Delineation and mapping of coastal shark habitat within a shallow lagoonal estuary

Estuaries function as important nursery and foraging habitats for many coastal species, including highly migratory sharks. Pamlico Sound, North Carolina, is one of the largest estuaries in the continental United States and provides a variety of potential habitats for sharks. In order to identify and spatially delineate shark habitats within Pamlico Sound, shark catch and environmental data were analyzed from the 2007–2014 North Carolina Division of Marine Fisheries (NCDMF) gillnet and longline surveys conducted within the estuary. Principal species were identified and environmental data recorded at survey sites (depth, temperature, salinity, dissolved oxygen, submerged aquatic vegetation (SAV) distance, and inlet distance) were interpolated across Pamlico Sound to create seasonal environmental grids with a 90-m2 cell size. Boosted Regression Tree (BRT) analysis was used to identify the most important environmental factors and ranges associated with presence of each principal species, and the resulting models were used to predict shark capture probability based on the environmental values within the grid cells. The Atlantic Sharpnose Shark (Rhizoprionodon terraenovae), Blacktip Shark (Carcharhinus limbatus), Bull Shark (Carcharhinus leucas), Sandbar Shark (Carcharhinus plumbeus), Smooth Dogfish (Mustelus canis), and Spiny Dogfish (Squalus acanthias) were the principal species in Pamlico Sound. Most species were associated with proximity to the inlet and/or high salinity, and warm temperatures, but the Bull Shark preferred greater inlet distances and the Spiny Dogfish preferred lower temperatures than the other species. Extensive Smooth Dogfish habitat overlap with seagrass beds suggests that seagrass may be a critical part of nursery habitat for this species. Spatial delineation of shark habitat within the estuary will allow for better protection of essential habitat and assessment of potential interactions with other species

Targeted neutralization of calmodulin in the nucleus blocks DNA synthesis and cell cycle progression

AbstractCalmodulin (CaM) is a major intracellular calcium binding protein which has been implicated in the regulation of cell proliferation. Previous studies using chemically synthesized CaM antagonists and anti-sense RNA indicated that CaM is important for initiation of DNA synthesis and cell cycle progression. However, these methods reduce total intracellular CaM and globally interfering with all the CaM-dependent processes. In order to explore the function of nuclear CaM during the cell cycle, a CaM inhibitor peptide was targeted to the nucleus of intact mammalian cells. Cell progression through S-phase was assessed by incorporation of the thymidine analogue, BrdU. Cells were transfected for 48 h with either the CaM inhibitor peptide gene or the control plasmid prior to analysis. Approx. 70% of the control cells incorporated BrdU. In striking contrast, double immunofluorescent labeling demonstrated that none of the cells expressing the CaM inhibitor peptide entered S-phase. This result indicates that neutralization of nuclear CaM by targeted expression of a CaM inhibitor peptide blocks DNA synthesis and cell cycle progression

Annexin A4 Reduces Water and Proton Permeability of Model Membranes but Does Not Alter Aquaporin 2–mediated Water Transport in Isolated Endosomes

Annexin A4 (Anx4) belongs to a ubiquitous family of Ca2+-dependent membrane-binding proteins thought to be involved in membrane trafficking and membrane organization within cells. Anx4 localizes to the apical region in epithelia; however, its physiological role is unclear. We show that Anx4 exhibited binding to liposomes (phosphatidylcholine:phosphatidylserine, 1:1) in the presence of Ca2+ and binding was reversible with EDTA. Anx4 binding resulted in liposome aggregation and a reduction in membrane water permeability of 29% (P < 0.001) at 25°C. These effects were not seen in the presence of Ca2+ or Anx4 alone and were reversible with EDTA. Measurements of membrane fluidity made by monitoring fluorescence anisotropy of 2-(12-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)dodecanoyl-1-hexadecanoyl-sn-glycero-3-phosphocholine (NBD-HPC) demonstrated that Anx4 binding rigidified the outer leaflet of the bilayer (P < 0.001), thus providing a molecular explanation for the inhibition of water flux. To determine whether Anx4 would produce similar effects on physiological membranes we constructed liposomes which recapitulated the lipid composition of the inner leaflet of the MDCK apical membrane. These membranes exhibited reductions to water permeability upon Anx4 binding (19.5% at 25°C, 31% at 37°C; P < 0.01 and P < 0.001, respectively) and to proton permeability (15% at 25°C, 19.5% at 37°C; P < 0.05). Since our in vitro experiments indicated an effect on membrane permeability, we examined localization of Anx4 in the kidney collecting duct, a region of the nephron responsible for concentrating urine through water reabsorbtion. Anx4 was shown to colocalize apically with aquaporin 2 (AQP2) in collecting duct epithelia. To test for the existence of a functional interaction between Anx4 and AQP2 we isolated AQP2-containing endosomes and exposed them to Anx4/Ca2+. Water flux rates were unchanged, indicating Anx4 does not directly regulate AQP2. We conclude that Anx4 can alter the physical properties of membranes by associating with them and regulate passive membrane permeability to water and protons. These properties represent important new functions for Anx4