RESUMO
A complementary DNA for the angiotensin II (AngII) type 1 (AT(1)) receptor from Mustela putorius furo (ferret) was isolated from a ferret atria cDNA library. The cDNA encodes a protein (fAT(1)) of 359 amino acids having high homologies (93-99%) to other mammalian AT(1) receptor counterparts. When fAT(1) was expressed in COS-7 cells and photoaffinity labeled with the photoactive analogue (125)I-¿Sar(1), Bpa(8)AngII, a protein of 100 kDa was detected by autoradiography. The formation of this complex was specific since it was abolished in the presence of the AT(1) non-peptidic antagonist L-158,809. Functional analysis indicated that the fAT(1) receptor efficiently coupled to phospholipase C as demonstrated by an increase in inositol phosphate production following stimulation with AngII. Binding studies revealed that the fAT(1) receptor had a high affinity for the peptide antagonist ¿Sar(1), Ile(8)AngII (K(d) of 5. 8+/-1.4 nM) but a low affinity for the AT(1) selective non-peptidic antagonist DuP 753 (K(d) of 91+/-15.6 nM). Interestingly, when we substituted Thr(163) with an Ala residue, which occupies this position in many mammalian AT(1) receptors, we restored the high affinity of this receptor for Dup 753 (11.7+/-5.13 nM). These results suggest that position 163 of the AT(1) receptor does not contribute to the overall binding of peptidic ligands but that certain non-peptidic antagonists such as Dup 753 are clearly dependent on this position for efficient binding.
Assuntos
Furões/genética , Losartan/metabolismo , Receptores de Angiotensina/genética , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , Alanina/genética , Alanina/metabolismo , Sequência de Aminoácidos , Substituição de Aminoácidos , Angiotensina II/análogos & derivados , Angiotensina II/metabolismo , Angiotensina II/farmacologia , Antagonistas de Receptores de Angiotensina , Animais , Sequência de Bases , Sítios de Ligação , Ligação Competitiva/efeitos dos fármacos , Células COS , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , DNA Recombinante/genética , DNA Recombinante/metabolismo , Imidazóis/farmacologia , Radioisótopos do Iodo , Losartan/farmacologia , Dados de Sequência Molecular , Plasmídeos , Ligação Proteica , Piridinas/farmacologia , Receptor Tipo 1 de Angiotensina , Receptor Tipo 2 de Angiotensina , Receptores de Angiotensina/metabolismo , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Treonina/genética , Treonina/metabolismoRESUMO
The present study was undertaken to investigate the role of angiotensin II (Ang II) in ovulation and ovarian steroidogenesis and prostaglandin (PG) production via the Ang II receptors in rabbit ovaries. In in vitro perfused rabbit ovaries, PD123319, a selective nonpeptide antagonist for AT2 receptors, reduced the Ang II-induced ovulation in a dose-dependent manner, whereas CV-11974, a selective nonpeptide antagonist for AT1 receptor, did not affect the Ang II-induced ovulation. Ang II also significantly stimulated the meiotic maturation of ovulated ova and follicular oocytes in the absence of gonadotropin. The addition of PD123319 at 10 (-6) M to the perfusate significantly inhibited the Ang II-induced oocyte maturation. Ang II did not stimulate the production of progesterone by perfused rabbit ovaries but significantly stimulated the production of estradiol (E2) and PGs. When PD123319 at 10(-6) M was added to the perfusate 30 min before the onset of Ang II administration, the Ang II-stimulated production of E2 and PGs was significantly blocked. Saralasin, a peptide analog of Ang II, inhibited the specific binding of [125I] iodo-[Sar1, Ile8] Ang II to rabbit ovarian membranes in a concentration-dependent manner, yielding an inhibitory constant (IC50) value of 1.58 x 10(-9) M. PD123319 and CV-11974 also inhibited the binding of [125I]iodo-[Sar1, Ile8] Ang II; however, PD123319 and CV-11974 were 15 and 40 times less potent than saralasin, respectively. Autoradiographic study revealed that an intense localization of Ang II receptors in the rabbit ovaries was present in the granulosa cell layers and the stroma of the preovulatory follicles. AT2 receptors were predominantly located in granulosa cells, whereas AT1 receptors were more concentrated in the stroma and thecal cell layers. In summary, Ang II induced ovulation and oocyte maturation and stimulated the production of E2 and PG by perfused rabbit ovary in vitro via the AT2 receptor. Thus, locally produced Ang II may be part of a novel intraovarian paracrine or autocrine control mechanism during the ovulatory process.
