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      Expression of calcification‐related ion transporters during blue mussel larval development

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          Abstract

          The physiological processes driving the rapid rates of calcification in larval bivalves are poorly understood. Here, we use a calcification substrate‐limited approach (low dissolved inorganic carbon, C T) and mRNA sequencing to identify proteins involved in bicarbonate acquisition during shell formation. As a secondary approach, we examined expression of ion transport and shell matrix proteins (SMPs) over the course of larval development and shell formation. We reared four families of Mytilus edulis under ambient (ca. 1865 µmol/kg) and low C T (ca. 941 µmol/kg) conditions and compared expression patterns at six developmental time points. Larvae reared under low C T exhibited a developmental delay, and a small subset of contigs was differentially regulated between ambient and low C T conditions. Of particular note was the identification of one contig encoding an anion transporter (SLC26) which was strongly upregulated (2.3–2.9 fold) under low C T conditions. By analyzing gene expression profiles over the course of larval development, we are able to isolate sequences encoding ion transport and SMPs to enhance our understanding of cellular pathways underlying larval calcification processes. In particular, we observe the differential expression of contigs encoding SLC4 family members (sodium bicarbonate cotransporters, anion exchangers), calcium‐transporting ATPases, sodium/calcium exchangers, and SMPs such as nacrein, tyrosinase, and transcripts related to chitin production. With a range of candidate genes, this work identifies ion transport pathways in bivalve larvae and by applying comparative genomics to investigate temporal expression patterns, provides a foundation for further studies to functionally characterize the proteins involved in larval calcification.

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          Most cited references73

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          An acidic matrix protein, Pif, is a key macromolecule for nacre formation.

          The mollusk shell is a hard tissue consisting of calcium carbonate crystals and an organic matrix. The nacre of the shell is characterized by a stacked compartment structure with a uniformly oriented c axis of aragonite crystals in each compartment. Using a calcium carbonate-binding assay, we identified an acidic matrix protein, Pif, in the pearl oyster Pinctada fucata that specifically binds to aragonite crystals. The Pif complementary DNA (cDNA) encoded a precursor protein, which was posttranslationally cleaved to produce Pif 97 and Pif 80. The results from immunolocalization, a knockdown experiment that used RNA interference, and in vitro calcium carbonate crystallization studies strongly indicate that Pif regulates nacre formation.
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            Calcium Homeostasis and Organelle Function in the Pathogenesis of Obesity and Diabetes.

            A number of chronic metabolic pathologies, including obesity, diabetes, cardiovascular disease, asthma, and cancer, cluster together to present the greatest threat to human health. As research in this field has advanced, it has become clear that unresolved metabolic inflammation, organelle dysfunction, and other cellular and metabolic stresses underlie the development of these chronic metabolic diseases. However, the relationship between these systems and pathological mechanisms is poorly understood. Here we discuss the role of cellular Ca(2+) homeostasis as a critical mechanism integrating the myriad of cellular and subcellular dysfunctional networks found in metabolic tissues such as liver and adipose tissue in the context of metabolic disease, particularly in obesity and diabetes.
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              Molluscan shell proteins: primary structure, origin, and evolution.

              In the last few years, the field of molluscan biomineralization has known a tremendous mutation, regarding fundamental concepts on biomineralization regulation as well as regarding the methods of investigation. The most recent advances deal more particularly with the structure of shell biominerals at nanoscale and the identification of an increasing number of shell matrix protein components. Although the matrix is quantitatively a minor constituent in the shell of mollusks (less than 5% w/w), it is, however, the major component that controls different aspects of the shell formation processes: synthesis of transient amorphous minerals and evolution to crystalline phases, choice of the calcium carbonate polymorph (calcite vs aragonite), organization of crystallites in complex shell textures (microstructures). Until recently, the classical paradigm in molluscan shell biomineralization was to consider that the control of shell synthesis was performed primarily by two antagonistic mechanisms: crystal nucleation and growth inhibition. New concepts and emerging models try now to translate a more complex reality, which is remarkably illustrated by the wide variety of shell proteins, characterized since the mid-1990s, and described in this chapter. These proteins cover a broad spectrum of pI, from very acidic to very basic. The primary structure of a number of them is composed of different modules, suggesting that these proteins are multifunctional. Some of them exhibit enzymatic activities. Others may be involved in cell signaling. The oldness of shell proteins is discussed, in relation with the Cambrian appearance of the mollusks as a mineralizing phylum and with the Phanerozoic evolution of this group. Nowadays, the extracellular calcifying shell matrix appears as a whole integrated system, which regulates protein-mineral and protein-protein interactions as well as feedback interactions between the biominerals and the calcifying epithelium that synthesized them. Consequently, the molluscan shell matrix may be a source of bioactive molecules that would offer interesting perspectives in biomaterials and biomedical fields.
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                Author and article information

                Contributors
                kirti.ramesh@bioenv.gu.se
                Journal
                Ecol Evol
                Ecol Evol
                10.1002/(ISSN)2045-7758
                ECE3
                Ecology and Evolution
                John Wiley and Sons Inc. (Hoboken )
                2045-7758
                29 May 2019
                June 2019
                : 9
                : 12 ( doiID: 10.1002/ece3.2019.9.issue-12 )
                : 7157-7172
                Affiliations
                [ 1 ] GEOMAR Helmholtz Centre for Ocean Research Kiel Germany
                [ 2 ] Department of Biological and Environmental Sciences, Sven Lovén Centre for Marine Infrastructure‐Kristineberg University of Gothenburg Fiskebäckskil Sweden
                [ 3 ] British Antarctic Survey Natural Environment Research Council Cambridge UK
                [ 4 ] Ashworth Laboratories, Institute of Evolutionary Biology University of Edinburgh Edinburgh UK
                [ 5 ] Ecological Chemistry Alfred‐Wegener‐Institut Helmholtz‐Zentrum für Polar‐und Meeresforschung Bremerhaven Germany
                [ 6 ] Helmholtz‐Institute for Functional Marine Biodiversity Oldenburg Germany
                Author notes
                [*] [* ] Correspondence

                Kirti Ramesh, GEOMAR Helmholtz Centre for Ocean Research, Kiel 24105, Germany.

                Email: kirti.ramesh@ 123456bioenv.gu.se

                Author information
                https://orcid.org/0000-0001-7889-6722
                https://orcid.org/0000-0002-3442-3824
                Article
                ECE35287
                10.1002/ece3.5287
                6662379
                31380040
                74ac3771-b4e1-4520-bec7-66af627d6dc1
                © 2019 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 30 October 2018
                : 06 May 2019
                : 08 May 2019
                Page count
                Figures: 3, Tables: 4, Pages: 16, Words: 12250
                Funding
                Funded by: European Union's Seventh Framework Programme
                Funded by: NERC Core Funding
                Categories
                Original Research
                Original Research
                Custom metadata
                2.0
                ece35287
                June 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.6.2 mode:remove_FC converted:29.07.2019

                Evolutionary Biology
                biomineralization,bivalves,gene expression,larvae
                Evolutionary Biology
                biomineralization, bivalves, gene expression, larvae

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