Donate Help Contact The AHA Sign In Home
American Heart Association
Hypertension
Search: search_blue_button Advanced Search
Table of Contents | Next Article »
Hypertension. 1995;26:595-601

This Article
Right arrow Abstract Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Koike, G.
Right arrow Articles by Dzau, V. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Koike, G.
Right arrow Articles by Dzau, V. J.

(Hypertension. 1995;26:595-601.)
© 1995 American Heart Association, Inc.

Articles

Investigation of the Phenylethanolamine N-Methyltransferase Gene as a Candidate Gene for Hypertension

George Koike; Howard J. Jacob; Jose E. Krieger; Claude Szpirer; Margret R. Hoehe; Masatsugu Horiuchi; Victor J. Dzau

From the Falk Cardiovascular Research Center, Stanford (Calif) University School of Medicine (G.K., J.E.K., M.H., V.J.D.); Cardiovascular Research Center, Massachusetts General Hospital–East, Charlestown (G.K., H.J.J.); Departement de Biologie Moleculaire, Université Libre de Bruxelles (Belgium) (C.S.); and Department of Genetics, Harvard Medical School, Boston, Mass (M.R.H.).

Correspondence to Victor J. Dzau, Falk Cardiovascular Research Center, Stanford University School of Medicine, 300 Pasteur Dr, Stanford, CA 94305-5246.


*    Abstract
up arrowTop
 *Abstract
down arrowIntroduction
 down arrowMethods
 down arrowResults
 down arrowDiscussion
 down arrowReferences
 
Abstract Genetic mapping studies have located a gene, Bp1, that accounts for approximately 30% of the genetic variation in the stroke-prone spontaneously hypertensive rat (SHRSP) to a region on chromosome 10 containing the angiotensin-converting enzyme gene. In humans, the gene encoding phenylethanolamine N-methyltransferase (PNMT) was localized near the angiotensin-converting enzyme gene on human chromosome 17. Since most of human chromosome 17 is known to be homologous to rat chromosome 10 and PNMT is known to play a role in blood pressure homeostasis, we reasoned (1) that the rat gene encoding PNMT (Pnmt) may reside on chromosome 10 within the confidence interval containing Bp1 and (2) that Pnmt is a good candidate gene for Bp1. With the use of a somatic cell hybrid panel and genetic mapping techniques, Pnmt mapped within the confidence interval that contains Bp1. To examine further this possibility of Pnmt as a candidate for Bp1, we cloned and characterized Pnmts of the original parental strains, the Wistar-Kyoto rat and SHRSP from the Heidelberg colony. We did not identify any sequence differences between the Wistar-Kyoto rats and SHRSP in the primary structure, in 1077 bp of the 5'-flanking region, or in the 256-bp 3'-end region, making Pnmt an unlikely gene for the genetic basis of salt-loaded hypertension.


Key Words: phenethanolamine N-methyltransferase • rats, inbred strains • DNA • cloning, molecular


*    Introduction
up arrowTop
 up arrowAbstract
 *Introduction
down arrowMethods
 down arrowResults
 down arrowDiscussion
 down arrowReferences
 
The terminal enzyme in the production of epinephrine that catalyzes the transmethylation of norepinephrine is PNMT (EC 2.1.1.28; S-adenosyl-L-methionine:phenylethanolamine N-methyltransferase).1 2 This enzyme is found predominantly in the chromaffin cells of the adrenal medulla and certain hypothalamic and brain stem neurons.3 4 5 Specifically, neurons containing PNMT are found in the area of the rostral ventrolateral medulla that plays a critical role in blood pressure homeostasis.6 Significant increases in PNMT activity and epinephrine content have been reported in various hypertensive animal models7 8 9 10 11 as well as in the brain stem of the SHR and SHRSP compared with the normotensive WKY.8 12 13 14 Inhibitors of PNMT reduce PNMT activity in the brain stem and lower blood pressure in the SHR and deoxycorticosterone acetate–salt models of hypertension,8 15 16 17 suggesting that PNMT activity might be involved in the development and maintenance of hypertension.

