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(Circulation. 2004;110:867-873.)
© 2004 American Heart Association, Inc.

Original Articles

Profiling of Aortic Smooth Muscle Cell Gene Expression in Response to Chronic Inhibition of Nitric Oxide Synthase in Rats

Morgan Dupuis, MS; Florent Soubrier, MD, PhD; Isabelle Brocheriou, MD; Ségolène Raoux, MS; Mounsif Haloui, PhD; Liliane Louedec, MS; Jean-Baptiste Michel, MD, PhD; Sophie Nadaud, PhD

From Unit 525 INSERM, Faculté de Médecine Pitié-Salpétrière (M.D., F.S., I.B., S.R., S.N.), and Unit 460 INSERM, CHU X. Bichat (M.H., L.L., J.-B.M.), Paris, France.

Correspondence to Florent Soubrier, MD, PhD, INSERM U525, Faculté de Médecine Pitié-Salpétrière, 91 boulevard de l’Hôpital, 75013 Paris, France. E-mail florent.soubrier{at}chups.jussieu.fr

Received December 9, 2003; revision received March 30, 2004; accepted April 13, 2004.

	Abstract

Top Abstract Introduction Methods Results and Discussion References

 
Background— Chronic inhibition of nitric oxide (NO) synthesis by N-nitro-L-arginine methyl ester (L-NAME) induces hypertension associated with remodeling of the arterial wall. In this study, we aimed at identifying genes and pathways involved in this process in aortic smooth muscle cells from Fischer 344 rats, which exhibit an accelerated hypertension after administration of L-NAME.

Methods and Results— We studied the transcriptional profile of aortic media after 15 days (moderate hypertension) and 30 days (accelerated hypertension) of L-NAME administration (50 mg · kg^–1 · d^–1) by using rat Affymetrix Genechips, and we present a large-scale validation of the DNA chip results by real-time reverse transcription–polymerase chain reaction (RT-PCR). We observed, in aortic media, a progressive increase in the number of modulated genes during L-NAME administration, with 53 genes significantly modulated after 15 days and 147 genes after 30 days. These expression changes were confirmed at 87% by RT-PCR. We found 28 known genes regulated at both 15 and 30 days (96% confirmation by RT-PCR). The functional classification of the regulated genes highlights 3 major biological pathways modulated in aortic media during L-NAME administration: genes regulating cell proliferation, genes involved in the extracellular matrix remodeling, and genes of the NO/cGMP signaling pathway.

Conclusions— As a consequence of the genomic approach, we observed a large increase in modulation of gene expression along the evolution of the model and the progressive implication of compensatory mechanisms, making expression profile analysis more complex.

Key Words: hypertension • aorta • muscle, smooth • transcription

	Introduction

Top Abstract Introduction Methods Results and Discussion References

 
Nitric oxide (NO) synthase inhibition by chronic administration of N-nitro-L-arginine methyl ester (L-NAME) induces a dose-dependent increase in blood pressure (BP)^1,2 and leads to arterial and organ injuries.^3,4 This model is characterized by thickening⁵ and inflammation⁶ of the arterial wall of large arteries and perivascular fibrosis in small arterioles of the heart,⁴ the central nervous system,⁷ and the kidney.⁸ These mechanisms constitute important steps in arterial remodeling occurring in different models of hypertension as well as in human hypertension.⁹ Therefore, this model provides a good opportunity to investigate the molecular basis of the arterial wall alterations in response to hypertension and endothelial dysfunction.

The renin-angiotensin system is one of the key effectors during L-NAME–induced hypertension, as attested by the beneficial effects of inhibitors of the renin-angiotensin system.^10,11 L-NAME administration also induces a nuclear factor-B–dependent vascular inflammation, provoking monocyte adhesion to the vessel wall through induction of monocyte chemotactic protein-1, interleukin-6, and macrophage colony–stimulating factor,^12,13 and triggers fibrosis of coronary arteries through an increased production of transforming growth factor (TGF)-ß₁.¹⁴

The aim of our study was to identify genes implicated in vascular alterations induced during moderate and accelerated hypertension, using the DNA chip technology. The design of our experiment allows a dynamic view of gene expression profiling in large arteries after L-NAME intoxication by studying 2 time points: 15 days of intoxication, at which hypertension is moderate and 30 days of intoxication, at which severe hypertension is observed. We studied aorta rather than small arteries because large arteries allowed us to isolate the media layer from the adventitia, and we focused our study on the aortic tunica media, essentially composed of vascular smooth muscle cells (VSMCs), which constitute the main target of endothelium-derived NO.

