doi: 10.1161/hy1001.092652
(Hypertension. 2001;38:1137.)
© 2001 American Heart Association, Inc.
Scientific Contributions |
Localization of the Endothelin System in Aldosterone-Producing Adenomas
From the INSERM Unit 36, Collège de France (G.E., P.C., F.P.), Paris, France; Service d’Anatamopathologie, Hopital Saint-Joseph (E.B.), Paris, France; and Catedra de Bioquimica Avanzada, Universidad Nacional de San Luis (G.C.), San Luis, Argentina.
Correspondence to Dr Florence Pinet, INSERM Unit 36, Collège de France, 11 place Marcellin Berthelol, 75005 Paris, France. E-mail florence.pinet{at}college-de-france.fr
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Abstract |
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Abstract—— Endothelin-1 (ET-1) could play a role in the regulation of aldosterone secretion of the human adrenal gland. The presence of the endothelin-converting enzyme 1 (ECE-1) and ET-1 suggests that there is a local ET system in the adrenal cortex, but the in situ synthesis of ET-1 remains to be confirmed. The cellular distribution of the whole ET system was evaluated in 20 cases of aldosterone-producing adenomas. Polymerase chain reaction studies gave strong signals for ECE-1 mRNA and the mRNAs for endothelin type A (ETA) and B (ETB) receptors and faint signals for prepro–ET-1 mRNA. In situ hybridization showed ETA receptors scattered throughout the adenoma, in both secretory cells and vascular structures (score, +). There were more ETB receptors (score, ++), but they were restricted mainly to the endothelium. ECE-1 mRNA and protein were ubiquitous and abundant in secretory cells (score, +++) and vascular structures (score, ++); the enzyme was active on big ET-1. There was no prepro–ET-1 mRNA in the cortex, except in the thickened precapillary arterioles present in only 30% of the aldosterone-producing adenomas studied. ET-1 immunoreactivity was detected in vascular structures (score, +), probably bound to receptors, suggesting that ET-1 has an endocrine action. The low concentrations of ET-1 could also indicate that it acts in a paracrine-autocrine fashion to control adrenal blood flow. The discrepancy between the concentrations of ECE-1 and its substrate suggests that ECE-1 has another role in the adrenal secretory cells. Our data indicate that ET probably is not a primary cause of the development or maintenance of the adenoma.
Key Words: endothelin • hypertension, endocrine • receptors, endothelin • regional blood flow • immunohistochemistry
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Introduction |
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Endothelin-1 (ET-1) is one of three 21–amino acid peptides, the ETs (ET-1, ET-2, and ET-3), that mediate a broad spectrum of biologic actions, such as the regulation of endocrine secretion.1 It also is involved in specific vascular lesions and inflammatory conditions (primary and malignant hypertension).2 The ETs act on 2 distinct high-affinity G protein–coupled receptors, endothelin type A (ETA)3 and type B (ETB),4 and are synthesized as large precursor polypeptides, the prepro-ETs, which are cleaved at 2 pairs of basic amino acids to generate intermediate peptides, the big ETs. The big ETs are then cleaved by an endothelin-converting enzyme (ECE), initially purified from bovine adrenal cortex,5 to produce the mature ETs. Two human ECE genes have been cloned, ECE-16 and ECE-2,7 and they are 59% identical.
