Biomedical subjects

F M Correa

Publications and source records attributed to F M Correa.

At least 19 recordsLinked to original sources

PubMed1999

Local renin-angiotensin system is involved in K+-induced aldosterone secretion from human adrenocortical NCI-H295 cells.

NCI-H295, a human adrenocarcinoma cell line, has been proposed as a model system to define the role of the renin-angiotensin system in the regulation of aldosterone production in humans. Because the precise cellular localization of the components of the renin-angiotensin system in human adrenal cortical cells remains unclear, we investigated their localization in this defined cell system. NCI-H295 cells expressed both angiotensinogen and renin as shown by reverse transcriptase polymerase chain reaction and immunohistochemistry. Human angiotensin-converting enzyme (ACE) was not detectable by immunocytochemistry, ACE binding, or reverse transcriptase polymerase chain reaction. However, 3.5 mmol/L K+ stimulated the formation of both angiotensin I and angiotensin II 1. 9- and 2.5-fold, respectively, and increased aldosterone release 3. 0-fold. The K+-induced stimulation of aldosterone release was decreased by captopril and enalaprilat (24% and 26%, respectively) and by the angiotensin type 1 (AT1)-receptor antagonist losartan (28%). Angiotensin II-induced stimulation of aldosterone release was abolished by losartan treatment. Specific [125I]Sar1-angiotensin II binding was detected by receptor autoradiography. The binding of [125I]Sar1-angiotensin II was completely displaced by the AT1 antagonist losartan but not by the AT2 receptor ligand PD 123319, confirming the expression of angiotensin II AT1 receptors in NCI-H295 cells. Our results demonstrate that NCI-H295 cells express most of the components of the renin-angiotensin system. Our failure to detect ACE, however, suggests that the production of angiotensin II in NCI-H295 cells may be ACE independent. NCI-H295 cells are able to produce angiotensin II, and K+ increases aldosterone secretion in part through an angiotensin-mediated pathway. The production of angiotensin II in NCI-H295 cells demonstrates that this human cell line can be useful to characterize the role of locally produced angiotensin II in the regulation of aldosterone release.

Adrenal Cortex↗

PubMed1998

Maturation and aging-related differences in responsiveness of rat aorta and carotid arteries to alpha1-adrenoceptor stimulation.

Functional correlates of alpha1-adrenoceptors were studied in thoracic aorta and carotid arteries from Wistar rats, at maturation (3- and 8-week-old) and aging (17- and 29-week-old) phases, using phenylephrine (PHE) and KCl. PHE and KCl effects on rat aorta did not differ with age. A reduction in maximal contractions to PHE or KCl occurred in carotid arteries during maturation. Concentration-inhibition curves to the alkylating agent chloroethylclonidine or WB4101 suggested at all ages studied a predominance of alpha1B-adrenoceptors in aorta. However, alpha1A-adrenoceptors may account for some activity in aortas of 29-week-old rats. In carotid arteries alpha1D-adrenoceptors account for functional correlates at all ages studied. The results indicate that maturation or aging affects vascular responsiveness or predominant receptor subtype differently in aorta and carotid arteries with intact endothelium.

Adrenergic alpha-Antagonists↗

PubMed1995

Kidney angiotensin II receptors and converting enzyme in neonatal and adult Wistar-Kyoto and spontaneously hypertensive rats.

The aim of the present study was to correlate the development of the renin angiotensin system (RAS) in the kidney of the rat with the development of genetic hypertension. Immature (1-week-old) and adult (12-week-old) normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive kidney rats (SHR) were used for quantification of angiotensin II (ANG II) receptors and angiotensin converting enzyme (ACE) binding sites using quantitative autoradiography. In both neonatal and adult animals of either strain, ANG II receptors were of AT1 subtype. In all kidney areas of 1-week-old rats. ANG II receptor density was higher in SHR than WKY. Binding density increased with age in WKY rats; thus, in the glomeruli and the outer stripe of the outer medulla of 12-week-old WKY, binding was significantly higher than that present in age-matched SHR. [125I]351A binding to ACE was highest in the outer medulla and not detectable in glomeruli. In 1-week-old rats, binding to ACE was higher in WKY than in SHR strain. No differences in ACE binding were found between adult SHR and WKY rats, with the exception of the inner stripe of the outer medulla, where no binding was detected in SHR. Our results support the hypothesis that the RAS in kidney is developmentally regulated and is involved in the development and maintenance of genetic hypertension in SHR.

Age Factors↗

PubMed1995

Characterization of endothelinA receptors in cerebral and peripheral arteries of the rat.

