Microtox. The method is faster and easier to perform than chemical quantification even when the sponge chemistry is known, and is appropriate for studies on variation in natural toxicity over a range of environmental conditions.