ET-1 was first found in the vascular endothelium,8 but it is also present in endocrine tissues, such as the adrenal gland,9 where it may mediate the adrenocortical response to ACTH.10 Large amounts of ECE-1 mRNA and protein have been found in the human adrenal gland, with the highest concentrations in the zona glomerulosa and fasciculata.11 Both ET receptor subtypes and ET peptides are found in the human adrenal gland.9 The mRNAs for ETA, ETB, and prepro–ET-1 (PPET-1) have also been detected by Northern blotting,9 RT-PCR,12 and immunohistochemistry.13 Quantitative analysis of the binding of 125I-ET-1 by autoradiography together with the use of selective ligands showed that ETB receptors are present through the entire adrenal cortex, whereas ETA receptors are restricted to the zona glomerulosa.14
Compelling evidence indicates that the ET components in the normal adrenal gland could play a role in the regulation of this gland. Several reports suggest that ET-1 is implicated in steroid hormone secretion. Exogenous ET-1 influenced adrenal function in human volunteers by selectively increasing the ACTH-induced secretion of aldosterone,15 and a limited study showed that patients with hypertension due to primary aldosteronism had abnormally high levels of plasma ET-1.16
Conn’s syndrome is a major cause of primary hyperaldosteronism due to an aldosterone-producing adenoma (APA) in 1 adrenal gland. The APA is histogenetically derived from the zona glomerulosa cells,14 which form the proliferative layer involved in the maintenance of growth of the entire adrenal cortex in mammals and the presence of both ET receptors in APAs.13,17 Thus, ET-1 may help to regulate the growth of the adrenal cortex.
This study was conducted to elucidate the pathophysiological significance of ET-1 in APA adrenal glands, as ET-1 is shown to be the predominant mature isoform in the human adrenal gland.12 We looked for mRNAs that encode all the components of the ET system in human APA and immunoreactive ET-1 and ECE-1 to determine whether there is a complete local ET system in the human adrenal cortex or whether ET-1 is supplied by the blood.
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Methods |
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Human Tissues
Human adrenal tissue was obtained from patients (n=20) undergoing unilateral adrenalectomy for Conn’s syndrome by the COMETE network in accordance with French laws. The patients included 11 middle-aged women and 9 middle-aged men. The adenomas weighed 3.1 to 16.8 g. A small piece of each specimen was frozen for RNA extraction, and the remainder was fixed in 4% buffered paraformaldehyde, for
24 hours, before processing.
RNA Isolation and RT-PCR
Total RNA was isolated from frozen adrenal cortex according to the protocol of Chomczynski and Sacchi.18 cDNA was prepared with 5x10-6 g total RNA and 15 pmol/L oligo(dT) with Moloney murine leukemia virus reverse transcriptase (GIBCO BRL) in 50 µL according to the manufacturer’s instructions. PCR was performed using 3 µL cDNA solution and specific primers for ECE-1,19 PPET-1,20 ETA,3 and ETB4 receptors and GAPDH,21 as previously described.22 Amplified products were analyzed on 2% agarose gel.
In Situ Hybridization
Radiolabeled riboprobes for PPET-1, ECE-1, ETA, and ETB mRNAs were prepared and used with deparaffinized sections (5 µm).22 The processed sections were exposed for 2 weeks with ECE-1, for 4 weeks with ETA and ETB probes, and for 6 weeks with PPET-1 probes and counterstained with toluidine blue.
Immunohistochemistry
Paraffin was removed from embedded sections (5 µm) with xylene, and the sections were rehydrated in a graded ethanol series. Their endogenous peroxidase was inactivated with 3% hydrogen peroxide in methanol for 10 minutes before incubation with monoclonal antibodies to CD31 (DAKO), 
-smooth muscle actin (-SMA) (DAKO), and ET-1 (IHC 6901; Peninsula Laboratories) according to the manufacturer’s instructions. The specific antiserum 473-17-A23 was used for ECE-1. Sections were then reacted with an avidin-biotin complex (ABC; DAKO), treated with 0.035% diaminobenzidine (Fluka) and counterstained with hematoxylin (according to Mayer). Control reactions without primary antibodies showed no nonspecific staining (not shown).
