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C Ojeda

Publications and source records attributed to C Ojeda.

At least 19 recordsLinked to original sources
PubMed2001

Characterization of a Na+-Ca2+ exchanger NCX1 isoform in bovine fasciculata cells of adrenal gland.

The Na+-Ca2+ exchanger (NCX) is well known to regulate intracellular Ca2+ concentration in many excitable cells. To date, three members of the NCX exchanger gene family have been identified: NCX1, NCX2 and NCX3. In zona fasciculata of the bovine adrenal gland, a Na+-Ca2+ exchanger has been recently found to display biochemical similarities with the NCX1 exchanger of renal basolateral cells. Although a Na+-Ca2+ exchanger has already been characterized in zona medulla of adrenal gland, such an exchanger has never been studied in the adrenal cortex. Thus, we have used a reverse transcriptase-polymerase chain reaction (RT-PCR) assay with specific NCX primers on fasciculata cell cultures. The sequencing of the amplified fragment revealed, for the first time, the presence of a NACA3 isoform of the NCX1 exchanger in the adrenal cortex, similar to the one found in adrenal chromaffin cells and in renal cortex. This isoform is 92 and 94% identical in the portion compared with kidney Na+-Ca2+ exchanger NACA3 of rat and pig, respectively.

Animals↗
PubMed2000

Characterization of Na/Ca exchange in plasmalemmal vesicles from zona fasciculata cells of the bovine adrenal gland.

The presence of an Na/Ca exchange system in fasciculata cells of the bovine adrenal gland was tested using isolated plasmalemmal vesicles. In the presence of an outwardly Na(+) gradient, Ca(2+) uptake was about 2-fold higher than in K(+) condition. Li(+) did not substitute for Na(+) and 5 mM Ni(2+) inhibited Ca(2+) uptake. Ca(2+) efflux from Ca(2+)-loaded vesicles was Na(+)-stimulated and Ni(2+)-inhibited. The saturable part of Na(+)-dependent Ca(2+) uptake displayed Michaelis-Menten kinetics. The relationship of Na(+)-dependent Ca(2+) uptake versus intravesicular Na(+) concentration was sigmoid (apparent K(0.5) approximately 24 mM; Hill number approximately 3) and Na(+) acted on V(max) without significant effect on K(m). Na(+)-stimulated Ca(2+) uptake was temperature-dependent (apparent Q(10) approximately 2.2). The inhibition properties of several divalent cations (Cd(2+), Sr(2+), Ni(2+), Ba(2+), Mn(2+), Mg(2+)) were tested and were similar to those observed in kidney basolateral membrane. The above results indicate the presence of an Na/Ca exchanger located on plasma membrane of zona fasciculata cells of bovine adrenal gland. This exchanger displays similarities with that of renal basolateral cell membrane.

Adrenal Glands↗
PubMed2000

Two types of pharmacologically distinct Ca(2+) currents with voltage-dependent similarities in zona fasciculata cells isolated from bovine adrenal gland.

Voltage-activated Ca(2+) currents, in zona fasciculata cells isolated from calf adrenal gland, were characterized using perforated patch-clamp recording. In control solution (Ca(2+): 2.5 mm) a transient inward current was followed, in 40% of the cells, by a sustained one. In 20 mm Ba(2+), 61% of the cells displayed an inward current, which consisted of transient and sustained components. The other cells produced either a sustained or a transient inward current. These different patterns were dependent upon time in culture. Current-voltage relationships show that both the transient and sustained components activated, peaked and reversed at similar potentials: -40, 0 and +60 mV, respectively. The two components, fully inactivated at -10 mV, were separated by double-pulse protocols from different holding potentials where the transient component could be inactivated or reactivated. The decaying phase of the sustained component was fitted by a double exponential (time constants: 1.9 and 20 sec at +10 mV); that of the transient component was fitted by a single exponential (time constant: 19 msec at +10 mV). Steady-state activation and inactivation curves of the two components were superimposed. Their half activation and inactivation potentials were similar, about -15 and -34 mV, respectively. The sustained component was larger in Ba(2+) than in Sr(2+) and Ca(2+). Ni(2+) (20 microm) selectively blocked the transient component while Cd(2+) (10 microm) selectively blocked the sustained one. (+/-)Bay K 8644 (0.5 microm) increased the sustained component and nitrendipine (0.5-1 microm) blocked it selectively. The sustained component was inhibited by calciseptine (1 microm). Both components were unaffected by omega-conotoxin GVIA and MVIIC (0.5 microm). These results show that two distinct populations of Ca(2+) channels coexist in this cell type. Although the voltage dependence of their activation and inactivation are comparable, these two components of the inward current are similar to T- and L-type currents described in other cells.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed2000

Functional expression of the L-type calcium channel in mice skeletal muscle during prenatal myogenesis.

