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  • 1
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    A sea urchin Na+K+2Cl- cotransporter is involved in the maintenance of calcification-relevant cytoplasmic cords in Strongylocentrotus droebachiensis larvae (2016)
    PANGAEA
    In:  Supplement to: Basse, Wiebke C; Gutowska, Magdalena A; Findeisen, Ulrike; Stumpp, Meike; Dupont, Sam; Jackson, Daniel J; Himmerkus, Nina; Melzner, Frank; Bleich, Markus (2015): A sea urchin Na+K+2Cl- cotransporter is involved in the maintenance of calcification-relevant cytoplasmic cords in Strongylocentrotus droebachiensis larvae. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 187, 184-192, https://doi.org/10.1016/j.cbpa.2015.05.005
    Details
    Publication Date: 2024-02-10
    Description: The cellular mechanisms of calcification in sea urchin larvae are still not well understood. Primary mesenchyme cells within the larval body cavity form a syncytium to secrete CaCO3 spicules from intracellular amorphous CaCO3 (ACC) stores. We studied the role of Na+K+2Cl- cotransporter (NKCC) in intracellular ACC accumulation and larval spicule formation of Strongylocentrotus droebachiensis. First, we incubated growing larvae with three different loop diuretics (azosemide, bumetanide, and furosemide) and established concentration-response curves. All loop diuretics were able to inhibit calcification already at concentrations that specifically inhibit NKCC. Calcification was most effectively inhibited by azosemide (IC50 = 6.5 µM), while larval mortality and swimming ability were not negatively impacted by the treatment. The inhibition by bumetanide (IC50 = 26.4 µM) and furosemide (IC50 = 315.4 µM) resembled the pharmacological fingerprint of the mammalian NKCC1 isoform. We further examined the effect of azosemide on the maintenance of cytoplasmic cords and on the occurrence of calcification vesicles using fluorescent dyes (calcein, FM1-43). Fifty micromolars of azosemide inhibited the maintenance of cytoplasmic cords and resulted in increased calcein fluorescence within calcification vesicles. The expression of NKCC in S. droebachiensis was verified by PCR and Western blot with a specific NKCC antibody. In summary, the pharmacological profile of loop diuretics and their specific effects on calcification in sea urchin larvae suggest that they act by inhibition of NKCC via repression of cytoplasmic cord formation and maintenance.
    Keywords: BIOACID; Biological Impacts of Ocean Acidification
    Type: Dataset
    Format: application/zip, 246 kBytes
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    DATA PANGAEA
  • 2
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    A shell regeneration assay to identify biomineralization candidate genes in mytilid mussels (2016)
    ELSEVIER SCIENCE BV
    In:  EPIC3Marine Genomics, ELSEVIER SCIENCE BV, 27, pp. 57-67, ISSN: 1874-7787
    Details
    Publication Date: 2017-01-06
    Description: Biomineralization processes in bivalve molluscs are still poorly understood. Here we provide an analysis of specifically expressed sequences from a mantle transcriptome of the blue mussel, Mytilus edulis. We then developed a novel, integrative shell injury assay to test, whether biomineralization candidate genes highly expressed in marginal and pallial mantle could be induced in central mantle tissue underlying the damaged shell areas. This experimental approach makes it possible to identify gene products that control the chemical micro-environment during calcification as well as organic matrix components. This is unlike existing methodological approaches that work retroactively to characterize calcification relevant molecules and are just able to examine organic matrix components that are present in completed shells. In our assay an orthogonal array of nine 1 mm holes was drilled into the left valve, and mussels were suspended in net cages for 20, 29 and 36 days to regenerate. Structural observations using stereo-microscopy, SEM and Raman spectroscopy revealed organic sheet synthesis (day 20) as the first step of shell-repair followed by the deposition of calcite crystals (days 20 and 29) and aragonite tablets (day 36). The regeneration period was characterized by time-dependent shifts in gene expression in left central mantle tissue underlying the injured shell, (i) increased expression of two tyrosinase isoforms (TYR3: 29-fold and TYR6: 5-fold) at day 20 with a decline thereafter, (ii) an increase in expression of a gene encoding a nacrein-like protein (max. 100-fold) on day 29. The expression of an acidic Asp-Ser-rich protein was enhanced during the entire regeneration process. This proof-of-principle study demonstrates that genes that are specifically expressed in pallial and marginal mantle tissue can be induced (4 out of 10 genes) in central mantle following experimental injury of the overlying shell. Our findings suggest that regeneration assays can be used systematically to better characterize gene products that are essential for distinct phases of the shell formation process, particularly those that are not incorporated into the organic shell matrix.
