Search PubMed⌕ Search

Biomedical subjects

D Schlondorff

Publications and source records attributed to D Schlondorff.

At least 37 records · Page 2Linked to original sources
PubMed1991

Preferential binding of oxidized LDL to rat glomeruli in vivo and cultured mesangial cells in vitro.

Hyperlipidemia may contribute to the pathogenesis of glomerular sclerosis. We therefore compared binding and uptake of native LDL and oxidized LDL (Ox-LDL) to cultured mesangial cells (MC) and the resulting effects on prostaglandin generation and cell proliferation. Ox-LDL, prepared from native LDL by incubation with copper, was bound to MC in a concentration dependent manner with a four- to fivefold increase in binding over LDL. In competition binding experiments Ox-LDL competed to 90% with LDL for binding sites, but LDL only displaced Ox-LDL to 15%. Furthermore polyinosinic acid, which blocks binding of Ox-LDL to macrophages, inhibited binding of Ox-LDL but not that of LDL to MC. Mesangial cells also preferentially took up Ox-LDL over LDL, and Ox-LDL resulted in higher [14C] oleate incorporation into cholesteryl esters than LDL, findings consistent with different handling of Ox-LDL and LDL by MC. LDL slightly stimulated mesangial cell proliferation at low concentration (10 to 50 micrograms/ml of LDL) returning to control levels at 100 and 250 micrograms/ml. In contrast Ox-LDL inhibited cell proliferation in a concentration-dependent manner, starting at concentrations as low as 10 to 25 micrograms/ml of Ox-LDL. Direct observations of mesangial cells by phase contrast microscopy confirmed the cytotoxic effects of Ox-LDL. Addition of Ox-LDL to mesangial cells resulted in a concentration-dependent increase in PGE2 synthesis within one hour, while at this time point LDL had no significant effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Regulation of Fc receptors for IgG on cultured rat mesangial cells.

Localization of immune complexes (IC) to the mesangium may contribute to glomerular disease. Recently, we and others characterized Fc receptors (Fc gamma R) for IgG-IC on mesangial cells (MC). This study examines regulation of Fc gamma R by cAMP, interferon gamma (IFN-gamma) and by macrophage colony stimulating factor (CSF-1), an agent controlling Fc gamma R in leukocytes and generated by MC. Preincubation of MC (3rd to 6th subculture) with CSF-1, db-cAMP or IFN-gamma for two to 48 hours resulted in a time dependent (maximal 24 to 48 hrs) two- to threefold increase of specific [125I] IgG-IC binding to MC at 4 degrees C. The increase of Fc receptors induced by CSF-1, db-cAMP or IFN-gamma was confirmed by enhanced binding of the monoclonal anti-Fc receptor antibody 2.4G2 to MC. Uptake of IgG-IC at 37 degrees C was also enhanced in MC pretreated with CSF-1, db-cAMP or IFN-gamma. This indicates that the increase in binding for IgG-IC is associated with functional receptors. Immunoprecipitation of extracts of [125I] surface labeled MC with polyclonal anti-Fc gamma R-Ab followed by SDS-PAGE also showed increased amounts of [125I] Fc gamma R protein after pretreatment with CSF-1, db-cAMP or IFN-gamma. The pretreatment also enhanced staining of MC with anti-Fc gamma R-Ab by immunogold-silver enhancement technique. We conclude that MC express Fc gamma R for IgG-IC that can be regulated by CSF-1, cAMP and IFN-gamma, factors that may be important in glomerular immune injury.

Animals↗
PubMed1991

Effects of PGE2 and a thromboxane A2 analogue on uptake of IgG complexes and LDL by mesangial cells.