The gene encoding human PNMT (PNMT) has been cloned, characterized,18 19 and initially assigned to human chromosome 17 with the use of mouse/human somatic cell hybrids.20 With the use of two- and three-generation reference pedigrees, the human PNMT gene was mapped to human chromosome 17q21-q22,21 which also contains the ACE gene. Since the rat ACE gene (Ace) is located within the confidence interval containing Bp1,22 23 we reasoned that the rat PNMT gene (Pnmt) also may be within the Bp1 interval and another potential candidate gene for the increase in blood pressure after a salt load. To fulfill this hypothesis, Pnmt must be within the Bp1 interval, and sequence differences in the coding or regulatory regions must exist between Pnmt of the SHRSP and WKY.

Here we report the genetic mapping of Pnmt and present the complete Pnmt (only the partial cDNA has been reported).24 25 We also investigate the possibility of Pnmt being Bp1. Although we cannot formally rule out Pnmt as Bp1, the lack of a single base pair difference in the coding region, 1077 bp of the 5'-flanking region or 256 bp of the 3'-flanking region, makes this gene a very unlikely candidate.


*    Methods
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 *Methods
down arrowResults
 down arrowDiscussion
 down arrowReferences
 
Animals
SHRSP and WKY used for this study were from colonies formerly maintained at the University of Heidelberg, Germany (currently maintained at the MDC-Berlin-Buch). LEW and F344 used were from Harlan Sprague Dawley (Indianapolis, Ind).

Probe Preparation
The probes for this study were prepared by PCR amplification of genomic DNA of the WKY. Primers (forward primer, 5'-GTGAGGTGTCTGGACAGGTC-3'; reverse primer, 5'-AAGGAGTTAGGGAGGCAAAT-3') were designed based on the partial cDNA sequence for rat PNMT24 and yielded a 0.85-kbp fragment (Fig 1). Partial sequencing of this fragment was performed and compared with the rat cDNA sequence with the use of an ABI 373A DNA sequencer (Applied Biosystems), confirming that the amplified fragment contained the identical sequence of the cDNA (data not shown). This probe was labeled with [{alpha}-32P]dCTP (3000 Ci/mmol, DuPont–New England Nuclear) by a random primer labeling kit (Life Technologies, Inc) and purified with a NICK column (Pharmacia LKB Biotechnology).



View larger version (9K):
[in this window]
[in a new window]
  		Figure 1. Diagram shows organization of Pnmt and restriction map of genomic clones. The top two lines with restriction enzyme sites represent the isolated genomic DNA from the WKY library ({lambda}GKW4), and the bottom two lines indicate the isolated genomic DNA from the SHRSP library ({lambda}GKS2). The middle line indicates the location of the exons ({square}) of Pnmt. The black bar indicates the probe amplified by PCR. B indicates BamHI; E, EcoRI; Hc, HincII; Hd, HindIII; S, Sal I; and X, XhoI.

Chromosomal Assignment of Pnmt
The chromosomal assignment of Pnmt was performed by PCR amplification of a mouse/rat somatic cell hybrid panel as previously described.22 Primers described above were used for this experiment. Primers of mouse D-100 (D8 MIT16-F&R, Research Genetics) were used for internal control of PCR amplification.

Isolation of Simple Sequence Repeat Near Pnmt
Two P1 clones were isolated (Genome Systems Inc) with the use of PCR primers flanking Pnmt (forward primer, 5'-TTGCCCATTGATGTGCAC-3'; reverse primer, 5'-AGAAGGAGATGACCCCCG-3'). DNA of these P1 clones was digested with the restriction enzyme Alu I, HaeIII, or Rsa I (New England Biolabs). These digested DNA fragments were pooled and ligated to linkers (annealed with primer 1, 5'-CTGAGCGGAATTCGTGAGACC-3'; and primer 2, 5'-phosphorylated-GGTCTCACGAATTCCGCTCAGTT-3'). After ligation of this linker, PCR was performed with a dU-containing primer (5'-CUACUACUACUACTGAGCGGAATTCGTGAGAC-3') with the following protocol: six cycles of 94°C for 45 seconds, 60°C for 45 seconds, and 72°C for 75 seconds. These PCR products were cloned with the use of the CLONEAMP pAMP10 System (Life Technologies, Inc). Clones were screened for SSRs by colony hybridization26 with the use of Colony/Plaque Screen (DuPont–New England Nuclear), with oligonucleotide (CA)15 end labeled with [{gamma}-32P]ATP (6000 Ci/mmol, DuPont–New England Nuclear). Hybridization was carried out at 55°C in Church's hybridization solution.27 Filters were washed at 55°C in 5x SSC (1x SSC is 150 mmol/L NaCl and 15 mmol/L sodium citrate) and 0.1% SDS, followed by autoradiography at -80°C with an intensifying screen. Positive clones were sequenced with an ABI 373A DNA sequencer. PCR primers flanking the repeat were designed with the computer program PRIMER (S.E. Lincoln et al, unpublished results, 1991).