The ET-1 antibody used was specific for human ET-1 but cross-reacted (76%) with human big ET-1 according to the manufacturer. However, we tested the antibody to determine whether it recognized receptor-bound ET-1 by using stable transfected Chinese hamster ovary (CHO) cell lines embedded in paraffin. These were CHO/PPET-1/ECE-1 cells, which secrete human ET-1, and CHO/ETB cells, which produce human ETB receptors. Briefly, cells were grown to confluence, scraped off, collected by centrifugation, fixed with 4% paraformaldehyde, and embedded in paraffin. Cultures of CHO/ETB grown to confluence were incubated with 20 nmol/L ET-1 for 30 minutes at 4°C and processed as described earlier.
The results of in situ hybridization (ISH) and immunohistochemistry (IHC) are summarized on a 4-point scale (Table), although the techniques used provide only semiquantitative data.
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ECE Extraction and Activity
Membranes were prepared at 4°C from 80 mg of APA.23 Big ET-1 was converted to ET-1 by incubation of the solubilized membranes (50 to 200 µg protein) with 0.1 µmol/L big ET-1 for 30 minutes at 37°C. The specificity of the ECE enzymatic activity was checked with phosphoramidon, a zinc metalloprotease inhibitor. The mixture was assayed using a commercial enzyme immunoassay kit (Cayman Chemical Co) to quantify ET-1.
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Results |
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ET System in APAs
Tissue samples from 20 patients of the COMETE network were analyzed, and the results shown are representative of all the cases. Histological examination of the specimens revealed a cortical adenoma in all cases; the appearance was heterogeneous, with a predominant hyperplasic zona glomerulosa (not shown).
RT-PCR of frozen tissue gave strong amplification of ECE-1 in all samples, low amplification of ETA and ETB mRNAs, and little or no amplification of PPET-1 (Figure 1).
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/HindIII, 
X174 DNA/HaeIII.
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The distribution of each component of the ET system in adrenocortical adenomas was determined by ISH. The results were in agreement with the RT-PCR data. Figure 2 is representative of the 20 APA samples studied. Dark-field and bright-field microphotographs of labeling with each riboprobe showed diffuse ETA receptor mRNA (Figure 2a and 2b), a high concentration of ETB receptor mRNA mainly on the framework of the secretory cells (Figure 2c and 2d), clumps of intense ECE-1 mRNA hybridization (Figure 2e and 2f), and no specific signal for PPET-1 mRNA (Figure 2g and 2h). ETA mRNA labeling was stronger in the compact cells than in the lipid-rich cells (Figure 2b), whereas ECE-1 mRNA was concentrated in glomerular lipid-rich cells but sparse in the adjacent compact zona glomerulosa–like cells (Figure 2f). ETB mRNA labeling was intense in the zona glomerulosa but weak in the zona fasciculata and mainly confined to capillaries running among tumor cells (Figure 2d). Sense probes yielded no labeling in any tissue (not shown).
Cellular Distribution of the ET System in APAs
Figure 3 shows the cellular distribution of receptor transcripts in APAs. Semiquantitative evaluation was performed on the 20 samples (Table). ETA mRNA (Figure 3a) was found in secretory cells (score, +), in the smooth muscle layer of arteries, and in a few smooth muscle cells (score, +) surrounding arterioles that stained for -SMA (Figure 3c). In contrast, ETB mRNA (Figure 3b) was found mainly within the network of capillary sinusoids, with high concentrations in endothelial cells (score, ++) that stained for CD-31 (Figure 3d) and in -SMA–positive cells adjacent to the vessels (score, ++) (Figure 3c). Secretory cells were occasionally labeled.
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We tested a specific anti–ET-1 antibody. The ET-1 antibody labeled CHO/PPET-1/ECE-1 cells producing ET-1 (Figure 4a) and CHO/ETB cells incubated with ET-1 (Figure 4c) under the same conditions as those used for the APA sections, but not CHO/ETB cells alone (Figure 4b). The antibody did not react nonspecifically with untransfected CHO cells (Figure 4d). Thus, the antibody labeled cells synthesizing ET-1 as well as cells binding ET-1.