We report here that the transcriptional profile of aortic media is deeply modified during accelerated hypertension after L-NAME administration, and we present a large-scale independent validation of the DNA chip technology by real-time reverse transcription–polymerase chain reaction (RT-PCR) applied to the study of vascular tissues from animal models.

	Methods

Top Abstract Introduction Methods Results and Discussion References

 
The Methods section is available in the online-only Data Supplement at http://www.circulationaha.org.

	Results and Discussion

Top Abstract Introduction Methods Results and Discussion References

 
Body Weight, Heart Weight, BP, and Aortic Media Thickness
Significant differences (–24%, P<0.05) in body weights are observed after 30 days of L-NAME administration (Table 1). Although heart weights are similar across the different groups, the ratio of heart weight to body weight increases gradually after L-NAME intoxication. Long-term administration of L-NAME results in a graduated rise in systolic BP, reaching 187±4.4 mm Hg (+22%, P<0.05) at 15 days of administration and 278±12 mm Hg (+74%, P<0.05) at 30 days. This high level of BP shows the high susceptibility of the Fischer 344 strain of rat to endothelial dysfunction compared with other strains, such as Wistar⁶ or Sprague-Dawley¹⁵ rats.

View this table: [in this window] [in a new window]   TABLE 1. Characteristics of Control and L-NAME–Treated Rats

By morphometric study, we observed that the aortic media thickness gains 40% (P<0.05) at day 15 and 35% (P<0.05) at day 30 of L-NAME administration (Table 1), compared with the control group, confirming previous results.^5,11,16 Interestingly, the thickening of the media is already significant at day 15 of administration, although BP is moderately increased at that time. Cell counting revealed that the number of VSMCs increases in media after L-NAME intoxication (+25% at day 15 and +21% at day 30, P<0.05), showing a proliferation of these cells (data not shown). Cell hypertrophy also occurs because the number of VSMCs by media surface decreases (–30% at day 15, P<0.05, and –25% at day 30). Thus, simultaneous proliferation and hypertrophy of VSMCs occur in the L-NAME model. Fifteen days later, the thickening is similar, whereas the rats present with severe hypertension. Thus, the moderate rise of BP associated with the NO synthase inhibition seems to be sufficient to deeply modify the aortic structure at day 15. Immunostaining for ED-1 revealed no infiltration of monocytes into aortic wall after L-NAME administration (data not shown).

Analysis of the DNA Chip Data
The arterial remodeling induced by L-NAME is associated with important phenotypic modifications of VSMCs, reinforcing the interest to characterize their transcription profile in our model. By studying the gene expression profile in rat aortic SMCs at 15 and 30 days of L-NAME administration, we showed that the number of modulated genes increases considerably with time and with BP increase. Statistical analysis indicated that the number of differentially expressed genes increases from 53 at day 15 (list available in Data Supplement Table 2) to 147 at day 30 (Data Supplement Table 3), and this number is probably inaccurate, because not all genes are present in the DNA chips used and because the sensitivity and specificity of the method are not complete. The significance of this increase is difficult to interpret, but it is likely that, at day 15, genes are directly modulated by BP increase as well as the absence of NO production. NO is known to modulate the transcription of genes owing to its action on transcription factor activity, such as activating protein-1 and nuclear factor-B.¹⁷ At day 30, the high level of BP, with its mechanical and biochemical consequences, is likely to be the major regulatory factor. The persistence of transcriptional regulatory signals throughout L-NAME administration is attested to by the modulation of 28 genes at both day 15 and day 30 (Table 2 and Data Supplement Table 4).

View this table: [in this window] [in a new window]   TABLE 2. Comparison of DNA Chips With Real-Time RT-PCR Results for Genes Regulated by 15 and 30 Days of L-NAME Administration