Mapping of the Genetic Marker for Pnmt
The experimental procedure in this section was previously described in detail.28 The PCR primers for Pnmt were characterized (allele sizes determined) for 12 different inbred rat strains. Since this genetic marker was not informative in the SHRSPxWKY intercross, the gene was mapped with an LEWxF344 intercross. This cross was genotyped with other genetic markers on rat chromosome 10, D10Mit1, D10Mgh5, D10Mgh7, D10Mgh8, and BAND3A.28 After genotyping, linkage analysis was performed with the MAPMAKER computer package29 with the use of the same criteria as previously described.28

Preparation and Screening of Genomic DNA Libraries
Genomic DNA was isolated as described26 from the spleens of WKY and SHRSP (spleens were kindly provided by Dr Detlev Ganten). Rat genomic libraries (prepared by Clontech Laboratories, Inc), which used partial Mbo I digests of genomic DNA cloned into the BamHI site of EMBL-3 (WKY genomic DNA library; independent plaques: 1.6x106; SHRSP genomic DNA library; independent plaques: 2.4x106). The genomic libraries were plated on Escherichia coli strain NM538 and screened by plaque hybridization30 with the use of Colony/Plaque Screen or Magna nylon membrane (Micron Separations Inc). Hybridization was carried out at 42°C in 50% formamide, 5x SSPE (1x SSPE is 150 mmol/L NaCl, 10 mmol/L NaH2PO4, and 1 mmol/L EDTA), 5x Denhardt's solution, 0.1% SDS, and 100 µg/mL denatured salmon sperm DNA with radiolabeled probe as described above. Filters were washed at 68°C in 0.1x SSC and 0.1% SDS for Colony/Plaque Screen or 0.2x SSC and 0.1% SDS for Magna nylon membrane. Positive clones, localized after autoradiography at -70°C with an intensifying screen, were isolated by repeated phage purification. Phage DNA was purified by the standard method26 with minor modification.

DNA Sequencing
Double-stranded DNA, prepared with Qiagen columns, was sequenced by the dideoxy chain termination method.31 Clones containing genomic DNA were sequenced in both directions with the T7 and T3 primers (Stratagene) or with internal primers and Sequenase (United States Biochemical).

Nomenclature
Rat strains, genes, and genetic markers were named in accordance with the rat nomenclature committee.28 32


*    Results
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMethods
 *Results
down arrowDiscussion
 down arrowReferences
 
Mapping of Pnmt
Pnmt was initially assigned to chromosome 10 with the use of a somatic cell hybrid panel (17 mouse/rat cell hybrids). The amplification pattern was concordant with chromosome 10 (Table 1). An internal control (mouse D-100) assured us that lack of an amplification product was not the result of a technical error in a specific lane.


View this table:
[in this window]
[in a new window]
  		Table 1. Cosegregation of Pnmt and Chromosome 10

Two P1 clones, which were confirmed to contain Pnmt by Southern blotting (data not shown), were screened to isolate (CA)n repeats. One clone containing a (CA)n repeat was identified and sequenced, and primers flanking the repeat were designed (forward primer, 5'-TCAAGTGTGCAGTGCCGT-3'; reverse primer, 5'-GCCC- GAGAACGTGTTTCTTA-3'). Allele sizes for this marker were determined for 12 rat strains (Table 2).


View this table:
[in this window]
[in a new window]
  		Table 2. Characterization of D10Mgh15 Allele Size

To map the marker for Pnmt, we used the progeny of an LEWxF344 intercross (the marker was not informative for the SHRSPxWKY cross). Fig 2 shows the genetic linkage map for the region containing Bp1 of rat chromosome 10 and illustrates that the marker for Pnmt (D10Mgh15) is within the interval containing Bp1.