We have characterized and quantified endothelin receptors in rat brain (anterior cerebral) and peripheral (aorta, carotid, and caudal) arteries, with the use of [125I]endothelin and quantitative autoradiography. Endothelin binding was saturable, of high affinity, and totally displaced by the selective endothelin ETA antagonist BQ 123. A single class of ETA receptors is located in the medial layer of peripheral and cerebral arteries, and its quantification by autoradiography allows study of their regulation and function.

Animals↗

PubMed1994

Expression of a novel non-angiotensin II [125I]CGP 42112 binding site in healing wounds of the rat brain.

We characterized a novel non-angiotensin II binding site that is recognized by the angiotensin II AT2 receptor ligand [125I]CGP 42112, in healing brain wounds of adult rats. The binding, which was highest at 3 days after injury, appears to be localized to activated microglia surrounding the wound. The novel CGP 42112 binding site may have a role in the function of microglia and in mechanisms of tissue repair in the brain.

Angiotensin II↗

PubMed1994

Specific, non-angiotensin, [125I]CGP 42112 binding sites in rat spleen macrophages.

We report the novel expression of a high affinity non-angiotensin II binding site to [125I]CGP 42112, predominantly localized to the red pulp of the rat spleen and to isolated rat spleen macrophages. Binding is not inhibited by angiotensin II, related peptides or AT1 or AT2 receptor ligands, and is not affected by several growth factors and cytokines involved in macrophage function. Thus, CGP 42112 recognizes binding sites other than angiotensin II AT2 sites in rat macrophages. CGP 42112 and related peptides might influence the regulation of macrophage function.

Amino Acid Sequence↗

PubMed1992

Enhanced angiotensin converting enzyme binding in arteries from spontaneously hypertensive rats.

AIM: To localize and measure angiotensin converting enzyme (ACE) in different vascular beds of genetically hypertensive rats. METHODS: Quantitative autoradiography using the angiotensin converting enzyme (E.C. 3.4.15.1) inhibitor [125I]351A. RESULTS: [125I]351A binding was significantly increased in the ascending aorta (both adventitia and intima), descending (abdominal) aorta, carotid artery and coronary arteries of adult, 12-week-old spontaneously hypertensive rats (SHR) compared with Wistar-Kyoto (WKY) rats. Increased [125I]351A binding was also present in the descending aorta of 1-week-old SHR compared with age-matched WKY rats, and both groups of young rats had much higher binding than adult rats. No difference in [125I]351A binding was found in the caudal (tail) artery of adult SHR compared with WKY rats. In both the atria and the ventricles of adult SHR, [125I]351A binding was very significantly reduced. CONCLUSIONS: Our results indicate that higher ACE concentrations occur in some arteries of genetically hypertensive rats, and support the hypothesis that local arterial concentrations of ACE affect the development and maintenance of genetic hypertension.

Animals↗

PubMed1992

Estrogens regulate angiotensin-converting enzyme and angiotensin receptors in female rat anterior pituitary.

We studied the effects of the estrous cycle, ovariectomy and estrogen replacement on angiotensin-converting enzyme (ACE) (kininase II, EC 3.4.15.1) and angiotensin II (AT) receptors in the pituitary gland of the female rat. Quantitative autoradiography, with the use of consecutive pituitary sections, allowed for simultaneous determination of changes in binding and in the potential AT synthetic ability of individual pituitaries, and for a correlation between these two phenomena. In the anterior pituitary, ACE activity and binding of the ACE inhibitor [125I]-351A were not changed during the estrous cycle. Ovariectomy produced a significant increase in ACE activity and binding, and both of these parameters returned to normal after estrogen replacement. There were no changes in ACE activity or binding in the posterior pituitary during the estrous cycle or after ovariectomy or hormone replacement. AT receptors were characterized as of the AT1 type, since they were displaced by the selective AT1 antagonist DuP 753 and not by the AT2 competitor PD 123177. There were marked changes in the concentration of AT1 receptors during the estrous cycle, with highest numbers in metestrus, lower in estrus and diestrus, and lowest during proestrus. Estrogen replacement in ovariectomized rats decreased AT1 receptor number in the anterior pituitary. Our results indicate a dual effect of estrogen on anterior pituitary AT, physiologically on AT receptor expression and pharmacologically on ACE activity.

Angiotensin II↗

PubMed1992

Pressor effects of acetylcholine microinjected into forebrain nuclei of unanesthetized rats.

