Electroreception in the euryhaline stingray, Dasyatis sabina.

This study quantified the electrosensitivity of a euryhaline elasmobranch, the Atlantic stingray (Dasyatis sabina) across a range of salinities. Specimens from a permanent freshwater (FW) population in the St Johns River system, FL, USA, were compared with stingrays from the tidally dynamic Indian River Lagoon in east Florida, USA. Behavioral responses of stingrays to prey-simulating electric stimuli were quantified in FW (0 p.p.t., rho=2026 Omega cm), brackish (15 p.p.t., rho=41 Omega cm) and full strength seawater (35 p.p.t., rho=19 Omega cm). This study demonstrated that the electrosensitivity of D. sabina is significantly reduced in FW. In order to elicit a feeding response, stingrays tested in FW required an electric field 200-300x greater than stingrays tested in brackish and saltwater (median FW treatments=1.4 microV cm(-1), median brackish-saltwater treatments=6 nV cm(-1)), and the maximum orientation distance was reduced by 35.2%, from 44.0 cm in the brackish and saltwater treatments to 28.5 cm in FW. The St Johns River stingrays did not demonstrate an enhanced electrosensitivity in FW, nor did they exhibit reduced sensitivity when introduced to higher salinities. Stingrays from both populations responded similarly to the prey-simulating stimulus when tested at similar salinities, regardless of their native environment. The reduction in electrosensitivity and detection range in FW is attributed to both an environmental factor (electrical resistivity of the water) and the physiological function of the ampullary canals. The plasticity of this sensory system to function across such a wide environmental range demonstrates its adaptive significance.

CD39L2, a gene encoding a human nucleoside diphosphatase, predominantly expressed in the heart.

E-NTPDases are extracellular enzymes that hydrolyze nucleotides. The human E-NTPDase gene family currently consists of five reported members (CD39, CD39L1, CD39L2, CD39L3, and CD39L4). Both membrane-bound and secreted family members have been predicted by encoded transmembrane and leader peptide motifs. In this report, we demonstrate that the human CD39L2 gene is expressed predominantly in the heart. In situ hybridization results from heart indicate that the CD39L2 message is expressed in muscle and capillary endothelial cells. We also show that the CD39L2 gene encodes an extracellular E-NTPDase. Flow cytometric experiments show that transiently expressed CD39L2 is present on the surface of COS-7 cells. Transfected cells also produce recombinant glycosylated protein in the medium, and this process can be blocked by brefeldin A, an inhibitor of the mammalian secretory pathway. The enzymology of CD39L2 shows characteristic features of a typical E-NTPDase, but with a much higher degree of specificity for NDPs over NTPs as enzymatic substrates. The kinetics of the ADPase activity exhibit positive cooperativity. The predominance of CD39L2 expression in the heart supports a functional role in regulating platelet activation and recruitment in this organ.

Biochemical characterization of CD39L4.

Nucleotides are involved in regulating a number of important processes ranging from inflammation to platelet aggregation. Enzymes that can modulate levels of nucleotides in the blood therefore represent important regulatory components in these physiological systems. CD39L4 is a soluble E-nucleoside triphosphate dephosphohydrolase (E-NTPDase) with specificity for nucleotide diphosphates (NDPs). In this study, stable mammalian and insect cell lines were generated expressing CD39L4 protein to purify and characterize the recombinant protein. We demonstrate that recombinant CD39L4 protein expressed in human embryonic carcinoma 293 cells is glycosylated by comparing the molecular masses before and after glycosidase treatment. Activity measurements of CD39L4 isolated from tunicamycin-treated, transiently transfected COS-7 cells indicate that glycosylation is not required for full ADPase activity. Recombinant human CD39L4 protein isolated from stable insect cells was glycosylated differently, but also demonstrated relative activity comparable to that of the mammalian protein. When denatured by SDS under nonreducing conditions, a fraction of the CD39L4 protein migrates as a 110 kDa disulfide-linked dimer. We determined that the monomer is the most active form of CD39L4 by measuring the activity of sucrose density gradient fractions of monomers and partially purified dimers. The physiological significance of the biochemical and enzymatic characterization is discussed.