Alexander A. Venn

Corals form their reef-building aragonite (CaCO₃) skeletons via transient precursor phases yet understanding of the dynamics of these early-stage transformations remains incomplete. Using time-independent myriad mapping (MM) at 50 nm resolution, we map five mineral phases near the skeleton surface of Stylophora pistillata corals grown in varying seawater pH. All precursors, crystalline and amorphous, exhibit a consistent exponential decay from the growth front, with a shared decay length of 0.7 ± 0.1 μm, independent of time, phase, or pH. This spatial decay, paired with the constant growth rate of the skeleton, reveals a decay time of 5.1 ± 0.5 minutes. The dominant precursor is not amorphous but crystalline: calcium carbonate hemihydrate (CCHH, CaCO₃·½H₂O). These results suggest that exponential crystallization kinetics govern coral biomineralization and may be a widespread feature in biogenic, geologic, and synthetic systems—traceable long after initial mineral deposition.

Biomineralization is the process by which corals build their exoskeletons, and ultimately the reef ecosystem framework. The current model of biomineralization in these animals involves the endocytosis of calcifying medium by the calcifying cells lining the exoskeleton, followed by the intracellular formation of amorphous calcium carbonate precursors. However, the intracellular trafficking pathways inside these cells are poorly described. In this study, we investigated the intracellular trafficking of calcifying medium at the growing edge of laterally extending Stylophora pistillata colonies. Using a combination of fluorescent probes and immunofluorescence, along with confocal microscopy in living and fixed samples, we observed that a portion of the endocytosed calcifying medium is directed towards the Rab11-dependent endosomal recycling system, and we suggest that this pathway can be involved in the process of biomineral formation.

Twinning occurs when two crystals share a coherent interface along which their lattices are mirror symmetric. Here, twinning is investigated in biogenic and geologic aragonite (CaCO ₃ ) in columnar nacre from the red abalone Haliotis rufescens , sheet nacre from the black‐lip pearl oyster Pinctada margaritifera , and geologic aragonite. All samples exhibit the expected single (110) twins, characterized by a 116.2° rotation of the aragonite a‐ axis. Surprisingly, multiple twins—double, triple, quadruple—are also observed, each involving successive 116.2° rotations of the a‐ axis around the c ‐axis. Multiple twinning is concomitant but distinct from the well‐known cyclic twinning of aragonite. Multiple twinning is most prevalent in columnar nacre, reaching up to quadruple twins, most frequently occurring between nacre tablets in the same column, whereas sheet nacre and geologic aragonite exhibit up to triple twins. The frequency of twins decreases with increasing twin multiple, consistent with a physical rather than biological origin. This interpretation is supported by observations in geologic aragonite and by simulations of columnar nacre growth, which incorporate only nucleation rate, growth rate, and basic geometric constraints, yet they reproduce the observed twinning behavior in columnar nacre, reinforcing the conclusion that multiple twinning arises from physical growth mechanisms.

Summary: The coral‐dinoflagellate photosymbiosis and coral calcification underpin shallow water, coral reef ecosystems. This review examines the pivotal role of pH regulation in the cell physiology of these processes. Despite simple tissue organization, photosymbiotic corals maintain a complex internal microenvironment, with distinct compartments exhibiting contrasting pH levels. For example, the acidic ‘symbiosome’ surrounds the algal symbionts, while the alkaline ‘extracellular calcifying medium’ occurs at the growing front of the skeleton. We discuss how pH regulation of these compartments is crucial to the functioning of coral photosymbiosis and calcification, as well as mitigating the internal acid–base imbalances that these processes create. The role of pH regulation in the interplay between photosymbiosis and calcification is also discussed, focusing on the influence of symbiont photosynthesis on transepithelial gradients and the distribution of energy sources in the coral colony. Throughout this review, insights into pH regulation derived from previous research on ocean acidification are integrated to deepen understanding. Finally, we propose research priorities to advance knowledge of coral resilience under changing ocean conditions, such as investigating inorganic carbon concentration within coral compartments, species‐specific differences and the impacts of thermal stress on pH regulation.