Vasoactive agents such as angiotensin (ANG) and eicosanoids can influence macromolecular deposition in the glomerular mesangium. We studied whether prostaglandin E2 (PGE2) and U-46619 (a stable analogue of thromboxane A2) could directly affect the uptake of immunoglobulin G (IgG) complexes or low-density lipoprotein (LDL) by cultured rat mesangial cells (MC). Preincubation of MC with PGE2 (10(-6) M) resulted in decreased uptake (in counts.min-1.well-1) of IgG complexes (PGE2, 2,589 +/- 72; control, 3,840 +/- 114; P less than 0.001) and LDL particles (PGE2, 23,176 +/- 1,145; control, 37,216 +/- 4,520; P less than 0.05). MC preincubated with forskolin (10(-5) M) also showed decreased uptake of IgG complexes (forskolin, 2,896 +/- 196; control, 3,840 +/- 114; P less than 0.005) and LDL particles (forskolin, 23,176 +/- 1,145; control, 37,216 +/- 4,520; P less than 0.05). In contrast, preincubation with ANG II (5 x 10(-7) M) showed significantly higher uptake of IgG complexes (ANG II, 4,475 +/- 282; control, 3,787 +/- 277; P less than 0.05) and LDL (ANG II, 48,573 +/- 1,079; control, 44,697 +/- 770; P less than 0.05). Similarly, preincubation with U-46619 (10(-6) M) resulted in significantly higher uptake of IgG complexes (U-46619, 5,370 +/- 300; control, 3,659 +/- 307; P less than 0.02) and LDL (U-46619, 48,298 +/- 1,418; control, 44,697 +/- 770; P less than 0.05). After preincubation with cyclooxygenase inhibitor meclofenamate (10(-6) M), ANG II (5 x 10(-7) M) resulted in a significantly higher uptake of IgG complexes compared with uptake by MC treated with ANG II alone (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

1-Methyl-3-isobutylxanthine↗
PubMed1991

Protective effects of specific platelet-activating factor receptor antagonists in experimental glomerulonephritis.

The present study was undertaken to evaluate the hypothesis that enhanced production of platelet-activating factor (PAF) contributes to the renal hemodynamic alterations and impaired glomerular permselectivity that characterize the heterologous phase of nephrotoxic serum nephritis. Intravenous administration of nephrotoxic gamma globulin to normal rats led to a rapid decline in glomerular filtration rate within 60 min from 1.08 +/- 0.08 to 0.45 +/- 0.12 ml/min/100 g b.wt., P less than .001, and in effective renal plasma flow rate from 2.56 +/- 0.15 to 1.35 +/- 0.21 ml/min/100 g b.wt., P less than .02. Concomitantly, the fractional excretion of protein rose from 3.4 +/- 0.6 X 10(-5) to 11.6 +/- 2.3 X 10(-5), P less than .03. Pretreatment with the specific PAF receptor antagonists WEB 2086 or WEB 2170 significantly ameliorated the impairment in glomerular filtration rate and in effective renal plasma flow rate induced by nephrotoxic globulin. Filtration rate declined by 0.63 +/- 0.12 ml/min/100 g b.wt. 60 min after administration of nephrotoxic globulin, but fell only 0.30 +/- 0.14 ml/min/100 g b.wt. in rats pretreated with WEB 2086, P less than .05 and 0.34 +/- 0.11 ml/min/100 g b.wt. in rats pretreated with WEB 2170, P less than .01. The decline in effective renal plasma flow rate 60 min after administration of nephrotoxic globulin was 1.21 +/- 0.30 ml/min/100 g b.wt., but only 0.86 +/- 0.23 ml/min/100 g b.wt. in rats pretreated with WEB 2086, NS, and 0.52 +/- 0.21 ml/min/100 g b.wt. in rats pretreated with WEB 2170, P less than .003.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Contributions of mesangial cells to glomerular immune functions.

