Termination of endothelin signaling: Role of nitric oxide

Michael S. Goligorsky, Hirokazu Tsukahara, Harold Magazine, Thomas T. Andersen, Asrar B. Malik, Wadie F. Bahou

Research output: Contribution to journalArticle

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Abstract

Cellular mechanisms responsible for the termination of ET-1 signal are poorly understood. In order to examine the hypothesis that nitric oxide serves as a physiological brake of ET-1 signaling, Chinese hamster ovary (CHO) cells stably transfected with the ET(A) receptor cDNA (CHO-ET) were studied. CHO-ET responded to ET-1 with robust [Ca2+](i) transients and developed a long-lasting homologous desensitization. Donors of nitric oxide (NO), 3-morpholino-sydnonimine HCl (SIN-1), or sodium nitroprusside (SNP) reduced the amplitude of these responses, accelerated the rate of [Ca2+](i) recovery, and counteracted the development of homologous desensitization by a cyclic GMP-independent mechanism, suggesting an alternative mode for NO modulation of ET-1 responses. Stimulation of CHO-ET cells with mastoparan, a wasp venom acting directly on G proteins (bypassing receptor activation), was inhibited by NO, revealing a postreceptoral target for NO-induced modulation of [Ca2+], mobilization. Using a lys9-biotinylated ET-1 (ET-1 [BtK9]), binding sites were 'mapped' in CHO-ET cells. Receptor-ligand complexes did not exhibit spontaneous dissociation during 60 min observations. Quantitative fluorescence microscopy revealed that SNP or SIN-1 caused a rapid, concentration-dependent, and reversible dissociation of biotinylated ET-1 from ET(A) receptor (EC50 = 75 μM and 6 μM, respectively), an effect that was not mimicked by 8-bromo-cyclic GMP. 'Sandwich' co-culture of endothelial cells with CHO-ET showed that activation of NO production by endothelial cells similarly resulted in dissociation of ET-1 [BtK9] from ET(A) receptors. We hypothesize that NO plays a role in physiological termination of ET-1 signalling by dual mechanisms: (1) displacement of bound ET-1 from its receptor, thus preventing homologous desensitization, and (2) interference with the postreceptoral pathway for [Ca2+](i) mobilization, hence inhibiting end-responses to ET-1.

Original languageEnglish
Pages (from-to)485-494
Number of pages10
JournalJournal of Cellular Physiology
Volume158
Issue number3
Publication statusPublished - Mar 1994
Externally publishedYes

Fingerprint

Endothelins
Cricetulus
Ovary
Nitric Oxide
Endothelial cells
Nitroprusside
Endothelial Cells
Chemical activation
Wasp Venoms
Modulation
Nitric Oxide Donors
Fluorescence microscopy
Cyclic GMP
Coculture Techniques
Fluorescence Microscopy
GTP-Binding Proteins
Cell culture
Brakes
Complementary DNA
Binding Sites

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

Cite this

Goligorsky, M. S., Tsukahara, H., Magazine, H., Andersen, T. T., Malik, A. B., & Bahou, W. F. (1994). Termination of endothelin signaling: Role of nitric oxide. Journal of Cellular Physiology, 158(3), 485-494.

Termination of endothelin signaling : Role of nitric oxide. / Goligorsky, Michael S.; Tsukahara, Hirokazu; Magazine, Harold; Andersen, Thomas T.; Malik, Asrar B.; Bahou, Wadie F.

In: Journal of Cellular Physiology, Vol. 158, No. 3, 03.1994, p. 485-494.

Research output: Contribution to journalArticle

Goligorsky, MS, Tsukahara, H, Magazine, H, Andersen, TT, Malik, AB & Bahou, WF 1994, 'Termination of endothelin signaling: Role of nitric oxide', Journal of Cellular Physiology, vol. 158, no. 3, pp. 485-494.
Goligorsky MS, Tsukahara H, Magazine H, Andersen TT, Malik AB, Bahou WF. Termination of endothelin signaling: Role of nitric oxide. Journal of Cellular Physiology. 1994 Mar;158(3):485-494.
Goligorsky, Michael S. ; Tsukahara, Hirokazu ; Magazine, Harold ; Andersen, Thomas T. ; Malik, Asrar B. ; Bahou, Wadie F. / Termination of endothelin signaling : Role of nitric oxide. In: Journal of Cellular Physiology. 1994 ; Vol. 158, No. 3. pp. 485-494.
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