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. 1995 Oct;116(3):1979–1984. doi: 10.1111/j.1476-5381.1995.tb16401.x

Actions of ADP, but not ATP, on cytosolic free Ca2+ in single rat hepatocytes mimicked by 2-methylthioATP.

C J Dixon 1, P H Cobbold 1, A K Green 1
PMCID: PMC1908934  PMID: 8640335

Abstract

1. Aequorin-injected, single rat hepatocytes generate series of repetitive transients in cytosolic free calcium concentration ([Ca2+]i) when stimulated with agonists acting through the phosphoinositide signalling pathway, including ADP and ATP. We have previously described differences in the [Ca2+]i responses of aequorin-injected hepatocytes to ADP and ATP. 2. The effects of the phosphorothioate analogue of ATP, 2-methylthioATP (2-meSATP), have been examined on single rat hepatocytes. This analogue is belived to be the most potent agonist at the P2Y1 subclass of purinoceptor. 3. The [Ca2+]i transients induced by 2-meSATP were indistinguishable from those induced by ADP, and in contrast to those induced by ATP. 4. At hig concentrations, 2-meSATP and ADP both induced transients at high frequency. In contrast, hepatocytes responded to high concentrations of ATP with an initial rapid rise in [Ca2+]i, followed by a slowly decaying fall. 5. The modulatory effects of elevated intracellular cyclic AMP concentration were the same on both 2-meSATP- and ADP-induced [Ca2+]i transients; the peak height and frequency of transients were enhanced. ATP-induced transients, however, underwent either an increase in duration or conversion into a sustained rise in [Ca2+]i. 6. ATP-induced transients were specifically potentiated by the co-addition of alpha, beta-methyleneATP, whereas 2-meSATP- and ADP-induced transients were unaffected by this treatment. 7. We conclude that 2-meSATP acts at the same receptor as ADP on rat hepatocytes, and that this is distinct from teh receptor(s) mediating the effects of ATP.

