Molostvov, Guerman, Fletcher, Simon, Bland, Rosemary and Zehnder, Daniel. (2008) Extracellular calcium-sensing receptor mediated signalling is involved in human vascular smooth muscle cell proliferation and apoptosis. Cellular Physiology and Biochemistry, Vol.22 (No.5-6). pp. 413-422. ISSN 1015-8987

Preview	PDF WRAP_Bland_0070700_230109_Molostvov.pdf - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader Download (206Kb)
Preview	PDF (Figures) WRAP_Bland_0070700-230109-Molostvov_figures.pdf - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader Download (328Kb)

Abstract

Calcium-sensing receptor (CaSR) plays key role in vascular calcification in patients with chronic kidney disease (CKD). We investigated the role of CaSR in regulating smooth muscle cell (SMC) proliferation and apoptosis. Incubation with 300µM neomycin (CaSR agonist) resulted in 7.5-fold (p<0.05) increase in ERK1,2 phosphorylation. It was reduced (p<0.01) by 10µM PD98059 (MEK1 inhibitor), indicating that CaSR agonist-induced effects were mediated via MEK1/ERK1,2 pathway. ERK1,2 phosphorylation was abolished by 5µM U73122 (PLC inhibitor), indicating that PLC signalling was crucial for MEK1/ERK1,2 activation. Confirming PLC activation, inositol triphosphate (IP3) production was increased by neomycin/gentamycin (p<0.05) and reduced by U73122. To confirm that ERK1,2 and PLC signalling were mediated via CaSR, Human Aortic SMC (HAoSMC) were transfected with CaSR siRNA. CaSR knockdown resulted in lower ERK1,2 neomycin response and IP3 production (p<0.01). Neomycin increased HAoSMC proliferation >3-fold, which was reduced in CaSR knockdown cells (p<0.01) and further inhibited by PD98059 and U73122 (p<0.05). Apoptosis was not affected by neomycin treatment. U73122 produced 3.5-fold increase in HAoSMC apoptosis, which was further increased by CaSR knockdown (5-fold, p<0.05). In conclusion, stimulation of CaSR leads to activation of MEK1/ERK1,2 and PLC pathways and up-regulation of cell proliferation. CaSR-mediated PLC activation is important for SMC survival and protection against apoptosis.

