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. 1995 May 1;307(Pt 3):775–782. doi: 10.1042/bj3070775

Desensitization and resensitization of human platelets to 5-hydroxytryptamine at the level of signal transduction.

P Roevens 1, D de Chaffoy de Courcelles 1
PMCID: PMC1136717  PMID: 7741708

Abstract

Desensitization of platelets to 5-hydroxytryptamine (5HT) (1 microM), during active removal of the agonist by the platelet 5HT-uptake system, was studied at the level of signal transduction. Desensitization to 5HT was dose-dependent and homologous. Without occupation of the 5HT2 receptor, neither an increase in cytosolic [Ca2+] (30 nM ionomycin), nor a separate or simultaneous activation of protein kinase C (by 10 microM 1-oleoyl-2-acetylglycerol), could induce desensitization to 5HT (1 microM). During the early phase of desensitization, the 5HT2 receptor was coupled to phospholipase C, whereas during the late phase of desensitization this coupling was disconnected. However, after disappearance of the agonist, the coupling in the resting platelet recovered quickly, and was nearly complete (82%) after 30 min. During this resensitization, the 5HT-inducibility of activation of phospholipase C, of the increase in cytosolic [Ca2+] and of stimulation of protein kinase C were restored in parallel. The time course for resensitization of the 5HT-induced increase in cytosolic [Ca2+] was independent of the presence of extracellular Ca2+. It is concluded that, after dissociation of 5HT from the platelet 5HT2-receptor, 5HT-induced responses rapidly resensitize. Because of its short duration and the parallelism in recovery between the different 'down-stream phospholipase C' intracellular transduction signals, it is considered that desensitization arises from a reversible change in the transduction mechanism at a step up to or including the activation of phospholipase C. Neither desensitization nor resensitization to 5HT is dependent on the presence of extracellular Ca2+.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Benya R. V., Kusui T., Shikado F., Battey J. F., Jensen R. T. Desensitization of neuromedin B receptors (NMB-R) on native and NMB-R-transfected cells involves down-regulation and internalization. J Biol Chem. 1994 Apr 22;269(16):11721–11728. [PubMed] [Google Scholar]
  2. Brass L. F. Homologous desensitization of HEL cell thrombin receptors. Distinguishable roles for proteolysis and phosphorylation. J Biol Chem. 1992 Mar 25;267(9):6044–6050. [PubMed] [Google Scholar]
  3. Briscoe C. P., Plevin R., Wakelam M. J. Rapid desensitization and resensitization of bombesin-stimulated phospholipase D activity in Swiss 3T3 cells. Biochem J. 1994 Feb 15;298(Pt 1):61–67. doi: 10.1042/bj2980061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burnstock G. Review lecture. Neurotransmitters and trophic factors in the autonomic nervous system. J Physiol. 1981;313:1–35. doi: 10.1113/jphysiol.1981.sp013648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crouch M. F., Lapetina E. G. A role for Gi in control of thrombin receptor-phospholipase C coupling in human platelets. J Biol Chem. 1988 Mar 5;263(7):3363–3371. [PubMed] [Google Scholar]
  6. Crouch M. F., Lapetina E. G. Dual mechanisms of platelet hormone receptor desensitization. Differential importance between agonists of protein kinase C-dependent and -independent pathways. J Biol Chem. 1989 Jan 5;264(1):584–588. [PubMed] [Google Scholar]
  7. Crouch M. F., Lapetina E. G. Spacial isolation of protein kinase C activation in thrombin stimulated human platelets. Biochem Biophys Res Commun. 1988 Oct 14;156(1):149–156. doi: 10.1016/s0006-291x(88)80817-9. [DOI] [PubMed] [Google Scholar]
  8. De Clerck F., Loots W., Somers Y., Beetens J., Wouters L., Wynants J., Janssen P. A. 5-Hydroxytryptamine and arachidonic acid metabolites modulate extensive platelet activation induced by collagen in cats in vivo. Br J Pharmacol. 1990 Apr;99(4):631–636. doi: 10.1111/j.1476-5381.1990.tb12982.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  10. Hug H., Sarre T. F. Protein kinase C isoenzymes: divergence in signal transduction? Biochem J. 1993 Apr 15;291(Pt 2):329–343. doi: 10.1042/bj2910329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kagaya A., Mikuni M., Kusumi I., Yamamoto H., Takahashi K. Serotonin-induced acute desensitization of serotonin2 receptors in human platelets via a mechanism involving protein kinase C. J Pharmacol Exp Ther. 1990 Oct;255(1):305–311. [PubMed] [Google Scholar]
