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. 1995 Jul;96(1):183–191. doi: 10.1172/JCI118019

Osmolar regulation of endothelin signaling in rat renal medullary interstitial cells.

M A Vernace 1, P F Mento 1, M E Maita 1, E P Girardi 1, M D Chang 1, E P Nord 1, B M Wilkes 1
PMCID: PMC185187  PMID: 7615788

Abstract

We tested the hypothesis that endothelin (ET) responsiveness in the renal medulla is modulated by ambient osmolarity. Cultured renal medullary interstitial cells (RMICs) were incubated from 3 to 24 h in isosmolar culture medium (300 mOsm/kg H2O) or media rendered hyperosmolar (600 mOsm/kg H2O) by the addition of urea. Under hyperosmolar conditions, the peak of ET-evoked Ca2+ transient was blunted by 45-58% (P < 0.02) and PGE2 accumulation decreased from 16- to 2-fold above basal values (P < 0.001). To explore whether hyperosmolar conditions blunt intracellular signaling via modulation of receptor number or expression, kinetics of ET binding and Northern blot analysis of ETA receptor mRNA was performed. Under hyperosmolar conditions, ETA receptor density was reduced by 84% versus isosmolar conditions (238 +/- 12 vs. 1450 +/- 184 fmol/mg) (P < 0.01). In contrast to the ligand binding studies, ETA receptor mRNA was increased by 58% (P < 0.05) in cells grown under hyperosmolar versus isosmolar media. These observations indicate that in the hyperosmolar setting, ET-evoked intracellular signaling is blunted in RMICs due to ET receptor downregulation. Since ETA receptor mRNA is increased under hyperosmolar conditions, we conclude that ET receptor downregulation is the consequence of either decreased translation of message, increased degradation of receptor peptide, or increased internalization of specific receptor sites.