View larger version (6K):
[in this window]
[in a new window]
  		Figure 2. Diagram shows the mapping position of D10Mgh15. The line indicates the position of and distance between genetic markers. Genetic distances are given in centimorgans computed with Kosambi's mapping function. The black box indicates the region containing Bp1.

Nucleotide Sequence of Pnmts
The structure of both Pnmts (SHRSP and WKY) is shown in Fig 1. The entire exon regions, the 1077-bp 5'-flanking region, the 256-bp 3'-end region, and the intron-exon boundaries of both genes were sequenced (Figs 3 and 4). Intron sizes were determined by restriction mapping and/or sequencing (Table 3). Pnmt is similar to the human and bovine homologues, having three exons interrupted by two introns,18 19 33 and spans approximately 2 kbp of DNA (Fig 1). All exon-intron junction sequences follow the GT/AG rule.34 The complete primary structure of the rat PNMT is deduced from exon sequences (Fig 3). The putative rat PNMT is composed of 285 amino acid residues, with an estimated molecular weight of 31.6 kD.



View larger version (78K):
[in this window]
[in a new window]
  		Figure 3. Putative primary structure of rat PNMT. The nucleotide sequence of the coding region presented in the top lines is numbered on the left, with +1 beginning at the first methionine. The deduced amino acid sequence presented in the bottom lines is numbered on the right. The termination codon TGA is denoted by asterisks (***). The italicized characters indicate the missing region of the partial cDNA for rat PNMT.24




View larger version (70K):
[in this window]
[in a new window]
  		Figure 4. Nucleotide sequence of the 5'-flanking region and 3'-end of Pnmt. A, Nucleotide sequence of the 5'-flanking region. The nucleotide sequence is numbered on the right. A TATA-box is outlined. The Sp1 binding sites (Sp1) and glucocorticoid responsive elements (GRE) are underlined. B, Nucleotide sequence of the 3'-end. The nucleotide sequence is numbered on the right. The polyadenylation (Poly A) signal is outlined.


View this table:
[in this window]
[in a new window]
  		Table 3. Nucleotide Sequence of the Intron-Exon Junction in Pnmt

There were no nucleotide differences in the regions sequenced between WKY and SHRSP. The 1077-bp 5'-flanking region of Pnmt contains consensus sequences for several known regulatory elements. These include the TATA box sequence,34 three potential Sp1 binding sites,35 and three homologous sequences to the glucocorticoid responsive element36 (Fig 4A), but no CAAT box sequence37 is found. The 256-bp 3'-end region contains the polyadenylation signal38 following a TGA translational stop codon (Fig 4B).


*    Discussion
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMethods
 up arrowResults
 *Discussion
down arrowReferences
 
Epidemiological and genetic studies have suggested that hypertension is a polygenic, heterogeneous disease. However, the specific genes causing hypertension are still unknown.39 40 41 To gain insight into human hypertension, researchers have studied rat models of genetic hypertension extensively. Recently, we and others performed genetic mapping studies on a cross of SHRSP and WKY22 23 and discovered a gene on rat chromosome 10 that we designated Bp1. This gene cosegregates with blood pressure after a salt load and accounts for nearly 30% of genetic variance.22 23

Ace mapped within the interval containing Bp1 and therefore was the first candidate gene. However, genetic mapping cannot prove the role for a candidate gene. To date, molecular biological studies have been unable to prove or refute Ace as Bp1. More recently, we (M.R.H.) genetically mapped the human PNMT gene to human chromosome 17q21-q22,21 which also contains the human ACE gene (ACE). Since PNMT seems to play a significant role in blood pressure homeostasis6 and previous physiological studies have demonstrated differences in PNMT activity and epinephrine content in the brain stems of SHR and SHRSP versus WKY,8 12 13 14 we hypothesize that Pnmt may be a candidate gene for Bp1. However, it is unclear whether the PNMT activity difference is primary or secondary. To examine this hypothesis, we set out to map Pnmt and clone it and compare its sequence between the WKY and SHRSP genes.

