© 1999 American Heart Association, Inc.
Original Contribution |
Structure of the Type B Human Natriuretic Peptide Receptor Gene and Association of a Novel Microsatellite Polymorphism With Essential Hypertension
From the Second Department of Internal Medicine, Nihon University School of Medicine, Tokyo, Japan.
Correspondence to Masayoshi Soma, MD, PhD, Second Department of Internal Medicine, Nihon University School of Medicine, 30-1 Ooyaguchi-kamimachi, Itabashi-ku, Tokyo 173, Japan. E-mail msoma{at}med.nihon-u.ac.jp
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Abstract |
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Abstract—The natriuretic peptide (NP) system may play a crucial role in development of essential hypertension (EH). C-type NP dilates arteries and lowers blood pressure and inhibits proliferation of vascular smooth muscle cells via the type B NP receptor (NPR-B). However, the association of the human NPR-B gene with EH has not been studied, because little is known about the genomic organization of this gene. We designed oligonucleotide primers based on the cDNA sequence of the human NPR-B gene, and long-range polymerase chain reaction (PCR) was performed. The amplified fragments were sequenced directly, and the exon/intron organization of the human NPR-B gene was determined. The gene, which spans
16.5 kbp, is composed of 22 exons,
and the intron-exon junctions follow the GT-AG rule. Seven hundred
fifty base pairs of the 5'-flanking region were sequenced using a
thermal asymmetric interlaced–PCR (TAIL-PCR) method. This region
contains 10 potential Sp1 binding sites and lacks a TATA box. Rapid
amplification of cDNA ends (RACE) revealed the transcriptional start
site at -14 bp. A CA/GT microsatellite repeat was identified with a
hybridization-based method and was converted to a sequence-tagged site
(STS). The GT microsatellite repeat was localized to intron 2 150 bp
downstream of the exon-intron junction. Two alleles, (GT)10 and
(GT)11, were detected in both EH patients and age-matched normotensive
(NT) controls. Multiple logistic linear regression analysis
indicated that the NPR-B genotype is associated significantly
with EH (odds ratio 1.55; 95% confidence interval, 1.02 to 2.35). The
(GT)11 frequency was 0.316 (65/206) for the EH group and 0.218 (44/202)
for the NT group and differed significantly between the EH and NT
groups (
2=4.97, P=0.026). The structural
organization of the human NPR-B gene was determined, and a novel GT
repeat polymorphism, which associated with EH, was identified.
These results suggest that one cause of EH is a mutation in this gene
or a closely related gene or region.
Key Words: natriuretic peptide • receptor, type B • gene structure • association • essential hypertension
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Introduction |
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The family of natriuretic peptides (NPs) elicits a number of vascular, renal, and endocrine effects, resulting in regulation of extracellular fluid volume and blood pressure.1 2 3 The NP family consists of 3 peptides: atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). Although ANP and BNP show high affinity for the type A NP receptor (NPR-A), CNP selectively binds to the type B NP receptor (NPR-B).4 CNP is secreted by endothelial cells5 and binds to NPR-B, which is abundant in the vascular smooth muscle cell membrane, and inhibits proliferation of vascular smooth muscle cells through the activation of the intracellular cGMP cascade.6 Intravenous administration of CNP led to reduced blood pressure in human subjects7 and increased forearm blood flow after intra-arterial injection.8 Abnormalities of NPR-B may therefore lead to hypertension. These receptor abnormalities were suggested by spontaneously hypertensive rats, which have plasma CNP concentrations similar to those in normotensive Wistar-Kyoto rats, but the vascular relaxing action of CNP was markedly attenuated.9
Genetic dissection is a powerful tool for determining the cause of a complex trait such as essential hypertension (EH)10 11 and for understanding the pathophysiological role of the protein encoded by the gene. We recently reported the association of the endothelial constitutive nitric oxide synthase gene and EH.12 13 As for human NPR-B, the gene was mapped to chromosome 9p21-p12.14 The cDNA and deduced amino acid sequences of human NPR-B have been described.15 The genomic structure, however, has not been determined.
