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      Detection of Prepro-ET-1 mRNA in Normal Rat Kidney by in situ RT-PCR

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          Background: Available evidence has shown that endothelin-1 (ET-1) acts in an autocrine/paracrine fashion rather than as a hormone or cytokine. Therefore, the analysis of local ET-1 production is a crucial step toward understanding its physiological and pathophysiological importance. In this study, in situ RT-PCR was utilized to detect tubular expression of prepro-ET-1 mRNA in normal rat kidney. Methods: Kidneys were taken from normal Sprague-Dawley rats weighing approximately 200 g. In situ RT-PCR was carried out using the preparations embedded in paraffin and cut at a thickness of 8 µm. Furthermore, we tried semiquantitation of the prepro-ET-1 mRNA expression along different nephron segments by densitometric analysis. Results: Prepro-ET-1 mRNA expression was detected in all tubular segments of the kidney from normal rats. Densitometric analysis demonstrated its highest expression in cortical collecting duct (CCD) and outer medullary collecting duct (OMCD). The expression was the lowest in thin descending limb of Henle’s loop (TDL). Conclusion: This study showed that all tubular segments have the ability to synthesize ET-1 in rat kidney. It would be worth evaluating the levels prepro-ET-1 mRNA expression in various diseases by in situ RT-PCR to understand its pathophysiological role in such settings.

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          Most cited references 6

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          Interaction of endothelin-3 with endothelin-B receptor is essential for development of epidermal melanocytes and enteric neurons.

          Defects in the gene encoding the endothelin-B receptor produce aganglionic megacolon and pigmentary disorders in mice and humans. We report that a targeted disruption of the mouse endothelin-3 ligand (EDN3) gene produces a similar recessive phenotype of megacolon and coat color spotting. A natural recessive mutation that results in the same developmental defects in mice, lethal spotting (ls), failed to complement the targeted EDN3 allele. The ls mice carry a point mutation of the EDN3 gene, which replaces the Arg residue at the C-terminus of the inactive intermediate big EDN3 with a Trp residue. This mutation prevents the proteolytic activation of big EDN3 by ECE-1. These findings indicate that interaction of EDN3 with the endothelin-B receptor is essential in the development of neural crest-derived cell lineages. We postulate that defects in the human EDN3 gene may cause Hirschsprung's disease.
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            Molecular characterization of endothelin receptors

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              Presence of immunoreactive endothelin in human plasma.

               Y Hirata,  T. Emori,  K Ando (1989)
              A highly specific and sensitive radioimmunoassay has been established for measurement of human endothelin (hET) in human plasma. After extraction of plasma with an octyl-silica column, this assay allowed for detection of immunoreactive (IR) hET as low as 0.2 fmol/ml. In 16 healthy subjects, the mean concentration of plasma IR-hET was 0.6 fmol/ml. Reverse-phase HPLC coupled with radioimmunoassay revealed two major IR-hET components, one corresponding to authentic hET(1-21) and another with more hydrophilicity than hET(1-21). These data indicate that ET is a circulating vasoconstrictor hormone in man.

                Author and article information

                Nephron Exp Nephrol
                Cardiorenal Medicine
                S. Karger AG
                October 2003
                17 November 2004
                : 95
                : 2
                : e55-e61
                aDepartment of Hygiene and Preventive Medicine, Faculty of Medicine, Saitama Medical School, and bSchool of Management, Tokyo University of Science, Saitama, Japan
                73672 Nephron Exp Nephrol 2003;95:e55–e61
                © 2003 S. Karger AG, Basel

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                Page count
                Figures: 3, Tables: 2, References: 27, Pages: 1
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                Original Paper

                Cardiovascular Medicine, Nephrology

                In situ RT-PCR, Endothelin-1, Kidney, Localization


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