Health  Vol.5 No.4 A , April 2013
Serotonin uptake rates in platelets from angiotensin II-induced hypertensive mice

Angiotensin II (Ang II) is a critical component of the reninangiotensin system that contributes to hypertension. Although platelets in blood from hypertensive subjects have an abnormal biological profile, it is unclear if circulating Ang II influences platelet aggregation or thrombus formation. One of the abnormalities presented to the platelets during hypertension is an elevated plasma concentration of serotonin (5-HT) caused by reduced 5-HT uptake secondary to loss of the 5-HT transporter (SERT) on the platelet plasma membrane. In the current study, we evaluated in vivo platelet function after 7 days of subcutaneous Ang II infusion to establish hypertension in mice and additionally assessed the biology of isolated platelets exposed to Ang II in vitro. The administration of Ang II elevated systolic blood pressure, but markers of platelet activation including P-selectin and PEJon/A staining were not changed. However, the aggregation response to collagen was reduced in isolated platelets from Ang II-infused mice, which also showed reduced 5-HT uptake by SERT. In vitro exposure of isolated platelets to Ang II also resulted in a loss of surface SERT associated with a reduced aggregation response to collagen. These abnormalities were reversed by increasing concentra tions of the Ang II receptor antagonist, valsartan. Interestingly, SERT KO mice failed to fully develop hypertension in response to Ang II infusion and isolated platelets from these animals were insensitive to the anti-aggregatory influence of Ang II. Thus, Ang II blunts the aggregation responses of platelets and the mechanism underlying this action may involve a loss of SERT on the platelet plasma membrane. The latter event depletes intracellular 5-HT in platelets, an event that is associated with reduced aggregation. The widespread use of antihypertensive drugs that target the renin-angiotensin system suggest the potential clinical utility of our findings and emphasize the importance of understanding the impact of Ang II on platelet function.

Cite this paper: Singh, P. , Fletcher, T. , Li, Y. , Rusch, N. and Kilic, F. (2013) Serotonin uptake rates in platelets from angiotensin II-induced hypertensive mice. Health, 5, 31-39. doi: 10.4236/health.2013.54A005.

[1]   Michelson, A.D. (2012) Platelets “interactions between platelets and the coagulation system” and “the role of platelets in fibrinolysis”. 3rd Edition, Academic Press, Boston, 425, 469.

[2]   Minuz, P., Patrignani, P., Gaino, S., Seta, F., Capone, M.L., Tacconelli, S., Degan, M., Faccini, G., Fornasiero, A., Talamini, G., Tommasoli, R., Arosio, E., Santonastaso, C.L., Lechi, A. and Patrono, C. (2004) Determinants of platelet activation in human essential hypertension. Hypertension, 43, 64-70. doi:10.1161/01.HYP.0000105109.44620.1B

[3]   Kerr, S., Brosnan, M.J., McIntyre, M., Reid, J.L., Dominiczak, A.F. and Hamilton, C.A. (1999) Superoxide anion production is increased in a model of genetic hypertension: Role of the endothelium. Hypertension, 33, 1353-1358. doi:10.1161/01.HYP.33.6.1353

[4]   Zalba, G., Beaumont, F.J., San Jose, G., Fortuno, A., Fortuno, M.A., Etayo, J.C. and Diez, J. (2000) Vascular NADH/NADPH oxidase is involved in enhanced superoxide production in spontaneously hypertensive rats. Hypertension, 35, 1055-1061. doi:10.1161/01.HYP.35.5.1055

[5]   Hernandez Hernandez, R., Carvajal, A.R. and Guerrero Pajuelo, J. (1991) The effect of doxazosin on platelet aggregation in normotensive subjects and patients with hypertension: An in vitro study. American Heart Journal, 121, 389-394. doi:10.1016/0002-8703(91)90877-K

[6]   Andrioli, G., Ortolani, R., Fontana, L., Gaino, S., Bellavite, P., Lechi, C., Minuz, P., Manzato, F., Tridente, G. and Lechi, A. (1996) Study of platelet adhesion in patients with uncomplicated hypertension. Journal of Hypertension, 14, 1215-1221. doi:10.1097/00004872-199610000-00010

[7]   El Haouari, M. and Rosado, J.A. (2009) Platelet function in hypertension. Blood Cells, Molecules and Diseases, 42, 38-43. doi:10.1016/j.bcmd.2008.07.003

[8]   Senchenkova, E.Y., Russell, J., Almeida-Paula, L.D., Harding, J.W. and Granger, D.N. (2010) Angiotensin II-mediated microvascular thrombosis. Hypertension, 56, 1089- 1095. doi:10.1161/HYPERTENSIONAHA.110.158220

[9]   Kalinowski, L., Matys, T., Chabielska, E., Buczko and W., Malinski, T. (2002) Angiotensin II AT1 receptor antagonists inhibit platelet adhesion and aggregation by nitric oxide release. Hypertension, 40, 521-527. doi:10.1161/01.HYP.0000034745.98129.EC

[10]   Brown, N.J. and Vaughan, D.E. (2000) Prothrombotic effects of angiotensin. Advances in Internal Medicine, 45, 419-429.