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , isiRev
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    PAPER (OA)
  • 3
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    Sea shell diversity and rapidly evolving secretomes: insights into the evolution of biomineralization. (2018)
    Details
    Publication Date: 2021-04-25
    Description: An external skeleton is an essential part of the body plan of many animals and is thought to be one of the key factors that enabled the great expansion in animal diversity and disparity during the Cambrian explosion. Molluscs are considered ideal to study the evolution of biomineralization because of their diversity of highly complex, robust and patterned shells. The molluscan shell forms externally at the interface of animal and environment, and involves controlled deposition of calcium carbonate within a framework of macromolecules that are secreted from the dorsal mantle epithelium. Despite its deep conservation within Mollusca, the mantle is capable of producing an incredible diversity of shell patterns, and macro- and micro-architectures. Here we review recent developments within the field of molluscan biomineralization, focusing on the genes expressed in the mantle that encode secreted proteins. The so-called mantle secretome appears to regulate shell deposition and patterning and in some cases becomes part of the shell matrix. Recent transcriptomic and proteomic studies have revealed marked differences in the mantle secretomes of even closely-related molluscs; these typically exceed expected differences based on characteristics of the external shell. All mantle secretomes surveyed to date include novel genes encoding lineage-restricted proteins and unique combinations of co-opted ancient genes. A surprisingly large proportion of both ancient and novel secreted proteins containing simple repetitive motifs or domains that are often modular in construction. These repetitive low complexity domains (RLCDs) appear to further promote the evolvability of the mantle secretome, resulting in domain shuffling, expansion and loss. RLCD families further evolve via slippage and other mechanisms associated with repetitive sequences. As analogous types of secreted proteins are expressed in biomineralizing tissues in other animals, insights into the evolution of the genes underlying molluscan shell formation may be applied more broadly to understanding the evolution of metazoan biomineralization.
    Keywords: Biomineralization; Mollusc; Mantle; Shell; Shell matrix proteins; Co-option; Lineage-specific novelties; Repetitive low complexity domain ; 551
    Language: English , English
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    CITATION GEO-LEO
  • 4
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    A rapidly evolving secretome builds and patterns a sea shell (2010)
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    Publication Date: 2021-03-29
    Description: Background: Instructions to fabricate mineralized structures with distinct nanoscale architectures, such as seashells and coral and vertebrate skeletons, are encoded in the genomes of a wide variety of animals. In mollusks, the mantle is responsible for the extracellular production of the shell, directing the ordered biomineralization of CaCO3 and the deposition of architectural and color patterns. The evolutionary origins of the ability to synthesize calcified structures across various metazoan taxa remain obscure, with only a small number of protein families identified from molluskan shells. The recent sequencing of a wide range of metazoan genomes coupled with the analysis of gene expression in non-model animals has allowed us to investigate the evolution and process of biomineralization in gastropod mollusks.
    Description: Results: Here we show that over 25% of the genes expressed in the mantle of the vetigastropod Haliotis asinina encode secreted proteins, indicating that hundreds of proteins are likely to be contributing to shell fabrication and patterning. Almost 85% of the secretome encodes novel proteins; remarkably, only 19% of these have identifiable homologues in the full genome of the patellogastropod Lottia scutum. The spatial expression profiles of mantle genes that belong to the secretome is restricted to discrete mantle zones, with each zone responsible for the fabrication of one of the structural layers of the shell. Patterned expression of a subset of genes along the length of the mantle is indicative of roles in shell ornamentation. For example, Has-sometsuke maps precisely to pigmentation patterns in the shell, providing the first case of a gene product to be involved in molluskan shell pigmentation. We also describe the expression of two novel genes involved in nacre (mother of pearl) deposition.
