Desensitization of type 1 angiotensin II receptor subtypes in the rat kidney.

Differences involving serine residues in the sequence of the carboxyl-terminal tail of type 1 angiotensin II (Ang II) receptor subtypes AT(1A) and AT(1B) suggest differences in desensitization ability. We examined the Ang II-induced homologous desensitization patterns of both receptor subtypes in freshly isolated renal structures: glomerulus (Glom), afferent arteriole, and cortical thick ascending limb (CTAL), whose content in each subtype mRNA is different, by measuring variations in intracellular calcium concentration. A preexposure to a maximal dose of Ang II, followed by a second application of the same concentration, induced: 1) a complete desensitization in Glom, where AT(1A) and AT(1B) mRNAs were expressed in similar proportions, and 2) no or partial desensitization in afferent arteriole and CTAL, where AT(1A) mRNA was predominant. In the absence of nephron structure containing only AT(1B) mRNA, we studied rat anterior pituitary cells that exhibit high content in this subtype and observed that desensitization was not complete. In Glom, CTAL, and pituitary cells, desensitization proceeded in a dose-dependent manner. In Glom and CTAL, desensitization occurred via a PKC-independent mechanism. These results suggest that desensitization does not depend on the nature of Ang II receptor subtype but either on the proportion of each subtype in a given cell and/or on cell specific type. This could allow adaptive biological responses to Ang II appropriate to the specific function of a given cell type.

Effects of angiotensin IV and angiotensin-(1-7) on basal and angiotensin II-stimulated cytosolic Ca2+ in mesangial cells.

This study analyzed the influence of two main metabolites of angiotensin II, angiotensin IV and angiotensin-(1-7), on basal and angiotensin II-dependent [Ca2+](i) in rat mesangial cells. Angiotensin IV behaved as a weak agonist. Its effects were abolished by angiotensin AT(1) receptor antagonists. Treatment with angiotensin II abolished the effect of a subsequent treatment with angiotensin IV whereas two successive angiotensin IV-dependent [Ca2+](i) peaks were obtained. Angiotensin II increased [Ca2+](i) in a Ca2+-free medium whereas angiotensin IV was inactive. Leucine-valine-valine-hemorphin 7, a hemorphin specific for the angiotensin AT(4) receptor, was devoid of any agonistic or antagonistic effect. In contrast, angiotensin-(1-7), if without influence on basal [Ca2+](i), inhibited angiotensin II- and angiotensin IV-dependent [Ca2+](i) increases. Total inhibition of the angiotensin IV effect was obtained whereas association of angiotensin-(1-7) to 8-(NN-diethylamino)-octyl-3,4,5-trimethoxybenzoate, an inhibitor of inositol phosphate-mediated Ca2+ release, was necessary to suppress the effect of angiotensin II. These results provide evidence that angiotensin II metabolites may participate in the control of [Ca2+](i) in mesangial cells at the initial stage of binding to the angiotensin AT(1) receptors.

Effects of angiotensin II and antagonists on AT(1) receptor expression in mesangial cells.

Rat mesangial cells were exposed to angiotensin II, angiotensin AT(1) receptor antagonists such as losartan, EXP 3174 and candesartan, or dexamethasone for increasing periods (1-24 h). Angiotensin AT(1A) and AT(1B) receptor mRNA were measured by reverse transcription-polymerase chain reaction (RT-PCR). Angiotensin II, losartan and EXP 3174 did not modify significantly angiotensin AT(1A) and AT(1B) receptor mRNA. Candesartan increased angiotensin AT(1B) receptor mRNA and, to a lesser extent, angiotensin AT(1A) receptor mRNA. In contrast, dexamethasone decreased mainly angiotensin AT(1B) receptor mRNA. As shown by Western blot analysis, exposure of mesangial cells to angiotensin II, losartan or EXP 3174 did not produce any change in angiotensin AT(1) receptor protein, whereas dexamethasone and candesartan exerted inhibitory effects. In conclusion, the angiotensin AT(1B) receptor subtype, the most abundantly distributed in rat mesangial cells, is inhibited by glucocorticoids. The effect of candesartan is more complex with a slight stimulation of angiotensin AT(1B) mRNA and a marked inhibition of angiotensin AT(1) receptor protein. In contrast, angiotensin II and the other angiotensin AT(1) receptor antagonists studied are inactive on angiotensin AT(1) mRNA and protein.

Functional evidence for an angiotensin IV receptor in rat resistance arteries.