1. The injection of 13.5-54 nmol/500 nl of acetylcholine (ACH) into different brain areas of unanesthetized freely-moving 200-250 g male Wistar rats caused only pressor responses. 2. In the prosencephalon, the lateral septal area was the site at which ACH was more effective, whereas injections into surrounding areas, such as the accumbens/bed nucleus striae terminalis, the medial septal area or the lateral ventricle were less effective. No blood pressure effects were observed after injection into the anterior amygdala. 3. In the diencephalon, the ventromedial hypothalamic nucleus was the most sensitive site, whereas injection of ACH into surrounding areas, such as the posterior and lateral hypothalamic or the dorsal and ventral premammillary nuclei was less effective. 4. At all sites tested, the local pretreatment with 138-276 nmol atropine abolished the pressor response to ACH, suggesting a mediation through muscarinic receptors. 5. The sites of injection were confirmed histologically. 6. The present data indicate the existence of a cholinergic-sensitive site involved in the control of blood pressure at the level of the lateral septal area.

Acetylcholine↗

PubMed1991

Changes in expression of angiotensin receptor subtypes in the rat aorta during development.

Quantitative autoradiography was used to characterize angiotensin AT1 and AT2 receptors, in the rat aorta at three developmental ages; embryonic day 18 (E18), and postnatal weeks 2 and 8. The expression of angiotensin receptors was higher in the aorta of E18 and 2-week-old rat. A major proportion of the angiotensin receptors expressed in the aorta at these two ages was AT2 (84 and 81% respectively). Conversely, in the aorta of 8-week-old rats, AT1 was the predominant angiotensin receptor subtype (71%). In 8-week-old rats, the AT2 subtype was also present (28%). In pre- and postnatal rats, [125I]Sar1-angiotensin II binding to AT1 receptors was sensitive to GTP gamma S whereas binding to AT2 receptors was not. AT2 receptors may serve an important role during stages of rapid growth of the aorta, and also have a significant function in the adult vasculature.

Aging↗

PubMed1991

Comparative quantification of rat brain and pituitary angiotensin-converting enzyme with autoradiographic and enzymatic methods.

Angiotensin-converting enzyme (ACE, kininase II, EC 3.4.15.1) was quantified in selected areas of the rat brain by 3 different methods. The enzyme activity was quantified by an enzymatic assay in homogenates of discrete brain areas with the use of the artificial substrate hippuryl-His-Leu-[14C]glycine. Binding of the specific ACE inhibitor [125I]351A was quantified after incubation of thin brain sections followed by quantitative autoradiography with comparison to [125I]standards. The antigenic sites of the enzyme were quantified by immunoautoradiography, using a polyclonal antibody to ACE in combination with [125I]protein A followed by quantitative autoradiography. There was a good correlation between the ACE activity, as determined by the quantitative enzymatic method, and both the enzyme inhibitor binding and the [125I]protein A binding, as determined by quantitative autoradiography, in brain areas and in anterior and posterior lobes of the pituitary gland. The present data demonstrate that ACE activity and ACE binding can be quantified in discrete tissue areas with different methods in a variety of conditions and with a high degree of accuracy.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed1990

Autoradiography of angiotensin-converting enzyme in fixed and unfixed rat brain using the specific enzyme inhibitor [125I]351A or a polyclonal antibody and [125I]staphylococcal protein A.

Angiotensin-converting enzyme (ACE, kininase II, EC 3.4.15.1) was visualized in unfixed rat brain sections by autoradiography after incubation with a polyclonal goat anti-rabbit lung converting-enzyme antibody and [125I]protein A. Paraformaldehyde fixation interfered with the recognition of ACE by its antibody in brain nuclei but not in the choroid plexus. Conversely, incubation of brain sections with the specific ACE inhibitor [125I]351A allowed ACE visualization in either unfixed and paraformaldehyde-fixed brain sections, since [125I]351A binding was not affected by our fixation conditions. Our results indicate that ACE could be visualized in unfixed brain sections directly by incubation with the specific inhibitor [125I]351A or indirectly by radioimmunohistochemistry and autoradiography. Different autoradiographic methods could be used to visualize and quantify ACE in unfixed or fixed tissues.

Animals↗

PubMed1990

Decreased angiotensin II receptors in subfornical organ of spontaneously hypertensive rats after chronic antihypertensive treatment with enalapril.

Angiotensin II (ANG II) receptor density was higher in many brain regions of untreated spontaneously hypertensive rats (SHR) compared to untreated Wistar-Kyoto (WKY) animals. Systemic inhibition of angiotensin converting enzyme with enalapril (25 mg/kg, per os for 14 days) produces a large decrease in ANG II receptors localized exclusively in the subfornical organ (SFO) of the SHR, and no alterations in ANG II receptors in the normotensive WKY rats. Selective decrease of ANG II receptors in the SFO of the genetically hypertensive rats with enalapril may be related to its therapeutic efficacy.