Assuntos
Angiotensina II/farmacologia , Oócitos/fisiologia , Ovário/efeitos dos fármacos , Indução da Ovulação , Receptores de Angiotensina/fisiologia , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/antagonistas & inibidores , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , Antagonistas de Receptores de Angiotensina , Animais , Autorradiografia , Benzimidazóis/farmacologia , Compostos de Bifenilo , Senescência Celular/efeitos dos fármacos , Feminino , Imidazóis/farmacologia , Piridinas/farmacologia , Coelhos , Receptores de Angiotensina/agonistas , Saralasina/farmacologia , Tetrazóis/farmacologiaRESUMO
The neuronal angiotensin II (Ang II) type 1 (AT1) receptor is coupled to the Ras-Raf-1-mitogen-activated protein (MAP) kinase signal-transduction pathway (Yang H, Lu D, Yu K, Raizada MK. Regulation of neuromodulatory actions of angiotensin II in the brain neurons by the Ras-dependent mitogen-activated protein kinase pathway. J Neurosci. 1996;16:4047-4058). In this study we compared the effects of angiotensin II (Ang II) on AT1 receptor phosphorylation and the ability of the phosphorylated receptor to bind Ang II in neuronal cultures of Wistar-Kyoto rat (WKY) and spontaneously hypertensive rat (SHR) brains to further our understanding of the Ang II signaling mechanism. Ang II caused a time-dependent phosphorylation of AT1 receptors in both WKY and SHR brain neurons. The level of phosphorylation was higher in the SHR brain neurons; this finding was consistent with increased AT1 receptors in these cells. MAP kinase was involved in this phosphorylation, a conclusion supported by the following evidence: (1) exogenous MAP kinase phosphorylated the AT1 receptor; (2) PD98059, a MAP kinase kinase inhibitor, attenuated Ang II-stimulated AT1 receptor phosphorylation; and (3) MAP kinase and AT1 receptors were coimmunoprecipitated in Ang II-stimulated neurons. Finally, MAP kinase phosphorylation was associated with the loss of 125I-[Sar1-Ile8]-Ang II binding ability of the AT1 receptor in both strains of neurons. These observations show that Ang II stimulates phosphorylation of the neuronal AT1 receptor by a mechanism involving MAP kinase and that the phosphorylated neuronal AT1 receptor does not exhibit Ang II binding activity in the brains of either WKY or SHR.
Assuntos
Angiotensina II/farmacologia , Encéfalo/metabolismo , Neurônios/metabolismo , Receptores de Angiotensina/metabolismo , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , Angiotensina II/metabolismo , Animais , Animais Recém-Nascidos , Encéfalo/citologia , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Células Cultivadas , Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Fosforilação/efeitos dos fármacos , Testes de Precipitina , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKYRESUMO
Peptidomimitism is applied to the medicinal chemistry in order to synthesize drugs that devoid of the disadvantages of peptides. AT1 antagonists constitute a new generation of drugs for the treatment of hypertension designed and synthesized to mimic the C-terminal segment of Angiotensin II and to block its binding action on AT1 receptor. An effort was made to understand the molecular basis of hypertension by studying the conformational analysis of Ang II and its derivatives as well as the AT1 antagonists belonging to SARTANs class of molecules. Such studies offer the possibility to reveal the stereoelectronic factors responsible for bioactivity of AT1 antagonists and to design and synthesize new analogs. An example will be given which proves that drugs with better pharmacological and financial profiles may arise based on this rational design.