To distinguish between the different effects of angiotensin IV (Ang IV) on resistance artery vasoreactivity, freshly isolated rat mesenteric arteries were perfused and the changes in their diameter were recorded under various conditions. Ang IV exerted vasoconstrictor effects on both normal vessels and vessels that had been precontracted with phenylephrine or serotonin. This effect was abolished by losartan or candesartan cilexetil, two type 1 angiotensin receptor antagonists, but not by PD 123319, a type 2 angiotensin receptor antagonist. No tachyphylaxis was observed for the vasoconstrictor effect of Ang IV. N(G)-nitro-L-arginine methyl ester, a nitric oxide synthase inhibitor, had no effect on Ang IV-induced vasoconstriction, whereas indomethacin, a cyclooxygenase inhibitor that was inactive by itself, influenced Ang IV-induced vasoconstriction, suggesting that Ang IV could stimulate the release of prostaglandins. Treatment of preconstricted vessels by candesartan cilexetil unraveled a vasodilator effect of Ang IV that was abolished by PD 123319, a type 2 angiotensin receptor antagonist. Unexpectedly, Ang IV still produced a vasoconstrictor effect on normal or preconstricted vessels after blockade of both type 1 and type 2 angiotensin receptors. Taken together, these results show that Ang IV influences resistance artery vasoreactivity via different mechanisms, one of which implicates a functionally active type 4 angiotensin receptor.

Genotype-phenotype relationships for the renin-angiotensin-aldosterone system in a normal population.

The renin-angiotensin-aldosterone system plays an important role in blood pressure regulation by influencing salt-water homeostasis and vascular tone. The purpose of the present study was to search for associations of single nucleotide polymorphisms on 3 major candidate genes of this system with the plasma concentrations of the corresponding renin-angiotensin-aldosterone system components considered as quantitative phenotypes. Genotyping was performed in 114 normotensive subjects for different variants of the angiotensinogen (AGT) gene (C-532T, G-6A, M235T), the angiotensin I-converting enzyme (ACE) gene [4656(CT)(2/3)], the aldosterone synthase (CYP11B2), and the type 1 angiotensin II receptor (AT1R) gene (A1166C) by hybridization with allele-specific oligonucleotides (ASO) or enzymatic digestion of polymerase chain reaction products. Plasma levels of AGT, ACE, angiotensin II (Ang II), aldosterone, and immunoreactive active renin were measured according to standard techniques. Platelet binding sites for Ang II were analyzed by the binding of radioiodinated Ang II to purified platelets. B(max) and K(D) values of the Ang II binding sites on platelets of each individual were calculated to examine a possible relationship between these parameters and the AT1R genotype. A highly significant association of the ACE 4656(CT)(2/3) variant with plasma ACE levels was observed (P<0.0001). ANOVA showed a significant effect of the AGT C-532T polymorphism on AGT plasma levels (P=0.017), but no significant effect was detectable with the other AGT polymorphisms tested, such as the G-6A or the M235T. A significant effect association was also found between the C-344T polymorphism of the CYP11B2 gene and plasma aldosterone levels, with the T allele associated with higher levels (P=0.02). No genotype effect of the AT1R A1166C polymorphism was detected either on the B(max) or the K(D) value of the Ang II receptors on platelets.

Mesangial AT1 receptors: expression, signaling, and regulation.

Mesangial cells are one of the main targets of angiotensin II (AngII) in the renal cortex. AngII receptors on mesangial cells are of high affinity (nanomolar range). They belong to the AT1 subtype as shown by the inhibitory effect of AT1 antagonists on [125I]-Sar1, Ala8 AngII binding and on all of the biologic effects mediated by AngII, such as cytosolic calcium stimulation, inositol phosphate formation, prostaglandin production, and cell contraction. AngII also exerts long-term effects on mesangial cells, including stimulation of cell growth and synthesis of a variety of proteins, essentially the components of the extracellular matrix (collagen, fibronectin) and the type 1 inhibitor of plasminogen activator. These effects are mediated, at least in part, by autocrine products, in particular endothelin, platelet-derived growth factor, and transforming growth factor-beta, whose synthesis is enhanced by AngII. Treatment by an AT1 receptor blocker of mice with experimental nephritis inhibits activation of type I collagen alpha2 chain promoter and prevents the development of glomerulosclerosis. AngII receptors in rat mesangial cells are equally distributed between the AT1A and AT1B isoforms. Treatment of these cells by AngII or losartan, an AT1 receptor blocker, has no effect on AT1A and AT1B receptor mRNA expression, whereas candesartan, another AT1 receptor blocker, increases and dexamethasone decreases this expression.