Item Type:	Journal Article
Subjects:	R Medicine > RC Internal medicine
Divisions:	Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010) Faculty of Medicine > Warwick Medical School > Clinical Sciences Research Institute (CSRI) Faculty of Medicine > Warwick Medical School > Education Development and Research Faculty of Medicine > Warwick Medical School > Metabolic and Vascular Health Faculty of Medicine > Warwick Medical School
Library of Congress Subject Headings (LCSH):	Cardiovascular system -- Calcification, Kidneys -- Diseases, Apoptosis
Journal or Publication Title:	Cellular Physiology and Biochemistry
Publisher:	S. Karger AG
ISSN:	1015-8987
Date:	9 December 2008
Volume:	Vol.22
Number:	No.5-6
Page Range:	pp. 413-422
Identification Number:	10.1159/000185484
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access
Funder:	Diabetes, Endocrine and Immersion Research Trust, Coventry Kidney Research Fund, Coventry and Warwickshire Kidney Patient Association
References:	1. Brown EM, MacLeod RJ: Extracellular calcium sensing and extracellular calcium signaling. Physiol Rev 2001; 81: 239-297. 2. Hofer AM, Brown EM: Extracellular calcium sensing and signaling. Nat Rev Mol Cell Biol 2003; 4: 530-538. 3. Brown EM, Gamba G, Riccardi D, Lombardi M, Butters R, Kifor O, Sun A, Hediger MA, Lytton J, Hebert SC: Cloning and characterisation of an extracellular Ca2+-sensing receptor from bovine parathyroid. Nature 1993; 366: 575-580. 4. Tfelt-Hansen J, Brown EM: The calcium-sensing receptor in normal physiology and pathophysiology: a review. Crit Rev Clin Lab Sci 2005; 42: 35-70. 5. Brown EM, Vassilev PM, Quinn S, Hebert SC: G-protein-coupled, extracellular Ca(2+)-sensing receptor: a versatile regulator of diverse cellular functions. Vitam Horm 1999; 55: 1-71. 6. Olszak IT, Poznansky MC, Evans RH, Olson D, Kos C, Pollak MR, Brown EM, Scadden DT: Extracellular calcium elicits a chemokinetic response from monocytes in vitro and in vivo. J Clin Invest 2000; 105: 1299-1305. 7. Smajilovic S, Tfelt-Hansen J: Calcium acts as a first messenger through the calcium-sensing receptor in the cardiovascular system. Cardiovasc Res 2007; 75: 457-467. 8. Tfelt-Hansen J, Hansen JL, Smajilovic S, Brown EM, Haunso S, Sheikh SP: The calcium receptor is functionally expressed in rat neonatal ventricular cardiomyocytes. Am J Physiol Heart Circ Physiol 2006; 290: H1165-H1171. 9. Wang R, Xu C, Zhao W, Zhang J, Cao K, Yang B, Wu L: Calcium and polyamine regulated calcium-sensing receptors in cardiac tissues. Eur J Biochem 2003; 270: 2680-2688. 10. Weston AH, Absi M, Ward DT, Ohanian J, Dodd RH, Dauban P, Petrel C, Ruat M, Edwards G: Evidence in favour of a calcium-sensing receptor in arterial endothelial cells. Studies with calindol and calhex 231. Circ Res 2005; 97: 391-398. 11. Ziegelstein RC, Xiong Y, He C, Hu Q: Expression of a functional extracellular calcium-sensing receptor in human aortic endothelial cells. Biochem Biophys Res Commun 2006; 342: 153-163. 12. Wonneberger K, Scofield MA, Wangemann P: Evidence for a calcium-sensing receptor in the vascular smooth muscle cells of the spiral modiolar artery. J Membrane Biol 2000; 175: 203-212. 13. Ohanian J, Gatfield KM, Ward DT, Ohanian V: Evidence for a functional calcium-sensing receptor that modulates myogenic tone in rat subcutaneous small arteries. Am J Physiol Heart Circ Physiol 2005; 288: H1756-H1762. 14. Smajilovic S, Hansen JL, Christoffersen TEH, Lewin E, Sheikh SP, Terwilliger EF, Brown EM, Haunso S, Tfelt-Hansen J: Extracellular calcium sensing in rat aortic vascular smooth muscle cells. Biochem Biophys Res Commun 2006; 348: 1215-1223. 15. Molostvov G, James S, Fletcher S, Bennett J, Lehnert H, Bland R, Zehnder D: Extracellular calcium-sensing receptor is functionally expressed in human artery. Am J Physiol Renal Physiol 2007; 293: F946-F955. 16. Giachelli CM: Vascular calcification mechanisms. J Am Soc Nephrol 2004; 15: 2959-2964. 17. Moe SM, Chen NX: Pathophysiology of vascular calcification in chronic kidney disease. Circ Res 2004; 95: 560-567. 18. Goodman WG, London G, Amann K, Block GA, Giachelli C, Hruska KA, Katteler M, Levin A, Massy Z, McCarron DA: Vascular calcification in chronic kidney disease. Am J Kidney Dis 2004; 43: 572-579. 19. Reynolds JL, Joannides AJ, Skepper JN, McNair R, Schurgers LJ, Proudfoot D, Jahnen-Dechent W, Weissberg PL, Shanahan CM: Human vascular smooth muscle cells undergo vesicle-mediated calcification in response to changes in extracellular calcium and phosphate concentrations: a potential mechanism for accelerated vascular calcification in ESRD. J Am Soc Nephrol 2004; 15: 2857-2867. 20. Block GA, Klassen PS, Lazarus JM, Ofsthun N, Lowrie EG, Chertow GM: Mineral metabolism, mortality, and morbidity in maintenance hemodialysis. J Am Soc Nephrol 2004; 15: 2208-2218. 21. Yang H, Curinga G, Giachelli CM: Elevated extracellular calcium levels induce smooth muscle cell matrix mineralization in vitro. Kidney Int 2004; 66: 2293-2299. 