Assuntos
Bloqueadores do Receptor Tipo 1 de Angiotensina II , Angiotensina II/análogos & derivados , Anti-Hipertensivos/química , Desenho de Fármacos , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/química , Angiotensina II/química , Angiotensina II/farmacologia , Anti-Hipertensivos/síntese química , Anti-Hipertensivos/farmacologia , Humanos , Hipertensão/tratamento farmacológico , Losartan/análogos & derivados , Losartan/química , Espectroscopia de Ressonância Magnética , Conformação Molecular , Mimetismo Molecular , Peptídeos/química , Peptídeos/farmacologia , Relação Estrutura-AtividadeRESUMO
The dipeptide mimic Val psi[CH(CONH2)NH]His (4) was incorporated into angiotensin II (AII) analogues to provide an octapeptide saralasin derivative (29) as well as tetrapeptide analogue 19. Three C-terminal tetrapeptides (21, 25, and 28) were also prepared. All compounds were tested for their ability to displace 3H-AII from rabbit adrenal gland homogenate and as antagonists of AII and AI on guinea pig ileum. The octapeptide analogue 29 was 700 times less active than the parent peptide 30. All the C-terminal fragments 19, 21, 25, and 28 have no measurable AII antagonist activity. Of the four tetrapeptide fragments, only 21 showed any appreciable binding activity.
Assuntos
1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , Angiotensina II/análogos & derivados , Dipeptídeos , Oligopeptídeos/síntese química , Glândulas Suprarrenais/metabolismo , Sequência de Aminoácidos , Angiotensina I/antagonistas & inibidores , Angiotensina II/antagonistas & inibidores , Angiotensina II/metabolismo , Antagonistas de Receptores de Angiotensina , Animais , Fenômenos Químicos , Físico-Química , Cobaias , Íleo/fisiologia , Dados de Sequência Molecular , Contração Muscular/efeitos dos fármacos , Oligopeptídeos/química , Oligopeptídeos/farmacologia , Fragmentos de Peptídeos/síntese química , Fragmentos de Peptídeos/farmacologia , Coelhos , Receptores de Angiotensina/metabolismo , Saralasina/análogos & derivados , Saralasina/química , Saralasina/farmacologiaRESUMO
Recently, it has become clear that many of the intracellular signals mediated by Ang II receptors are similar to the signaling pathways activated by receptor tyrosine kinases. In the present paper, we are reporting a full characterization of Ang II receptors in rat fetal membranes. We assayed binding of the Ang II antagonist [125I]Sar1Ile8Ang II and the AT2 specific competitor [(125)1]CGP42112. Both ligands exhibited a rapid equilibrium and a high specificity for Ang II receptors. Competition studies confirmed the presence of both receptor subtypes, with a predominance of AT2 receptors and the following order of potency: CGP42112>Ang II>Losartan>PD123177. Immunoblotting studies of tyrosine phosphoproteins showed that Ang II (10(-6)M) mediates a rapid reduction in tyrosine phosphorylation of several proteins with apparent molecular masses in the range of 30-45 kDa. Increasing concentrations of Ang II (10(-9) - 10(-6)M) showed a dose-response behavior, suggesting a clear physiological role of the observed effect. The response, blocked by Losartan and PD123177, seems to be mediated by both receptor subtypes. These results clearly indicate that both Ang II receptors mediate tyrosine dephosphorylation in early stages of development and support a role of these receptors in growth and development.
Assuntos
Receptores de Angiotensina/metabolismo , Tirosina/metabolismo , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , Inibidores da Enzima Conversora de Angiotensina , Animais , Ligação Competitiva , Feminino , Radioisótopos do Iodo , Cinética , Fosforilação , Ratos , Ratos Endogâmicos WKY , Receptor Tipo 2 de AngiotensinaRESUMO
The potential for imaging the angiotensin II receptor was evaluated using the radioiodinated peptide antagonist [125I][Sar1, Ile8)angiotensin II. The radioligand provides a receptor-mediated signal in several tissues in rat (kidneys, adrenal and liver). The receptor-mediated signal of 3% ID/g kidney cortex should be sufficient to permit imaging, at least via SPECT. The radiotracer is sensitive to reductions in receptor concentration and can be used to define in vivo dose-occupancy curves of angiotensin II receptor ligands. Receptor-mediated images of [123I][Sar1, Ile8]angiotensin II were obtained in the rat kidney and Rhesus monkey liver.