[Active metabolites derived from angiotensin II]. / Les métabolites actifs dérivés de l'angiotensine II.

It has been recently shown that angiotensin II (Ang II) is not the only active peptide of the renin-angiotensin system. Several of its degradation products including Ang III (obtained by deletion of the N terminal amino acids), Ang IV (obtained by deletion of the two N terminal amino acids), and Ang II (1-7) (obtained by deletion of the C terminal amino acid), also possess biological functions. These peptides are formed via the activity of several enzymes: angiotensin--converting enzyme, aminopeptidases A and N, neutral endopeptidase and prolylendopeptidase. Ang III possesses most of the properties of Ang II and shares the same receptors AT1 and AT2. In addition this peptide is particularly important in brain physiology and plays a major role in the secretion of arginine vasopressine. Ang IV possesses its own receptors distinct from AT1 and AT2. Some of its effects (for example, stimulation of the synthesis of the type 1 inhibitor of plasminogen activator by endothelial cells) were previously attributed to Ang II. Others effects, like renal and cerebral vasodilatation, are opposed to Ang II effects. The role of Ang IV in renal physiology remains to be determined. Ang II (1-7) exhibits direct and indirect effects, the latter resulting from Ang II (1-7)-dependent formation of nitric oxide and vasodilatory prostaglandins. Ang II (1-7) potentiates the hypotensive effect of bradykinin and plays also a major role in the control of the hydroelectrolytic balance. It possesses its own receptor: AT1-7, recognizable by (sar1-thr8) Ang II or Sarthran. Finally Ang II (1-7) is converted into Ango II (1-5), by angiotensin-converting enzyme. This peptide is inactive. All of these enzymes, peptides and receptors are present in kidney. Thus the renin-angiotensin system appears to be much more complicated than thought a few years ago, setting the problem of new therapeutic tools for the treatment of hypertension and glomerulosclerosis.

Hemorphins inhibit angiotensin IV binding and interact with aminopeptidase N.

[125I]-Ang IV binding to rabbit collecting duct cell membranes was inhibited by hemorphins (H), a class of endogenous peptides obtained by hydrolysis of the beta chain of hemoglobin. The most potent competitors were those with a valine in their N-terminal part such as LVV-H7 and VV-H7 (IC50 = 1.3 nM) followed by VV-H8 and K6VV-H7 (5.1 nM). The same H, like Ang IV, interacted with aminopeptidase N (APN) as shown by their inhibitory effect (28-36%) on APN activity. HPLC analysis showed that only H with a N-terminal valine or leucine were hydrolyzed. Since H are detected in the body fluids, they are likely to act as endogenous competitors of Ang IV.

Characterization of angiotensin IV-degrading enzymes and receptors on rat mesangial cells.

Because mesangial cells (MC) are a target and a degradation site for angiotensin II (ANG II), we characterized the degrading enzymes and receptors of ANG IV, a metabolite of ANG II, on these cells. ANG IV was metabolized into its NH2-terminal deleted peptides, ANG II-(4-8), ANG II-(5-8), and ANG II-(6-8) by rat MC. Total protection of ANG IV was obtained only when PC-18, a specific aminopeptidase N (APN) inhibitor, and JFH-27A, a mixed inhibitor of dipeptidylaminopeptidase (DAP) and neutral endopeptidase (NEP), were simultaneously added. In contrast, thiorphan, an NEP inhibitor, was inactive. These results demonstrate the exclusive role of APN and DAP in ANG IV degradation. 125I-labeled ANG IV binding was studied in the presence of PC-18 and JFH-27A to suppress ligand degradation. Under these conditions, ANG IV-specific receptors could be demonstrated with a KD of 1.8 nM and a density of 55 fmol/mg. In contrast with MC, no evidence for ANG IV receptors could be obtained in freshly isolated glomeruli. ANG IV stimulated cytosolic calcium concentration in MC, whereas its NH2-terminal deleted metabolites were inactive. Therefore, ANG IV must be protected from degradation by APN and DAP in studies on the nonimmediate biological effects of this peptide.

Evidence for angiotensin IV receptors in human collecting duct cells.