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Selected References

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  1. Abbracchio M. P., Burnstock G. Purinoceptors: are there families of P2X and P2Y purinoceptors? Pharmacol Ther. 1994;64(3):445–475. doi: 10.1016/0163-7258(94)00048-4. [DOI] [PubMed] [Google Scholar]
  2. Bailey S. J., Hourani S. M. A study of the purinoceptors mediating contraction in the rat colon. Br J Pharmacol. 1990 Aug;100(4):753–756. doi: 10.1111/j.1476-5381.1990.tb14087.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barnard E. A., Burnstock G., Webb T. E. G protein-coupled receptors for ATP and other nucleotides: a new receptor family. Trends Pharmacol Sci. 1994 Mar;15(3):67–70. doi: 10.1016/0165-6147(94)90280-1. [DOI] [PubMed] [Google Scholar]
  4. Berridge M. J. Calcium oscillations. J Biol Chem. 1990 Jun 15;265(17):9583–9586. [PubMed] [Google Scholar]
  5. Brown H. A., Lazarowski E. R., Boucher R. C., Harden T. K. Evidence that UTP and ATP regulate phospholipase C through a common extracellular 5'-nucleotide receptor in human airway epithelial cells. Mol Pharmacol. 1991 Nov;40(5):648–655. [PubMed] [Google Scholar]
  6. Burnstock G., Kennedy C. Is there a basis for distinguishing two types of P2-purinoceptor? Gen Pharmacol. 1985;16(5):433–440. doi: 10.1016/0306-3623(85)90001-1. [DOI] [PubMed] [Google Scholar]
  7. Carter T. D., Hallam T. J., Cusack N. J., Pearson J. D. Regulation of P2y-purinoceptor-mediated prostacyclin release from human endothelial cells by cytoplasmic calcium concentration. Br J Pharmacol. 1988 Dec;95(4):1181–1190. doi: 10.1111/j.1476-5381.1988.tb11754.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cobbold P. H., Rink T. J. Fluorescence and bioluminescence measurement of cytoplasmic free calcium. Biochem J. 1987 Dec 1;248(2):313–328. doi: 10.1042/bj2480313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cobbold P., Woods N., Wainwright J., Cuthbertson R. Single cell measurements in research on calcium-mobilising purinoceptors. J Recept Res. 1988;8(1-4):481–491. doi: 10.3109/10799898809049006. [DOI] [PubMed] [Google Scholar]
  10. Dixon C. J., Cobbold P. H., Green A. K. Adenosine 5'-[alpha beta-methylene]triphosphate potentiates the oscillatory cytosolic Ca2+ responses of hepatocytes to ATP, but not to ADP. Biochem J. 1993 Aug 1;293(Pt 3):757–760. doi: 10.1042/bj2930757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dixon C. J., Woods N. M., Cuthbertson K. S., Cobbold P. H. Evidence for two Ca2(+)-mobilizing purinoceptors on rat hepatocytes. Biochem J. 1990 Jul 15;269(2):499–502. doi: 10.1042/bj2690499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dubyak G. R., el-Moatassim C. Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides. Am J Physiol. 1993 Sep;265(3 Pt 1):C577–C606. doi: 10.1152/ajpcell.1993.265.3.C577. [DOI] [PubMed] [Google Scholar]
  13. Fredholm B. B., Abbracchio M. P., Burnstock G., Daly J. W., Harden T. K., Jacobson K. A., Leff P., Williams M. Nomenclature and classification of purinoceptors. Pharmacol Rev. 1994 Jun;46(2):143–156. [PMC free article] [PubMed] [Google Scholar]
  14. Gibb C. A., Singh S., Cook D. I., Poronnik P., Conigrave A. D. A nucleotide receptor that mobilizes Ca2+ in the mouse submandibular salivary cell line ST885. Br J Pharmacol. 1994 Apr;111(4):1135–1139. doi: 10.1111/j.1476-5381.1994.tb14863.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gordon J. L. Extracellular ATP: effects, sources and fate. Biochem J. 1986 Jan 15;233(2):309–319. doi: 10.1042/bj2330309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Green A. K., Cobbold P. H., Dixon C. J. Elevated intracellular cyclic AMP exerts different modulatory effects on cytosolic free Ca2+ oscillations induced by ADP and ATP in single rat hepatocytes. Biochem J. 1994 Sep 15;302(Pt 3):949–955. doi: 10.1042/bj3020949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Iredale P. A., Hill S. J. Increases in intracellular calcium via activation of an endogenous P2-purinoceptor in cultured CHO-K1 cells. Br J Pharmacol. 1993 Dec;110(4):1305–1310. doi: 10.1111/j.1476-5381.1993.tb13960.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Iredale P. A., Martin K. F., Alexander S. P., Hill S. J., Kendall D. A. Inositol 1,4,5-trisphosphate generation and calcium mobilisation via activation of an atypical P2 receptor in the neuronal cell line, N1E-115. Br J Pharmacol. 1992 Dec;107(4):1083–1087. doi: 10.1111/j.1476-5381.1992.tb13410.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Keppens S., De Wulf H. Characterization of the biological effects of 2-methylthio-ATP on rat hepatocytes: clear-cut differences with ATP. Br J Pharmacol. 1991 Oct;104(2):301–304. doi: 10.1111/j.1476-5381.1991.tb12426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Keppens S., De Wulf H. Characterization of the liver P2-purinoceptor involved in the activation of glycogen phosphorylase. Biochem J. 1986 Dec 1;240(2):367–371. doi: 10.1042/bj2400367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Keppens S. The complex interaction of ATP and UTP with isolated hepatocytes. How many receptors? Gen Pharmacol. 1993 Mar;24(2):283–289. doi: 10.1016/0306-3623(93)90304-g. [DOI] [PubMed] [Google Scholar]