In the present study, we determined the genomic structure of human NPR-B and found a novel GT repeat polymorphism that is associated with EH.
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Materials and Methods |
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Examination of Genomic Sequences Including Protein Coding Region
Genomic DNA was extracted from peripheral blood leukocytes by standard methods. On the basis of the information for the human NPR-B cDNA, we designed oligonucleotide primers; specific primers were designed to amplify the intronic regions (Table 1
). Long and accurate polymerase chain
reaction (LA-PCR)16 was performed using these primers, and
a single PCR product was generated in each case. More than 25
primers were designed and synthesized (Sawaday Technology, Tokyo,
Japan) to cover the genomic sequence from the start codon to the
termination codon (Table 1). Total human genomic DNA from
several individuals was used as a template to confirm identity with the
structure of the PCR products. After an initial denaturing step of
3 minutes at 94°C, the PCR conditions were 30 cycles of denaturation
for 25 seconds at 98°C, annealing and extension for 20 seconds at
68°C, and a final extension at 72°C for 10 minutes. LA-PCR was
performed according to the manufacturer's instructions (LA-PCR kit
version No. 2, Takara Syuzo, Tokyo, Japan). LA-PCR products were
electrophoresed on 0.4% agarose gels. For sequencing, the bands were
cut out of the gel and purified on a column (Microcon 100, Amicon Inc,
Beverly, Mass).
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Isolation of the 5'-Flanking Region
To isolate the 5'-flanking region of the NPR-B gene, thermal
asymmetric interlaced (TAIL)–PCR was used. The PCR conditions were
described previously,17 except that the annealing
temperature was 68°C. Five hundred nanograms of genomic DNA was used
as the template in the primary PCR reaction. Four arbitrary degenerate
(AD) primers were used: AD1, AD2, AD3, and AD4. Three specific primers,
which are complementary to the NPR-B gene, were synthesized (Table 2), and the PCR products were
electrophoresed on 1.5% agarose gels. For sequencing, bands were cut
out of the gel and purified as described above.
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Sequencing
PCR products were sequenced directly using a Thermo
Sequenase kit (Amersham) after purification over a column. More than 40
nucleotides of the coding region of the NPR-B gene were
sequenced together with the intron-exon boundaries.
Rapid Amplification of 5'-cDNA Ends (5'-RACE)
The 5'-RACE experiments were conducted to map the
transcriptional start site. 5'-RACE was performed using the 5'-full
RACE core set (Takara Syuzo Co Ltd, Tokyo, Japan) according to the
manufacturer's instructions. cDNA was synthesized using avian
myeloblastosis virus (AMV) reverse transcriptase XL and a
5'-phosphorylated 15-nucleotide antisense
primer (base pair +266 to +252 with respect to the ATG codon) and
having the sequence 5'-(P) AGC TTG AGG TCC ACA-3'. One microgram of
human pituitary gland poly-(A) selected +RNA (Clontech) was
reverse-transcribed using 5 U of AMV reverse transcriptase and 200 pmol
of the antisense primer. After purification of the single-stranded
cDNA, a ring structure was made (concatenating) using T4 RNA ligase. A
primary PCR was performed using a sense primer F1 5'-TGG ACC TGC GGT
TTG TCA GCT C (+185 to +206) and antisense primer R1 5'-TGC CAG GGC TGC
CAC CAA CA (+40 to +23). An aliquot of the initial PCR reaction served
as the template for a secondary PCR with sense primer F2 5'-AAC TGG AAG
GCG CCT GCT CTG A (+209 to +230) and antisense primer R2 5'-AGC AGA AGT
GAT GGC AGC GCC ATG G (+23 to -2), which yields a 98-bp product.
PCR products were analyzed on a 1.5% agarose gel and
sequenced directly.