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Figure 5 shows the cellular distribution of ECE-1 (Figure 5a and 5b) and PPET-1 (Figure 5c to 5f). There was a strong signal for ECE-1 mRNA in several cell types, mainly secretory cells (score, +++) (Figure 5a) and some endothelial cells (score, ++) (Table). ECE-1 immunoreactivity correlated perfectly with the ISH results, confirming the high concentration of the enzyme (Figure 5b). No signal for PPET1 mRNA was found in any structure (Figure 5c). However, ET-1 immunoreactivity was found in endothelial cells (score, ++) (Figure 5d), showing a discrepancy between the de novo synthesis of PPET-1 and the presence of ET-1 peptide.
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Only 6 samples of APA adrenal cortex presented a strong labeling for PPET-1 mRNA (Figure 5e), which localized to the thickened wall of some precapillary arterioles characterized by Lack et al.24 The high concentration of PPET-1 mRNA was correlated with ET-1 immunoreactivity (Figure 5f). The thickened capillary arterioles also had ECE-1 in their endothelial and perivascular cells and ETA in the smooth muscle cells (not shown).
ECE-1 Activity in the APA Adrenal Cortex
Solubilized membranes were prepared from a sample of APA adrenal cortex, and the enzymatic properties of ECE-1 were measured. The enzyme activity converted 0.1 µmol/L big ET-1 at a rate of 3.7 pg · min-1 · mg protein-1 and was >90% inhibited by phosphoramidon.
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Discussion |
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We examined the role of the ET system in endocrine glands. Patients with Conn’s syndromes were studied because their tumors arise from the zona glomerulosa of the adrenal cortex. The high concentrations of human ECE-1 mRNA and its protein in the adrenal gland11 suggest that ECE-1 is involved in other systems, such as the regulation of hormone secretion, rather than simply generating ET-1 from its precursor.
We tested this notion by determining the amounts and the cell-specific distributions of the ET system in human APAs. The adrenal cortex of APAs contains high concentrations of ECE-1 and ET receptor mRNAs, but PPET-1 mRNA is not easily detected. The Table summarizes the cellular localization of the components of the ET system, showing that secretory cells contain ETA mRNA and ECE-1 mRNA and protein, whereas the vascular structures contain both receptors, ECE-1 mRNA and protein, as well as ET-1 peptide probably associated with the receptors. To our knowledge, this is the first report of the cellular distributions of PPET-1, ETA, and ETB mRNAs in the human adrenal.
The distribution of ET receptor mRNAs determined by ISH is in good agreement with previous results of autoradiography that could not elucidate the type of cells showing the binding.14 ET-1 acts as secretagogue for calf adrenal zona glomerulosa cells25 and is trophic for rat adrenal zona glomerulosa cells.26 These authors suggested that ET-1 could be the factor that causes enhanced aldosterone secretion in APAs. The decrease in ETB receptors reported by Rossi et al27 and Zeng et al17 could cause the higher plasma ET-1 concentrations described by some,16 as ETB may also function as a physiological clearance receptor for extracellular ET-1.28 Lariviere et al29 suggested that the increased ET-1 content is due to its involvement in the maintenance of high blood pressure in mineralocorticoid hypertension. The vascular distribution of both ET receptors, plus the presence of ETA in some secretory cells, supports the proposal of Hinson et al10 that ET-1 regulates aldosterone secretion in the adrenal glands in a paracrine fashion.