Assuntos
1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , Radioisótopos do Iodo , Receptores de Angiotensina/metabolismo , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , 1-Sarcosina-8-Isoleucina Angiotensina II/farmacologia , Glândulas Suprarrenais/metabolismo , Glândulas Suprarrenais/ultraestrutura , Antagonistas de Receptores de Angiotensina , Animais , Anti-Hipertensivos/metabolismo , Anti-Hipertensivos/farmacologia , Autorradiografia , Feminino , Imidazóis/metabolismo , Imidazóis/farmacologia , Rim/metabolismo , Rim/ultraestrutura , Fígado/metabolismo , Fígado/ultraestrutura , Macaca mulatta , Masculino , Ensaio Radioligante , Ratos , Ratos Sprague-Dawley , Tetrazóis/metabolismo , Tetrazóis/farmacologiaRESUMO
KR31080 (2-butyl-5-methyl-6-(1-oxopyridin-2-yl)-3-[[2'-(1H-tetrazol- 5-yl) biphenyl-4-yl]methyl]-3H-imidazo[4,5-b] pyridine) is a potent inhibitor of angiotensin type 1 (AT1) receptors in rabbit aorta and human recombinant AT1 receptors. In the isolated rabbit thoracic aorta, KR31080 caused a nonparallel shift to the right of the concentration-response curves to angiotensin II (AII) with decreased maximal response (pD'2 = 10.1 +/- 0.1), but had no effect on the contractile response induced by norepinephrine. KR31080 inhibited specific [125I]AII binding to rabbit aortic membranes (AT1 receptors) and [125I][Sar1, Ile8]AII binding to human recombinant AT1 receptors in a concentration-dependent manner with IC50 values of 0.84 +/- 0.08 nM and 1.92 +/- 0.15 nM, respectively, but did not inhibit specific [125I]AII binding to bovine cerebellum membranes (AT2 receptors). In the Scatchard analysis, KR31080 interacted with rabbit aortic AT1 receptors in a competitive manner, similar to losartan. These results demonstrate that KR31080 is a potent and AT1 selective angiotensin receptor antagonist which exerts a competitive antagonism in the [125I]AII binding assay and insurmountable AT1 receptor antagonism in the functional study.
Assuntos
Antagonistas de Receptores de Angiotensina , Piridinas/farmacologia , Tetrazóis/farmacologia , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , Angiotensina II/metabolismo , Angiotensina II/farmacologia , Animais , Aorta Torácica/metabolismo , Aorta Torácica/fisiologia , Bovinos , Cerebelo/efeitos dos fármacos , Cerebelo/metabolismo , Humanos , Imidazóis/farmacologia , Técnicas In Vitro , Radioisótopos do Iodo , Losartan/farmacologia , Membranas , Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Coelhos , Receptor Tipo 1 de Angiotensina , Receptor Tipo 2 de Angiotensina , Proteínas Recombinantes/metabolismoRESUMO
1H-NMR spectra for the angiotensin agonist sarcosine-(Sar)Arg-Val-Tyr-Ile-His-Sar-Phe [( Sar1,Sar7]Ang II) and the antagonist Sar-Arg-Val-Tyr-Ile-His-Sar-Ile in dimethylsulfoxide-d6 were examined at 400 MHz. Splitting of the resonances for Tyr, His, and Sar protons revealed that the His6-Sar7 peptide bond existed in both cis and trans forms, with one isomer predominating in the ratio 5:1 in both peptides. Comparison of the chemical shifts for the His6 and Phe8 ring protons in these peptides suggested a His/Phe stacking interaction in [Sar1,Sar7]Ang II which is important for agonist activity.
Assuntos
1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , Angiotensina II/análogos & derivados , Histidina , Espectroscopia de Ressonância Magnética , Fenilalanina , Conformação Proteica , EstereoisomerismoRESUMO
In addition to its well-characterized renal hemodynamic effects, angiotensin II (Ang II) promotes growth of cultured glomerular and tubular cells, suggesting a possible role in renal development. To better define potential developmental effects of Ang II, we examined the expression of Ang II receptors in embryonic (E19) and postnatal (1, 2, 3, 10 days, 6 weeks, 3 and 9 months) rat kidneys, using in situ autoradiography and the nonpeptide antagonists losartan and PD-123177 to identify receptor subtypes. At E19, 125I-[Sar1, Ile8]Ang II binding was equally reduced by losartan and PD-123177, indicating the presence of both AT1 and AT2 receptors. A progressive increase in Ang II receptor density occurred after birth, reaching a plateau at day 10. At that time, the AT1 subtype predominated and was virtually the sole subtype present thereafter. Ang II receptor density and AT1 mRNA levels decreased in aging rats. Total AT1 receptor mRNA levels in both kidney and liver were determined by Northern hybridization analysis using a radiolabeled AT1 anti-sense cRNA probe. In both tissues, AT1 mRNA levels increased rapidly following birth, reached a maximum on day 10 and decreased thereafter. To further characterize the ontogenic effects on AT1 gene expression, renal AT1A and AT1B receptor mRNA isoforms were determined by reverse transcription and the polymerase chain reaction. No significant differences were observed during maturation between the relative levels of AT1A and AT1B mRNAs, with the AT1A isoform accounting for approximately 78% at any time point. Thus, renal AT1 receptor density increases rapidly after birth, in association with an increase in both AT1A and AT1B receptor gene expression. As the predominant receptor isoform in the adult kidney, the AT1A receptor may account for the majority of the effects of Ang II on glomerular and tubular function.