Detection and Identification of a Microsatellite
Polymorphism
A CA/TG repeat microsatellite was identified in LA-PCR
products with a hybridization method described
previously.18 This microsatellite was converted to a
sequence-tagged site (STS) using the following primers: NPR-B-STS-F1
(GT strand) 5'-GGAGAGCTCTAAGTCTTAAGGCAC and NPR-B-STS-R1 (CA
strand) 5'-GGTCTCCCCCTCTTCTAATCCTG. The PCR products were separated
on a 6% polyacrylamide sequencing gel and visualized by
autoradiography (Figure 1). To establish a control for the number
of dinucleotide repeats, amplified fragments were subcloned
into a pCR II plasmid vector (Invitrogen, Carlsbad, Calif) and
sequenced. Plasmid clones containing known numbers of GT repeats were
used as standard templates in the PCR (Figures 1 and 2).
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Subjects
The study population included 103 patients with EH (EH
group; 48.5±9.1 years, mean±SD) who were given a diagnosis according
to the criteria of the World Health Organization. These criteria
include a sitting systolic blood pressure (SBP) of >160
mm Hg or diastolic blood pressure (DBP) of >95
mm Hg on 3 occasions spanning 2 months from the first medical
examination, without administration of antihypertensive drugs. Subjects
given a diagnosis of secondary hypertension were excluded. As a control
group, 101 normotensive healthy subjects (NT group; 48.2±7.1 years)
were also studied. The NT subjects had no family history of
hypertension, and, in all instances, their SBPs were <140 mm Hg
and their DBPs were <85 mm Hg. A positive family history was
defined as hypertension diagnosed in grandparents, parents, or
siblings. Both the patients and the control group were recruited from
the northern part of Tokyo, and informed consent was obtained from each
individual according to a protocol approved by the Human Studies
Committee at Nihon University.
Biochemical Analysis
The plasma concentration of total cholesterol
and the serum concentrations of creatinine and uric acid
were measured by standard methods in the clinical laboratory department
of the Nihon University hospital.
Statistical Analysis
Data are presented as mean±SD. Allele frequencies
were calculated from the genotypes of all subjects.
Hardy-Weinberg equilibrium was assessed by 2
analysis. Significant differences between the total number of
alleles on all chromosomes for the EH and NT groups were assessed
by 2 analysis with one degree of
freedom. The association between genotypes and hypertension was
evaluated by logistic linear regression analysis. EH was
regarded as the dependent variable and genotype, sex, age,
body mass index (BMI), and the concentrations of plasma total
cholesterol, serum, creatinine, and uric acid
were entered as independent variables. Differences in the clinical
data between the EH and NT groups and between genotypes were
assessed by ANOVA followed by the Fisher protected least significant
difference (PLSD) test. A value of P<0.05 was considered
significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Structure of the NPR-B Gene
The structural organization of the human NPR-B gene was determined by analysis of genomic clones and total genomic DNA by PCR amplification across introns, followed by sequencing of the intron-exon junction. In the present study, we determined the genomic structure of human NPR-B by direct sequencing of the long PCR products, not by screening a genomic library. The LA-PCR can amplify more than 20 kbp of DNA. Confirmation of whether the PCR products amplified the NPR-B gene was performed by nucleotide sequencing of more than 40 nucleotides within the cDNA region of the 5'-portion and 3'-portion of the exon. The sizes and positions of the exons, together with the codons and corresponding amino acids interrupted by introns, are listed in Table 3
. The genomic DNA contains 21 introns of
variable size. The NPR-B gene is 16.5 kbp in length, which is
more than 30 times longer than the cDNA. The size range for the exons
is 69 bp (exon 18) to 667 bp (ATG-exon 1) and for the introns is
87 bp (intron 14) to 6.5 kbp (intron 3). All the intron-exon junction
sequences follow the GT-AG rule shown in Table 4 and conform to the
consensus.19 A GT/CA microsatellite repeat was found in
intron 2 150 bp downstream of the exon-intron junction (Figure 3).
