Bone morphogenetic protein 4 enhances canonical transient receptor potential expression , store-operated Ca 2 entry , and basal [ Ca 2 ] i in rat distal pulmonary arterial smooth muscle cells

Lu W, Ran P, Zhang D, Lai N, Zhong N, Wang J. Bone morphogenetic protein 4 enhances canonical transient receptor potential expression, store-operated Ca entry, and basal [Ca ]i in rat distal pulmonary arterial smooth muscle cells. Am J Physiol Cell Physiol 299: C1370–C1378, 2010. First published September 15, 2010; doi:10.1152/ajpcell.00040.2010.—Recent advances have identified an important role of bone morphogenetic protein 4 (BMP4) in pulmonary vascular remodeling, yet the underlying mechanisms remain largely unexplored. We have previously found that Ca influx through store-operated calcium channels (SOCC), which are mainly thought to be composed of canonical transient receptor potential (TRPC) proteins, likely contribute to the pathogenic development of chronic hypoxic pulmonary hypertension. In this study, we investigated the effect of BMP4 on expression of TRPC and store-operated Ca entry (SOCE) in pulmonary arterial smooth muscle cells (PASMCs). Real-time quantitative PCR and Western blotting revealed that treatment with BMP4 (50 ng/ml, 60 h) increased TRPC1, TRPC4, and TRPC6 mRNA and protein expression in growth-arrested rat distal PASMCs. Moreover, in comparison to vehicle control, cells treated with BMP4 also exhibited enhanced SOCE, and elevated basal intracellular calcium concentration ([Ca ]i) as determined by fluorescent microscopy using the Ca indicator Fura-2 AM. Perfusing cells with Ca -free Krebs-Ringer bicarbonate solution (KRBS) or KRBS containing SOCC antagonists SKF-96365 or NiCl2 attenuated the increases in basal [Ca ]i caused by BMP4. Specific knockdown of BMP4 by small interference RNA significantly decreased the mRNA and protein expression of TRPC1, TRPC4, and TRPC6 and reduced SOCE and basal [Ca ]i in serum-stimulated PASMCs. We conclude that BMP4 regulates calcium signaling in PASMCs likely via upregulation of TRPC expression, leading to enhanced SOCE and basal [Ca ]i in PASMCs, and by this mechanism contributes to pulmonary vascular remodeling during pulmonary arterial hypertension.

[1]  C. Ince,et al.  Microcirculation , 2012, Springer US.

[2]  L. Shimoda,et al.  Differences in STIM1 and TRPC expression in proximal and distal pulmonary arterial smooth muscle are associated with differences in Ca2+ responses to hypoxia. , 2008, American journal of physiology. Lung cellular and molecular physiology.

[3]  G. Hansmann,et al.  An antiproliferative BMP-2/PPARgamma/apoE axis in human and murine SMCs and its role in pulmonary hypertension. , 2008, The Journal of clinical investigation.

[4]  H. Beppu,et al.  Bone Morphogenetic Protein (BMP) Type II Receptor Is Required for BMP-mediated Growth Arrest and Differentiation in Pulmonary Artery Smooth Muscle Cells* , 2008, Journal of Biological Chemistry.

[5]  O. Eickelberg,et al.  Transforming growth factor beta/bone morphogenic protein signaling in pulmonary arterial hypertension: remodeling revisited. , 2007, Trends in cardiovascular medicine.

[6]  H. Beppu,et al.  Repulsive Guidance Molecule RGMa Alters Utilization of Bone Morphogenetic Protein (BMP) Type II Receptors by BMP2 and BMP4* , 2007, Journal of Biological Chemistry.

[7]  P. Insel,et al.  Pulmonary artery smooth muscle cells from normal subjects and IPAH patients show divergent cAMP-mediated effects on TRPC expression and capacitative Ca2+ entry. , 2007, American journal of physiology. Lung cellular and molecular physiology.

[8]  W. Seeger,et al.  Dysregulated Bone Morphogenetic Protein Signaling in Monocrotaline-Induced Pulmonary Arterial Hypertension , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[9]  L. Shimoda,et al.  Ca2+ Channels and Chronic Hypoxia , 2006, Microcirculation.

[10]  C. V. Remillard,et al.  TRP Channels, CCE, and the Pulmonary Vascular Smooth Muscle , 2006, Microcirculation.

[11]  A. Hata,et al.  BMP-dependent activation of caspase-9 and caspase-8 mediates apoptosis in pulmonary artery smooth muscle cells. , 2006, American journal of physiology. Lung cellular and molecular physiology.