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Selected References

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  1. Aboolian A., Nord E. P. Bradykinin increases cytosolic free [Ca2+] in proximal tubule cells. Am J Physiol. 1988 Sep;255(3 Pt 2):F486–F493. doi: 10.1152/ajprenal.1988.255.3.F486. [DOI] [PubMed] [Google Scholar]
  2. Aboolian A., Vander Molen M., Nord E. P. Differential effects of phorbol esters on PGE2 and bradykinin-induced elevation of [Cai2+] in MDCK cells. Am J Physiol. 1989 Jun;256(6 Pt 2):F1135–F1143. doi: 10.1152/ajprenal.1989.256.6.F1135. [DOI] [PubMed] [Google Scholar]
  3. Arai H., Hori S., Aramori I., Ohkubo H., Nakanishi S. Cloning and expression of a cDNA encoding an endothelin receptor. Nature. 1990 Dec 20;348(6303):730–732. doi: 10.1038/348730a0. [DOI] [PubMed] [Google Scholar]
  4. Awazu M., Parker R. E., Harvie B. R., Ichikawa I., Kon V. Down-regulation of endothelin-1 receptors by protein kinase C in streptozotocin diabetic rats. J Cardiovasc Pharmacol. 1991;17 (Suppl 7):S500–S502. doi: 10.1097/00005344-199100177-00142. [DOI] [PubMed] [Google Scholar]
  5. Barnett R. L., Ruffini L., Hart D., Mancuso P., Nord E. P. Mechanism of endothelin activation of phospholipase A2 in rat renal medullary interstitial cells. Am J Physiol. 1994 Jan;266(1 Pt 2):F46–F56. doi: 10.1152/ajprenal.1994.266.1.F46. [DOI] [PubMed] [Google Scholar]
  6. Barnett R., Ortiz P. A., Blaufox S., Singer S., Nord E. P., Ramsammy L. Atrial natriuretic factor alters phospholipid metabolism in mesangial cells. Am J Physiol. 1990 Jan;258(1 Pt 1):C37–C45. doi: 10.1152/ajpcell.1990.258.1.C37. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Cozza E. N., Vila M. C., Gomez-Sanchez C. E. ET-1 receptors in C-6 cells: homologous down-regulation and modulation by protein kinase C. Mol Cell Endocrinol. 1990 Apr 17;70(2):155–164. doi: 10.1016/0303-7207(90)90155-2. [DOI] [PubMed] [Google Scholar]
  9. Daukas G., Zigmond S. H. Inhibition of receptor-mediated but not fluid-phase endocytosis in polymorphonuclear leukocytes. J Cell Biol. 1985 Nov;101(5 Pt 1):1673–1679. doi: 10.1083/jcb.101.5.1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Denton K. M., Anderson W. P. Vascular actions of endothelin in the rabbit kidney. Clin Exp Pharmacol Physiol. 1990 Dec;17(12):861–872. doi: 10.1111/j.1440-1681.1990.tb01289.x. [DOI] [PubMed] [Google Scholar]
  11. Fontoura B. M., Nussenzveig D. R., Pelton K. M., Maack T. Atrial natriuretic factor receptors in cultured renomedullary interstitial cells. Am J Physiol. 1990 Apr;258(4 Pt 1):C692–C699. doi: 10.1152/ajpcell.1990.258.4.C692. [DOI] [PubMed] [Google Scholar]
  12. Garcia-Perez A., Burg M. B. Renal medullary organic osmolytes. Physiol Rev. 1991 Oct;71(4):1081–1115. doi: 10.1152/physrev.1991.71.4.1081. [DOI] [PubMed] [Google Scholar]
  13. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  14. King A. J., Brenner B. M., Anderson S. Endothelin: a potent renal and systemic vasoconstrictor peptide. Am J Physiol. 1989 Jun;256(6 Pt 2):F1051–F1058. doi: 10.1152/ajprenal.1989.256.6.F1051. [DOI] [PubMed] [Google Scholar]
  15. Kohan D. E. Endothelin synthesis by rabbit renal tubule cells. Am J Physiol. 1991 Aug;261(2 Pt 2):F221–F226. doi: 10.1152/ajprenal.1991.261.2.F221. [DOI] [PubMed] [Google Scholar]
  16. Kohan D. E., Padilla E. Osmolar regulation of endothelin-1 production by rat inner medullary collecting duct. J Clin Invest. 1993 Mar;91(3):1235–1240. doi: 10.1172/JCI116286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  18. Mironneau C., Rakotoarisoa L., Sayet I., Mironneau J. Modulation of [3H]dihydropyridine binding by activation of protein kinase C in vascular smooth muscle. Eur J Pharmacol. 1991 Nov 13;208(3):223–230. doi: 10.1016/0922-4106(91)90099-4. [DOI] [PubMed] [Google Scholar]
  19. Muirhead E. E., Germain G., Leach B. E., Pitcock J. A., Stephenson P., Brooks B., Brosius W. L., Daniels E. G., Hinman J. W. Production of renomedullary prostaglandins by renomedullary interstitial cells grown in tissue culture. Circ Res. 1972 Sep;31(9 Suppl):161–172. [PubMed] [Google Scholar]
  20. Nadler S. P., Zimpelmann J. A., Hébert R. L. Endothelin inhibits vasopressin-stimulated water permeability in rat terminal inner medullary collecting duct. J Clin Invest. 1992 Oct;90(4):1458–1466. doi: 10.1172/JCI116013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nunez D. J., Brown M. J., Davenport A. P., Neylon C. B., Schofield J. P., Wyse R. K. Endothelin-1 mRNA is widely expressed in porcine and human tissues. J Clin Invest. 1990 May;85(5):1537–1541. doi: 10.1172/JCI114601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Oka J. A., Christensen M. D., Weigel P. H. Hyperosmolarity inhibits galactosyl receptor-mediated but not fluid phase endocytosis in isolated rat hepatocytes. J Biol Chem. 1989 Jul 15;264(20):12016–12024. [PubMed] [Google Scholar]
  23. Pachter J. A., Mayer-Ezell R., Cleven R. M., Fawzi A. B. Endothelin (ETA) receptor number and calcium signalling are up-regulated by protein kinase C-beta 1 overexpression. Biochem J. 1993 Aug 15;294(Pt 1):153–158. doi: 10.1042/bj2940153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sakurai T., Morimoto H., Kasuya Y., Takuwa Y., Nakauchi H., Masaki T., Goto K. Level of ETB receptor mRNA is down-regulated by endothelins through decreasing the intracellular stability of mRNA molecules. Biochem Biophys Res Commun. 1992 Jul 15;186(1):342–347. doi: 10.1016/s0006-291x(05)80813-7. [DOI] [PubMed] [Google Scholar]
  25. Sakurai T., Yanagisawa M., Takuwa Y., Miyazaki H., Kimura S., Goto K., Masaki T. Cloning of a cDNA encoding a non-isopeptide-selective subtype of the endothelin receptor. Nature. 1990 Dec 20;348(6303):732–735. doi: 10.1038/348732a0. [DOI] [PubMed] [Google Scholar]
  26. Schnermann J., Lorenz J. N., Briggs J. P., Keiser J. A. Induction of water diuresis by endothelin in rats. Am J Physiol. 1992 Sep;263(3 Pt 2):F516–F526. doi: 10.1152/ajprenal.1992.263.3.F516. [DOI] [PubMed] [Google Scholar]
  27. Schramek H., Gstraunthaler G., Willinger C. C., Pfaller W. Hyperosmolality regulates endothelin release by Madin-Darby canine kidney cells. J Am Soc Nephrol. 1993 Aug;4(2):206–213. doi: 10.1681/ASN.V42206. [DOI] [PubMed] [Google Scholar]
  28. Ullian M. E., Linas S. L. Role of receptor cycling in the regulation of angiotensin II surface receptor number and angiotensin II uptake in rat vascular smooth muscle cells. J Clin Invest. 1989 Sep;84(3):840–846. doi: 10.1172/JCI114244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wilkes B. M., Ruston A. S., Mento P., Girardi E., Hart D., Vander Molen M., Barnett R., Nord E. P. Characterization of endothelin 1 receptor and signal transduction mechanisms in rat medullary interstitial cells. Am J Physiol. 1991 Apr;260(4 Pt 2):F579–F589. doi: 10.1152/ajprenal.1991.260.4.F579. [DOI] [PubMed] [Google Scholar]
  30. Wilkes B. M., Susin M., Mento P. F., Macica C. M., Girardi E. P., Boss E., Nord E. P. Localization of endothelin-like immunoreactivity in rat kidneys. Am J Physiol. 1991 Jun;260(6 Pt 2):F913–F920. doi: 10.1152/ajprenal.1991.260.6.F913. [DOI] [PubMed] [Google Scholar]
  31. Yanagisawa M., Kurihara H., Kimura S., Tomobe Y., Kobayashi M., Mitsui Y., Yazaki Y., Goto K., Masaki T. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988 Mar 31;332(6163):411–415. doi: 10.1038/332411a0. [DOI] [PubMed] [Google Scholar]
  32. Yang T., Terada Y., Nonoguchi H., Ujiie K., Tomita K., Marumo F. Effect of hyperosmolality on production and mRNA expression of ET-1 in inner medullary collecting duct. Am J Physiol. 1993 Apr;264(4 Pt 2):F684–F689. doi: 10.1152/ajprenal.1993.264.4.F684. [DOI] [PubMed] [Google Scholar]

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