  22. Keppens S., Vandekerckhove A., De Wulf H. Characterization of the effects of adenosine 5'-[beta-thio]-diphosphate in rat liver. Br J Pharmacol. 1993 Mar;108(3):663–668. doi: 10.1111/j.1476-5381.1993.tb12858.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Keppens S., Vandekerckhove A., De Wulf H. Extracellular ATP and UTP exert similar effects on rat isolated hepatocytes. Br J Pharmacol. 1992 Feb;105(2):475–479. doi: 10.1111/j.1476-5381.1992.tb14278.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lustig K. D., Shiau A. K., Brake A. J., Julius D. Expression cloning of an ATP receptor from mouse neuroblastoma cells. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5113–5117. doi: 10.1073/pnas.90.11.5113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Merten M. D., Breittmayer J. P., Figarella C., Frelin C. ATP and UTP increase secretion of bronchial inhibitor by human tracheal gland cells in culture. Am J Physiol. 1993 Nov;265(5 Pt 1):L479–L484. doi: 10.1152/ajplung.1993.265.5.L479. [DOI] [PubMed] [Google Scholar]
  26. Milligan G. Mechanisms of multifunctional signalling by G protein-linked receptors. Trends Pharmacol Sci. 1993 Jun;14(6):239–244. doi: 10.1016/0165-6147(93)90019-g. [DOI] [PubMed] [Google Scholar]
  27. Murgo A. J., Contrera J. G., Sistare F. D. Evidence for separate calcium-signaling P2T and P2U purinoceptors in human megakaryocytic Dami cells. Blood. 1994 Mar 1;83(5):1258–1267. [PubMed] [Google Scholar]
  28. Murgo A. J., Sistare F. D. K562 leukemia cells express P2T (adenosine diphosphate) purinergic receptors. J Pharmacol Exp Ther. 1992 May;261(2):580–585. [PubMed] [Google Scholar]
  29. Murrin R. J., Boarder M. R. Neuronal "nucleotide" receptor linked to phospholipase C and phospholipase D? Stimulation of PC12 cells by ATP analogues and UTP. Mol Pharmacol. 1992 Mar;41(3):561–568. [PubMed] [Google Scholar]
  30. Needham L., Cusack N. J., Pearson J. D., Gordon J. L. Characteristics of the P2 purinoceptor that mediates prostacyclin production by pig aortic endothelial cells. Eur J Pharmacol. 1987 Feb 10;134(2):199–209. doi: 10.1016/0014-2999(87)90166-x. [DOI] [PubMed] [Google Scholar]
  31. O'Connor S. E., Dainty I. A., Leff P. Further subclassification of ATP receptors based on agonist studies. Trends Pharmacol Sci. 1991 Apr;12(4):137–141. doi: 10.1016/0165-6147(91)90530-6. [DOI] [PubMed] [Google Scholar]
  32. Palea S., Corsi M., Pietra C., Artibani W., Calpista A., Gaviraghi G., Trist D. G. ADP beta S induces contraction of the human isolated urinary bladder through a purinoceptor subtype different from P2X and P2Y. J Pharmacol Exp Ther. 1994 Apr;269(1):193–197. [PubMed] [Google Scholar]
  33. Raha S., de Souza L. R., Reed J. K. Intracellular signalling by nucleotide receptors in PC12 pheochromocytoma cells. J Cell Physiol. 1993 Mar;154(3):623–630. doi: 10.1002/jcp.1041540322. [DOI] [PubMed] [Google Scholar]
  34. Reimer W. J., Dixon S. J. Extracellular nucleotides elevate [Ca2+]i in rat osteoblastic cells by interaction with two receptor subtypes. Am J Physiol. 1992 Nov;263(5 Pt 1):C1040–C1048. doi: 10.1152/ajpcell.1992.263.5.C1040. [DOI] [PubMed] [Google Scholar]
  35. Robb S., Cheek T. R., Hannan F. L., Hall L. M., Midgley J. M., Evans P. D. Agonist-specific coupling of a cloned Drosophila octopamine/tyramine receptor to multiple second messenger systems. EMBO J. 1994 Mar 15;13(6):1325–1330. doi: 10.1002/j.1460-2075.1994.tb06385.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vigne P., Feolde E., Breittmayer J. P., Frelin C. Characterization of the effects of 2-methylthio-ATP and 2-chloro-ATP on brain capillary endothelial cells: similarities to ADP and differences from ATP. Br J Pharmacol. 1994 Jul;112(3):775–780. doi: 10.1111/j.1476-5381.1994.tb13146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Webb T. E., Simon J., Krishek B. J., Bateson A. N., Smart T. G., King B. F., Burnstock G., Barnard E. A. Cloning and functional expression of a brain G-protein-coupled ATP receptor. FEBS Lett. 1993 Jun 14;324(2):219–225. doi: 10.1016/0014-5793(93)81397-i. [DOI] [PubMed] [Google Scholar]
  38. Wilkinson G. F., McKechnie K., Dainty I. A., Boarder M. R. P2Y purinoceptor and nucleotide receptor-induced relaxation of precontracted bovine aortic collateral artery rings: differential sensitivity to suramin and indomethacin. J Pharmacol Exp Ther. 1994 Feb;268(2):881–887. [PubMed] [Google Scholar]
  39. Woods N. M., Cuthbertson K. S., Cobbold P. H. Agonist-induced oscillations in cytoplasmic free calcium concentration in single rat hepatocytes. Cell Calcium. 1987 Feb;8(1):79–100. doi: 10.1016/0143-4160(87)90038-8. [DOI] [PubMed] [Google Scholar]
  40. Woods N. M., Cuthbertson K. S., Cobbold P. H. Repetitive transient rises in cytoplasmic free calcium in hormone-stimulated hepatocytes. Nature. 1986 Feb 13;319(6054):600–602. doi: 10.1038/319600a0. [DOI] [PubMed] [Google Scholar]

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