Because angiotensin II (Ang II) has been found at high concentrations in the proximal tubule fluid and because tubular brush border membranes exhibit a marked capacity for degrading Ang II, we thought it of interest to examine the binding sites for Ang II (3-8) (referred to as Ang IV), a metabolite of Ang II, downstream in the nephron. We studied the binding of [125I]-Ang IV and also of [125I]-Sar1, Ala8, Ang II to SV-40 transformed human collecting duct cell (HCD) membranes. No specific binding site for [125I]-Sar1, Ala8, Ang II and no Ang II-dependent cytosolic calcium response could be observed. Moreover, no signal for the human type I Ang II receptor (hAT1) mRNA was present in HCD cells. In contrast, [125I]-Ang IV bound specifically to HCD cell membranes. Mean Kd and Bmax values derived from saturation binding studies were 5.6 +/- 2.0 nM and 1007.6 +/- 140.2 fmol/mg protein, respectively. The rank order of affinity for competitive Ang II-related peptides was: Ang IV > Ang III > Ang II > Ang II (4-8) > Ang II (1-7). [125I]-Ang IV binding was not modified by nonpeptide AT1 (losartan) or AT2 (PD123177) antagonists. GTP gamma S and dithiotreitol did not affect [125I]-Ang IV binding. Ang IV stimulated cAMP production by intact HCD cells in the presence of forskolin but did not modify cGMP production or cytosolic calcium concentration. Taken together, these results indicate that HCD cells represent a target site for Ang IV but do not possess Ang II receptors.

Regulation of angiotensin II receptor subtypes by dexamethasone in rat mesangial cells.

The objective of this study was to examine the role of dexamethasone on the expression of angiotensin II (Ang II) receptors in cultured rat mesangial cells. Dexamethasone caused concentration- and time-dependent decreases in 125I-[Sar1,Ala8]Ang II binding that were prevented by glucocorticoid receptor inhibition with mifepristone. A lag time of 24 hours and a dexamethasone concentration of at least 10 nmol/L were necessary for this effect to occur. Dexamethasone-induced reduction of 125I-[Sar1,Ala8]Ang II binding resulted from decreased Ang II type 1 (AT1) receptor density. No change in the apparent dissociation constant was observed. Dexamethasone also markedly inhibited Ang II-dependent inositol phosphate accumulation. Both reverse transcription-polymerase chain reaction and Northern blot analysis using specific short probes from the 3' noncoding region of the cDNA demonstrated the presence of AT1A and AT1B receptor mRNAs in rat mesangial cells, with a slight predominance of AT1B. Therefore, we studied the effect of dexamethasone on the expression of these two subtypes in rat mesangial cells. Dexamethasone produced a time-dependent decrease of AT1B receptor mRNA that was apparent after 6 hours of incubation, whereas AT1A receptor mRNA did not change. Mifepristone also suppressed the dexamethasone-induced decrease in AT1B receptor mRNA. In conclusion, glucocorticoids diminish Ang II receptor density at the mesangial cell surface through a mechanism that implies successive interaction with the glucocorticoid receptor and specific reduction in AT1B receptor mRNA expression. This differential regulation of both AT1 receptor subtypes might allow glucocorticoids to exert adjusted effects in their various target tissues.

[Angiotensin IV, a new component of the renin-angiotensin system, which acts on kidney cells]. / L'angiotensine IV, un nouveau composant du système rénine-angiotensine actif sur les cellules rénales.

Angiotensin IV (Ang IV), the hexapeptide obtained from angiotensin II (Ang II) by deletion of the first two N terminal amino acids, possesses specific receptors in various tissues. Our aim was to search for such receptors in two types of renal cells, rat mesangial cells and principal cells of the human collecting duct. [125I]-Ang IV specifically bound to mesangial cell surface and to membranes prepared from the principal cells. In both cases, affinity was approximately 5 nmol/L and receptor density was close to 1000 fmol/mg protein. The order of potency of different competitors was as follows: Ang IV > Ang III > Ang II > Ang II (4-8) > Ang II (1-7). Binding sites were distinct from those of Ang II since type 1 or type 2 Ang II receptor nonpeptide antagonists produced no displacement. Reciprocally, Ang IV did not displace Ang II from its binding sites. Ang IV inhibited the vasoconstrictor effect of Ang II on rat mesangial cells and increases cyclic AMP production in principal cells, but only when it had been previously stimulated. Taken together, these results demonstrate that the glomerulus and the collecting duct represent target sites for Ang IV and suggest that Ang IV could influence the renal functions.

Differential regulation of angiotensin II and losartan binding sites in glomeruli and mesangial cells.