ET-1 is involved in adrenal steroid secretion, but its cellular origin remains unclear. RT-PCR of whole APA cortex resulted in a faint band for PPET-1, in accordance with published reports,9,27 although PPET-1 mRNA has been detected in dispersed and purified human adrenocortical cells.30,31 However, we detected no PPET-1 signal on secretory cells containing ECE-1 by ISH. One possible explanation is that the low level of transcription is not detected by ISH. Otherwise, RT-PCR amplified PPET-1 mRNA from a contaminant vessel of the capsule, or the technique used to disperse adrenocortical cells activated the transcription of PPET-1. Immunohistochemical studies are somewhat inconsistent about the distribution of ET-1 immunoreactivity. Some reports are in agreement with the present report, with ET-1 immunoreactivity confined to endothelial cells.12 Others found it in parenchymal cells,32,13 although it was recently demonstrated that the immunoreactive protein is ET-1–like and is a different size than PPET-1 or ET-1.33
Our experiments with cell lines showed that the anti–ET-1 antibody we used recognized both endogenous ET-1 and ET-1 bound to its receptor, suggesting that the discrepancy between ISH and IHC is due to the detection of ET-1 bound to ETB. A recent report showed that internalized ET-1–stimulated ETA receptors are rapidly recycled to the membrane, whereas internalized ETB receptors with almost irreversibly bound ET-1 are degraded more slowly.34 This would explain why receptor-bound ET-1 immunoreactivity was seen only on ETB-labeled cells and not in ETA receptors.
The presence of PPET-1 in the thickening of the walls of the precapillary arterioles of patients with a lymphocytic vascularitis24 is not surprising because this gene is very sensitive to shear stress and hypoxia and its transcription is enhanced in the majority of vascular diseases (see Miyauchi and Masaki2 for a recent review). The presence of ECE-1 in secretory cells in APAs might not be a hallmark of ET-1 biosynthesis but indicates an alternative function of ECE-1. Johnson et al35 recently showed that ECE-1 has a broad enzymatic activity for substrates other than big ETs, some of which are present in the adrenal cortex.
We conclude that ET-1 is not synthesized de novo in the human adrenal. The presence of ET within endothelial cells of adrenal capillaries could also indicate very low levels of newly synthesized ET-1. This would be consistent with a paracrine-autocrine role for ET-1 in controlling adrenal blood flow, as proposed by Davenport et al.12 However, several questions remain unanswered, such as the function of the surprisingly large amount of ECE-1.
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Acknowledgments |
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We thank Prof P.-F. Plouin and the COMETE network, and we thank Dr C. Parnot for providing CHO/PPET-1/ECE-1 cells. Dr Egidy was supported by funding from "La Fondation pour la Recherche Médicale." The English text was edited by Dr Owen Parkes.
Received December 15, 2000; first decision December 28, 2000; accepted April 23, 2001.
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References |
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1. Nussdorfer G, Rossi GP, Malendowicz LK, Mazzochi G. Autocrine-paracrine endothelin system in the physiology and pathology of steroid-secreting tissues. Pharmacol Rev. 1999; 51: 403–437.
2. Miyauchi T, Masaki T. Pathophysiology of endothelin in the cardiovascular system. Annu Rev Physiol. 1999; 61: 391–415.[Medline] [Order article via Infotrieve]
3. Hosoda K, Nakao K, Hiroshi S, Suga S, Ogawa Y, Mukoyama M, Shirami G, Saito Y, Nakanishi S, Imura H. Cloning and expression of human endothelin-1 receptor cDNA. FEBS Lett. 1991; 287: 23–26.[Medline] [Order article via Infotrieve]
4. Sakamoto A, Yanagisawa T, Sakurai T, Takuwa Y, Yanagisawa H, Masaki T. Cloning and functional expression of human cDNA for the ETB endothelin receptor. Biochem Biophys Res Commun. 1991; 178: 656–663.[Medline] [Order article via Infotrieve]
5. Xu D, Emoto N, Giaid A, Slaughter C, Kaw S, deWit D, Yanagisawa M. ECE-1: a membrane-bound metalloprotease that catalyzes the proteolytic activation of big endothelin-1. Cell. 1994; 78: 473–485.[Medline] [Order article via Infotrieve]
6.
Valdenaire O, Rohrbacher E, Mattei M. Organization of the gene encoding the human endothelin-converting enzyme (ECE-1). J Biol Chem. 1995; 270: 29794–29798.