Assuntos
Rim/química , Fígado/química , Receptores de Angiotensina/genética , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , 1-Sarcosina-8-Isoleucina Angiotensina II/farmacologia , Angiotensina II/metabolismo , Animais , Expressão Gênica/fisiologia , Hibridização In Situ , Radioisótopos do Iodo , Isomerismo , Masculino , RNA Mensageiro/análise , Ensaio Radioligante , Ratos , Ratos Sprague-Dawley , Receptores de Angiotensina/química , Receptores de Angiotensina/metabolismoRESUMO
1. A high density of angiotensin II receptors was observed in the rat carotid body by in vitro autoradiography employing 125I-[Sar1, Ile8]-angiotensin II as radioligand. Displacement studies demonstrated that the receptors were of the AT1 subtype. 2. The binding pattern indicated that the AT1 receptors occurred over clumps of glomus cells, the principal chemoreceptor cell of the carotid body. Selective lesions of the sympathetic or afferent innervation of the carotid body had little effect on the density of receptor binding, demonstrating that the majority of AT1 receptors were intrinsic to the glomus cells. 3. To determine the direct effect of angiotensin II on chemoreceptor function, without the confounding effects of the vasoconstrictor action of angiotensin II, carotid sinus nerve activity was recorded from the isolated carotid body in vitro. The carotid body was superfused with Tyrode solution saturated with carbogen (95 % O2, 5 % CO2), maintained at 36 C, and multi-unit nerve activity recorded with a suction electrode. 4. Angiotensin II elicited a dose-dependent excitation of carotid sinus nerve activity (maximum increase of 36 +/- 11 % with 10 nM angiotensin II) with a threshold concentration of 1 nM. The response was blocked by the addition of an AT1 receptor antagonist, losartan (1 microM), but not by the addition of an AT2 receptor antagonist, PD123319 (1 microM). 5. In approximately 50 % of experiments the excitation was preceded by an inhibition of activity (maximum decrease of 24 +/- 8 % with 10 nM angiotensin II). This inhibitory response was markedly attenuated by losartan but not affected by PD123319. 6. These observations demonstrate that angiotensin II, acting through AT1 receptors located on glomus cells in the carotid body, can directly alter carotid chemoreceptor afferent activity. This provides a means whereby humoral information about fluid and electrolyte homeostasis might influence control of cardiorespiratory function.