[12]  N. Morrell.  Pulmonary hypertension due to BMPR2 mutation: a new paradigm for tissue remodeling? , 2006, Proceedings of the American Thoracic Society.

[13]  G. Semenza,et al.  Hypoxia Inducible Factor 1 Mediates Hypoxia-Induced TRPC Expression and Elevated Intracellular Ca2+ in Pulmonary Arterial Smooth Muscle Cells , 2006, Circulation research.

[14]  Y. Fukuchi,et al.  Downregulation of type II bone morphogenetic protein receptor in hypoxic pulmonary hypertension. , 2006, American journal of physiology. Lung cellular and molecular physiology.

[15]  A. Pozzi,et al.  Bone Morphogenetic Protein 4 Promotes Pulmonary Vascular Remodeling in Hypoxic Pulmonary Hypertension , 2005, Circulation research.

[16]  L. Shimoda,et al.  Inhibition of hypoxic pulmonary vasoconstriction by antagonists of store-operated Ca2+ and nonselective cation channels. , 2005, American journal of physiology. Lung cellular and molecular physiology.

[17]  Dejun Sun,et al.  Acute hypoxia increases intracellular [Ca2+] in pulmonary arterial smooth muscle by enhancing capacitative Ca2+ entry. , 2005, American journal of physiology. Lung cellular and molecular physiology.

[18]  N. Rudarakanchana,et al.  Dysfunctional Smad Signaling Contributes to Abnormal Smooth Muscle Cell Proliferation in Familial Pulmonary Arterial Hypertension , 2005, Circulation research.

[19]  J. Putney,et al.  Store-operated calcium channels. , 2005, Physiological reviews.

[20]  P. Thistlethwaite,et al.  Bosentan inhibits transient receptor potential channel expression in pulmonary vascular myocytes. , 2004, American journal of respiratory and critical care medicine.

[21]  I. Fantozzi,et al.  ATP-induced mitogenesis is mediated by cyclic AMP response element-binding protein-enhanced TRPC4 expression and activity in human pulmonary artery smooth muscle cells. , 2004, American journal of physiology. Cell physiology.

[22]  Ying Yu,et al.  Overexpression of TRPC1 enhances pulmonary vasoconstriction induced by capacitative Ca2+ entry. , 2004, American journal of physiology. Lung cellular and molecular physiology.

[23]  I. Fantozzi,et al.  Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[24]  C. Tse,et al.  Chronic Hypoxia–Induced Upregulation of Store-Operated and Receptor-Operated Ca2 Channels in Pulmonary Arterial Smooth Muscle Cells: A Novel Mechanism of Hypoxic Pulmonary Hypertension , 2004, Circulation research.

[25]  L. Shimoda,et al.  Capacitative calcium entry and TRPC channel proteins are expressed in rat distal pulmonary arterial smooth muscle. , 2004, American journal of physiology. Lung cellular and molecular physiology.

[26]  P. Aaronson,et al.  Capacitative calcium entry as a pulmonary specific vasoconstrictor mechanism in small muscular arteries of the rat , 2003, British journal of pharmacology.

[27]  K. Yip,et al.  ET-1 activates Ca2+ sparks in PASMC: local Ca2+ signaling between inositol trisphosphate and ryanodine receptors. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[28]  I. Fantozzi,et al.  Bone morphogenetic proteins induce apoptosis in human pulmonary vascular smooth muscle cells. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[29]  M. Berridge,et al.  Calcium signalling: dynamics, homeostasis and remodelling , 2003, Nature reviews. Molecular cell biology.

[30]  Ying Yu,et al.  PDGF stimulates pulmonary vascular smooth muscle cell proliferation by upregulating TRPC6 expression. , 2003, American journal of physiology. Cell physiology.

[31]  Ying Yu,et al.  Inhibition of endogenous TRP1 decreases capacitative Ca2+ entry and attenuates pulmonary artery smooth muscle cell proliferation. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[32]  N. Rudarakanchana,et al.  Functional analysis of bone morphogenetic protein type II receptor mutations underlying primary pulmonary hypertension. , 2002, Human molecular genetics.

[33]  R. Trembath,et al.  Primary Pulmonary Hypertension Is Associated With Reduced Pulmonary Vascular Expression of Type II Bone Morphogenetic Protein Receptor , 2002, Circulation.

[34]  A. Gurney,et al.  Store-Operated Channels Mediate Ca2+ Influx and Contraction in Rat Pulmonary Artery , 2001, Circulation research.