[11]   Unger, T., Culman, J. and Gohlke, P. (1998) Ang II receptor blockade and end-organ protection: Pharmacological rationale and evidence. Journal of Hypertension, 16, S3-S9.

[12]   Gkaliagkousi, E., Ritter, J. and Ferro, A. (2007) Platelet-derived nitric oxide signaling and regulation. Circulation Research, 101, 654-662. doi:10.1161/CIRCRESAHA.107.158410

[13]   Taddei, S., Virdis, A., Ghiadoni, L., Mattei, P., Salvetti, A. (1998) Effects of angiotensin converting enzyme inhibition on endothelium-dependent vasodilatation in essential hypertensive patients. Journal of Hypertension, 16, 447- 456. doi:10.1097/00004872-199816040-00006

[14]   Muthalif, M.M., Karzoun, N.A., Gaber, L., Khandekar, Z., Benter, I.F., Saeed, A.E., Parmentier, J.H., Estes, A. and Malik, K.U. (2000) Angiotensin II-induced hypertension: Contribution of Ras GTPase/Mitogen-activated protein kinase and cytochrome P450 metabolites. Hypertension, 36, 604-609. doi:10.1161/01.HYP.36.4.604

[15]   Rosenblum, W.I., El-Sabban, F. and Hirsh, P.D. (1986). Angiotensin delays platelet aggregation after injury of cerebral arterioles. Stroke, 17, 1203-1205. doi:10.1161/01.STR.17.6.1203

[16]   Ziu, E., Freyaldenhoven S., Mercado, C., Lensing S., Ware, J., Ahmed, B. and Kilic, F. (2012) Down-regulation of the serotonin transporter in hyperreactive platelets counteracts the pro-thrombotic effect of serotonin. Journal of Molecular and Cellular Cardiology, 5, 1112-1121. doi:10.1016/j.yjmcc.2012.02.004

[17]   Brenner, B., Harney, J.T., Ahmed, B.A., Jeffus, B.C., Unal, R., Mehta, J.L. and Kilic, F. (2007) Plasma serotonin level and the platelet serotonin transporter. Journal of Neurochemistry, 102, 206-215. doi:10.1111/j.1471-4159.2007.04542.x

[18]   Ottervanger, J.P., Stricker, B.H., Huls, J. and Weeda, J.N. (1994) Bleeding attributed to the intake of paroxetine. American Journal of Psychiatry, 151, 781-782.

[19]   Carneiro, A.M., Cook, E.H., Murphy, D.L. and Blakely, R.D. (2008) Interactions between integrin alphaIIbbeta3 and the serotonin transporter regulate serotonin transport and platelet aggregation in mice and humans. Journal of Clinical Investigation, 118, 1544-1552. doi:10.1172/JCI33374

[20]   Kelm, S. and Schauer, R. (1997) Sialic acids in molecular and cellular interactions. International Review of Cytology, 175, 137-240. doi:10.1016/S0074-7696(08)62127-0

[21]   Tanaka, H., Ishida, Y., Kaneko, T. and Matsumoto, N. (1989) Isolation of human megakaryocytes by immunomagnetic beads. British Journal of Haematology, 73, 18-22. doi:10.1111/j.1365-2141.1989.tb00212.x

[22]   Sato, N., Kiyokawa, N., Takada, K., Itagaki, M., Saito, M., Sekino, T., Suzuki, T., Taguchi, T., Mimori, K., Lanza, F. and Fujimoto, J. (2000) Characterization of monoclonal antibodies against mouse and rat platelet glycoprotein V (CD42d). Hybridoma, 19, 455-461. doi:10.1089/027245700750053940

[23]   Berry, C.N., Lorrain, J., Lochot, S., Delahaye, M., Lalé, A., Savi, P., Lechaire, I., Ferrari, P., Bernat, A., Schaeffer, P., Janiak, P., Duval, N., Grosset, A., Herbert, J.M. and O’Connor, S.E. (2001) Antiplatelet and antithrombotic activity of SL65.0472, a mixed 5-HT1B/5-HT2A receptor antagonist. Journal of Thrombosis and Haemostasis, 85, 521-528.

[24]   Ottervanger, J.P., Stricker, B.H., Huls, J. and Weeda, J.N. (1994) Bleeding attributed to the intake of paroxetine. American Journal of Psychiatry, 151, 781-782.