The aim of the present report was to examine the effect of several agents on angiotensin II (ANG II) and losartan receptors using 125I-[Sar1,Ala8]ANG II and [3H]losartan as radiolabeled ligand, respectively. ANG II receptors were downregulated in glomeruli from rats infused with ANG II during 3 wk or rats receiving losartan orally during 1 wk. The number of sites (Bmax) was reduced, but the dissociation constant (Kd) value was unchanged. Losartan receptors were downregulated in glomeruli from rats receiving losartan, but remained unchanged in glomeruli from rats infused with ANG II. Since in vivo administration of losartan results in increase of plasma ANG II and formation of metabolites, in vitro studies using human mesangial cells were performed to better analyze the present findings. Treatment of mesangial cells during 4 days by ANG II, losartan, or its metabolite, EXP-3174, also produced downregulation of 125I-[Sar1,Ala8]ANG II binding sites with a decreased Bmax and unchanged Kd value. Only treatment of mesangial cells by ANG II or EXP-3174 produced downregulation of [3H]losartan binding sites. In contrast, exposure of these cells to losartan resulted in upregulation of [3H]losartan binding sites. Under all conditions, only Bmax was modified. Whereas internalization of [3H]losartan in mesangial cells was negligible under all experimental conditions, there was an increase of the percentage of internalized 125I-[Sar1,Ala8]ANG II after exposure of the cells to ANG II or AT1 antagonists. No change was observed in mesangial cell AT1 receptor mRNA levels. This study demonstrates that 1) AT1 mRNA is expressed in human mesangial cells; 2) the characteristics of 125I-[Sar1,Ala8]ANG II and [3H]losartan binding sites in rat glomeruli and human mesangial cells are different, with Kd and Bmax values greater in both preparations when [3H]losartan was utilized; 3) both types of binding sites obey different regulations, and the effects of losartan in vivo are due in part to the associated increase in plasma ANG II levels and the transformation of the drug into its metabolite, EXP-3174; 4) downregulation of AT1 receptors does not depend on changes in mRNA expression but is associated with increased relative internalization.

Characterization of [3H]losartan receptors in isolated rat glomeruli.

[3H]Losartan bound specifically to isolated rat glomeruli. Scatchard analysis revealed a single class of losartan binding sites with an apparent dissociation constant (KD) of 6.2 nM and a density of receptor sites (Bmax) of 1.2 pmol/mg protein. In comparison, [3H][Sar1,Ala8]angiotensin II binding sites exhibited the same KD value (4.3 nM), but a considerably lower Bmax (52 fmol/mg protein). Moreover whereas [125I][Sar1,Ala8]angiotensin II was almost equally displaced by angiotensin II, [Sar1,Ala8] angiotensin II and losartan, [3H]losartan was potently displaced by losartan only. Finally, [125I][Sar1,Ala8]angiotensin II but not [3H]losartan binding sites were sensitive to guanosine triphosphate (GTP) gamma S and Dithiothreitol. These data, together with the recent demonstration of intrinsic effects of losartan, support the view that [3H]losartan does not label only the angiotensin II type 1 receptor (AT1).

Intrinsic properties of the nonpeptide angiotensin II antagonist losartan in glomeruli and mesangial cells at high concentrations.

Intrinsic activities of the nonpeptide angiotensin II antagonist losartan were examined in a number of in vitro assays. Losartan produced contraction of rat isolated glomeruli at 100 mumol/l and of human mesangial cells at 1 to 100 mumol/l. Cell surface reduction was associated with disorganization of the alpha actin microfilament bundles. Losartan also stimulated cytosolic calcium concentration in cultured human mesangial cells at high concentrations (10-100 mumol/l). Losartan-dependent cytosolic free calcium concentration increase was not affected by nicardipine or 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxy-benzoate hydrochloride, whereas it was abolished in a calcium-free medium. There was a marked homologous desensitization response to losartan which was also obtained after pretreatment by EXP 3174 (2-n-butyl-4-chloro-1-[(2'-(1H-tetrazol-5-yl) biphenyl-4-yl)methyl]imidazole-5-carboxylic acid), the metabolite of losartan. The search for other agonistic effects of losartan in human mesangial cells including inositoltriphosphate formation, prostaglandin E2 production, [3H]leucine or [3H]thymidine incorporation was negative. Losartan and EXP 3174 were not toxic for human mesangial cells at the concentrations studied as judged by the absence of release of lactate dehydrogenase and the normal uptake of neutral red. These studies demonstrate that losartan exhibits glomerular effects in vitro only at high concentrations. Their relevance to in vivo situations is still questionable.