The somatic cell hybrid panel analysis demonstrated that the chromosomal location of Pnmt is chromosome 10, which is homologous to human chromosome 17. Our initial mapping attempts consisted of searching for restriction fragment length polymorphisms with 16 restriction enzymes (Alu I, BamHI, Bfa I, Bgl II, Dpn II, EcoRI, Hae III, HindIII, Kpn I, Msp I, Pst I, Pvu II, Rsa I, {alpha}-Taq I, Xba I, Xho I) and single-strand conformational polymorphisms (data not shown) that would yield a polymorphism between SHRSP and WKY. Neither strategy yielded a polymorphism, so we searched for SSRs in genomic {lambda}-clones containing Pnmt; however, none were identified. Subsequently, a (CA)n repeat for the Pnmt was identified in a P1 clone, which generally carries a 75- to 100-kbp insert.42 As the genetic distance between Pnmt and this (CA)n repeat is less than 0.1 cM, it could be used as a genetic marker for Pnmt. Unfortunately, this SSR was not polymorphic between SHRSP and WKY. Pnmt was finally mapped to within the interval containing Bp1 with the use of an LEWxF344 intercross.

The mapping data suggested that Pnmt was a candidate gene for Bp1; however, our inability to identify a polymorphism between the SHRSP and WKY implied that the genes may not be different. We next cloned it from the SHRSP and WKY to see whether we could identify any sequence variation.

Screening of genomic DNA libraries of WKY and SHRSP yielded four clones from the WKY and two from the SHRSP. The structure of the gene was determined by restriction mapping and sequencing of these clones. As shown in Fig 1, Pnmt spans approximately 2 kbp of DNA, containing three exons and two introns. The organization of the rat gene is similar to those of the human and bovine genes.18 19 33 Rat PNMT is composed of 285 amino acid residues as deduced from the nucleotide sequence of entire exon regions. The estimated molecular weight is 31.6 kD. The primary structure of rat PNMT is similar to those of human PNMT (282 amino acid residues, 30.9 kD)20 and bovine PNMT (284 amino acid residues, 31.1 kD).43 Rat PNMT is 80% to 90% homologous to these other species. We also sequenced and characterized the regulatory regions of Pnmt, ie, 1077 bp of the 5'-flanking region and 256 bp of the 3'-end region (Fig 4). There are consensus sequences for several known regulatory elements, such as the TATA box, Sp1 binding site, glucocorticoid responsive element, and polyadenylation signal. The structure of the regulatory region of the rat gene is similar to those of human18 19 and bovine33 genes.

We compared Pnmt between WKY and SHRSP. No differences were found for the entire coding region, the 5'-flanking region, and the 3'-end region. Introns 1 and 2 were also sequenced, and again there were no differences; however, a 200-bp region of intron 1 could not be sequenced. PCR primers flanking the region of intron 1 that could not be sequenced were designed and amplified; however, regular polyacrylamide gel analysis and single-strand conformational polymorphism analysis failed to identify polymorphisms.

If there is no difference between the genes, how do we account for the expression differences reported between the SHRSP and WKY? One possibility is that the expression differences observed are a secondary consequence of the hypertension in the SHRSP. Another possibility that plagues the rat community is the variation among strains with the same name. We favor the former because we obtained the SHRSP and WKY from the same group which produced the original cross that showed cosegregation of blood pressure with the interval that contains Pnmt.

Given our inability to detect any difference at the restriction mapping level down to the sequence of Pnmt between the SHRSP and WKY, we believe it is unlikely that Pnmt itself contributes to the genetic basis of salt-loaded hypertension. However, we cannot exclude the possibilities that differences exist in the region that we did not analyze in this report or that Bp1 encodes a trans-acting factor regulating Pnmt expression.


*    Selected Abbreviations and Acronyms
 
ACE = angiotensin-converting enzyme
F344 = Fischer rat(s)
LEW = Lewis rat(s)
PCR = polymerase chain reaction
PNMT = phenylethanolamine N-methyltransferase
SDS = sodium dodecyl sulfate
SHR = spontaneously hypertensive rat(s)
SHRSP = stroke-prone spontaneously hypertensive rat(s)
SSR = simple sequence repeat
WKY = Wistar-Kyoto rat(s)


*    Acknowledgments
 
This work was supported in part by National Institutes of Health grants HL-46631, HL-35610, and HL-35252 to V.J.D.; grants from National Institute of Diabetes, Digestive and Kidney Diseases, and Bristol-Myers Squibb to H.J.J.; and grants from the Fund for Scientific Medical Research (FRSM, Belgium) and the Belgian programme on interuniversity attraction poles initiated by the Belgian State Prime Minister's Office (SSTC/DWTC) to C.S. G.K. is the recipient of a Postdoctoral Fellowship Award from the American Heart Association, California Affiliate. C.S. is a Research Director of the FNRS, Belgium. We extend special thanks to Ester Sternberg and Sam Listwak for providing the DNAs of the cross of LEWxF344 and to Donna M. Brown, Anna Pettersson, Michele Riviere, Jason S. Simon, Eric S. Winer, and Armand J. MacMurray for help in mapping Pnmt. Nucleotide sequence data from this article have been deposited in the GenBank data base under accession numbers U11275 and U11694.