Angiotensin II↗

PubMed1989

Enalapril decreases angiotensin II receptors in subfornical organ of SHR.

We studied brain angiotensin II (ANG II) receptors by quantitative autoradiography in adult normotensive Wistar-Kyoto (WKY) rats and in spontaneously hypertensive rats (SHR) after treating the rats with the converting-enzyme inhibitor enalapril, 25 mg/kg, po daily for 14 days. Enalapril treatment decreased blood pressure in only SHR, inhibited plasma angiotensin-converting enzyme activity by 85%, and increased plasma ANG I concentration and renin activity in both WKY and SHR. In the untreated SHR animals, ANG II receptor concentrations were higher in the subfornical organ, the area postrema, the nucleus of the solitary tract, and the inferior olive when compared with the untreated WKY rats. Enalapril treatment produced a large decrease in only subfornical organ ANG II receptors of SHR. The selective reversal of the alteration in subfornical organ ANG II receptors in SHR may indicate a decreased central response to ANG II and may be related to the mode of action of angiotensin-converting enzyme inhibitors in this model.

Acetylcholinesterase↗

PubMed1988

Effect of chronic administration of the converting enzyme inhibitor enalapril (MK 421) on brain atrial natriuretic peptide receptors in Wistar-Kyoto and spontaneously hypertensive rats.

Spontaneously hypertensive rats (SHR) showed lower brain ANP binding density when compared with normotensive Wistar-Kyoto (WKY) rats. In the WKY, angiotensin converting enzyme inhibitor, enalapril (25 mg/kg, p.o. for 14 days), decreased the number of ANP binding sites selectively in the subfornical organ and area postrema. Conversely, enalapril increased ANP binding density in the SHR, but only in the area postrema. Enalapril has central effects on ANP binding sites, specific to the circumventricular organs.

Animals↗

PubMed1988

A radioimmunohistochemical method for autoradiographic visualization of cell antigens using 125I-staphylococcal protein A.

125I-Staphylococcal protein A was used to visualize immunoreactive cell antigen in rat brain and pituitary by autoradiography. Autoradiograms of rat brain sections generated with 125I-protein A were clear and showed low background signals. We were able to visualize neural structures containing tyrosine hydroxylase or methionine-enkephalin-like immunoreactivities in the brain, and vasopressin-like immunoreactivity in the pituitary gland. Our results suggest that 125I-protein A can be used for the radioimmunohistochemical visualization of cell antigens in tissue sections.

Animals↗

PubMed1988

A simple two-step immunocytochemical method using protein A-peroxidase to stain immunoreactive cell antigens.

We report a simple two-step immunocytochemical method that uses staphylococcal protein A conjugated to horseradish peroxidase and the H2O2-3,3'-diaminobenzidine chromogenic reaction to stain neurons and fibers containing immunoreactive cell antigens. Our protein A-peroxidase method was found to produce stains of equal or superior quality to those of the classical 3-step peroxidase-anti-peroxidase (PAP) technique. Since the chromogenic reaction with protein A-peroxidase has low non-specific tissue binding, the reaction can be carried out at acidic pH with higher horseradish peroxidase activity and no significant increase in background. Using this method, we were able to produce clear stains of neurons and fibers containing tyrosine hydroxylase or fibers containing methionine-enkephalin. The use of protein A-peroxidase conjugate substantially simplified the immunocytochemistry procedure, and reduced both the time and cost of experiments.

Animals↗

PubMed1988

Pharmacological effects and metabolism of neurotensin and bradykinin in the isolated rat uterus.

Neurotensin (NT) and bradykinin (BK) were found to cause contractions of isolated rat uterus preparations. In 97% of the experiments, acute tachyphylaxis followed soon after the initial administration of NT. Interrelation between the oxytocic effects of NT and BK was not observed. Among the NT fragments studied, only NT-(9-13) had an oxytocic effect (less than 1.0%). All NT fragments tested induced tachyphylaxis to NT regardless of their efficacy. Using HPLC analysis, NT but not BK was found to be degraded by the intact rat uterus. A major involvement of a carboxydipeptidase cleaving at Tyr11-Ile12 is suggested. Carboxyl-blocked neurotensinamide (NT-NH2) was found to be resistent to proteolysis and not develop tachyphylaxis. No cross-tachyphylaxis was observed with NT or NT-(1-11). The results suggest the existence of different receptors for NT and BK in the uterus, as well as the existence of different receptors or receptor states that interact with NT or NT-NH2 in the rat uterus.

Animals↗