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A TAIL-PCR method using the AD3 primer detected a single clear band.
This band was 1 kbp, and the sequence was determined after
purification with a column. Seven hundred fifty base pairs of the
5'-flanking region were sequenced (Figure 4). This region did not include a TATA
box. The transcriptional start site was identified by sequencing the
concatenation product and is shown in Figure 4.
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Association of a Microsatellite Polymorphism With
Essential Hypertension
Clinical characteristics of the EH and NT subjects are shown in
Table 5. SBP, DBP, BMI, pulse rate, and
plasma concentration of total cholesterol were
significantly higher in the EH group than in the NT group. Age and
serum concentrations of creatinine or uric acid were not
different between the 2 groups.
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We found a novel GT/CA microsatellite repeat in intron 2, 150 bp
downstream of the exon-intron junction. Two alleles, (GT)10 and
(GT)11, were detected in both the EH group and the NT group. The
observed and expected heterozygosities were 0.16 and 0.15,
respectively, which were in good agreement with Hardy-Weinberg
equilibrium (2=0.19, P=0.98). The
frequencies of the (GT)10/(GT)10, (GT)10/(GT)11, and (GT)11/(GT)11
genotypes were 0.624 (63/101), 0.317 (32/101), and 0.059
(6/101) for the NT group and 0.515 (53/103), 0.340 (35/103), and 0.146
(15/103) for the EH group, respectively. Logistic linear regression
analysis adjusted by age and sex revealed that NPR-B
genotype was associated significantly with EH (odds ratio 1.55;
95% confidence interval, 1.02 to 2.35).The frequencies of the (GT)10
and (GT)11 alleles were 0.782 (158/202) and 0.218 (44/202) for the
NT group and 0.684 (141/206) and 0.316 (65/206) for the EH group,
respectively, and differed significantly between the 2 groups
(2=4.97, P=0.026). The
genotype or allele frequency was not associated with BMI,
pulse rate, or plasma levels of total cholesterol.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In the present study, the genomic structure of the human NPR-B gene was determined by analysis of genomic clones and total genomic DNA by PCR amplification across the intron-exon junction followed by sequencing. The long-range PCR and direct sequencing of PCR products were useful for determining the genomic structure as described previously.20 Advantages of this method are that radioisotope facilities and culture techniques for genomic library screening are not required. The difficulty in using this method is that even in long-range PCR, the genome cannot be amplified when there is an intron in the gene that is >20 to 30 kbp long. The identification of the human NPR-B PCR products was done by sequencing and gave more than 40 bp of identity to the human cDNA.15 The genomic structure is very similar to that of the rat21 and the human NPR-A gene,21A which contains 22 exons. The 5'-flanking region of the human NPR-B gene lacks a TATA box but contains 10 potential Sp1 binding sites.
The significance of NP family genes in hypertension in animals has been reported, and the targeted deletion of the pro-ANP gene or NPR-A gene yielded mice with high blood pressure.22 23 However, the genetic studies of human ANP gene are conflicting. A study of 2 ANP restriction fragment length polymorphisms in healthy Norwegian twins revealed no association with SBP, DBP, or blood pressure variability.24 Rutledge et al25 reported that the allele frequency of an Hpa II mutation in intron 2 was significantly higher in the hypertensive individuals compared with normotensive African Americans, but this observation was not confirmed in Australians.26 The Hpa II polymorphism was not associated with salt-sensitive hypertension in Caucasians.27 In contrast to the ANP gene, there is no available data on NP receptors. In the present study, we found a novel GT repeat in intron 2 of the NPR-B gene and demonstrated a significant association with EH. No differences in sex, age, pulse rate, BMI, or the plasma concentration of total cholesterol were apparent between subjects who were homozygous for the (GT)10 allele and those who had at least one (GT)11 allele. These results suggest that the GT repeat polymorphism may be in linkage disequilibrium with the actual cause of EH, which may be the NPR-B gene or a locus near this gene.