Mesangial cells play an important role in the maintenance of the structure and function of the glomerulus. In addition mesangial cells are capable of macromolecular uptake, and generation of autocoids and cytokines. Our studies have explored the mechanism of IgG uptake by cultured rat mesangial cells. Mesangial cells were found to express Fc receptors for IgG as demonstrated by specific binding studies, indirect immunofluorescence microscopy, immunoblotting and Northern blot analysis. The number of Fc receptors for IgG on mesangial cells was upregulated by interferon gamma, cyclic AMP and monocyte-macrophage specific colony stimulating factor--CSF-1. In addition uptake of IgG complexes was acutely increased by angiotensin II and inhibited by cAMP. These latter effects are independent of receptor number, but are mediated by changes in the cytoskeleton. These observations may be of significance for hemodynamically independent effects of vasoactive agents on mesangial function. Finally we examined the potential of mesangial cells for generation of CSF-1. Cultured mesangial cells produced radioimmunoassayable CSF-1 and expressed mRNA for CSF-1. Interferon gamma stimulated, whereas cAMP and agents increasing cAMP, such as forskolin and PGE2, inhibited CSF-1 production. This was due to decreased transcription resulting in lower levels of mRNA for CSF-1. The interactions of CSF-1, other cytokines, prostaglandins and cAMP may be important in regulating immune-like functions of mesangial cells, and especially their response to immunecomplexes.

Angiotensin II↗
PubMed1990

Mouse mesangial cells produce colony-stimulating factor-1 (CSF-1) and express the CSF-1 receptor.

CSF-1 stimulates the survival, proliferation, and differentiation of mononuclear phagocytes and may also play a role in placental development. The expression of CSF-1 and the CSF-1 receptor (CSF-1R) and their regulation were examined in cultures of mouse mesangial cells (MC). The concentration of CSF-1 in the medium of cultured MC increased linearly with time over 24 h. IFN-gamma stimulated and dibutyryl cyclic AMP inhibited CSF-1 production in a dose-dependent manner. MC expression of CSF-1 mRNA was shown by Northern blot analysis, and CSF-1 mRNA levels were increased within 4 h of IFN-gamma addition and inhibited within 4 h of dibutyryl cyclic AMP addition. Indirect immunofluorescence indicated that 90% of the untreated cultured MC expressed CSF-1. In addition, CSF-1R expression by MC was demonstrated by immunofluorescence with anti-receptor antibody, specific binding of [125I] CSF-1, and expression of the CSF-1R mRNA by Northern blot analysis. Thus, mouse MC, specialized pericytes of non-bone marrow origin, not only produce CSF-1 but also express receptors for CSF-1. The effects of CSF-1 on MC may be important in the control of immune function in the glomerulus.

Animals↗
PubMed1990

Identification of specific glomerular cell types in culture by use of lectin and antibody binding.

To characterize the outgrowth of cells from cultured rat glomeruli, we examined the cell morphology in relation to the expression of various antigenic determinants (factor VIII-related antigen, rat Thy 1.1, Heyman nephritogenic antigen, common leucocyte and Ia antigens) and lectin binding patterns. For this purpose, early (3-7 days) to late (4-5 weeks) glomerulus cultures, as well as subcultures of mesangial cells were used. In the early samples, the predominance of small round (type I) cells growing in a cobblestone pattern was noted. These cells failed to express specific binding for any of the antibodies or lectins tested. 5-10% of the outgrowing cells appeared single and morphologically of stellate shape (type II). These cells were reactive for Concanavalia ensiformis (ConA), Triticum vulgaris (WGA) and Ricinus communis (RCA I), and stained for rat Thy 1.1 antigen. Large round, single cells (type III), found mostly at the edges of the outgrowth bound only Bandeiraea simplicifolia (BSI-B4) and FVIII antibodies, a pattern consistent with endothelial cells. A fourth (type IV) cell type, elongated, with cells mostly found in bundles and growing on a layer consisting of other cells, was reactive for ConA, WGA, Limax flavus (LFA) and Maclura pomifera (MPA) lectins. In the late outgrowth (3-5 weeks after explant), the lectin and antibody binding patterns of each cell type remained essentially unchanged. However, considerable changes in the prevalence of the morphologic cell types were noted. Cells that had been subcultured to obtain pure 'mesangial' cells, exhibited the same morphology as the type II cells and expressed identical staining patterns with all probes used. The binding of lectins was likewise confirmed by staining frozen sections of rat kidneys.