7.
Emoto N, Yanagisawa M. Endothelin-converting enzyme-2 is a membrane-bound, phosphoramidon-sensitive metalloprotease with acidic pH optimum. J Biol Chem. 1995; 270: 15262–15268.
8. Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayishi Y, Mitsui Y, Yazaki Y, Goto K, Masaki T. A novel potent, vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988; 332: 411–415.[Medline] [Order article via Infotrieve]
9. Imai T, Hirata Y, Eguchi S, Kanno K, Ohta K, Emori T, Sakamoto A, Yanagisawa M, Masaki T, Marumo F. Concomitant expression of receptor subtype and isopeptide of endothelin by human adrenal gland. Biochem Biophys Res Commun. 1992; 182: 1115–1121.[Medline] [Order article via Infotrieve]
10.
Hinson J, Vinson G, Kapas S, Teja R. The role of endothelin in the control of adrenocortical function: stimulation of endothelin release by ACTH and the effects of endothelin-1 and endothelin-3 on steroidogenesis in rat and human adrenocortical cells. J Endocrinol. 1991; 128: 275–280.
11. Korth P, Bohle R, Corvol P, Pinet F. Cellular distribution of endothelin-converting enzyme-1 in human tissues. J Hystochem Cytochem. 1999; 47: 447–462.
12. Davenport A, Hoskins S, Kuc R, Plumpton C. Differential distribution of endothelin peptides and receptors in human adrenal gland. Histochem J. 1996; 28: 779–789.[Medline] [Order article via Infotrieve]
13. Hiraki H, Hoshi N, Hasegawa H, Tanigawa T, Emura I, Seito T, Yamaki T, Fukuda T, Watanabe K, Suzuki T. Regular immunohistochemical localization of endothelin-1 and endothelin-B receptor in normal, hyperplasic and neoplastic human adrenocortical cells. Pathol Int. 1997; 47: 117–125.[Medline] [Order article via Infotrieve]
14. Rossi G, Albertin G, Belloni A, Zanin L, Biasolo M, Prayer-Galeti T, Bader M, Nussdorfer G, Palu G, Pessina A. Gene expression, localization, and characterization of endothelin A and B receptors in the human adrenal cortex. J Clin Invest. 1994; 94: 1226–1234.
15. Vierhapper H, Nowotny P, Waldhausl W. Effect of endothelin-1 in man: impact on basal and adrenocorticotropin-stimulated concentrations of aldosterone. J Clin Endocrinol Metab. 1995; 80: 948–951.[Abstract]
16. Letizia C, DeToma G, Cerci S, Scuro L, DeCiocchis A, D’Ambrosio C, Massa R, Cavallaro A, Scavo D. Plasma endothelin-1 levels in patients with aldosterone-producing adenoma and pheochromocytoma. Clin Exp Hypertens. 1996; 18: 921–931.
17. Zeng Z, Tang X, Yang D, Li H, Zhang R, Zeng Q, Li M, Chen J, Lu Z, Demura H, Naruse M, Shi Y. Immunoreactive endothelin-1 and its receptors in human adrenal tissues. J Cardiovasc Pharmacol. 1998; 31: S212–S214.
18. Chomczynski P, Sacchi N. Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987; 162: 156–159.[Medline] [Order article via Infotrieve]
19. Shimada K, Matsushita Y, Wakabayashi K, Takahashi M, Matsubar A, Iijima Y, Tanzawa K. Cloning and functional expression of human endothelin-converting enzyme cDNA. Biochem Biophys Res Commun. 1995; 207: 807–812.[Medline] [Order article via Infotrieve]
20. Itoh Y, Yanagisawa M, Ohkubo S, Kimura C, Tosaka T, Inoue A, Ishida N, Mitsui Y, Onda H, Fujino M. Cloning and sequence analysis of cDNA encoding the precursor of a human endothelium-derived vasoconstrictor peptide, endothelin: identity of human and porcine endothelin. FEBS Lett. 1988; 231: 440–444.[Medline] [Order article via Infotrieve]
21.