Received May 25, 1994; first decision July 26, 1994; accepted July 10, 1995.


*    References
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMethods
 up arrowResults
 up arrowDiscussion
 *References
 

  1. Kirshner N, Goodall M. The formation of adrenaline from noradrenaline. Biochim Biophys Acta. 1957;24:658-659.
  2. Axelrod J. Purification and properties of phenylethanolamine-N-methyl transferase. J Biol Chem. 1962;237:1657-1660. [Free Full Text]
  3. Hökfelt T, Fuxe K, Goldstein M, Johansson O. Evidence for adrenaline neurons in the rat brain. Acta Physiol Scand. 1973;89:286-288. [Medline] [Order article via Infotrieve]
  4. Hökfelt T, Fuxe K, Goldstein M, Johansson O. Immunohistochemical evidence for the existence of adrenaline neurons in the rat brain. Brain Res. 1974;66:235-251.
  5. Saavedra JM, Palkovits M, Brownstein MJ, Axelrod J. Localisation of phenylethanolamine N-methyl transferase in the rat brain nuclei. Nature. 1974;248:695-696. [Medline] [Order article via Infotrieve]
  6. Reis DJ, Morrison S, Ruggiero DA. The C1 area of the brainstem in tonic and reflex control of blood pressure. Hypertension. 1988;11(suppl I):I-8-I-13.
  7. DeQuattro V, Nagatsu T, Maronde R, Alexander N. Catecholamine synthesis in rabbits with neurogenic hypertension. Circ Res. 1969;24:545-555. [Abstract/Free Full Text]
  8. Saavedra JM, Grobecker H, Axelrod J. Adrenaline-forming enzyme in brainstem: elevation in genetic and experimental hypertension. Science. 1976;191:483-484. [Abstract/Free Full Text]
  9. Petty MA, Reid JL. Catecholamine synthesizing enzymes in brain stem and hypothalamus during the development of renovascular hypertension. Brain Res. 1979;163:277-288. [Medline] [Order article via Infotrieve]
  10. Saavedra JM. Brain catecholamines during development of DOCA-salt hypertension in rats. Brain Res. 1979;179:121-127. [Medline] [Order article via Infotrieve]
  11. Fuxe K, Agnati LF, Kitayama I, Zoli M, Janson AM, Härfstrand A, Vincent M, Kalia M, Goldstein M, Sassard J. Evidence for discrete alterations in central cardiovascular catecholamine and neuropeptide Y immunoreactive neurons in aged male rats and in genetically hypertensive male rats of the Lyon strain. Eur Heart J. 1987;8:139-145.
  12. Wijnen HJLM, Versteeg DHG, Palkovits M, De Jong W. Increased adrenaline content of individual nuclei of the hypothalamus and the medulla oblongata of genetically hypertensive rats. Brain Res. 1977;135:180-185. [Medline] [Order article via Infotrieve]
  13. Wijnen HJLM, Palkovits M, De Jong W, Versteeg DHG. Elevated adrenaline content in nuclei of the medulla oblongata and the hypothalamus during the development of spontaneous hypertension. Brain Res. 1978;157:191-195. [Medline] [Order article via Infotrieve]
  14. Chalmers JP, Minson J, Denoroy L, Stead B, Howe PRC. Brainstem PNMT neurons and experimental hypertension in the rat. Clin Exp Hypertens. 1984;6:243-258.
  15. Saavedra JM. Adrenaline levels in brain stem nuclei and effects of a PNMT inhibitor on spontaneously hypertensive rats. Brain Res. 1979;166:283-292. [Medline] [Order article via Infotrieve]
  16. Black J, Waeber B, Bresnahan MR, Gavras I, Gavras H. Blood pressure response to central and/or peripheral inhibition of phenylethanolamine N-methyltransferase in normotensive and hypertensive rats. Circ Res. 1981;49:518-524. [Abstract/Free Full Text]
  17. Chatelain RE, Manniello MJ, Dardik BN, Rizzo M, Brosnihan KB. Antihypertensive effects of CGS 19281A, an inhibitor of phenylethanolamine-N-methyltransferase. J Pharmacol Exp Ther. 1990;252:117-125. [Abstract/Free Full Text]