Glomerular effects of angiotensin II: a reappraisal based on studies with non-peptide receptor antagonists.

AIM: To examine the glomerular effects of angiotensin II (Ang II). METHOD: A survey of recent studies that have provided relevant information. RESULTS: Glomeruli and mesangial cells of murine and human origin have receptors for Ang II (AT receptors) that are linked to phospholipase C. The dissociation constant (Kd) for these receptors is in the nanomolar range, and they are denser in freshly isolated glomeruli than in cultured mesangial cells. Pharmacological studies with the AT1 receptor, using losartan and its metabolite E3174, and with the AT2 receptor using PD 123177 and CGP 42112A, have shown that glomerular receptors for Ang II belong to the AT1 type. Furthermore, all the functional effects of Ang II in glomeruli and mesangial cells, including a rise in intracellular calcium, the stimulation of prostaglandin and protein synthesis, and glomerular vasoreactivity, are mediated by the AT1 receptor. [3H]-losartan binds specifically to mesangial cells but with different parameters from those observed for [125I]-Ang II. Losartan and E 3174 behave as non-competitive antagonists. Losartan exhibits some intrinsic effects but only at high concentrations not likely to be reached in vivo at normal doses. AT1 receptors in glomeruli are downregulated by plasma concentrations of Ang II and losartan. CONCLUSIONS: These results demonstrate that the cellular mode of action of Ang II in glomeruli is mediated by the AT1 receptor type.

[Mechanism of action and catabolism of atrial natriuretic factor in cultured human and animal kidney cells]. / Mode d'action et catabolisme du facteur auriculaire natriurétique dans les cellules rénales humaines et animales en culture.

Cultures of renal cells from human or animal origins have allowed the modes of action and the degradation pathways of atrial natriuretic factor (ANF) to be characterized. Human glomerular mesangial and epithelial cells possess ANF receptors of both types, only clearance receptors (C) in mesangial cells, receptors with guanylate cyclase activity (A) and C receptors in epithelial cells which are, in addition, equipped with ectoenzymes rapidly degrading extracellular ANF. Epithelial cells which have been stimulated by ANF secrete cyclic guanosine monophosphate (cGMP) at their apical side. Vascular smooth muscle cells prepared from the rabbit renal cortex also possess A receptors of high affinity and C receptors. Neutral endopeptidase (NEP), an enzyme of which ANF is a specific substrate in the kidney, is expressed at the cell surface. Its expression is inhibited by factors present in the serum and is increased by glucocorticoids. Principal cells of the collecting duct are also a target for ANF via A and C receptors. Taken together, these studies demonstrate that the kidneys are sites both for the physiological effects and the degradation of ANF. Production of cGMP results in vasodilation in the renal cortex, increase of glomerular filtration rate and decrease of sodium reabsorption in the collecting duct. Degradation of ANF occurs via two different ways, its conversion into inactive peptides by NEP and its internalization after binding to C receptors.

Cell surface receptors and ectoenzymes in mesangial cells.

Mesangial cells possess a variety of receptors for hormones and autacoids. They are also equipped with ectoenzymes whose function may be to control the availability of autacoids and hormones at their receptor sites. Several examples are considered. Receptors for angiotensin II (AII) are present both on murine and human mesangial cells. One single group of receptors has been demonstrated in each of these preparations. Mesangial cell AII receptors are linked to phospholipase C via a G protein. They belong to the AT1 subtype because (125I)AII is displaced from its binding sites preferentially by AT1 antagonists such as DUP 753 and EXP 3,174, whereas AT2 antagonists are much less potent. AT1 antagonists suppress the biological effects of AII in mesangial cells, including the stimulation of intracellular calcium concentration and the increase of prostaglandin synthesis and of (3H)leucine incorporation. Mesangial cells also have receptors for atrial natriuretic factor, but the distribution between B receptors with guanylate cyclase activity and clearance (C) receptors varies with the species. Both types are present in murine mesangial cells, whereas only C receptors are found in human mesangial cells. In contrast, human epithelial cells possess both B and C receptors. Ecto-5'-nucleotidase activity results in the production of adenosine, which acts on mesangial cells through A1 and A2 receptors. This enzyme is markedly induced in rat mesangial cells by interleukin-1, whose effect is mediated in part by prostaglandin E2 and cAMP. Various other cAMP-stimulating agents also induce 5'-nucleotidase expression in rat mesangial cells. Ectopeptidases are present in all glomerular cell types but essentially in epithelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)