[35]  Ying Yu,et al.  Capacitative Ca(2+) entry in agonist-induced pulmonary vasoconstriction. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[36]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[37]  Y. Hara,et al.  The Transient Receptor Potential Protein Homologue TRP6 Is the Essential Component of Vascular &agr;1-Adrenoceptor–Activated Ca2+-Permeable Cation Channel , 2001, Circulation research.

[38]  C. L. Bailey,et al.  Upregulated TRP and enhanced capacitative Ca(2+) entry in human pulmonary artery myocytes during proliferation. , 2001, American journal of physiology. Heart and circulatory physiology.

[39]  M. Humbert,et al.  Sporadic primary pulmonary hypertension is associated with germline mutations of the gene encoding BMPR-II, a receptor member of the TGF-β family , 2000, Journal of medical genetics.

[40]  S. Hodge,et al.  Familial primary pulmonary hypertension (gene PPH1) is caused by mutations in the bone morphogenetic protein receptor-II gene. , 2000, American journal of human genetics.

[41]  R. Trembath,et al.  Heterozygous germline mutations in BMPR2, encoding a TGF-β receptor, cause familial primary pulmonary hypertension , 2000, Nature Genetics.

[42]  S. Vukicevic,et al.  Bone morphogenetic protein‐7 (osteogenic protein‐1) inhibits smooth muscle cell proliferation and stimulates the expression of markers that are characteristic of SMC phenotype in vitro , 2000, Journal of cellular physiology.

[43]  P. Aaronson,et al.  Voltage‐independent calcium entry in hypoxic pulmonary vasoconstriction of intrapulmonary arteries of the rat , 2000, The Journal of physiology.

[44]  V. Golovina.  Cell proliferation is associated with enhanced capacitative Ca2+ entry in human arterial myocytes. , 1999, American journal of physiology. Cell physiology.

[45]  R. Marthan,et al.  Cellular mechanisms and role of endothelin-1-induced calcium oscillations in pulmonary arterial myocytes. , 1998, American journal of physiology. Lung cellular and molecular physiology.

[46]  K. Miyazono,et al.  Inhibition of rat vascular smooth muscle proliferation in vitro and in vivo by bone morphogenetic protein-2. , 1997, The Journal of clinical investigation.

[47]  R. Hurst,et al.  Cloning and Expression of a Novel Mammalian Homolog ofDrosophila Transient Receptor Potential (Trp) Involved in Calcium Entry Secondary to Activation of Receptors Coupled by the Gq Class of G Protein* , 1997, The Journal of Biological Chemistry.

[48]  M Hori,et al.  Calcium movements, distribution, and functions in smooth muscle. , 1997, Pharmacological reviews.

[49]  M. Berridge,et al.  Calcium signalling and cell proliferation , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[50]  N. Zhong,et al.  Sildenafil inhibits chronically hypoxic upregulation of canonical transient receptor potential expression in rat pulmonary arterial smooth muscle. , 2010, American journal of physiology. Cell physiology.

[51]  L. Shimoda,et al.  Differences in STIM 1 and TRPC expression in proximal and distal pulmonary arterial smooth muscle are associated with differences in Ca 2 responses to hypoxia , 2008 .

[52]  G. Semenza,et al.  Hypoxia Inducible Factor 1 Mediates Hypoxia-Induced TRPC Expression and Elevated Intracellular Ca 2 in Pulmonary Arterial Smooth Muscle Cells , 2006 .

[53]  Ying Yu,et al.  Overexpression of TRPC 1 enhances pulmonary vasoconstriction induced by capacitative Ca 2 entry , 2004 .

[54]  R. Derynck,et al.  Smad-dependent and Smad-independent pathways in TGF-beta family signalling. , 2003, Nature.

[55]  张锺儒,et al.  对急性哮喘加剧的成年患者两种不同教育干预的评价[英]／Co^^té J…∥Am J Respir Crit Care Med , 2002 .

[56]  J. Massagué,et al.  Controlling TGF-beta signaling. , 2000, Genes & development.

[57]  M. Horibe,et al.  Capacitative Ca2+ entry and tyrosine kinase activation in canine pulmonary arterial smooth muscle cells , 2000 .

[58]  M. Horibe,et al.  Capacitative Ca(2+) entry and tyrosine kinase activation in canine pulmonary arterial smooth muscle cells. , 2000, American journal of physiology. Lung cellular and molecular physiology.

[59]  V. Golovina.  Cell proliferation is associated with enhanced capacitative Ca(2+) entry in human arterial myocytes. , 1999, The American journal of physiology.