Animals↗
PubMed1990

Effects of vasoactive agents on uptake of immunoglobulin G complexes by mesangial cells.

Mesangial handling of immune complexes may be important in immune injury. We examined whether vasoactive agents, independent of hemodynamic effects, could directly influence uptake of immunoglobulin G (IgG) complexes by mesangial cells (MC). Under basal conditions, MC took up more gold particles coated with IgG2b (P less than 0.05) than coated with IgG2a. Preincubation of MC with angiotensin II (ANG II) (5 x 10(-7) M) resulted in enhancement of uptake (P less than 0.05) of IgG2b-gold [7,578 +/- 968 counts per minute (cpm) per well], IgG2a-gold (4,566 +/- 295 cpm/well; P less than 0.01), and bovine serum albumin (BSA)-gold (2,532 +/- 66; P less than 0.05) when compared with respective controls (IgG2b-gold, 5,513 +/- 762 cpm/well; IgG2a-gold, 3,282 +/- 439; BSA-gold, 2,279 +/- 97). Cytochalasin B produced a significant reduction of uptake of IgG2b-gold (2,224 +/- 88 cpm/well) when compared with the uptake by control cells (3,711 +/- 287 cpm/well) (P less than 0.05). Pretreatment of MC with atrial natriuretic peptide (ANP, 10(-9) M) slightly decreased uptake of IgG-gold under basal conditions (P less than 0.05) and decreased the response to ANG II (P less than 0.05). Similarly, dopamine (DA, 10(-6) M) attenuated uptake of IgG2b-gold under basal (P less than 0.01) as well as ANG II-stimulated (P less than 0.01) states. ANP increased mesangial guanosine 3',5'-cyclic monophosphate (cGMP), DA, adenosine 3',5'-cyclic monophosphate (cAMP) content.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗
PubMed1990

Renal vasoconstriction caused by short-term cholesterol feeding is corrected by thromboxane antagonist or probucol.

Recent studies indicate that short-term cholesterol feeding causes vascular hyperreactivity and/or increased tone in certain vascular beds. The present study in rats examined the effect of 3 wk of cholesterol-supplemented diet (CSD) on renal hemodynamics. We tested the hypothesis that LDL oxidized in vivo is causally related to increased renal vascular tone by adding the antioxidant drug probucol to the CSD (CSD + P). Micropuncture of surface nephrons in the CSD rats demonstrated that single nephron glomerular filtration rate (SNGFR) and single nephron afferent plasma flow (QA) were markedly lower than in normal rats, whereas glomerular capillary pressure (PGC), afferent arteriolar resistance (RA), and single nephron filtration fraction (SNFF) were higher. In the CSD + P animals, almost all of these hemodynamic abnormalities were absent. TXB2 and PGE2 were increased in proximal tubule fluid and urine in the CSD rats, but normal in the CSD + P group. Infusion of a TXA2 receptor antagonist into the suprarenal aorta of CSD rats caused a rapid return to normal of RBF (renal blood flow), GFR (glomerular filtration rate), SNGFR, QA, RA, PGC, and Kf (ultrafiltration coefficient). Our observations demonstrate that cholesterol feeding leads to renal vasoconstriction, which appears to be mediated largely by increased TXA2 production. The fact that probucol prevented the hemodynamic abnormalities as well as the increased TX production is consistent with the hypothesis that LDL oxidized in vivo initiates events leading to TX mediated vasoconstriction.

Animals↗
PubMed1990

Thromboxane mediates the renal hemodynamic effects of platelet activating factor.