In conclusion, we determined the genomic structure of the human NPR-B gene and found a novel GT repeat polymorphism that is associated with EH. This allele may be in linkage disequilibrium with the actual causal mutation, suggesting that one cause of EH is a mutation in the NPR-B gene or a closely located gene. The genomic structure of the human NPR-B gene may contribute to our understanding of the actual cause of EH.
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Acknowledgments |
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This work was supported by a grant from the Ministry of Education, Science, and Culture of Japan. We wish to thank Hideko Tobe for her technical assistance and Drs. A. Kubo, M. Kunimoto, N. Fukuda, and Y. Watanabe for collecting samples.
Received April 21, 1998; accepted January 3, 1999.
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References |
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1. Maack T, Camargo MJF, Kleinert HD, Laragh JH, Atlas SA. Atrial natriuretic factor: structure and functional properties. Kidney Int. 1985;27:607–615.[Medline] [Order article via Infotrieve]
2.
Needleman P, Adams SP, Cole BR, Currie MG, Geller DM,
Michener ML, Saper CB, Schwartz D Standaert DG. Atriopeptins as cardiac
hormones. Hypertension. 1985;7:469–482.
3.
Cantin M, Genest J. The heart and the atrial
natriuretic factor. Endocr Rev. 1985;6:107–127.
4.
Koller KJ, Lowe DG, Bennett GL, Minamino N, Kangawa K,
Matsuo H, Goeddel DV. Selective activation of the B
natriuretic peptide receptor by C-type
natriuretic peptide (CNP). Science. 1991;252:120–123.
5. Suga S, Nakao K, Itoh H, Komatsu Y, Ogawa Y, Hama N, Imura H. Endothelial production of C-type natriuretic peptide and its marked augmentation by transforming growth factor-ß: possible existence of a vascular natriuretic peptide system. J Clin Invest. 1992;90:1145–1149.
6.
Komatsu Y, Itoh H, Suga S, Ogawa Y, Hama N, Kishimoto
I, Nakagawa O, Igaki T, Doi K, Yoshimasa T, Nkao K. Regulation of
endothelial production of C-type
natriuretic peptide in coculture with vascular smooth
muscle cells: role of the vascular natriuretic peptide
system in vascular growth inhibition. Circ Res. 1996;78:606–614.
7. Igaki T, Itoh H, Hama N, Ogawa Y, Komatsu Y, Mukoyama M, Sugawara A, Yoshimasa T, Tanaka I, Nakao K. C-type natriuretic peptides in chronic renal failure and its action in human. Kidney Int. 1996;49:S144–S147.
8.
Davidson NC, Barr CS, Struthers AD. C-type
natriuretic peptide: an endogenous
inhibitor of vascular angiotensin-converting
enzyme activity. Circulation. 1996;93:1155–1159.
9. Wei C-M, Kim CH, Khraibi AA, Miller VM, Burnett JC Jr. Atrial natriuretic peptide and C-type natriuretic peptide in spontaneously hypertensive rats and vasorelaxing action in vivo. Hypertension. 1994;23(pt 2):903–907.
10.
Lander ES, Shork NJ. Genetic dissection of complex
traits. Science. 1994;265:2037–2048.