The densities of skeletal muscle intramembrane charge movement and macroscopic L-type Ca(2+) current have been shown to increase during prenatal development. In the present work, the electrophysiological characteristics of L-type Ca(2+) channels were analyzed over the embryonic period E14 to E19 using the whole-cell and cell-attached procedures. At the macroscopic level, the whole-cell L-type Ca(2+) conductance increased 100% between E14 and E19. This enhancement was accompanied by a small negative shift of the voltage dependence and a marked acceleration of the inactivation kinetics. At the single-channel level, the unitary conductance decreased significantly from 13.2 +/- 0.1 pS (n = 8) at E14 to 10.7 +/- 0.3 pS (n = 7) at E18 and the open probability was multiplied by 2. No significant change of the density of functional channels was observed during the same period. In contrast to the density of intramembrane charge movement, which, under the same conditions, has been shown to increase between 16 and 19 days, L-type Ca(2+) channels properties change mostly between 14 and 16 days. Taken together, these results suggest that the two functions of the dihydropyridine receptor are carried by two different proteins which could be differentially regulated by subunit composition and/or degree of phosphorylation.

Analysis of Variance↗
PubMed1998

Activation by angiotensin II of Ca(2+)-dependent K+ and Cl- currents in zona fasciculata cells of bovine adrenal gland.

The effects of angiotensin II (100 nM) on the electrical membrane properties of zona fasciculata cells isolated from calf adrenal gland were studied using the whole cell patch recording method. In current-clamp condition, angiotension II induced a biphasic membrane response which began by a transient hyperpolarization followed by a depolarization more positive than the control resting potential. These effects were abolished by Losartan (10(-5) M), an antagonist of angiotensin receptors of type 1. The angiotensin II-induced transient hyperpolarization was characterized in voltage-clamp condition from a holding potential of -10 mV. Using either the perforated or the standard recording method, a transient outward current accompanied by an increase of the membrane conductance was observed in response to the hormonal stimulation. This outward current consisted of an initial fast peak followed by an oscillating or a slowly decaying plateau current. In Cl(-)-free solution, the outward current reversed at -78.5 mV, a value close to EK. It was blocked by external TEA (20 mM) and by apamin (50 nM). In K(+)-free solution, the transient outward current, sensitive to Cl- channel blocker DPC (400 microM), reversed at -52 mV, a more positive potential than ECl. Its magnitude changed in the same direction as the driving force for Cl-. The hormone-induced transient outward current was never observed when EGTA (5 mM) was added to the pipette solution. The plateau current was suppressed in nominally CA(2+)-free solution (47% of cells) and was reversibly blocked by Cd2+ (300 microM) but not by nisoldipine (0.5-1 microM) which inhibited voltage-gated Ca2+ currents identified in this cell type. The present experiments show that the transient hyperpolarization induced by angiotensin II is due to Ca(2+)-dependent K+ and Cl- currents. These two membrane currents are co-activated in response to an internal increase of [Ca2+]i originating from intra- and extracellular stores.

Angiotensin II↗
PubMed1998

Influence of the transmission route and disease duration in the histopathology of chronic hepatitis C: a study of 101 patients.

OBJECTIVE: Different studies have demonstrated that factors such as transmission route, disease duration and age at the time of infection can influence the histological evolution of chronic hepatitis by the hepatitis C virus (HCV). The aim of this study was to determine if epidemiological factors such as disease duration and transmission route influence the severity of the histological lesions of patients with chronic hepatitis by HCV. DESIGN: A prospective study. METHODS: The hepatic biopsies of 101 patients diagnosed with chronic hepatitis by HCV were studied. The patients were divided into three groups according to transmission mode: (1) post-transfusional (n = 28), (2) associated with the use of drugs by parenteral route or intravenous drug use (n = 28), and (3) sporadic hepatitis (n = 45). RESULTS: We found more severe forms of hepatopathy in post-transfusional hepatitis and sporadic groups than in the intravenous drug user group of patients. The disease evolution time was significantly higher in patients diagnosed as having chronic active hepatitis with or without cirrhosis (13.8 +/- 9 years) than in patients with chronic persistent hepatitis (8 +/- 4 years), P < 0.01. We found a significant correlation between the evolution time of the infection by HCV and the Histology Activity Index (P < 0.01). The multivariate analysis showed that only the transmission route and the disease evolution time are predictive variables of Histology Activity Index in chronic hepatitis C. CONCLUSION: These results suggest that the post-transfusional and sporadic transmission routes and a greater evolution time of the disease are epidemiological variables that are associated with the presence of more severe histological lesions in chronic hepatitis C.

