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.

Radiocarbon age, and sedimentation and accumulation rate of core JM05-G001 obtained during Jan Mayen cruise JM050704 to the Barents Sea

Instrumental monitoring of the climate at high northern latitudes has documented the ongoing warming of the last few decades. Climate modelling has also demonstrated that the global warming signal will be amplified in the polar region. Such temperature increases would have important implications on the ecosystem and biota of the Barents Sea. This study therefore aims to reconstruct the climatic changes of the Barents Sea based on benthic foraminifera over approximately the last 1400 years at the decadal to sub-decadal scale. Oxygen and carbon isotope analysis and benthic foraminiferal species counts indicate an overall warming trend of approximately 2.6°C through the 1400-year record. In addition, the well-documented cooling period equating to the 'Little Ice Age' is evident between c. 1650 and 1850. Most notably, a series of highly fluctuating temperatures are observed over the last century. An increase of 1.5°C is shown across this period. Thus for the first time we are able to demonstrate that the recent Arctic warming is also reflected in the oceanic micro-fauna.