Despite its prominent role as an intracellular messenger in all organisms, cytosolic free calcium ([Ca2+]i) has never been quantified in corals or cnidarians in general. Ratiometric calcium dyes and cell imaging have been key methods in successful research on [Ca2+]i in model systems, and could be applied to corals. Here, we developed a procedure to quantify [Ca2+]i in isolated cells from the model coral species Stylophora pistillata using Indo-1 and confocal microscopy. We quantified [Ca2+]i in coral cells with and without intracellular dinoflagellate symbionts, and verified our procedure on cultured mammalian cells. We then used our procedure to measure changes in [Ca2+]i in coral cells exposed to a classic inhibitor of [Ca2+]i regulation, thapsigargin, and also used it to record elevations in [Ca2+]i in coral cells undergoing apoptosis. Our procedure paves the way for future studies into intracellular calcium in corals and other cnidarians.

Coral reefs, the largest bioconstruction on Earth, are formed by calcium carbonate skeletons of corals. Coral skeleton formation commonly referred to as calcification occurs in a specific compartment, the extracellular calcifying medium (ECM), located between the aboral ectoderm and the skeleton. Calcification models often assume a direct link between the surrounding seawater and the ECM. However, the ECM is separated from the seawater by several tissue layers and the cœlenteron, which contains the cœlenteric fluid found in both polyps and cœnosarc (tissue connecting the polyps). Symbiotic dinoflagellate-containing cells line the cœlenteron and their photosynthetic activity contributes to changes in the chemistry of the cœlenteric fluid, particularly with respect to pH. The aim of our study is to compare cœlenteron pH between the cœnosarc and polyps and to compare areas of high or low dinoflagellate density based on tissue coloration. To achieve this, we use liquid ion exchange (LIX) pH microsensors to profile pH in the cœlenteron of polyps and the cœnosarc in different regions of the coral colony in light and darkness. We interpret our results in terms of what light and dark exposure means for proton gradients between the ECM and the coelenteron, and how this could affect calcification.

Calcium carbonate (CaCO₃) is abundant on Earth, is a major component of marine biominerals and thus of sedimentary and metamorphic rocks and it plays a major role in the global carbon cycle by storing atmospheric CO₂ into solid biominerals. Six crystalline polymorphs of CaCO₃ are known—3 anhydrous: calcite, aragonite, vaterite, and 3 hydrated: ikaite (CaCO₃·6H₂O), monohydrocalcite (CaCO₃·1H₂O, MHC), and calcium carbonate hemihydrate (CaCO₃·½H₂O, CCHH). CCHH was recently discovered and characterized, but exclusively as a synthetic material, not as a naturally occurring mineral. Here, analyzing 200 million spectra with Myriad Mapping (MM) of nanoscale mineral phases, we find CCHH and MHC, along with amorphous precursors, on freshly deposited coral skeleton and nacre surfaces, but not on sea urchin spines. Thus, biomineralization pathways are more complex and diverse than previously understood, opening new questions on isotopes and climate. Crystalline precursors are more accessible than amorphous ones to other spectroscopies and diffraction, in natural and bio-inspired materials.

The calcifying cells of corals are responsible for skeleton formation, a process that is the basis for reef building. Their cell biology is therefore of primary interest, but current knowledge is limited because direct in vivo investigation of this cell type is challenging. Studies at the growing edge of laterally extending coral colonies allow for direct observations of actively calcifying cells in living samples. In the current study, we used this approach to study lipid droplets, which are considered storage organelles that fuel cells for physiological processes. Using the fluorescent lipid probe Nile Red and in vivo confocal microscopy, we observed lipid droplets in calcifying cells of the coral Stylophora pistillata and we suggest that they play a key role in coral calcification.

In corals, pH regulation of the extracellular calcifying medium (ECM) by the calcifying cell layer is a crucial step in the calcification process and is potentially important to influencing how corals respond to ocean acidification. Here, we analyzed the growing edge of the reef coralStylophora pistillatato make the first characterization of the proton gradient across the coral calcifying epithelium. At seawater pH 8 we found that while the calcifying epithelium elevates pH in the ECM on its apical side above that of seawater, pH on its basal side in the mesoglea is markedly lower, highlighting that the calcifying cells are exposed to a microenvironment distinct from the external environment. Coral symbiont photosynthesis elevates pH in the mesoglea, but experimental ocean acidification and decreased seawater inorganic carbon concentration lead to large declines in mesoglea pH relative to the ECM, which is maintained relatively stable. Together, our results indicate that the coral calcifying epithelium is functionally polarized and that environmental variation impacts pH_ECMregulation through its effects on the basal side of the calcifying cells.