Tso J, Sun X, Kao T, Reece K, Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucl Acids Res. 1985; 13: 2485–2502.
22. Egidy G, Juillerat-Jeanneret L, Korth P, Bosman F, Pinet F. The endothelin system in human normal colon. Am J Physiol. 2000; 279: G211–G222.
23. Korth P, Egidy G, Parnot C, LeMoullec J, Corvol P, Pinet F. Construction, expression and characterization of a soluble form of human endothelin-converting enzyme. FEBS Lett. 1997; 417: 365–370.[Medline] [Order article via Infotrieve]
24. Lack EE. Tumors of the Adrenal Gland and Extra-Adrenal Paraganglia: Atlas of Tumor Pathology. Washington, DC: Armed Forces Institute of Pathology; 1997; 19: 91–122.
25. Cozza E, Gomez-Sanchez C, Foecking M, Chiou S. Endothelin binding to cultured calf adrenal zona glomerulosa cells and stimulation of aldosterone secretion. J Clin Invest. 1989; 84: 1032–1035.
26.
Mazzochi G, Rossi G, Rebuffat P, Malendowicz L, Markowska A, Nussdorfer G. Endothelins stimulate deoxyribonucleic acid synthesis and cell proliferation in rat adrenal zona glomerulosa, acting through an endothelin A receptor coupled with protein kinase C- and tyrosine kinase-dependent signaling pathways. Endocrinology. 1997; 138: 2333–2337.
27.
Rossi G, Belloni A, Albertin G, Zanin L, Biasolo M, Nussdorfer G, Palu G, Pessina A. Endothelin-1 and its receptors A and B in human aldosterone-producing adenomas. Hypertension. 1995; 25: 842–847.
28.
Dupuis J, Stewart D, Cernacek P, Gosselin G. Human pulmonary circulation is an important site for both clearance and production of endothelin-1. Circulation. 1996; 94: 1578–1584.
29.
Lariviere R, Thibault G, Schiffrin E. Increased endothelin-1 content in blood vessels of deoxycorticosterone acetate-salt hypertensive but not in spontaneously hypertensive rats. Hypertension. 1993; 21: 294–300.
30.
Rossi G, Albertin G, Neri G, Andreis P, Hofmann S, Pessina A, Nussdorfer G. Endothelin-1 stimulates steroid secretion of human adrenocortical cells ex vivo via both ETA and ETB receptor subtypes. J Clin Endocrinol Metab. 1997; 82: 3445–3449.
31. Mathieu M, Hamroun D, Pouzeratte Y, Guerineau N, Guillon G, Chevillard C. Adrenal steroid-secreting cells express endothelin-1. Endocrin Res. 1998; 24: 743–747.[Medline] [Order article via Infotrieve]
32. Li Q, Grimelius L, Gröndal S, Höög A, Johansson H. Immunohistochemical localization of endothelin-1 in non-neoplasic adrenal gland tissue. Virchows Arch Pathol Anat. 1994; 425: 259–264.
33. Li Q, Grimelius L, Höög A, Johansson H, Kjellman M, Larsson A. Characterization of endothelin-1-related protein in human adrenal cortex and in cortical lesions. Histochem Cell Biol. 1999; 111: 33–38.[Medline] [Order article via Infotrieve]
34.
Bremnes T, Paasche J, Mehlum A, Sandberg C, Bremnes B, Attramadal H. Regulation and intracellular trafficking pathways of the endothelin receptors. J Biol Chem. 2000; 275: 17596–17604.
35.
Johnson G, Stevenson T, Ahn K. Hydrolysis of peptide hormones by endothelin-converting enzyme-1: a comparison with neprilysin. J Biol Chem. 1999; 274: 4053–4058.
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