Anthropogenically increased atmospheric carbon dioxide (pCO2) leads to ocean acidification, disrupting calcification in marine calcifiers by reducing the saturation state of calcium carbonate. Calcium is not only a crucial component in the shell and skeleton structure but also serves as an essential second messenger for regulating biomineralization across many species. Ocean acidification is well-studied as causing shell dissolution in a diversity of bivalve species by disordering calcium deposition. However, it remains unclear whether the calcium-mediated signaling pathway regulating biomineralization is also affected. This study assessed eastern oyster (Crassostrea virginica) to determine how calcium signaling responds to elevated pCO₂ and influences shell formation. Under elevated pCO2, increased intracellular calcium concentration was found in primary epithelial cell cultures from oyster mantle. Meanwhile, we observed upregulation of calmodulin, a primary sensor of intracellular calcium, while its downstream effector, calcineurin, was downregulated. In addition, four conserved shell matrix proteins (SMPs), representing shell construction conditions, were significantly upregulated in the CO2-exposed mantle cells. In vivo, larval C. virginica exhibited developmental stage-dependent alterations in calcium signaling and SMPs disarrangement stimulated by pCO2. We hypothesize that dysregulation of calcium signaling disrupts the expressions of SMPs and causes oyster shell deformation. Pharmaceutical blockage of the calcium-calmodulin binding induced abnormal expression of related genes and shell matrix changes consistent with those caused by elevated pCO2, both in vivo and in vitro. Importantly, calcineurin restored SMPs expression in CO2-treated mantle cells. These findings suggest that shell deformities under ocean acidification are related to disruption of the calcium-calmodulin signaling pathway, inhibiting calcineurin activity and affecting SMPs production.
Xu W., Huang C., Matt J., Hollenbeck C. & Martin L., 2025. Ocean acidification disrupts the biomineralization process in the oyster Crassostrea virginica via intracellular calcium signaling dysregulation. Research Square. Article.



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