11. Kurtz TW, Spence MA. Genetics of essential hypertension. Am J Med. 1993;94:77–84.[Medline] [Order article via Infotrieve]
12. Nakayama T, Soma M, Takahashi Y, Izumi Y, Kanmatsuse K, Esumi M. Association analysis of CA repeat polymorphism of the endothelial nitric oxide synthase gene with essential hypertension in Japanese. Clin Genet. 1997;51:26–30.[Medline] [Order article via Infotrieve]
13. Uwabo J, Soma M, Nakayama T, Kanmatsuse K. Association of a variable number of tandem repeats in the endothelial constitutive nitric oxide synthase gene with essential hypertension. Am J Hypertens.. 1998;11:125–128.[Medline] [Order article via Infotrieve]
14. Lowe DG, Klisak I, Sparkes RS, Mohandas T, Goeddel DV. Chromosomal distribution of three members of the human natriuretic peptide receptor/guanylyl cyclase gene family. Genomics. 1990;8:304–312.[Medline] [Order article via Infotrieve]
15. Chang M-S, Lowe DG, Lewis M, Hellmiss R, Chen E, Goeddel DV. Differential activation by atrial and brain natriuretic peptides of two different receptor guanylate cyclases. Nature. 1989;341:68–72.[Medline] [Order article via Infotrieve]
16.
Cheng S, Fockler C, Barnes WM, Higuchi R. Effective
amplification of long targets from cloned inserts and human genomic
DNA. Proc Natl Acad Sci U S A. 1994;91:5695–5699.
17. Liu Y, Whittier RF. Thermal asymmetric interlaced PCR: Automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking. Genomics. 1995;25:674–681.[Medline] [Order article via Infotrieve]
18. Iizuka M, Makino R, Sekiya T, Hayashi K. Selective isolation of highly polymorphic (dC- dA)n, (dG-dT)n microsatellites by stringent hybridization. Genet Anal Tech Appl. 1993;10:2–5.[Medline] [Order article via Infotrieve]
19.
Mount SM. A catalog of splice junction sequences.
Nucleic Acids Res. 1982;10:459–472.
20. Nakayama T, Soma M, Izumi Y, Kanmatsuse K. Organization of the human prostacyclin synthase gene. Biochem Biophys Res Commun. 1996;221:803–806.[Medline] [Order article via Infotrieve]
21.
Yamaguchi M, Rutledge LJ, Garbers DL. The primary
structure of the rat guanylyl cyclase A/atrial natriuretic
peptide gene. J Biol Chem. 1990;265:20414–20420.
21. Takahashi Y, Nakayama T, Soma M, Izumi Y, Kanmatsuse K. Organization of the human natriuretic peptide receptor A gene. Biochem Biophys Res Commun. 1998;246:736–739.[Medline] [Order article via Infotrieve]
22.
John SW, Krege JH, Oliver PM, Hagaman JR, Hodgin JB,
Pang SC, Flynn TG, Smithies O. Genetic decreases in atrial
natriuretic peptide and salt-sensitive hypertension.
Science. 1995;267:679–681.
23. Lopez MJ, Wong SK-F, Kishimoto I, Dubois S, Mach V, Friesen J, Garbers DL, Beuve A. Salt-resistant hypertension in mice lacking the guanylyl cyclase-A receptor for atrial natriuretic peptide. Nature. 1995;378:65–68.[Medline] [Order article via Infotrieve]
24. Berge KE, Berg K. No effect on blood pressure level or variability of polymorphism in DNA at the locus for atrial natriuretic factor. Clin Genet. 1994;46:433–435.[Medline] [Order article via Infotrieve]
25. Rutledge DR, Sun Y, Ross EA. Polymorphisms within the atrial natriuretic peptide gene in essential hypertension. J Hypertens. 1995;13:953–955.[Medline] [Order article via Infotrieve]
26. Morris BJ, Zee RYL, Ying L-H, Griffiths LR. Independent, marked associations of alleles of the insulin receptor and dipeptidyl carboxypeptidase-1 genes with essential hypertension. Clin Sci (Colch). 1993;85:189–195.[Medline] [Order article via Infotrieve]
27. Schorr U, Beige J, Ringel J, Turan S, Kreutz R, Distler A, Sharma AM. Hpa II polymorphism of the atrial natriuretic peptide gene and the blood pressure response to salt intake in normotensive men. J Hypertens. 1997;15:715–718.[Medline] [Order article via Infotrieve]
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