  12. Kanner B. I., Schuldiner S. Mechanism of transport and storage of neurotransmitters. CRC Crit Rev Biochem. 1987;22(1):1–38. doi: 10.3109/10409238709082546. [DOI] [PubMed] [Google Scholar]
  13. Lefkowitz R. J., Hausdorff W. P., Caron M. G. Role of phosphorylation in desensitization of the beta-adrenoceptor. Trends Pharmacol Sci. 1990 May;11(5):190–194. doi: 10.1016/0165-6147(90)90113-m. [DOI] [PubMed] [Google Scholar]
  14. Muallem S., Pandol S. J., Beeker T. G. Hormone-evoked calcium release from intracellular stores is a quantal process. J Biol Chem. 1989 Jan 5;264(1):205–212. [PubMed] [Google Scholar]
  15. Neer E. J., Clapham D. E. Roles of G protein subunits in transmembrane signalling. Nature. 1988 May 12;333(6169):129–134. doi: 10.1038/333129a0. [DOI] [PubMed] [Google Scholar]
  16. Pollock W. K., MacIntyre D. E. Desensitization and antagonism of vasopressin-induced phosphoinositide metabolism and elevation of cytosolic free calcium concentration in human platelets. Biochem J. 1986 Feb 15;234(1):67–73. doi: 10.1042/bj2340067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rasmussen H. The cycling of calcium as an intracellular messenger. Sci Am. 1989 Oct;261(4):66–73. doi: 10.1038/scientificamerican1089-66. [DOI] [PubMed] [Google Scholar]
  18. Rink T. J., Pozzan T. Using quin2 in cell suspensions. Cell Calcium. 1985 Apr;6(1-2):133–144. doi: 10.1016/0143-4160(85)90040-5. [DOI] [PubMed] [Google Scholar]
  19. Roevens P., De Clerck F., de Chaffoy de Courcelles D. The synergistic effect of 5-hydroxytryptamine and epinephrine on the human platelet is related to the activation of phospholipase C. Eur J Pharmacol. 1993 May 15;245(3):273–280. doi: 10.1016/0922-4106(93)90107-k. [DOI] [PubMed] [Google Scholar]
  20. Siess W. Molecular mechanisms of platelet activation. Physiol Rev. 1989 Jan;69(1):58–178. doi: 10.1152/physrev.1989.69.1.58. [DOI] [PubMed] [Google Scholar]
  21. Watson S. P., McNally J., Shipman L. J., Godfrey P. P. The action of the protein kinase C inhibitor, staurosporine, on human platelets. Evidence against a regulatory role for protein kinase C in the formation of inositol trisphosphate by thrombin. Biochem J. 1988 Jan 15;249(2):345–350. doi: 10.1042/bj2490345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wojcikiewicz R. J., Tobin A. B., Nahorski S. R. Desensitization of cell signalling mediated by phosphoinositidase C. Trends Pharmacol Sci. 1993 Jul;14(7):279–285. doi: 10.1016/0165-6147(93)90131-3. [DOI] [PubMed] [Google Scholar]
  23. Yakel J. L., Lagrutta A., Adelman J. P., North R. A. Single amino acid substitution affects desensitization of the 5-hydroxytryptamine type 3 receptor expressed in Xenopus oocytes. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5030–5033. doi: 10.1073/pnas.90.11.5030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. de Chaffoy de Courcelles D., Leysen J. E., De Clerck F., Van Belle H., Janssen P. A. Evidence that phospholipid turnover is the signal transducing system coupled to serotonin-S2 receptor sites. J Biol Chem. 1985 Jun 25;260(12):7603–7608. [PubMed] [Google Scholar]
  25. de Chaffoy de Courcelles D., Roevens P., Van Belle H. 1-Oleoyl-2-acetyl-glycerol (OAG) stimulates the formation of phosphatidylinositol 4-phosphate in intact human platelets. Biochem Biophys Res Commun. 1984 Sep 17;123(2):589–595. doi: 10.1016/0006-291x(84)90270-5. [DOI] [PubMed] [Google Scholar]
  26. de Chaffoy de Courcelles D., Roevens P., Van Belle H., Kennis L., Somers Y., De Clerck F. The role of endogenously formed diacylglycerol in the propagation and termination of platelet activation. A biochemical and functional analysis using the novel diacylglycerol kinase inhibitor, R 59 949. J Biol Chem. 1989 Feb 25;264(6):3274–3285. [PubMed] [Google Scholar]
  27. de Chaffoy de Courcelles D., Roevens P., Wynants J., Van Belle H. Serotonin-induced alterations in inositol phospholipid metabolism in human platelets. Biochim Biophys Acta. 1987 Feb 18;927(2):291–302. doi: 10.1016/0167-4889(87)90146-7. [DOI] [PubMed] [Google Scholar]
  28. de Chaffoy de Courcelles D., Roevens P., van Belle H. Stimulation by serotonin of 40 kDa and 20 kDa protein phosphorylation in human platelets. FEBS Lett. 1984 Jun 11;171(2):289–292. doi: 10.1016/0014-5793(84)80506-2. [DOI] [PubMed] [Google Scholar]
  29. de Clerck F., David J. L., Janssen P. A. Inhibition of 5-hydroxytryptamine-induced and -amplified human platelet aggregation by ketanserin (R 41 468), a selective 5-HT2-receptor antagonist. Agents Actions. 1982 Jul;12(3):388–397. doi: 10.1007/BF01965409. [DOI] [PubMed] [Google Scholar]

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