Adolescent↗
PubMed1998

Severe and early alteration of action potential during acute cardiac rejection in rats.

INTRODUCTION: Alteration of cardiac action potential and its adaptation to heart rate could contribute to cardiac dysfunction and arrhythmias during acute cardiac rejection. METHODS AND RESULTS: Heterotopic heart transplantation was performed in allogeneic and syngeneic rats in which the action potentials of right and left ventricles were measured at 1, 2.5, 3.3, and 5.7 Hz successively using standard microelectrode techniques and compared with nontransplanted hearts. For each frequency, we measured action potential amplitude, action potential duration, transmembrane resting potential, and Vmax. In the right ventricle, at 1 Hz in the presence of rejection (n = 40), a significant increase was observed in action potential duration at 20%, 50%, and 70% repolarization (82.5%, 75.6%, and 70.8%, respectively) and in action potential amplitude (+17.9 mV), and the resting potential was decreased (-5.3 mV). A lack of adaptation of action potential duration to the driving frequency was observed in the rejecting heart group in contrast to controls (n = 20) and nonrejecting hearts (n = 13). Similar results were observed in the left ventricle and surprisingly in the native hearts (n = 11) of recipients with allografted rejecting hearts in the abdominal position. CONCLUSION: Action potential and its adaptation to the driving frequency is considerably altered during acute rejection. A humoral factor could contribute to cardiac dysfunction.

Action Potentials↗
PubMed1996

The transient outward current of isolated bovine adrenal zona fasciculata cells: comparison between standard and perforated patch recording methods.

Voltage-clamp experiments were performed on single bovine adrenal fasciculata cells in short-term primary culture using either standard (broken membrane) or perforated whole-cell patch clamp recording. The membrane current measured with the perforated method was dominated by a very stable transient outward current. By contrast, the transient outward current recorded using the standard method was unstable. The reversal potential of the transient outward current varied linearly with the logarithm of [K+]e with a slope of 47 mV per decade. The onset of activation was sigmoidal and was fitted with a power function where n = 4. Time constants ranged from 1 to 4 msec with a maximum at -25 mV. The steady-state activation curve spanned the voltage range -50 +80 mV without reaching a clear maximum. During a pulse, the current decayed in a biexponential manner. Time constants tau 1 and tau 2 were voltage-dependent and ranged from 50 to 200 msec respectively for a voltage step at +50 mV. The steady-state inactivation was dependent on the conditioning pulse duration. Using short conditioning pulses (1.2 sec), the curve which spanned the voltage range -40 to -20 mV, was 15 mV more positive than that obtained with longer conditioning pulses (60 sec). Time constants of this "very slow inactivation" process (tau vs) determined for voltage steps at -60 and -50 mV were 15 and 10 sec respectively. A "facilitation process" of the peak current was observed when the duration or the amplitude of conditioning pulses were increased in the voltage range -100 to -50 mV. Recovery from inactivation followed a biexponential time course which seemed a mixture of both inactivation processes. In some experimental conditions, isolated cells were able to produce overshooting action potentials. These results are discussed in relation with the membrane electrogenesis of this cell type.

Adrenal Cortex↗
PubMed1995

Transverse-axial tubular system in guinea pig ventricular cardiomyocyte: 3D reconstruction, quantification and its possible role in K+ accumulation-depletion phenomenon in single cells.