22. Bostrom K, Watson KE, Horn S, Wortham C, Herman IM, Demer LL: Bone morphogenetic protein expression in human atherosclerotic lesions. J Clin Invest 1993; 91: 1800-1809. 23. Shioi A, Nishizawa Y, Jono S, Koyama H, Hosoi M, Morii H: Betaglycerophosphate accelerates calcification in cultured bovine vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 1995; 15: 2003-2009. 24. Tyson KL, Reynolds JL, McNair R, Zhang Q, Weissberg PL, Shanahan CM: Osteo/chondrocytic transcription factors and their target genes exhibit distinct patterns of expression in human arterial calcification. Arterioscler Thromb Vasc Biol 2003; 23: 489-494. 25. Chattopadhyay N, Yano S, Tfelt-Hansen J, Rooney P, Kanuparthi D, Bandyopadhyay S, Ren X, Terwilliger E, Brown EM: Mitogenic action of calcium-sensing receptor on rat calvarial osteoblasts. Endocrinology 2004; 145: 3451-3462. 26. Shroff R, McNair R, Figg N, Donald A, Deanfield J, Rees L, Shanahan C: Accelerated vascular calcification in dialysis patients is a vesicle-mediated process associated with apoptosis and aosteoblastic differentiation in a model of intact human arteries. JASN 19, Abstract Issue, November, 2007. 27. Alam M, Kirton JP, Wilkinson FL, Towers E, Rouhi M, Rucka A, Alexander YM, Martin D, Riccardi D, Canfield AE: The calcimimetic R-568 inhibits mineral deposition by vascular smooth muscle cells. JASN 19, Abstract Issue, November, 2007. 28. Hills CE, Bland R, Wheelans DC, Bennett J, Ronco PM, Squires PE: Glucose-evoked alterations in connexin 43-mediated cell-to-cell communication in human collecting duct: a possible role in diabetic nephropathy. Am J Physiol Renal Physiol 2006; 291: F1045-F1051. 29. Tanoue T, Nishida E: Molecular recognitions in the MAP kinase cascades. Cell Signal 2003; 15: 455-462. 30. Kifor O, MacLeod RJ, Diaz R, Bai M, Yamaguchi T, Yao T, Kifor I, Brown EM: Regulation of MAP kinase by calcium-sensing receptor in bovine parathyroid and CaR-transfected HEK293 cells. Am J Physiol Renal Physiol 2001; 280: F291-F302. 31. Huang Z, Cheng SL, Slatopolsky E: Sustained activation of the extracellular signal-regulated kinase pathway is required for extracellular calcium stimulation of human osteoblast proliferation. J Biol Chem 2001; 276: 21351- 21358. 32. Ward DT, McLarnon SJ, Riccardi D: Aminoglycosides increase intracellular calcium levels and ERK activity in proximal tubular OK cells expressing the extracellular calcium-sensing receptor. J Am Soc Nephrol 2002; 13: 1481-1489. 33. Schnaper HW: Cell signal transduction through the mitogen-activated protein kinase pathway. Pediatr Nephrol 1998; 12: 790-795. 34. Cho HM, Choi SH, Hwang KC, Oh SY, Kim HG, Yoon DH, Choi MA, Lim SY, Song H, Jang Y, Kim TW: The Src/PLC/PKC/MEK/ERK signaling pathway is involved in aortic smooth muscle cell proliferation induced by glycated LDL. Mol Cell 2004; 19: 60-66. 35. Riccardi D, Park J, Lee W, Gamba G, Brown EM, Hebert SC: Cloning and functional expression of a rat kidney extracellular calcium/polyvalent cation-sensing receptor. Proc Natl Acad Sci USA 1995; 92: 131-135. 36. Kifor O, Diaz R, Butters R, Brown EM: The Ca2+-sensing receptor (CaR) activates phospholipases C, A2 and D in bovine parathyroid and CaR-transfected human embryonic kidney (HEK293) cells. J Bone Miner Res 1997; 12: 715-725. 37. Bai M, Trivedi S, Lane CR, Yang Y, Quinn SJ, Brown EM: Protein kinase C phosphorylation of threonine at position 888 in Ca2+-sensing receptor (CaR) inhibits coupling to Ca2+ store release. J Biol Chem 1998; 273: 21267-21275. 38. Ward DT, Maldonado-Perez D, Hollins L, Riccardi D: Aminoglycosides induce acute cell signaling and chronic cell death in renal cells that express the calcium-sensing receptor. J Am Soc Nephrol 2005; 16: 1236-1244. 39. Wellendorph P, Brauner-Osborne H: Molecular cloning, expression and sequence analysis of GPRC6A, a novel family C G-protein-coupled receptor. Gene 2004; 335: 37-46. 40. Lin KI, Chattopadhyay N, Bai M, Alvarez R, Dang CV, Baraban JM, Brown EM, Ratan RR: Elevated extracellular calcium can prevent apoptosis via the calcium-sensing receptor. Biochem Biophys Res Commun 1998; 249: 325-331. 41. Kauffmann-Zeh A, Rodriguez-Viciana P, Ulrich E, Gilbert C, Coffer P, Downward J, Evan G: Suppression of c-Myc-induced apoptosis by Ras signalling through PI(3)K and PKB. Nature 1997; 385: 544-548. 42. Cornelissen J, Armstrong J, Holt CM: Mechanical stretch induces phosphorylation of p38-MAPK and apoptosis in human saphenous vein. Arterioscler Thromb Vasc Biol 2004; 24: 451-456. 43. Ichijo H, Nishida E, Irie K, ten Dijke P, Saitoh M, Moriguchi T, Takagi M, Matsumoto K, Miyazono K, Gotoh Y: Induction of apoptosis by ASK1, a mammalian MAPKKK that activates SAPK/JNK and p38 signaling pathways. Science 1997; 275: 90-94. 44. Sun Y, Liu M, Li H, Shi S, Zhao Y, Wang R, Xu C: Calcium-sensing receptor induces rat neonatal ventricular cardiomyocyte apoptosis. Biochem Biophys Res Commun 2006; 350: 942-948.
URI:	http://wrap.warwick.ac.uk/id/eprint/394

Data sourced from Thomson Reuters' Web of Knowledge

Request changes to a record

Actions (login required)

View Item