Two Distinct P2 Purinergic Receptors, P2Y and P2U, Are Coupled to Phospholipase C in Mouse Pineal Gland Tumor Cells |
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Authors: | Byung-Chang Suh Jin H. Son Tong H. Joh Kyong-Tai Kim |
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Affiliation: | Department of Life Science and Basic Science Research Institute, Pohang University of Science and Technology, Pohang, Republic of Korea;and; Laboratory of Molecular Neurobiology, Cornell University Medical College at the W. M. Burke Medical Research Institute, White Plains, New York, U.S.A. |
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Abstract: | Abstract: We found that extracellular ATP can increase the intracellular Ca2+ concentration ([Ca2+]i) in mouse pineal gland tumor (PGT-β) cells. Studies of the [Ca2+]i rise using nucleotides and ATP analogues established the following potency order: ATP, adenosine 5′-O-(3-thiotriphosphate) ≥ UTP > 2-chloro-ATP > 3′-O-(4-benzoyl)benzoyl ATP, GTP ≥ 2-methylthio ATP, adenosine 5′-O-(2-thiodiphosphate) (ADPβS) > CTP. AMP, adenosine, α,β-methyleneadenosine 5′-triphosphate, β,γ-methyleneadenosine 5′-triphosphate, and UMP had little or no effect on the [Ca2+]i rise. Raising the extracellular Mg2+ concentration to 10 mM decreases the ATP-and UTP-induced [Ca2+]i rise, because the responses depend on the ATP4? and UTP4? concentrations, respectively. The P2U purinoceptor-selective agonist UTP and the P2Y purinoceptor-selective agonist ADPβS induce inositol 1,4,5-trisphosphate generation in a concentration-dependent manner with maximal effective concentrations of ~100 µM. In sequential stimulation, UTP and ADPβS do not interfere with each other in raising the [Ca2+]i. Costimulation with UTP and ADPβS results in additive inositol 1,4,5-trisphosphate generation to a similar extent as is achieved with ATP alone. Pretreatment with pertussis toxin inhibits the action of UTP and ATP by maximally 45–55%, whereas it has no effect on the ADPβS response. Treatment with 1 µM phorbol 12-myristate 13-acetate inhibits the ADPβS-induced [Ca2+]i rise more effectively than the ATP- and UTP-induced responses. These results suggest that P2U and P2Y purinoceptors coexist on PGT-β cells and that both receptors are linked to phospholipase C. |
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Keywords: | Mouse pineal gland tumor (PGT-β) cells Phospholipase C ATP Nucleotides Ca2+ mobilization Inositol 1,4,5-trisphosphate P2 purinoceptor |
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