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Title: Endothelin-1 stimulates prostaglandin E2 production in an extracellular calcium-independent manner in cultured rat mesangial cells

Abstract

To study a possible role of endothelin-1 (ET-1) in the regulation of glomerular function, we examined the presence of receptors for, and the biological action of, ET-1 in cultured rat mesangial (M) cells. The first-subcultured M cells prepared from isolated glomeruli of Sprague-Dawley rats were used. ET-1 binding was assayed by using {sup 125}I-ET-1. Inositol 1,4,5-trisphosphate (IP3) was determined by IP3-specific binding assay. Intracellular calcium (iCa{sup 2}{sup +}) was measured in fura-2 loaded cells. Prostaglandin E2 (PGE2) was measured by radioimmunoassay. In M cells there existed two classes of binding sites specific for ET-1 (Kd was 0.24 and 4.4 nmol/L, Bmax was 130 and 1070 fmol/mg, respectively). ET-1 (10{sup {minus} 7} mol/L) induced a rapid and transient increase in IP3, followed by transient and sustained increases in iCa{sup 2}{sup +}. Nicardipine (10{sup {minus} 6} mol/L) inhibited only the sustained increase in iCa{sup 2}{sup +}. ET-1 (10{sup {minus} 9} mol/L to 10{sup {minus} 7} mol/L) significantly stimulated PGE2 production with the concentration dependency. Nicardipine (10{sup {minus} 6} mol/L) and diltiazem (10{sup {minus} 6} mol/L) did not inhibit the PGE2 production. We conclude that M cells have specific ET-1 receptors linked to phosphoinositide turnover and PGE2 production, and PGE2 production by ET-1more » may be through an extra-cellular calcium-independent mechanism. Our results suggest that ET-1 plays an important role in the regulation of glomerular functions by modulating PGE2 production in M cells.« less

Authors:
; ; ; ; ; ;  [1]
  1. Osaka Univ. Medical School, (Japan)
Publication Date:
OSTI Identifier:
5642815
Resource Type:
Journal Article
Journal Name:
American Journal of Hypertension; (USA)
Additional Journal Information:
Journal Volume: 4:2 Pt 1; Journal ID: ISSN 0895-7061
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; PEPTIDES; BIOCHEMICAL REACTION KINETICS; PROSTAGLANDINS; BIOSYNTHESIS; ANIMAL CELLS; CALCIUM COMPOUNDS; DOSE-RESPONSE RELATIONSHIPS; IODINE 125; RADIOIMMUNOASSAY; RATS; RECEPTORS; ALKALINE EARTH METAL COMPOUNDS; ANIMALS; BETA DECAY RADIOISOTOPES; BIOASSAY; DAYS LIVING RADIOISOTOPES; DIAGNOSTIC TECHNIQUES; ELECTRON CAPTURE RADIOISOTOPES; IMMUNOASSAY; IMMUNOLOGY; INTERMEDIATE MASS NUCLEI; INTERNAL CONVERSION RADIOISOTOPES; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; KINETICS; MAMMALS; MEMBRANE PROTEINS; NUCLEI; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; PROTEINS; RADIOASSAY; RADIOIMMUNODETECTION; RADIOIMMUNOLOGY; RADIOISOTOPES; REACTION KINETICS; RODENTS; SYNTHESIS; TRACER TECHNIQUES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Fukunaga, M, Ochi, S, Takama, T, Yokoyama, K, Fujiwara, Y, Orita, Y, and Kamada, T. Endothelin-1 stimulates prostaglandin E2 production in an extracellular calcium-independent manner in cultured rat mesangial cells. United States: N. p., 1991. Web.
Fukunaga, M, Ochi, S, Takama, T, Yokoyama, K, Fujiwara, Y, Orita, Y, & Kamada, T. Endothelin-1 stimulates prostaglandin E2 production in an extracellular calcium-independent manner in cultured rat mesangial cells. United States.
Fukunaga, M, Ochi, S, Takama, T, Yokoyama, K, Fujiwara, Y, Orita, Y, and Kamada, T. Fri . "Endothelin-1 stimulates prostaglandin E2 production in an extracellular calcium-independent manner in cultured rat mesangial cells". United States.
@article{osti_5642815,
title = {Endothelin-1 stimulates prostaglandin E2 production in an extracellular calcium-independent manner in cultured rat mesangial cells},
author = {Fukunaga, M and Ochi, S and Takama, T and Yokoyama, K and Fujiwara, Y and Orita, Y and Kamada, T},
abstractNote = {To study a possible role of endothelin-1 (ET-1) in the regulation of glomerular function, we examined the presence of receptors for, and the biological action of, ET-1 in cultured rat mesangial (M) cells. The first-subcultured M cells prepared from isolated glomeruli of Sprague-Dawley rats were used. ET-1 binding was assayed by using {sup 125}I-ET-1. Inositol 1,4,5-trisphosphate (IP3) was determined by IP3-specific binding assay. Intracellular calcium (iCa{sup 2}{sup +}) was measured in fura-2 loaded cells. Prostaglandin E2 (PGE2) was measured by radioimmunoassay. In M cells there existed two classes of binding sites specific for ET-1 (Kd was 0.24 and 4.4 nmol/L, Bmax was 130 and 1070 fmol/mg, respectively). ET-1 (10{sup {minus} 7} mol/L) induced a rapid and transient increase in IP3, followed by transient and sustained increases in iCa{sup 2}{sup +}. Nicardipine (10{sup {minus} 6} mol/L) inhibited only the sustained increase in iCa{sup 2}{sup +}. ET-1 (10{sup {minus} 9} mol/L to 10{sup {minus} 7} mol/L) significantly stimulated PGE2 production with the concentration dependency. Nicardipine (10{sup {minus} 6} mol/L) and diltiazem (10{sup {minus} 6} mol/L) did not inhibit the PGE2 production. We conclude that M cells have specific ET-1 receptors linked to phosphoinositide turnover and PGE2 production, and PGE2 production by ET-1 may be through an extra-cellular calcium-independent mechanism. Our results suggest that ET-1 plays an important role in the regulation of glomerular functions by modulating PGE2 production in M cells.},
doi = {},
journal = {American Journal of Hypertension; (USA)},
issn = {0895-7061},
number = ,
volume = 4:2 Pt 1,
place = {United States},
year = {1991},
month = {2}
}