Real-Time PCR Validation of DNA Chip Expression Data
To confirm the changes in gene expression determined by the statistical analysis of the chip data, we performed real-time RT-PCR on a large number of selected genes. Validation by RT-PCR was performed on 2 independent experimental series of rats, a first series used for both the DNA chip and RT-PCR experiments (series 1) and a second series, used only for the RT-PCR experiments (series 2). First, we checked the expression of the genes in rat samples that were also used for the Affymetrix chip experiments to evaluate the validity of Affymetrix data. We measured the expression of all genes modulated at both periods of intoxication (15 and 30 days) and of some genes detected only at 15 or 30 days of intoxication. All known genes modulated at days 15 and 30 are confirmed (Table 2), except for 1, the insulin-like growth factor-2 receptor (96% confirmation rate). Thus, there is a close correlation between qualitative changes in mRNA expression detected at 2 time points with the DNA chip and real-time RT-PCR measurements, although the magnitude of the modulation observed varies according to the method used. Genes modulated at just 1 of the 2 intoxication durations according to DNA chip measurements were also confirmed for 87% of them by RT-PCR (Table 3). For some genes, RT-PCR measurements showed that they were modulated at both experimental times, suggesting a better sensitivity of this method in these cases.

View this table: [in this window] [in a new window]   TABLE 3. Comparison of DNA Chips With Real-Time RT-PCR Results for Genes Regulated After 15 or 30 Days of L-NAME Administration

Second, we studied by RT-PCR a large series of genes on an independent series of rats (series 2), not used for the DNA chip experiments, to estimate interexperimental variations. Of all the known genes and unidentified expressed sequence tags modulated at days 15 and 30, 78% are validated by real-time RT-PCR (Table 2). All members of the HSP family that are modulated in the first series of rats are unchanged by L-NAME in the second series. Except for this set of genes, the results corroborate those obtained with the previous rats. This result illustrates one of the drawbacks of the genomic approach, which allows a large number of genes to be studied, even those not related to the experiment and whose expression can be altered by intercurrent experimental factors unavoidable with in vivo experiments. Thus, confirmation on an independent series of animals is mandatory for the genomic approach of in vivo experiments.

Functional Classification of the Regulated Genes
Genes modulated by L-NAME in the aortic media were categorized into 7 general categories (cell division, cell signaling/cell communication, cell structure/motility, cell/organism defense, gene/protein expression, metabolism, and unclassified) based on the role of known genes, as proposed by Hwang et al¹⁸ (Figures 1 to 3 ).

View larger version (27K): [in this window] [in a new window]   Figure 1. Functional distribution of genes regulated in rat aortic media after 15 days of L-NAME administration.

View larger version (25K): [in this window] [in a new window]   Figure 2. Functional distribution of genes regulated in rat aortic media after 30 days of L-NAME administration.

View larger version (29K): [in this window] [in a new window]   Figure 3. Functional distribution of genes regulated in rat aortic media after 15 and 30 days of L-NAME administration.

The modulation of effectors belonging to these functional groups highlights 3 main biological pathways that are regulated in aortic media after L-NAME administration (Figure 4) and that are discussed in more detail below.

View larger version (19K): [in this window] [in a new window]   Figure 4. Major biological pathways modulated after L-NAME administration in aortic media.

Regulation of Cell Proliferation
We detected, by immunohistochemistry, an increase in the number of proliferating cell nuclear antigen–positive cells in aortic media during L-NAME intoxication (Figure 5), highlighting proliferation of VSMCs. This result explains the increase of the number of VSMCs observed in media, and these changes are concomitant with the modulation of several genes regulating cell proliferation. At 15 days of L-NAME intoxication, upregulation of an important set of genes occurs (CDC-2, CKS-2, cyclin A), which may favor proliferation of VSMCs observed during hypertension.¹³ At 30 days, we observe a shift in the upregulation of cyclin A to cyclin G₁ and downregulation of an antiproliferative antigen (B-cell translocation gene 2).

View larger version (120K): [in this window] [in a new window]   Figure 5. Immunohistochemistry of aorta. Aorta sections from control rat, rat receiving L-NAME for 15 days, and rat receiving L-NAME for 30 days were stained immunohistochemically for proliferating cell nuclear antigen (PCNA), bone sialoprotein II (BSPII), and osteopontin (OPN). Bar=30 µm.