  18. Baetge EE, Behringer RR, Messing A, Brinster RL, Palmiter RD. Transgenic mice express the human phenylethanolamine N-methyltransferase gene in adrenal medulla and retina. Proc Natl Acad Sci U S A. 1988;85:3648-3652. [Abstract/Free Full Text]
  19. Sasaoka T, Kaneda N, Kurosawa Y, Fujuta K, Nagatsu T. Structure of human phenylethanolamine N-methyltransferase gene: existence of two types of mRNA with different transcription initiation sites. Neurochem Int. 1989;15:555-565.
  20. Kaneda N, Ichinose H, Kobayashi K, Oka K, Kishi F, Nakazawa A, Kurosawa Y, Fujita K, Nagatsu T. Molecular cloning of cDNA and chromosomal assignment of the gene for human phenylethanolamine N-methyltransferase, the enzyme for epinephrine biosynthesis. J Biol Chem. 1988;263:7672-7677. [Abstract/Free Full Text]
  21. Hoehe MR, Plaetke R, Otterud B, Stauffer D, Holik J, Byerley WF, Baetge EE, Gershon ES, Lalouel J-M, Leppert M. Genetic linkage of the human gene for phenylethanolamine N-methyltransferase (PNMT), the adrenaline-synthesizing enzyme, to DNA markers on chromosome 17q21-q22. Hum Mol Genet. 1992;1:175-178. [Abstract/Free Full Text]
  22. Jacob HJ, Lindpaintner K, Lincoln SE, Kusumi K, Bunker RK, Mao Y-P, Ganten D, Dzau VJ, Lander ES. Genetic mapping of a gene causing hypertension in the stroke-prone spontaneously hypertensive rat. Cell. 1991;67:213-224.[Medline] [Order article via Infotrieve]
  23. Hilbert P, Lindpaintner K, Beckmann JS, Serikawa T, Soubrier F, Dubay C, Cartwright P, De Gouyon B, Julier C, Takahashi S, Vincent M, Ganten D, Georges M, Lathrop GM. Chromosomal mapping of two genetic loci associated with blood-pressure regulation in hereditary hypertensive rats. Nature. 1991;353:521-529. [Medline] [Order article via Infotrieve]
  24. Mezey É. Cloning of the rat adrenal medullary phenylethanolamine-N-methyltransferase. Nucleic Acids Res. 1989;17:2125. [Free Full Text]
  25. Ross ME, Evinger MJ, Hyman SE, Carroll JM, Mucke L, Comb M, Reis DJ, Joh TH, Goodman HM. Identification of a functional glucocorticoid response element in the phenylethanolamine N-methyltransferase promoter using fusion genes introduced into chromaffin cells in primary culture. J Neurosci. 1990;10:520-530. [Abstract]
  26. Sambrook J, Fritsch EF, Maniatis T, eds. Molecular Cloning: A Laboratory Manual. 2nd ed. New York, NY: Cold Spring Harbor Laboratory Press; 1989.
  27. Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984;81:1991-1995. [Abstract/Free Full Text]
  28. Jacob HJ, Brown DM, Bunker RK, Daly MJ, Dzau VJ, Goodman A, Koike G, Kren V, Kurtz T, Lernmark Å, Levan G, Mao Y-P, Pettersson A, Pravenec M, Simon JS, Szpirer C, Szpirer J, Trolliet MR, Winer ES, Lander ES. A genetic linkage map of the laboratory rat, Rattus norvegicus. Nature Genet. 1995;9:63-69. [Medline] [Order article via Infotrieve]
  29. Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L. MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics. 1987;1:174-187. [Medline] [Order article via Infotrieve]
  30. Benton WD, David RW. Screening {lambda}gt recombinant clones by hybridization to single plaques in situ. Science. 1977;196:180-182. [Abstract/Free Full Text]
  31. Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977;74:5463-5467. [Abstract/Free Full Text]
  32. Definition, nomenclature, and conservation of rat strains. ILAR News. 1993;34:S1-S26.