K+ accumulation-depletion (AD) phenomena were found in single guinea pig ventricular myocytes using the patch-clamp method in whole cell configuration. We suggest that the cardiomyocyte transverse-axial tubular system (TATS) lumen is the restricted extracellular space where the K+ AD could take place. A three-dimensional (3D) reconstruction of the TATS in a cardiomyocyte segment from serial ultrafine sections was made by three-dimensional isosurface rendering and quantitative data were obtained from the image processing. This original approach of the TATS intricated network gave a new vision of this membrane system; moreover, quantitative data about the tubular membrane importance (52.6% of the total plasma membrane) and its surface area versus the tubular volume fraction (STATS/VTATS = 13.5 micron 2/micron 3) would fit in the electrophysiological results. The hypothesis whereby this 'extracellular' compartment could play, in single cells, a role as important as that of narrow clefts in the whole heart is discussed.

Animals↗
PubMed1992

Comparative inhibitory effects of dihydropyridines on platelet aggregation, calcium uptake and cyclic AMP concentration.

We studied the in vitro effects of several calcium channel blockers from the dihydropyridine (DHP) family on platelet aggregation and endogenous serotonin secretion, calcium uptake and cyclic AMP (cAMP) concentration using washed rat platelets. We found that, after 1 min incubation, nifedipine (Nif), nitrendipine (Nit) and nisoldipine (Nis) inhibited the thrombin-induced platelet aggregation and serotonin secretion with IC50 of about 140, 5 and 2 mumol/l, respectively. Nis and Nit are thus much more active than Nif. We also found that the thrombin-induced Ca2+ uptake amounted to 2,600 +/- 326 pmol Ca2+/10(9) platelets in control conditions. In the presence of 10 mumol/l of the DHP, this uptake was decreased by 19, 49 or 77%, with Nif, Nit or Nis, respectively. Compound BAY K 8644 (BK) with known agonistic properties on the calcium channel had inhibitory effects on the studied parameters. These compounds were in the order of Nif < BK < Nit < Nis. When added to previously aggregated platelets, Nit caused them to deaggregate. These results seem to be similar to those obtained with cAMP analogues or adenylate cyclase activators. The platelet resting cAMP concentration was therefore measured in the presence of the DHP. A nonsignificant increase was found with 20 mumol/l Nif whereas significant increases of 20 and 68% as compared with controls were obtained with 20 mumol/l Nit and Nis, respectively. Partition studies between platelets and plasma lipoproteins indicated that the effects might be related to the lipophilicity of the compounds. These data suggest that these agents work on platelet activity by multiple effects located intracellularly or at the membrane level.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Risk of cancer in patients with gastric dysplasia. Follow-up study of 67 patients.

This work is based on the follow-up evaluation of 67 moderate (MD) or severe gastric dysplasias (SD) diagnosed by endoscopic biopsy. Forty-one patients had moderate gastric dysplasia, 22 (53.65%) had regression of MD, 14 patients (34.4%) had persistence, three (7.31%) had progression to SD, and two (4.87%) had transformation in gastric adenocarcinoma. Twenty-six patients were diagnosed with severe gastric dysplasia: in 12 patients (46.15%) gastric lesions regressed to normality (five cases), mild (six cases) or moderate dysplasia (one case); six patients (23.07%) showed persistent histologic changes of SD in the subsequent biopsy specimens; eight patients (30.7%) presented progression of lesions to gastric adenocarcinoma after 1 to 79 months of follow-up evaluation. The authors conclude that moderate and severe gastric dysplasias are preneoplastic lesions and a valuable marker of gastric cancer risk; the risk of gastric cancer after moderate or severe dysplasia is of 9.52%, excluding those cases with short follow-up. The authors claim that these patients may receive a conservative clinical treatment with frequent endoscopic studies until the appearance of either early carcinoma to indicate gastrectomy, or no dysplasia at all or mild dysplasia in specimens from at least two consecutive biopsies.

Adult↗
PubMed1990

Microinjection of a conserved peptide sequence of p34cdc2 induces a Ca2+ transient in oocytes.

The product of the yeast cell cycle control gene cdc2, and its homologs in higher eukaryotes (p34cdc2), all contain a perfectly conserved sequence of 16 amino acids that has not been found in any other protein sequence. Microinjection of this peptide triggers a specific increase in the concentration of intracellular free Ca2+ that originates from intracellular stores in both starfish and Xenopus oocytes. Thus, p34cdc2 might interact through its conserved peptide domain with some component of the Ca2(+)-regulatory system.

Amino Acid Sequence↗
PubMed1987

Platelet aggregation and endogenous 5-HT secretion in presence of Ca2+, Sr2+ and Ba2+. Effects of calcium antagonists.