Moreover, the upregulation at days 15 and 30 of transcription inhibitors Id1, Id2, and Id3 is of particular interest because Id2 and Id3 promote VSMC proliferation.^19,20

Extracellular Matrix Remodeling
Among elements of the extracellular matrix, a group of genes is of interest because they are known to be effectors of the vascular calcification process. At days 15 and 30, osteoadherin, which is a member of the small leucine-rich proteoglycan (SLRP) family described in regulation of mineralization, is downregulated in aortic media. Second, bone sialoprotein II and periostin are transcriptionally activated at days 15 and 30, and their expression is known to be correlated with calcification: the former is upregulated during medial calcification,²¹ whereas the latter was originally isolated as an osteoblast-specific factor.²² We observed, by immunohistochemistry, expression of bone sialoprotein II in aortic media (Figure 5), but we did not detect modulation of its protein level by this method. At day 30 of L-NAME administration, an increased expression of osteopontin, detected at the mRNA level, may act as a counterregulatory mechanism. Osteopontin is a potent inhibitor of vascular calcification²³ identified as being upregulated in aortic smooth muscle cells showing a calcifying phenotype.²⁴ We observed an increase in the immunostaining against osteopontin at days 15 and 30 (Figure 5), showing that stabilization of this effector at a protein level is an earlier event than the increase of its mRNA level, which was detected at day 30. Induction of this inhibitor could explain why we did not detect calcifying lesions in media by Von Kossa staining (data not shown). These results highlight the complexity of the regulation of vascular calcification during severe hypertension.

A second type of modulation involving genes coding for elements of the extracellular matrix occurs, which may actively participate in the fibrosis process. Indeed, vascular fibrosis occurs into the coronary vessels after 4 to 8 weeks of L-NAME intoxication, and this remodeling is associated with an overproduction of TGF-ß,¹⁴ known to induce the synthesis of extracellular matrix proteins. We show that the TGF-ß pathway is regulated in media with the induction at days 15 and 30 of factors activating the pathway, such as thrombospondin-1, which is an extracellular protein able to bind and activate latent TGF-ß,²⁵ and the latent transforming growth factor-ß binding protein-2 (LTBP2), which regulates TGF-ß signaling. Sinha et al²⁶ showed that expression of LTBP2 parallels TGF-ß₁ after an arterial injury. After 30 days, several members of the SLRP family, in addition to osteoadherin, are downregulated: fibromodulin, decorin, and lumican. These compounds may act as natural repressors of fibrosis by their ability to interact with TGF-ß and to inhibit its activity.²⁷ Decorin was shown to inhibit the increased production of extracellular matrix induced by TGF-ß in the kidney.²⁸ In addition, genes induced by TGF-ß during fibrosis,¹⁴ such as fibronectin, are upregulated. However, the induction of SMAD6 and SMAD7, which inhibit TGF-ß signaling, counteracts the signaling of this pathway at day 30.

Regulation of Smooth Muscle Tone
Several modulated genes belonging to the cell signaling and communication class constitute potent regulators of smooth muscle tone. The mRNA level of soluble guanylate cyclase, an important NO target in VSMCs, is already decreased by L-NAME at day 15, corroborating the results obtained in aorta from spontaneously hypertensive rats.²⁹ Thus, the lack of activation of the enzyme by NO seems to trigger a downregulation of the gene expression. The NO pathway is therefore inhibited both at the level of NO synthesis by L-NAME and by decreased expression of the effector normally activated by NO binding in aortic media. In addition, N^G,N^G-dimethylarginine dimethylaminohydrolase, which hydrolyzes methylarginine, an NO synthase inhibitor, is downregulated at day 30 by L-NAME.

At day 30, downregulation of RGS-2 expression could participate in the BP rise, because disruption of this gene results in hypertensive animals.³⁰ RGS-2 is a target of cGMP-dependent protein kinase I- (an NO effector), which attenuates G protein–coupled receptor signaling, such as the thrombin receptor PAR-1.³¹ RGS-2 downregulation and PAR-1 mRNA upregulation after 30 days of NO inhibition could potentiate the vasoconstrictive effect of thrombin in the vascular wall.

Conclusions
Modulation of gene expression, in particular of some effectors we identified, is of primary importance in the arterial remodeling process that occurs during hypertension. The expression profile determined at 2 steps of the evolution of hypertension under L-NAME allows us to observe that the worsening of hypertension is associated with an increasing recruitment of genes, which belong to several biological pathways, explaining the pathological phenotype of the aortic wall.

Second, the interest of the genomic approach is to give an extended view on expression modulation for several functional systems of the cell and avoids the bias of the study of a series of selected genes. In this study, we have demonstrated the complexity of gene regulation in this in vivo model, because the variations in gene expression show the existence of counterregulatory loops or physiological compensatory mechanisms that might allow a limitation of the pathological processes.