    Description: research
    Keywords: 560 ; VV 000 ; Paläontologie
    Language: English
    Type: article , publishedVersion
    Format: 10 S.
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    CITATION GEO-LEO
  • 5
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    Transposable Elements: From DNA Parasites to Architects of Metazoan Evolution (2018)
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    Publication Date: 2021-04-27
    Description: One of the most unexpected insights that followed from the completion of the human genome a decade ago was that more than half of our DNA is derived from transposable elements (TEs). Due to advances in high throughput sequencing technologies it is now clear that TEs comprise the largest molecular class within most metazoan genomes. TEs, once categorised as "junk DNA", are now known to influence genomic structure and function by increasing the coding and non-coding genetic repertoire of the host. In this way TEs are key elements that stimulate the evolution of metazoan genomes. This review highlights several lines of TE research including the horizontal transfer of TEs through host-parasite interactions, the vertical maintenance of TEs over long periods of evolutionary time, and the direct role that TEs have played in generating morphological novelty.
    Keywords: transposable element; junk DNA; molecular parasite; molecular domestication; functionalisation; exonisation; exaptation; SINE; LINE ; 551
    Language: English
    Type: article , publishedVersion
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  • 6
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    Formin Is Associated with Left-Right Asymmetry in the Pond Snail and the Frog (2018)
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    Publication Date: 2021-03-29
    Description: While components of the pathway that establishes left-right asymmetry have been identified in diverse animals, from vertebrates to flies, it is striking that the genes involved in the first symmetry-breaking step remain wholly unknown in the most obviously chiral animals, the gastropod snails. Previously, research on snails was used to show that left-right signaling of Nodal, downstream of symmetry breaking, may be an ancestral feature of the Bilateria [1, 2]. Here, we report that a disabling mutation in one copy of a tandemly duplicated, diaphanousrelated formin is perfectly associated with symmetry breaking in the pond snail. This is supported by the observation that an anti-formin drug treatment converts dextral snail embryos to a sinistral phenocopy, and in frogs, drug inhibition or overexpression by microinjection of formin has a chirality-randomizing effect in early (pre-cilia) embryos. Contrary to expectations based on existingmodels [3–5],wediscovered asymmetric gene expression in 2- and 4-cell snail embryos, preceding morphological asymmetry. As the formin-actin filament has been shown to be part of an asymmetry-breaking switch in vitro [6, 7], together these results are consistent with the view that animals with diverse body plans may derive their asymmetries from the same intracellular chiral elements [8].
    Keywords: Formin; Left-Right Asymmetry; Pond Snail; Frog ; 551
    Language: English
    Type: article , publishedVersion
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    CITATION GEO-LEO
  • 7
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    The Holo-Transcriptome of a Calcified Early Branching Metazoan (2018)
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    Publication Date: 2021-04-25
    Description: djackso@uni-goettingen.de Specialty section: This article was submitted to Aquatic Microbiology, a section of the journal Frontiers in Marine Science Received: 31 October 2016 Accepted: 10 March 2017 Published: 28 March 2017 Citation: Germer J, Cerveau N and Jackson DJ (2017) The Holo-Transcriptome of a Calcified Early Branching Metazoan. Front. Mar. Sci. 4:81. doi: 10.3389/fmars.2017.00081 The Holo-Transcriptome of a Calcified Early Branching Metazoan Juliane Germer, Nicolas Cerveau and Daniel J. Jackson* Department of Geobiology, Georg-August University of Göttingen, Göttingen, Germany Symbiotic interactions are widespread throughout the animal kingdom and are increasingly recognized as an important trait that can shape the evolution of a species. Sponges are widely understood to be the earliest branching clade of metazoans and often contain dense, diverse yet specific microbial communities which can constitute up to 50% of their biomass. These bacterial communities fulfill diverse functions influencing the sponge’s physiology and ecology, and may have greatly contributed to the evolutionary success of the Porifera. Here we have analyzed and characterized the holo-transcriptome of the hypercalcifying demosponge Vaceletia sp. and compare it to other sponge transcriptomic and genomic data. Vaceletia sp. harbors a diverse and abundant microbial community; by identifying the underlying molecular mechanism of a variety of lipid pathway components we show that the sponge seems to rely on the supply of short chain fatty acids by its bacterial community. Comparisons to other sponges reveal that this dependency may be more pronounced in sponges with an abundant microbial community. Furthermore, the presence of bacterial polyketide synthase genes suggests bacteria are the producers of Vaceletia’s abundant mid-chain branched fatty acids, whereas demospongic acids may be produced by the sponge host via elongation and desaturation of short-chain precursors. We show that the sponge and its microbial community have the molecular tools to interact through different mechanisms including the sponge’s immune system, and the presence of eukaryotic-like proteins in bacteria. These results expand our knowledge of the complex gene repertoire of sponges and show the importance of metabolic interactions between sponges and their endobiotic microbial communities.