The mechanism by which platelet-activating factor affects renal hemodynamics is controversial. In the present study we examined the hypothesis that thromboxane mediates the renal hemodynamic response to platelet activating factor (PAF) by determining the effects of pretreatment with a selective thromboxane receptor antagonist. Infusion of platelet-activating factor into the left renal artery of normal rats in a dose that failed to alter mean arterial pressure or blood hematocrit values reduced urinary flow rate from 25.9 +/- 6.3 to 12.6 +/- 1.5 microliters/min (P less than .01), reduced effective renal plasma flow rate from 14.01 +/- 2.09 to 5.42 +/- 2.38 ml/min/kg body weight (P less than .01), reduced glomerular filtration rate from 4.81 +/- 1.00 to 1.68 +/- 0.34 ml/min/kg (P less than .03), and increased the fractional excretion of protein from 0.021 +/- 0.008 to 0.094 +/- 0.005 percent (P less than .05). Pretreatment with the selective thromboxane receptor antagonist SKF 96148 (10 mg/kg) not only prevented PAF-induced reductions in effective renal plasma flow rate and glomerular filtration rate but increased these values (effective renal plasma flow rate: 14.09 +/- 2.76 vs. 16.84 +/- 2.08 ml/min/kg, P less than .05; glomerular filtration rate: 4.42 +/- 0.43 vs. 5.50 +/- 0.59 ml/min/kg, P less than .03) and aborted proteinuria. Pretreatment with indomethacin (2 mg/kg) ameliorated PAF-induced alterations in renal hemodynamics and glomerular permselectivity in the final collection period. Infusion of lyso-PAF, a biologically inactive precursor of PAF, or of indomethacin alone or SKF 96148 alone had no significant effect on measured parameters. These observations suggest that thromboxane mediates the renal hemodynamic and permselective effects of PAF.

Animals↗
PubMed1990

A fish oil diet preserves renal function in nephrotoxic serum nephritis.

Fish oil diets preserve renal function in murine lupus, but we have found that these diets accelerate renal deterioration in renoprival nephropathy. In this study we examined the effects of dietary fish oil in accelerated nephrotoxic serum nephritis. For 1 month, 14 female rats were fed diets that differed only in fat composition, containing either menhaden (fish) oil or beef tallow (control). Rats were then preimmunized with rabbit IgG and, 5 days later, were injected with nephrotoxic serum. Glomerular filtration rate (GFR) was measured continuously in conscious animals by means of intraperitoneal 14C-labeled inulin minipumps. Fish oil-containing diets markedly attenuated the nephrotoxic serum-induced decline in GFR and the rise in proteinuria and significantly reduced glomerular prostaglandin E2 and thromboxane A2. The results of tests of renal histology showed no differences between the two groups. Five days after preimmunization, rats fed fish oil had more rabbit IgG remaining in their serum and had mounted less of an antibody response to the rabbit IgG. Fish oil diets also resulted in an attenuated disappearance of injected 14C-labeled rabbit IgG. In vitro, peritoneal macrophages from rats fed fish oil took up less rabbit IgG than macrophages from rats fed control diets. Thus the beneficial effects of a fish oil diet may result from defective immune surveillance and from alterations in eicosanoids.

Animals↗
PubMed1990

Effects of fish oil on glomerular function in rats with diabetes mellitus.

