Abstract
The calcium carbonate skeletons of corals provide the underlying structure of coral reefs; however, the cellular mechanisms responsible for coral calcification remain poorly understood. In osteoblasts from vertebrate animals, a Na+/Ca2+ exchanger (NCX) present in the plasma membrane transports Ca2+ to the site of bone formation. The aims of this study were to establish whether NCX exists in corals and its localization within coral cells, which are essential first steps to investigate its potential involvement in calcification. Data mining identified genes encoding for NCX proteins in multiple coral species, a subset of which were more closely related to NCXs from vertebrates (NCXA). We cloned NCXA from Acropora yongei (AyNCXA), which, unexpectedly, contained a peptide signal that targets proteins to vesicles from the secretory pathway. AyNCXA subcellular localization was confirmed by heterologous expression of fluorescently tagged AyNCXA protein in sea urchin embryos, which localized together with known markers of intracellular vesicles. Finally, immunolabeling of coral tissues with specific antibodies revealed AyNCXA was present throughout coral tissue. AyNCXA was especially abundant in calcifying cells, where it exhibited a subcellular localization pattern consistent with intracellular vesicles. Altogether, our results demonstrate AyNCXA is present in vesicles in coral calcifying cells, where potential functions include intracellular Ca2+ homeostasis and Ca2+ transport to the growing skeleton as part of an intracellular calcification mechanism.
Highlights
Coral reef ecosystems are valuable ecological [1] and economic resources [2] centered around the calcium carbonate (CaCO3) exoskeletons deposited by scleractinian corals
We cloned two full-length transcripts encoding for putative A. yongei Na+/Ca2+ exchanger (NCX) (AyNCXA and AyNCXB)
An alignment of AyNCXA and a mammalian NCX1 is provided in S1 Fig. In addition, AyNCXA contains a peptide signal with very high (0.995) probability to localize the protein to vesicles of the secretory pathway [25]
Summary
Coral reef ecosystems are valuable ecological [1] and economic resources [2] centered around the calcium carbonate (CaCO3) exoskeletons deposited by scleractinian corals. The aboral ectodermis ( known as the calicoblastic epithelium or calicodermis) is directly above the subcalicoblastic medium (SCM) and the skeleton, and is the tissue layer with the most direct role in calcification ([3]; reviewed in [4]). The cellular mechanisms for coral calcification are poorly understood (reviewed in [5]). Recent research indicates corals exert strong biological control on skeleton formation through intracellular calcification mechanisms. Calicoblastic cells express HCO3- transporting proteins that likely supply dissolved inorganic carbon [5,6,7], as well as coral acidic rich.
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