- Nitric oxide (NO) and endothelin-1 (ET-1) are known to play a major role in renal and vascular pathophysiology and exhibit a close interaction with ET-1, stimulating NO production; NO in turn inhibits ET-1 expression. Our objectives were (1) to establish a novel transgenic mouse model facilitating ET-1 expression assessment in vivo, (2) to validate this model by assessing prepro-ET-1 promoter activity in mice embryos by means of our novel model and comparing expression sites to well-established data on ET-1 in fetal development and (3) to investigate renal ET-NO interaction by assessing prepro-ET-1 promoter activity in different structures of the renal cortex in the setting of blocked NO synthases via L-NAME administration. We established transgenic mice carrying a lacZ reporter gene under control of the human prepro-ET-1 gene promoter sequence (8 kb of 5′ sequences).
- Bluo-Gal staining of tissue sections revealed intracellular blue particles as indicators of prepro-ET-1 promoter activity. In mouse embryos, we detected high prepro-ET-1 promoter activity in the craniofacial region, as well as in bone and cartilage consistent with the literature. In order to investigate the interaction of ET-1 and NO in the kidney in vivo, transgenic mice at the age of 3-4 months were treated with a single dose of the NO synthase inhibitor L-NAME (25 mg (kg bw)(-1) i.p.) 12 h before kidney removal. Bluo-Gal staining of kidney sections revealed intracellular blue particles as indicators of prepro-ET-1 promoter activity in tubular and vascular endothelium and glomerular cells.
- Particle count was closely correlated to kidney tissue ET-1 content (R=0.918, P<0.001). Comparison of counts revealed an increase by 135+/-53% in L-NAME treated (n=12) compared to non-treated mice (n=10, P=0.001). Cell-type specific evaluation revealed an increase of 136+/-51% in tubular (P=0.001) and 105+/-41% in glomerular cells (P=0.046), but no significant increase in vascular endothelium. In conclusion, our study revealed a close interaction of renal endothelin and the NO system in a cell-type specific manner. Our new transgenic model provides a unique opportunity to analyse regulation of the ET system on a cellular level in vivo.
<em>Endothelin</em>-<em>1</em> promotes cell survival in renal cell carcinoma through the <em>ET</em>(A) receptor.
<em>Endothelin</em>-<em>1</em> and its receptors <em>ET</em>(A) and <em>ET</em>(B) in drug-induced gingival overgrowth.
Regulation and expression of <em>endothelin</em>-<em>1</em> (<em>ET</em>-<em>1</em>) and <em>ET</em>-receptors in rat epithelial cells of renal and intestinal origin.
- The hormone endothelin-1 (ET-1) is involved in many functions of the kidney and intestine. In addition to its vasoactive and proliferative effects, ET-1 is involved in the maintenance of water and salt balance, and in drug excretion by influencing the activity of different transporters in the epithelial cells of these two organs. To study ET-1 function and its role in pathophysiological processes in epithelial cells in vitro, we investigated ET-1 and ET-receptor expression and inducibility of ET-1 excretion by cytokines in three rat cell lines of intestinal (IEC-6) and renal (NRK-52E and GERP) origin.
- Immunocytochemistry showed that all three cell lines express ET-1 and the ET-A and ET-B receptor. ET-1 was expressed intracellularly, and also the ET-A receptor showed a punctate intracellular staining pattern. The ET-B receptor was localized in the membrane, which was confirmed by Western blot analysis. Real-time RT-PCR and ELISA showed that exposure of IEC-6 cells to the cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha), induced ET-1 mRNA expression and excretion, while IL-2 was ineffective.
- In NRK-52E cells, IL-1beta and TNFalpha induced ET-1 excretion as well. In GERP cells, adequate measurement of cytokine effects on ET-1 excretion was not possible, since ET-1 excretion under non-stimulated conditions was around the lowest level of detection. In conclusion, we showed ET-1 and ET-receptor expression, and inducibility of ET-1 by cytokines in IEC-6, NRK-52E, and GERP cells.