The mechanisms responsible for hyperfiltration in diabetes mellitus (DM) as well as for the initiation and progression of diabetic nephropathy are not fully elucidated. Enhanced prostaglandin E2 (PGE2) production has been invoked in the former and thromboxane (TXB2) and hyperlipidemia in the latter. Fish oil (FO)-enriched diets can favorably alter eicosanoid synthesis and serum lipid profiles. We therefore examined the effects of a FO-enriched diet on glomerular filtration (GFR), proteinuria, glomerular eicosanoid production, and serum lipids in rats with streptozotocin-induced DM (STZ-DM). Groups of 5-8 rats with STZ-DM were maintained on low insulin and then pair-fed with isocaloric diets enriched with either FO (20% w/w) or beef tallow (BT; 20% w/w). GFR was determined in the same animals at onset of diet and after 8 and 20 weeks on the respective diets by [14C]inulin clearance using implanted osmotic minipumps each time. Significant hyperfiltration was present initially and GFR did not change on either diet for 20 weeks, in spite of a significant and greater than 50% decrease in all prostaglandins (PGE2, TXB2, PGF2 alpha, 6-keto, PGF1 alpha) produced by glomeruli isolated from DM/FO as compared to DM/BT or control rats. FO diet completely corrected the hypertriglyceridemia of diabetes and significantly reduced the mild and early proteinuria of DM. The decrease in proteinuria and the correction of hyperlipidemia of DM by a FO-enriched diet may be beneficial in the long term not only for the development of diabetic glomerulopathy, but also for the accelerated atherosclerosis of DM.

Animals↗
PubMed1989

A specific Fc gamma receptor on cultured rat mesangial cells.

Mesangial cells represent specialized pericytes in the renal glomerulus that contribute to the regulation of a variety of glomerular functions. Recently we and others have shown that cultured mesangial cells bind and take up immune complexes in an Fc-dependent manner leading in turn to generation of PGE2, reactive oxygen, and platelet-activating factor. The present studies were designed to further characterize potential Fc-gamma R on mesangial cells. Binding assays with either monomeric or heat aggregated (HA) [125I] labeled rat subclass-specific IgG were performed at 4 degrees C for 2 h on subcultured rat mesangial cells. Monomeric rat IgG2a, IgG2b, IgG1 and HA IgG2a bound only nonspecifically. Saturable Fc-dependent binding occurred for HA IgG2b and HA IgG1 though maximal binding and affinity were much higher for IgG2b. The presence of an Fc-gamma R was confirmed by surface protein iodination of mesangial cells (MC) and immunoprecipitation with either a polyclonal or mAb 2.4G2 prepared against murine Fc-gamma R. Both antibodies precipitated a 45-kDa iodinated protein band from cultured rat MC that comigrated with that from murine macrophage J774 cells on SDS-PAGE. This protein band also reacted with the polyclonal anti Fc-gamma R antibody on immunoblots. In contrast rat renal papillary epithelial cells were negative. The 45-kDa protein recognized by the rat anti-Fc-gamma R antibody 2.4G2 probably represents the binding site for HA IgG2b, as the 2.4G2 antibody also blocked binding of HA IgG2b. By immunofluorescence microscopy all MC stained positively with the polyclonal anti-Fc-gamma R antibody. A cDNA probe for the Fc-gamma RII-alpha on murine macrophages hybridized to mRNA from cultured rat MC which was of the same size (though less abundant) as that from J774 macrophages. These results further characterize the Fc-gamma R on cultured rat MC, and raise the possibility that the mesangial Fc-gamma R may play a role in the handling of immune-complexes by the renal glomerulus.

Animals↗
PubMed1989

Interactions of low density lipoprotein with rat mesangial cells.

Hyperlipidemia may contribute to the pathogenesis of glomerular sclerosis. We therefore studied binding and uptake of low density lipoprotein (LDL) by cultured rat mesangial cells. In addition effects of LDL on PGE2 synthesis and cell proliferation were determined. At 4 degrees C mesangial cells bound [125I] LDL in a time- and concentration-dependent manner with half-maximal binding observed at 5 micrograms/ml of LDL protein. Binding was blocked by excess unlabeled LDL and by heparin. Uptake (binding plus internalization) of LDL at 37 degrees C markedly exceeded binding at 4 degrees C, continued to increase even with longer periods of incubation, and showed no saturability, consistent with uptake of LDL by mesangial cells. Further evidence for LDL uptake by mesangial cells was obtained by use of the fluorescent probe 1,1'-dioactadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate-labeled LDL (Dil-LDL). Incubation of mesangial cells with Dil-LDL at 37 degrees C showed positive fluorescence for all mesangial cells, indicating uptake of the Dil-LDL. LDL had a biphasic effect on mesangial cell proliferation as determined by [3H] thymidine incorporation. LDL at 10 micrograms/ml enhanced [3H] thymidine uptake modestly, but significantly, whereas a progressive and marked inhibition occurred at LDL concentration from 100 to 500 micrograms/ml. While LDL at 10 and 100 micrograms/ml significantly stimulated PGE2 production, inhibition of PGE2 by meclofenamate did not influence the effects of LDL on [3H] thymidine incorporation. We conclude that mesangial cells show specific binding and uptake of LDL and that high concentrations of LDL markedly decrease mesangial cell proliferation. These findings may pertain to the pathogenesis of glomerular lesions in hyperlipidemia of renal disease.