Washed rat platelets aggregation and endogenous serotonin release were studied after thrombin stimulation in the presence of different concentrations of Ca2+, Sr2+ or Ba2+. The extent of platelet aggregation and release was found to depend upon the external concentration of these cations. For all of them, an optimum concentration could be defined. Higher concentrations were shown to inhibit both aggregation and release. Efficiency to support thrombin-induced aggregation was in the order Ca2+ greater than Sr2+ greater than Ba2+. Complete inhibition of aggregation and release induced by thrombin was obtained after a 30 second preincubation with 38 uM nitrendipine, 1 mM Cd2+ or 1 mM Mn2+. Inhibition was obtained in the presence of Ca2+, Sr2+ or Ba2+. These results are consistent with the hypothesis that Sr2+ and Ba2+ are able to support platelet activation acting as Ca2+ substitutes. Following thrombin stimulation, they could penetrate the platelets and mimick a rise in cytoplasmic Ca2+.

Animals↗
PubMed1987

The effect of calcium channel blockers on blood platelet function, especially calcium uptake.

The effects of organic and inorganic calcium antagonists on washed platelets from rat and human have been studied. Platelet aggregation was assessed by turbidimetry. Endogenous serotonin release was measured on the same sample by means of electrochemically treated carbon fiber electrodes. The organic calcium antagonist, nitrendipine, and the inorganic calcium channel blockers (Co2+, Mn2+, Cd2+, La3+) drastically inhibited rat and human platelet aggregation induced by thrombin, ADP or adrenaline in the presence of 0.32 mM Ca2+. In our conditions, the thrombin-induced release of endogenous serotonin was found to be external Ca2+-dependent and completely inhibited by 20 microM nitrendipine or 1 mM Cd2+. In addition, Ba2+ or Sr2+ ions can be substituted for Ca2+ to bring about platelet aggregation as well as endogenous serotonin secretion. In Ba2+ or Sr2+-containing media, rat platelet aggregation and/or serotonin secretion can be inhibited by either nitrendipine or Cd2+. Finally, we have also studied the thrombin- and external Ca2+-dependence of radiolabeled calcium uptake by rat platelets. We found that the thrombin-induced 45Ca uptake was inhibited by either 18 microM nitrendipine or 1 mM Cd2+. These results provide strong evidence for the existence of an influx of divalent cations (Ca2+, Sr2+, Ba2+) triggering platelet function. They also suggest, although they do not prove, that the translocation of these cations occurs through an agonist-operated channel as proposed by Hallam and Rink (FEBS Lett. 186 (1986) 175-179).

Animals↗
PubMed1983

Block by Cs of K current iK1 and of carbachol induced K current iCch in frog atrium.

1. In frog atrium, Cs ions block both the inward rectifier iK1 and the carbachol induced K current iCch. 2. Both iK1 and iCch display a high affinity for Cs with a K0.5 of 4 X 10(-5) M for iK1 and of 8 X 10(-5) M for iCch at V = -50 mV. 3. Block of both iK1 and iCch is strongly voltage dependent. When fitted by the block model of Woodhull (1973), delta is greater than 1 for the two currents. 4. From these similarities, action of Cch on frog atrium K permeability could be interpreted as a modification of iK1.

Animals↗
PubMed1983

Inhibition by Ba of background current and of its modifications by carbachol in frog atrium.

In previous work on frog atrium [11], we have shown that the carbachol-induced K current (iCCh) and the inward rectifying K channel (iK1) are both inhibited by cesium ions. A more detailed study [1, 2] indicates that the dose-response and the voltage dependence of the inhibition by Cs of iK1 and iCCh are similar. These results are in favor of a modulation by acetylcholine of the iK1 channels. Such a conclusion has been put forward by [3] and by [9]. However, Cs ions can also block the pacemaker currents iK2 [8] and if [5]. An 'if like' component seems to be present in frog atrium (Bonvallet and Ojeda, unpublished results). For these reasons, we have studied the action of barium which can be considered as a more specific inhibitor of the inward rectifying K channels in skeletal muscle [13], [14] and in heart muscle [4]. We report here results obtained on two types of preparations: one in which carbachol increases the background conductance (typical effect) and another one (four fibres from the same frog heart) where carbachol decreases the background conductance (atypical effect) as observed by Carmeliet and Ramon (1980) in sheep Purkinje fibres. In both cases, the results show that, in presence of 5 mM barium, carbachol has no effect on the background current.

Animals↗