[25]   Przyklenk, K. (2010) Targeted inhibition of the serotonin 5-HT2A receptor improves coronary patency in an in vivo model of recurrent thrombosis. Journal of Thrombosis and Haemostasis, 8, 331-340. doi:10.1111/j.1538-7836.2009.03693.x

[26]   Nishihira, K. (2006) Inhibition of 5-hydroxytryptamine receptor prevents occlusive thrombus formation on neointima of the rabbit femoral artery. Journal of Thrombosis and Haemostasis, 4, 247-255. doi:10.1111/j.1538-7836.2005.01702.x

[27]   Walther, D.J., Peter, J.U., Winter, S., Höltje, M., Paulmann, N., Grohmann, M., Vowinckel, J., Alamo-Bethencourt, V., Wilhelm, C.S., Ahnert-Hilger, G. and Bader, M. (2003) Serotonylation of small GTPases is a signal transduction pathway that triggers platelet alpha-granule release. Cell, 115, 851-862. doi:10.1016/S0092-8674(03)01014-6

[28]   Shirakawa, R., Yoshioka, A., Horiuchi, H., Nishioka, H., Tabuchi, A. and Kita, T. (2000) Small GTPase Rab4 regulates Ca2+-induced α-granule secretion in platelets. The Journal of Biological Chemistry, 275, 33844-33849. doi:10.1074/jbc.M002834200

[29]   Ahmed, B.A., Jeffus, B.C., Bukhari, S.I., Harney, J.T., Unal, R., Lupashin, V.V., van der Sluijs, P. and Kilic, F. (2008) Serotonin transamidates Rab4 and facilitates its binding to the C terminus of serotonin transporter. The Journal of Biological Chemistry, 283, 9388-9398. doi:10.1074/jbc.M706367200

[30]   Kharade, S., Sonkusare, S., Srivastava, A., Thakali, K., Fletcher, T.W., Rhee, S.W. and Rusch N.J. (2013) The β3 subunit contributes to vascular calcium channel upregulation and hypertension in angiotensin II-infused C57BL/6 mice. Hypertension, 61, 137-142. doi:10.1161/HYPERTENSIONAHA.112.197863

[31]   Nasjletti, A. and Malik, K.U. (1982) Interrelations between prostaglandins and vasoconstrictor hormones: Contribution to blood pressure regulation. Federation Proceedings, Al, 2394-2399.

[32]   Ratliff, B.B., Sekulic, M., Rodebaugh, J. and Solhaug, M.J. (2010) Angiotensin II regulates NOS expression in afferent arterioles of the developing porcine kidney. Pediatric Research, 68, 29-34. doi:10.1203/PDR.0b013e3181e12770

[33]   Freedman, J.E., Loscalzo, J., Barnard, M.R., Alpert, C., Keaney, J.F. and Michelson, A.D. (1997) Nitric oxide released from activated platelets inhibits platelet recruitment. Journal of Clinical Investigation, 100, 350-356. doi:10.1172/JCI119540

[34]   Kilic, F. and Rudnick, G. (2000) Oligomerization of the serotonin transporter and its functional consequences. Proceedings of the National Academy of Sciences USA, 97, 3106-3111. doi:10.1073/pnas.97.7.3106

[35]   Homberg, J.H., Mudde, J.M., Braam, B., Ellenbroek, B. and Cuppen, E. (2006) Blood pressure in mutant rats lacking the 5-hydroxytryptamine transporter. Hypertension, 48, e115-e116. doi:10.1161/01.HYP.0000246306.61289.d8

[36]   Bengel, D., Murphy, D.L. and Andrews, A.M. (1998) Altered brain serotonin homeostasis and locomotor insensitivity to 3,4-methylenedioxymethamphetamine (“ecstasy”) in serotonin transporter deficient mice. Molecular Pharmacology, 53, 649-655.

[37]   Guy, L.R. (2006) Platelet secretion. In: Michelson, A.D., Ed., Platelets, Academic Press, Boston, 309-319.

[38]   James, G. (2006) White platelet structure. In: Michelson, A.D., Ed., Platelets, Academic Press, New York, 45-75.

[39]   Saxena, P.R. and Villalón, C.M. (1990) Cardiovascular effects of serotonin agonists and antagonists. Journal of Cardiovascular Pharmacology, 7, S17-S34.

[40]   Watts, S.W. (2005) 5-HT in systemic hypertension: Foe, friend or fantasy? Clinical Science (London), 108, 399- 412. doi:10.1042/CS20040364

[41]   Doggrell, S.A. (2003) The role of 5-HT on the cardiovascular and renal systems and the clinical potential of 5- HT modulation. Expert Opinion on Investigational Drugs, 12, 805-823. doi:10.1517/13543784.12.5.805

[42]   Kilic, F., Ziu, E. and Freyaldenhoven, S. (2009) Plasma serotonin and platelet serotonin transporter: Molecular and cellular aspects in cardiovascular research. Chapter IX. In: Brataas, J. and Nanstveit, V., Eds., Handbook of Cardiovascular Research, Nova Science Publishers, Inc., Hauppauge, 792-797.

[43]   Mercado, C. and Kilic, F. (2010) The impact of plasma serotonin on platelet biology. Molecular Interventions, 10, 338-349.

[44]   Mercado, C., Ziu, E. and Kilic, F. (2011) Communication between 5-HT and small GTPases. Current Opinion in Pharmacology, 11, 23-28. doi:10.1016/j.coph.2011.01.006

[45]   Freedman, J.E., Loscalzo, J., Barnard, M.R., Alpert, C., Keaney, J.F. and Michelson, A.D. (1997) Nitric oxide released from activated platelets inhibits platelet recruitment. The Journal of Clinical Investigation, 100, 350-356. doi:10.1172/JCI119540