No abstract

Cidaroid sea urchins are the sister clade to all other extant echinoids and have numerous unique features, including unusual primary spines. These lack an epidermis when mature, exposing their high‐magnesium calcite skeleton to seawater and allowing the settlement of numerous epibionts. Cidaroid spines are made of an inner core of classical monocrystalline skeleton and an outer layer of polycrystalline magnesium calcite. Interestingly, cidaroids survived the Permian‐Triassic crisis, which was characterized by severe acidification of the ocean. Currently, numerous members of this group inhabit the deep ocean, below the saturation horizon for their magnesium calcite skeleton. This suggests that members of this taxon may have characteristics that may allow them to resist ongoing ocean acidification linked to global change. We compared the effect of acidified seawater (pH 7.2, 7.6, or 8.2) on mature spines with a fully developed cortex to that on young, growing spines, in which only the stereom core was developed. The cortex of mature spines was much more resistant to etching than the stereom of young spines. We then examined the properties of the cortex that might be responsible for its resistance compared to the underlying stereomic layers, namely morphology, intramineral organic material, magnesium concentration, intrinsic solubility of the mineral, and density. Our results indicate that the acidification resistance of the cortex is probably due to its lower magnesium concentration and higher density, the latter reducing the amount of surface area in contact with acidified seawater. The biofilm and epibionts covering the cortex of mature spines may also reduce its exposure to seawater.

Echinoderms are considered particularly sensitive to ocean acidification (OA) as their skeleton is made of high-magnesium calcite, one of the most soluble forms of calcium carbonate. Recent studies have investigated effects of OA on the skeleton of "classical" sea urchins (euechinoids), but the impact of etching on skeleton mechanical properties is almost unknown. Furthermore, the integrity of the skeleton of cidaroids has never been assessed, although their extracellular fluid is under-saturated with respect to their skeleton, and the skeleton of their primary spines is in direct contact with seawater. In this study, we compared the dissolution of test plates and spines as well as the spine mechanical properties (two-points bending tests) in a cidaroid (Eucidaris tribuloides) and a euechinoid (Tripneustes ventricosus) submitted to a 5 week acidification experiment (pH of 8.1, 7.7, and 7.4). Test plates of both species were not affected by dissolution. The spines of E. tribuloides showed no mechanical effects at pH 7.4 despite having traces of corrosion on secondary spines. On the contrary, spines of the T. ventricosus were significantly etched at both pH 7.7 and 7.4 and their fracture force reduced by 16 to 35%, respectively. This increased brittleness is probably of little significance with regards to predation protection but has consequences in terms of energy allocation.

BackgroundFluorochrome staining is among the most widely used techniques to study growth dynamics of echinoderms. However, it fails to detect fine-scale increments because produced marks are commonly diffusely distributed within the skeleton. In this paper we investigated the potential of trace element (manganese) labeling and subsequent cathodoluminescence (CL) imaging in fine-scale growth studies of echinoderms.ResultsThree species of sea urchins (Paracentrotus lividus, Echinometra sp. and Prionocidaris baculosa) were incubated for different periods of time in seawater enriched in different Mn2+ concentrations (1 mg/L; 3 mg/L; 61.6 mg/L). Labeling with low Mn2+ concentrations (at 1 mg/L and 3 mg/L) had no effect on behavior, growth and survival of sea urchins in contrast to the high Mn2+ dosage (at 61.6 mg/L) that resulted in lack of skeleton growth. Under CL, manganese produced clearly visible luminescent growth fronts in these specimens (observed in sectioned skeletal parts), which allowed for a determination of the average extension rates and provided direct insights into the morphogenesis of different types of ossicles. The three species tend to follow the same patterns of growth. Spine growth starts with the formation of microspines which are simultaneously becoming reinforced by addition of thickening layers. Spine septa develop via deposition of porous stereom that is rapidly (within less than 2 days) filled by secondary calcite. Development of the inner cortex in cidaroids begins with the formation of microspines which grow at ~3.5 μm/day. Later on, deposition of the outer polycrystalline cortex with spinules and protuberances proceeds at ~12 μm/day. The growth of tooth can be rapid (up to ~1.8 mm/day) and starts with the formation of primary plates (pp) in plumula. Later on, during the further growth of pp in aboral and lateral directions, secondary extensions develop inside (in chronological order: lamellae, needles, secondary plate, prisms and carinar processes), which are increasingly being solidified towards the incisal end. Interradial growth in the ambital interambulacral test plates exceeds meridional growth and inner thickening.ConclusionsMn2+ labeling coupled with CL imaging is a promising, low-cost and easily applicable method to study growth dynamics of echinoderms at the micro-length scale. The method allowed us to evaluate and refine models of echinoid skeleton morphogenesis.

Growth dynamics of the primary spine of the cidaroid sea urchin Phyllacanthus imperialis was assessed for the first time using pulsed (26) Mg-labeling and NanoSIMS isotopic imaging. The sea urchin was incubated twice (for 48 h) in artificial seawater with elevated level of (26) Mg. After each labeling event, the sea urchin was returned for 72 h to seawater with natural isotopic abundance of (26) Mg. NanoSIMS ion microprobe was subsequently used to visualize the labeled regions of the spine with submicrometer lateral resolution. The growth of the new skeleton was restricted to the distalmost and peripheral portions of the spine. Skeletogenesis involved mostly the deposition of continuous thickening layers and lateral growth involving bridges between previously formed trabeculae. The timescale of formation of individual thickening layers (ca. 1 µm in width) on the stereom trabeculae was on the order of 1 day. Longitudinal growth occurred mainly at the periphery in the form of small portions of the thickening deposits or more massive microspines that appeared to branch and fuse with those above and below. These microspines were found to grow at about 10 µm/day. These results reveal that the skeletal growth of a juvenile cidaroid spine is complex and highly heterogeneous, with different extension rates depending on the stage of the stereom development and/or direction of the growth fronts. The growth pattern observed here at the submicrometer scale provides direct evidence supporting the earlier suggestions that the lamellar structure of echinoderm stereom is formed by periodic deposition of continuous mineral layers.