Abstract

Carbonic anhydrases (CA) play an important role in biomineralization from invertebrates to vertebrates. Previous experiments have investigated the role of CA in coral calcification, mainly by pharmacological approaches. This study reports the molecular cloning, sequencing, and immunolocalization of a CA isolated from the scleractinian coral Stylophora pistillata, named STPCA. Results show that STPCA is a secreted form of alpha-CA, which possesses a CA catalytic function, similar to the secreted human CAVI. We localized this enzyme at the calicoblastic ectoderm level, which is responsible for the precipitation of the skeleton. This localization supports the role of STPCA in the calcification process. In symbiotic scleractinian corals, calcification is stimulated by light, a phenomenon called "light-enhanced calcification" (LEC). The mechanism by which symbiont photosynthesis stimulates calcification is still enigmatic. We tested the hypothesis that coral genes are differentially expressed under light and dark conditions. By real-time PCR, we investigated the differential expression of STPCA to determine its role in the LEC phenomenon. Results show that the STPCA gene is expressed 2-fold more during the dark than the light. We suggest that in the dark, up-regulation of the STPCA gene represents a mechanism to cope with night acidosis.

Highlights

  • Carbonic anhydrases (CA)4 are ubiquitous metalloenzymes that catalyze the reversible hydration of carbon dioxide into bicarbonate: CO2 ϩ H2O % HCO3Ϫ ϩ Hϩ

  • Several studies have demonstrated the involvement of CA in coral calcification [7, 12,13,14,15,16,17,18]: CA could help in supplying dissolved inorganic carbon (DIC) or in removing carbonic acid from the site of skeletogenesis

  • A GenBankTM comparison using BLASTX confirmed that the partial sequence derived from S. pistillata corresponds to a carbonic anhydrase

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Summary

Carbonic Anhydrase in the Scleractinian Coral Stylophora pistillata

Results show that STPCA is a secreted form of ␣-CA, which possesses a CA catalytic function, similar to the secreted human CAVI We localized this enzyme at the calicoblastic ectoderm level, which is responsible for the precipitation of the skeleton. One specific feature of calcification in symbiotic corals is that calcium carbonate deposition in skeleton is higher in light than in dark conditions This phenomenon, called “light-enhanced calcification” (LEC), is due to the symbiosis that corals establish with their photosynthetic Dinoflagellates. We have cloned, sequenced, and localized an ␣-CA from the coral Stylophora pistillata and determined the differential gene expression between light and dark to generate more information on the function of CA in coral calcification and in the light-enhanced calcification phenomenon. Our results allow us to propose a model to explain the role of CA in the coral calcification process

EXPERIMENTAL PROCEDURES
RESULTS
Bootstrap values for ML and MR
Moderate Very high Very low
DISCUSSION
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