  33. Batter DK, D'Mello SR, Turzai LM, Hughes HB III, Gioio AE, Kaplan BB. The complete nucleotide sequence and structure of the gene encoding bovine phenylethanolamine N-methyltransferase. J Neurosci Res. 1988;19:367-376. [Medline] [Order article via Infotrieve]
  34. Breathnach R, Chambon P. Organization and expression of eukaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349-383. [Medline] [Order article via Infotrieve]
  35. Dynan WS, Saffer JD, Lee WS, Tjian R. Transcription factor Sp1 recognizes promoter sequences from the monkey genome that are similar to the simian virus 40 promoter. Proc Natl Acad Sci U S A. 1985;82:4915-4919. [Abstract/Free Full Text]
  36. Jantzen H-M, Strähle U, Gloss B, Stewart F, Schmid W, Boshart M, Miksicek R, Schütz G. Cooperativity of glucocorticoid responseelements located far upstream of the tyrosine aminotransferase gene. Cell. 1987;49:29-38. [Medline] [Order article via Infotrieve]
  37. Efstratiadis A, Posakony JW, Maniatis T, Lawn RM, O'Connell C, Spritz RA, DeRiel JK, Forget BG, Weissman SM, Slightom JL, Blechl AE, Smithies O, Baralle FE, Shoulders CC, Proudfoot NJ. The structure and evolution of the human ß-globin gene family. Cell. 1980;21:653-668. [Medline] [Order article via Infotrieve]
  38. Proudfoot NJ, Brownlee GG. 3' Non-coding region sequences in eukaryotic messenger RNA. Nature. 1976;263:211-214. [Medline] [Order article via Infotrieve]
  39. Havlik RJ, Garrison RJ, Feinleib M, Kannel WB, Castelli WP, McNamara PM. Blood pressure aggregation in families. Am J Epidemiol. 1979;110:304-312. [Abstract/Free Full Text]
  40. Higgins M, Keller J, Moore F, Ostrander L, Metzner H, Stock L. Studies of blood pressure in Tecumseh, Michigan, I: blood pressure in young people and its relationship to personal and familial characteristics and complications of pregnancy in mothers. Am J Epidemiol. 1980;111:142-155. [Abstract/Free Full Text]
  41. Levine RS, Hennekens CH, Perry A, Cassady J, Gelband H, Jesse MJ. Genetic variance of blood pressure levels in infant twins. Am J Epidemiol. 1982;116:759-764. [Abstract/Free Full Text]
  42. Smoller DA, Petrov D, Hartl DL. Characterization of bacteriophage P1 library containing inserts of Drosophila DNA of 75-100 kilobase pairs. Chromosoma. 1991;100:487-494. [Medline] [Order article via Infotrieve]
  43. Baetge EE, Suh YH, Joh TH. Complete nucleotide and deduced amino acid sequence of bovine phenylethanolamine N-methyltransferase: partial amino acid homology with rat tyrosine hydroxylase. Proc Natl Acad Sci U S A. 1986;83:5454-5458.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 HypertensionHome page
Y. Saad, S. Yerga-Woolwine, J. Saikumar, P. Farms, E. Manickavasagam, E. J. Toland, and B. Joe
Congenic Interval Mapping of RNO10 Reveals a Complex Cluster of Closely-Linked Genetic Determinants of Blood Pressure
Hypertension, November 1, 2007; 50(5): 891 - 898.
[Abstract] [Full Text] [PDF]

Home page
 HypertensionHome page
R. L. Jaworski, M. Jirout, S. Closson, L. Breen, P. L. Flodman, M. A. Spence, V. Kren, D. Krenova, M. Pravenec, and M. P. Printz
Heart Rate and Blood Pressure Quantitative Trait Loci for the Airpuff Startle Reaction
Hypertension, February 1, 2002; 39(2): 348 - 352.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Koike, G.
Right arrow Articles by Dzau, V. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Koike, G.
Right arrow Articles by Dzau, V. J.

Hypertension Home | Subscriptions | Archives | Feedback | Authors | Help | AHA Journals Home | Search
Copyright © 1995 American Heart Association, Inc. All rights reserved. Unauthorized use prohibited.