    Description: Open-Access-Publikationsfonds 2017
    Keywords: sponge, microbes, symbiosis, transcriptome, immunity, metabolism, fatty acid, signaling pathway ; 551
    Language: English
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  • 8
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    An Antarctic molluscan biomineralisation tool-kit (2018)
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    Publication Date: 2021-04-27
    Description: The Antarctic clam Laternula elliptica lives almost permanently below 0 °C and therefore is a valuable and tractable model to study the mechanisms of biomineralisation in cold water. The present study employed a multidisciplinary approach using histology, immunohistochemistry, electron microscopy, proteomics and gene expression to investigate this process. Thirty seven proteins were identified via proteomic extraction of the nacreous shell layer, including two not previously found in nacre; a novel T-rich Mucin-like protein and a Zinc-dependent metalloprotease. In situ hybridisation of seven candidate biomineralisation genes revealed discrete spatial expression patterns within the mantle tissue, hinting at modular organisation, which is also observed in the mantle tissues of other molluscs. All seven of these biomineralisation candidates displayed evidence of multifunctionality and strong association with vesicles, which are potentially involved in shell secretion in this species.
    Keywords: mollusc; biomineralisation ; 551
    Language: English
    Type: article , publishedVersion
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    CITATION GEO-LEO
  • 9
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    Combining independent de novo assemblies optimizes the coding transcriptome for nonconventional model eukaryotic organisms (2018)
    BioMed Central
    Details
    Publication Date: 2021-03-29
    Keywords: 551
    Language: English
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    CITATION GEO-LEO
  • 10
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    Variation in rates of early development in Haliotis asinina generate competent larvae of different ages (2018)
    Details
    Publication Date: 2021-03-29
    Description: Introduction Inter-specific comparisons of metazoan developmental mechanisms have provided a wealth of data concerning the evolution of body form and the generation of morphological novelty. Conversely, studies of intra-specific variation in developmental programs are far fewer. Variation in the rate of development may be an advantage to the many marine invertebrates that posses a biphasic life cycle, where fitness commonly requires the recruitment of planktonically dispersing larvae to patchily distributed benthic environments. Results We have characterised differences in the rate of development between individuals originating from a synchronised fertilisation event in the tropical abalone Haliotis asinina, a broadcast spawning lecithotrophic vetigastropod. We observed significant differences in the time taken to complete early developmental events (time taken to complete third cleavage and to hatch from the vitelline envelope), mid-larval events (variation in larval shell development) and late larval events (the acquisition of competence to respond to a metamorphosis inducing cue). We also provide estimates of the variation in maternally provided energy reserves that suggest maternal provisioning is unlikely to explain the majority of the variation in developmental rate we report here. Conclusions Significant differences in the rates of development exist both within and between cohorts of synchronously fertilised H. asinina gametes. These differences can be detected shortly after fertilisation and generate larvae of increasingly divergent development states. We discuss the significance of our results within an ecological context, the adaptive significance of mechanisms that might maintain this variation, and potential sources of this variation.
    Description: Open-Access-Publikationsfonds 2012
    Keywords: 551
    Language: English
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