	Acknowledgments

 
This work was supported by INSERM (Institut National de la Santé et de la Recherche Médicale) and Pierre and Marie Curie University and by grants from the Ministère de la Recherche, the Leducq foundation, the région Ile de France, and the Fondation de Recherche Aventis. Morgan Dupuis is a recipient of a fellowship from the French Society of Hypertension. This work is dedicated to Ségolène Raoux, who died on May 6, 2003.

	Footnotes

 
The online-only Data Supplement is available at http://www.circulationaha.org.

Deceased.

	References

Top Abstract Introduction Methods Results and Discussion References

 
1. Arnal JF, Warin L, Michel JB. Determinants of aortic cyclic guanosine monophosphate in hypertension induced by chronic inhibition of nitric oxide synthase. J Clin Invest. 1992; 90: 647–652.[Medline] [Order article via Infotrieve]

2. Ribeiro MO, Antunes E, de Nucci G, et al. Chronic inhibition of nitric oxide synthesis: a new model of arterial hypertension. Hypertension. 1992; 20: 298–303.[Abstract/Free Full Text]

3. Baylis C, Mitruka B, Deng A. Chronic blockade of nitric oxide synthesis in the rat produces systemic hypertension and glomerular damage. J Clin Invest. 1992; 90: 278–281.[Medline] [Order article via Infotrieve]

4. Numaguchi K, Egashira K, Takemoto M, et al. Chronic inhibition of nitric oxide synthesis causes coronary microvascular remodeling in rats. Hypertension. 1995; 26: 957–962.[Abstract/Free Full Text]

5. Ikegaki I, Hattori T, Yamaguchi T, et al. Involvement of Rho-kinase in vascular remodeling caused by long-term inhibition of nitric oxide synthesis in rats. Eur J Pharmacol. 2001; 427: 69–75.[CrossRef][Medline] [Order article via Infotrieve]

6. Tomita H, Egashira K, Kubo-Inoue M, et al. Inhibition of NO synthesis induces inflammatory changes and monocyte chemoattractant protein-1 expression in rat hearts and vessels. Arterioscler Thromb Vasc Biol. 1998; 18: 1456–1464.[Abstract/Free Full Text]

7. Blot S, Arnal JF, Xu Y, et al. Spinal cord infarcts during long-term inhibition of nitric oxide synthase in rats. Stroke. 1994; 25: 1666–1673.[Abstract]

8. Xu Y, Arnal JF, Hinglais N, et al. Renal hypertensive angiopathy: comparison between chronic NO suppression and DOCA-salt intoxication. Am J Hypertens. 1995; 8: 167–176.[CrossRef][Medline] [Order article via Infotrieve]

9. Intengan HD, Schiffrin EL. Vascular remodeling in hypertension: roles of apoptosis, inflammation, and fibrosis. Hypertension. 2001; 38: 581–587.[Abstract/Free Full Text]

10. Michel JB, Xu Y, Blot S, et al. Improved survival in rats administered NG-nitro L-arginine methyl ester due to converting enzyme inhibition. J Cardiovasc Pharmacol. 1996; 28: 142–148.[CrossRef][Medline] [Order article via Infotrieve]

11. Takemoto M, Egashira K, Tomita H, et al. Chronic angiotensin-converting enzyme inhibition and angiotensin II type 1 receptor blockade: effects on cardiovascular remodeling in rats induced by the long-term blockade of nitric oxide synthesis. Hypertension. 1997; 30: 1621–1627.[Abstract/Free Full Text]

12. Gonzalez W, Fontaine V, Pueyo ME, et al. Molecular plasticity of vascular wall during N^G-nitro-L-arginine methyl ester–induced hypertension: modulation of proinflammatory signals. Hypertension. 2000; 36: 103–109.[Abstract/Free Full Text]

13. Kitamoto S, Egashira K, Kataoka C, et al. Increased activity of nuclear factor-B participates in cardiovascular remodeling induced by chronic inhibition of nitric oxide synthesis in rats. Circulation. 2000; 102: 806–812.[Abstract/Free Full Text]

14. Tomita H, Egashira K, Ohara Y, et al. Early induction of transforming growth factor-ß via angiotensin II type 1 receptors contributes to cardiac fibrosis induced by long-term blockade of nitric oxide synthesis in rats. Hypertension. 1998; 32: 273–279.[Abstract/Free Full Text]

15. Tomida T, Numaguchi Y, Nishimoto Y, et al. Inhibition of COX-2 prevents hypertension and proteinuria associated with a decrease of 8-iso-PGF2alpha formation in L-NAME-treated rats. J Hypertens. 2003; 21: 601–609.[CrossRef][Medline] [Order article via Infotrieve]