Assuntos
Angiotensina II/fisiologia , Corpo Carotídeo/fisiologia , Neurônios Aferentes/fisiologia , Receptores de Angiotensina/fisiologia , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , Antagonistas de Receptores de Angiotensina , Animais , Autorradiografia , Estimulação Elétrica , Eletrofisiologia , Imidazóis/farmacologia , Radioisótopos do Iodo , Ligantes , Losartan/farmacologia , Masculino , Potenciais da Membrana/fisiologia , Técnicas de Patch-Clamp , Piridinas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor Tipo 1 de Angiotensina , Receptor Tipo 2 de Angiotensina , Receptores de Angiotensina/agonistasRESUMO
The carboxyl-terminal cytoplasmic domain of the angiotensin II type 1 (AT1) receptor has recently been shown to interact with several classes of cytoplasmic proteins that regulate different aspects of AT1 receptor physiology. Employing yeast two-hybrid screening of a mouse kidney cDNA library with the carboxyl-terminal cytoplasmic domain of the murine AT1a receptor as a bait, we have isolated a novel protein with a predicted molecular mass of 18 kDa, which we have named ATRAP (for AT1 receptor-associated protein). ATRAP interacts specifically with the carboxyl-terminal domain of the AT1a receptor but not with those of angiotensin II type 2 (AT2), m3 muscarinic acetylcholine, bradykinin B2, endothelin B, and beta2-adrenergic receptors. The mRNA of ATRAP was abundantly expressed in kidney, heart, and testis but was poorly expressed in lung, liver, spleen, and brain. The ATRAP-AT1a receptor association was confirmed by affinity chromatography, by specific co-immunoprecipitation of the two proteins, and by fluorescence microscopy, showing co-localization of these proteins in intact cells. Overexpression of ATRAP in COS-7 cells caused a marked inhibition of AT1a receptor-mediated activation of phospholipase C without affecting m3 receptor-mediated activation. In conclusion, we have isolated a novel protein that interacts specifically with the carboxyl-terminal cytoplasmic domain of the AT1a receptor and affects AT1a receptor signaling.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal , Proteínas de Transporte/metabolismo , Receptores de Angiotensina/metabolismo , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/metabolismo , Sequência de Aminoácidos , Angiotensina II/farmacologia , Animais , Sequência de Bases , Proteínas de Transporte/genética , Clonagem Molecular , DNA Complementar/genética , Relação Dose-Resposta a Droga , Ativação Enzimática , Camundongos , Dados de Sequência Molecular , Ligação Proteica , Receptor Tipo 1 de Angiotensina , Receptor Tipo 2 de Angiotensina , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Distribuição Tecidual , Fosfolipases Tipo C/metabolismoRESUMO
Mas oncogene has been shown to have focus-inducing ability in NIH 3T3 cells which are tumorigenic in vivo in nude mice. Its stable expression in a variety of cell lines conferred some angiotensin responsiveness. To understand why mas-transfected cells exhibit a transformed phenotype and if angiotensin responsiveness plays any role in this process, we studied the growth characteristics of mas-transfected 3T3 cells and demonstrated that they lose contact inhibition, exhibit foci formation, and increased DNA synthesis even in absence of serum. Our results suggest that the transformed phenotype is due to the production of a mas receptor ligand distinct from angiotensin.
Assuntos
Células 3T3/patologia , Proteínas de Neoplasias/fisiologia , Oncogenes , Proteínas Proto-Oncogênicas/fisiologia , 1-Sarcosina-8-Isoleucina Angiotensina II/análogos & derivados , 1-Sarcosina-8-Isoleucina Angiotensina II/farmacologia , Angiotensina II/farmacologia , Angiotensina III/farmacologia , Antagonistas de Receptores de Angiotensina , Animais , Cálcio/metabolismo , Captopril/farmacologia , Linhagem Celular Transformada , Transformação Celular Neoplásica/genética , Inibição de Contato , Replicação do DNA , Enalaprilato/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Neoplasias/biossíntese , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas/biossíntese , Receptores Acoplados a Proteínas G , Proteínas Recombinantes/biossínteseRESUMO
The effects of the newly-synthesized AT II analogue (Sar1 azaVal3 Ile8) AT II were investigated in comparison with the octapeptide AT II and the analogue saralazin (Sar1 Ala8) AT II, using intracerebroventricular administration, with respect to the following parameters: the level of biogenic monoamines (DA, NA and 5-HT) and the metabolites HVA and 5-HIAA in mouse forebrain; the behaviour of the animals--cataleptogenic actions of mice, PTZ convulsive--seizure threshold in mice, apomorphine stereotypy in rats and behaviour of rats in a conflict situation. The analogue (Sar1 azaVal3 Ile8) AT II, unlike saralazin and AT II, was found to induce a rise in the NA and 5-HT levels, causing also catalepsy that is different from the catalepsy induced by saralazine, AT II and haloperidol, because of its rapid onset and decline; it increases the PTZ convulsive--seizure threshold and reduces the number of punished responses to the conflict drinking test (anxiomimetic effect) in a dose 20 times lower than the dose inducing the remaining effects. This effect was antagonized by saralazine. It is concluded that the newly-synthesized analogue (Sar1 azaVal3 Ile8) AT II induces effects similar to those caused by AT II, being at the same time different to a certain extent from the effects (quantitative and qualitative) of octapeptide AT II.