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Microinjection of human Jurkat T-lymphocytes with nicotinic acid adenine dinucleotide phosphate (NAADP(+)) dose-dependently stimulated intracellular Ca(2+)-signaling. At a concentration of 10 nM NAADP(+) evoked repetitive and long-lasting Ca(2+)-oscillations of low amplitude, whereas at 50 and 100 nM, a rapid and high initial Ca(2+)-peak followed by trains of smaller Ca(2+)-oscillations was observed. Higher concentrations of NAADP(+) (1 and 10 microM) gradually reduced the initial Ca(2+)-peak, and a complete self-inactivation of Ca(2+)-signals was seen at 100 microM. The effect of NAADP(+) was specific as it was not observed with nicotinamide adenine dinucleotide phosphate. Both inositol 1,4, 5-trisphosphate- and cyclic adenosine diphosphoribose-mediated Ca(2+)-signaling were efficiently inhibited by coinjection of a self-inactivating concentration of NAADP(+). Most importantly, microinjection of a self-inactivating concentration of NAADP(+) completely abolished subsequent stimulation of Ca(2+)-signaling via the T cell receptor/CD3 complex, indicating that a functional NAADP(+) Ca(2+)-release system is essential for T-lymphocyte Ca(2+)-signaling.

Type

Journal article

Journal

J Cell Biol

Publication Date

07/08/2000

Volume

150

Pages

581 - 588

Keywords

Adenosine Diphosphate Ribose, Antigens, CD3, Calcium Signaling, Cyclic ADP-Ribose, Dose-Response Relationship, Drug, Humans, Image Processing, Computer-Assisted, Inositol 1,4,5-Trisphosphate, Jurkat Cells, Lymphocyte Activation, Microinjections, NADP, Receptors, Antigen, T-Cell, Ryanodine Receptor Calcium Release Channel, T-Lymphocytes