Animals↗
PubMed1989

Atrial natriuretic peptide and nitroprusside cause relaxation of cultured rat mesangial cells.

These studies evaluate the effects of atrial natriuretic peptide (ANP) and nitroprusside (NP) on cultured mesangial cells (MC) grown on a flexible silicone rubber surface. The basal tone of the MC produced wrinkles on the silicone rubber surface. A decrease in number or magnitude of wrinkles was considered to represent cell relaxation, whereas an increase represented cell contraction. ANP (10(-9) M) produced a relaxation in greater than 60% of the cells by 10 min. The percentage of cells showing a decrease of wrinkles was significantly higher (P less than 0.05 at 5 min and P less than 0.001 at 10 min) during the ANP-treated period than during the control period. NP (10(-5) M) caused a decrease of wrinkles in greater than 80% of cells (P less than 0.02 at 5 min and P less than 0.01 at 10 min) compared with a 5% decrease in the control period. Dibutyryl guanosine 3',5'-cyclic monophosphate (DBcGMP; 10(-4) M) also produced a decrease of wrinkles in 81% of the cells (P less than 0.02) compared with a 9% decrease in the control period. MC treated with ANP, NP, or DBcGMP and then labeled with 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)-phallacidin did not show obvious alteration of morphology of actin filaments compared with untreated cells. ANP could inhibit as well as partially reverse the agonist (angiotensin II)-induced contractile response. ANP (10(-10)-10(-8) M) as well as NP (10(-5) M) increased intracellular cGMP content of MC (P less than 0.005) compared with control cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Amanitins↗
PubMed1989

Relationship of GTP-binding proteins, phospholipase C, and PGE2 synthesis in rat glomerular mesangial cells.

We evaluated the role of GTP-binding proteins in the activation of phospholipase C, release of arachidonic acid, and synthesis of prostaglandin (PG) E2 in response to platelet-activating factor (PAF) and angiotensin II (ANG II) in cultured rat mesangial cells. Pretreatment with pertussis toxin (PT) decreased PGE2 formation and arachidonic acid release in response to PAF and ANG II but not that to A 23187. PT pretreatment also inhibited formation of inositol trisphosphate (IP3) in response to ANG II or PAF but did not significantly alter the rise in intracellular calcium detected by fura-2. PT catalyzed ADP ribosylation of two proteins of molecular mass approximately 40 and 41 kDa. Further evidence for involvement of GTP-binding protein in phospholipase C activation was that GTP-gamma S stimulated IP3 generation. Immunoblots with antibodies directed against different inhibitory alpha subunits of GTP-binding proteins showed that the major 40-kDa PT substrate reacted with an antibody directed against a decapeptide of the G protein subunit alpha i2 that is also found in leukocytes. This was further confirmed by Northern blot that showed the existence of mRNA in mesangial cells that hybridized with a cDNA probe for G alpha i2. In addition lesser amounts of mRNA hybridized with a restriction fragment cDNA probe for G alpha i3, which corresponds to the 41-kDa substrate for PT ribosylation. These results show that phospholipase C activation by PAF and ANG II in mesangial cells involves a specific G protein, most likely G alpha i2.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