TAILIEUCHUNG - Báo cáo khoa học: Cyclic ADP-ribose requires CD38 to regulate the release of ATP in visceral smooth muscle

It is well established that the intracellular second messenger cADP-ribose (cADPR) activates Ca 2+ release from the sarcoplasmic reticulum through ryanodine receptors. CD38 is a multifunctional enzyme involved in the for-mation of cADPR in mammals. | IFEBS Journal Cyclic ADP-ribose requires CD38 to regulate the release of ATP in visceral smooth muscle Leonie Durnin and Violeta N. Mutafova-Yambolieva Department of Physiology and CellBiology University of Nevada Schoolof Medicine Reno NV USA Keywords ATP bladder cADP-ribose CD38 NAD purinergic neurotransmission Correspondence V. N. Mutafova-Yambolieva Department of Physiology and CellBiology University of Nevada Schoolof Medicine Center for Molecular Medicine MS 575 Reno NV 89557-0575 USA Fax 1 775 784 6903 Tel 1 775 784 6274 E-mail vmutafova@ Received 30 April 2011 revised 24 June 2011 accepted 30 June 2011 doi It is well established that the intracellular second messenger cADP-ribose cADPR activates Ca2 release from the sarcoplasmic reticulum through ryanodine receptors. CD38 is a multifunctional enzyme involved in the formation of cADPR in mammals. CD38 has also been reported to transport cADPR in several cell lines. Here we demonstrate a role for extracellular cADPR and CD38 in modulating the spontaneous but not the electrical field stimulation-evoked release of ATP in visceral smooth muscle. Using a small-volume superfusion assay and an HPLC technique with fluorescence detection we measured the spontaneous and evoked release of ATP in bladder detrusor smooth muscles isolated from CD38 and CD38 mice. cADPR 1 nM enhanced the spontaneous overflow of ATP in bladders isolated from CD38 mice. This effect was abolished by the inhibitor of cADPR receptors on sarcoplasmic reticulum 8-bromo-cADPR 80 im and by ryanodine 50 im but not by the nonselective P2 puriner-gic receptor antagonist pyridoxal phosphate 6-azophenyl-2 4 -disulfonate 30 M . cADPR failed to facilitate the spontaneous ATP overflow in bladders isolated from CD38 mice indicating that CD38 is crucial for the enhancing effects of extracellular cADPR on spontaneous ATP release. Contractile responses to ATP were potentiated by cADPR suggesting that the two

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