- These rat intestinal and renal cell lines appear to be suitable for further characterisation of ET-1 function and its role in pathophysiological processes in epithelial cells.
Endothelin 1 (ET-1) Antibody |
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abx024086-100ug | Abbexa | 100 ug | 1128 EUR |
Endothelin-1 (ET-1), big (Rat) |
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023-32 | PHOENIX PEPTIDE | 100 μg | 206.28 EUR |
ET-1(Endothelin-1) ELISA Kit |
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EKF60092-48T | Biomatik Corporation | 48T | 396.9 EUR |
ET-1(Endothelin-1) ELISA Kit |
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EKF60092-5x96T | Biomatik Corporation | 5x96T | 2693.25 EUR |
ET-1(Endothelin-1) ELISA Kit |
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EKF60092-96T | Biomatik Corporation | 96T | 567 EUR |
ET-1(Endothelin-1) ELISA Kit |
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EU0205 | FN Test | 96T | 628.92 EUR |
Endothelin-1 (ET-1), big (Human) |
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023-10 | PHOENIX PEPTIDE | 100 μg | 154.44 EUR |
Porcine endothelin-1 (ET-1) ELISA Kit |
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QY-E40122 | Qayee Biotechnology | 96T | 480 EUR |
Rat Endothelin 1 (ET-1) CLIA Kit |
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abx195562-96tests | Abbexa | 96 tests | 990 EUR |
Rat ET-1 -Endothelin 1- CLIA Kit |
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E-CL-R0121-24Tests | Elabscience Biotech | 24 Tests | 180 EUR |
Rat ET-1 -Endothelin 1- CLIA Kit |
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E-CL-R0121-48Tests | Elabscience Biotech | 48 Tests | 546 EUR |
Rat ET-1 -Endothelin 1- CLIA Kit |
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E-CL-R0121-96Tests | Elabscience Biotech | 96 Tests | 682 EUR |
Rat ET-1 -Endothelin 1- CLIA Kit |
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E-CL-R0121-96Tests10 | Elabscience Biotech | 96 Tests *10 | 6820 EUR |
Rat ET-1 -Endothelin 1- CLIA Kit |
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E-CL-R0121-96Tests5 | Elabscience Biotech | 96 Tests *5 | 3410 EUR |
Endothelin-1 (ET-1) ELISA Kit (1 Plate) |
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K045-H1 | Arbor Assays | 1x96 well plate | 644 EUR |
Endothelin-1 (ET-1), big (Human) - Antibody |
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H-023-10 | PHOENIX PEPTIDE | 50 μl | 238.68 EUR |
Dog Endothelin 1,ET-1 Elisa Kit |
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EK762013 | AFG Bioscience LLC | 96 Wells | 0.93 EUR |
Rat ET-1(Endothelin 1) ELISA Kit |
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EKF57841-48T | Biomatik Corporation | 48T | 396.9 EUR |
Rat ET-1(Endothelin 1) ELISA Kit |
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EKF57841-5x96T | Biomatik Corporation | 5x96T | 2693.25 EUR |
Rat ET-1(Endothelin 1) ELISA Kit |
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EKF57841-96T | Biomatik Corporation | 96T | 567 EUR |
Rat Endothelin 1(ET-1) Elisa Kit |
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EK720590 | AFG Bioscience LLC | 96 Wells | 0.58 EUR |
rat Endothelin 1,ET-1 ELISA Kit |
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EKC38988-48T | Biomatik Corporation | 48T | 535.99 EUR |
rat Endothelin 1,ET-1 ELISA Kit |
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EKC38988-5x96T | Biomatik Corporation | 5x96T | 3637.08 EUR |
rat Endothelin 1,ET-1 ELISA Kit |
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EKC38988-96T | Biomatik Corporation | 96T | 765.7 EUR |
Dog Endothelin 1,ET-1 ELISA Kit |
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EKC32040-48T | Biomatik Corporation | 48T | 535.99 EUR |
Dog Endothelin 1,ET-1 ELISA Kit |
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EKC32040-5x96T | Biomatik Corporation | 5x96T | 3637.08 EUR |