A sea urchin Na+K+2Cl- cotransporter is involved in the maintenance of calcification-relevant cytoplasmic cords in Strongylocentrotus droebachiensis larvae

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.

Stomach content records of albacore (Thunnus alalunga) and bluefin tuna (Thunnus thynnus) in the North Atlantic Drift Region focusing on the Bay of Biscay between 2004 and 2011

The tuna stomach database from AZTI-Tecnalia corresponds to 7 years of sampling from 2004 to 2011. Due to the absence of continuity in the different projects dealing with the feeding ecology of tunas, the sampling could not be performed every year for both species, and no sample was collected in 2008. However, the fish stomach content record contents composition - by prey weight - of 1525 albacore caught in the Bay of Biscay and surrounding waters of the North Atlantic Drift Region in 2005 (n=397), 2006 (n=196), 2007 (n=37), 2009 (n=95), 2010 (n=566) and 2011 (n=234) ; and of 686 bluefin tunas caught in the Southeastern Bay of Biscay in 2004 (n=32), 2005 (n=36), 2006 (n=3), 2009 (n=257), 2010 (n=233) and 2011 (n=125). Samples have been obtained from scientific research surveys (using a variety of different fishing gears), from commercial fisheries catches, from individual fish voluntarily sampled by recreational fishermen and from fish accidentally stranded on coastlines. Each predator is identified by an ID and its length and wet weight are given. In case the wet weight could not be measured, it was estimated through a length-weight relationship equation and is indicated in the comment for the Predator mass column. The total weight of each prey is given, as well as the weight of each prey taxonomic group in each stomach.

(Table 1) Number of alleles per locus and heterozygosity of Arctic charr (Salvelinus alpinus) populations in Iceland

The ecological theory of adaptive radiation predicts that the evolution of phenotypic diversity within species is generated by divergent natural selection arising from different environments and competition between species. Genetic connectivity among populations is likely also to have an important role in both the origin and maintenance of adaptive genetic diversity. Our goal was to evaluate the potential roles of genetic connectivity and natural selection in the maintenance of adaptive phenotypic differences among morphs of Arctic charr, Salvelinus alpinus, in Iceland. At a large spatial scale, we tested the predictive power of geographic structure and phenotypic variation for patterns of neutral genetic variation among populations throughout Iceland. At a smaller scale, we evaluated the genetic differentiation between two morphs in Lake Thingvallavatn relative to historically explicit, coalescent-based null models of the evolutionary history of these lineages. At the large spatial scale, populations are highly differentiated, but weakly structured, both geographically and with respect to patterns of phenotypic variation. At the intralacustrine scale, we observe modest genetic differentiation between two morphs, but this level of differentiation is nonetheless consistent with strong reproductive isolation throughout the Holocene. Rather than a result of the homogenizing effect of gene flow in a system at migration-drift equilibrium, the modest level of genetic differentiation could equally be a result of slow neutral divergence by drift in large populations. We conclude that contemporary and recent patterns of restricted gene flow have been highly conducive to the evolution and maintenance of adaptive genetic variation in Icelandic Arctic charr.