16. Levy BI, Michel JB, Salzmann JL, et al. Effects of chronic inhibition of converting enzyme on mechanical and structural properties of arteries in rat renovascular hypertension. Circ Res. 1988; 63: 227–239.[Abstract/Free Full Text]

17. Marshall HE, Merchant K, Stamler JS. Nitrosation and oxidation in the regulation of gene expression. FASEB J. 2000; 14: 1889–1900.[Abstract/Free Full Text]

18. Hwang DM, Dempsey AA, Wang RX, et al. A genome-based resource for molecular cardiovascular medicine: toward a compendium of cardiovascular genes. Circulation. 1997; 96: 4146–4203.[Abstract/Free Full Text]

19. Matsumura ME, Lobe DR, McNamara CA. Contribution of the helix-loop-helix factor Id2 to regulation of vascular smooth muscle cell proliferation. J Biol Chem. 2002; 277: 7293–7297.[Abstract/Free Full Text]

20. Mueller C, Baudler S, Welzel H, et al. Identification of a novel redox-sensitive gene, Id3, which mediates angiotensin II–induced cell growth. Circulation. 2002; 105: 2423–2428.[Abstract/Free Full Text]

21. Shanahan CM, Proudfoot D, Tyson KL, et al. Expression of mineralisation-regulating proteins in association with human vascular calcification. Z Kardiol. 2000; 89 (suppl 2): 63–68.[CrossRef][Medline] [Order article via Infotrieve]

22. Takeshita S, Kikuno R, Tezuka K, et al. Osteoblast-specific factor 2: cloning of a putative bone adhesion protein with homology with the insect protein fasciclin I. Biochem J. 1993: 294 (pt 1): 271–278.[Medline] [Order article via Infotrieve]

23. Wada T, McKee M, Steitz S, et al. Calcification of vascular smooth muscle cell cultures: inhibition by osteopontin. Circ Res. 1999; 84: 166–178.[Abstract/Free Full Text]

24. Steitz SA, Speer MY, Curinga G, et al. Smooth muscle cell phenotypic transition associated with calcification: upregulation of Cbfa1 and downregulation of smooth muscle lineage markers. Circ Res. 2001; 89: 1147–1154.[Abstract/Free Full Text]

25. Schultz-Cherry S, Ribeiro S, Gentry L, et al. Thrombospondin binds and activates the small and large forms of latent transforming growth factor-beta in a chemically defined system. J Biol Chem. 1994; 269: 26775–26782.[Abstract/Free Full Text]

26. Sinha S, Heagerty AM, Shuttleworth CA, et al. Expression of latent TGF-beta binding proteins and association with TGF-beta 1 and fibrillin-1 following arterial injury. Cardiovasc Res. 2002; 53: 971–983.[Abstract/Free Full Text]

27. Iozzo RV. The biology of the small leucine-rich proteoglycans: functional network of interactive proteins. J Biol Chem. 1999; 274: 18843–18846.[Free Full Text]

28. Border WA, Noble NA, Yamamoto T, et al. Natural inhibitor of transforming growth factor-beta protects against scarring in experimental kidney disease. Nature. 1992; 360: 361–364.[CrossRef][Medline] [Order article via Infotrieve]

29. Ruetten H, Zabel U, Linz W, et al. Downregulation of soluble guanylyl cyclase in young and aging spontaneously hypertensive rats. Circ Res. 1999; 85: 534–541.[Abstract/Free Full Text]

30. Heximer SP, Knutsen RH, Sun X, et al. Hypertension and prolonged vasoconstrictor signaling in RGS2-deficient mice. J Clin Invest. 2003; 111: 444–452.

31. Tang M, Wang G, Lu P, et al. Regulator of G-protein signaling-2 mediates vascular smooth muscle relaxation and blood pressure. Nat Med. 2003; 9: 1506–1512.[CrossRef][Medline] [Order article via Infotrieve]

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This Article Abstract Full Text (PDF) Data Supplement All Versions of this Article: 110/7/867    most recent 01.CIR.0000138850.72900.FEv1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Dupuis, M. Articles by Nadaud, S. Search for Related Content PubMed PubMed Citation Articles by Dupuis, M. Articles by Nadaud, S. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Related Collections Remodeling Animal models of human disease Other hypertension