PubMed Health⌕ Search

Biomedical subjects

D Redon

Publications and source records attributed to D Redon.

11 recordsLinked to original sources
PubMed1998

Calibration of intracellular Ca transients of isolated adult heart cells labelled with fura-2 by acetoxymethyl ester loading.

A procedure for calibration of fluorescence signals from adult rat heart cells loaded with the -AM ester of fura-2 is described. Calibration is complicated by dye compartmentation and potentially incomplete dye hydrolysis. These problems were overcome by subtracting from fluorescence transients the non-cytosolic (mitochondrial) component of fura-2 fluorescence plus any Ca-insensitive component of dye fluorescence, after selectively and sequentially quenching cytosolic and non-cytosolic dye with Mn. The Kd of fura-2 in cells loaded by the -AM ester, in cells depleted of ATP and equilibrated with Ca buffers, was found to be 371 +/- 39 nM at 37 degrees C. We found that calibration values for RMAX and RMIN derived from previously measured cells were of general validity, removing the need to measure RMAX and RMIN on every cell. Once these calibration values are determined, the calibration procedure to measure cytosolic Ca on any cell is a five minute procedure to determine compartmentation, using just one non-toxic and inexpensive solution. Finally, we have calculated how the errors intrinsic to the measurements translate into errors of the calculated Ca concentration and transient peak heights. These calculations allow reasonable parameters for data acquisition to be set.

Adenosine Triphosphate↗
PubMed1998

Ca uptake by heart cells: I. Ca uptake by the sarcoplasmic reticulum of intact heart cells in suspension.

Electric field stimulation of adult rat heart cells suspended in medium with 0.2 mM Ca and isoproterenol caused 45Ca uptake at a rate (5.25 pmol/mg/beat) proportional to stimulation frequency. Uptake was strongly inhibited by verapamil or thapsigargin. 45Ca autoradiography showed that stimulation dependent verapamil sensitive uptake was associated with the rod shaped cells, while the uptake by round cells was unaffected by stimulation and was verapamil-insensitive. 45Ca efflux measurements revealed a caffeine-sensitive component of uptake which was abolished by thapsigargin, and a caffeine-insensitive component. Part of the latter was sensitive to thapsigargin but not to 30 s of stimulation; another part was sensitive to such stimulation but not to thapsigargin. With longer times of stimulation, the caffeine-insensitive pool increased in size, part of which appeared to be mitochondrial Ca uptake via a thapsigargin-sensitive pool. The caffeine-sensitive pool labelled quickly in stimulated cells and its size and rate of labelling was increased by stimulation frequency (3.87 pmol/mg/beat), while the caffeine-insensitive pool labelled more slowly and was relatively insensitive to stimulation (0.77 pmol/mg/beat). We conclude that essentially all of the SR Ca pool, as defined by its involvement in excitation-contraction coupling, is released by caffeine.

Action Potentials↗
PubMed1998

Ca uptake by heart cells: II. Most entering Ca appears to leave without mixing with the sarcoplasmic reticulum Ca pool.

The rate of verapamil-sensitive uptake of 45Ca by rat heart cells stimulated to beat in suspension with 0.2 mM Ca and isoproterenol was increased > 2-fold by cell loading with the chelator Quin-2. No effect of Quin-2 loading was observed on the rate of uptake of trace levels of 54Mn, present in addition to Ca, which was used as an index of Ca channel activity. Quin-2 loading also had little effect on the rate of 45Ca uptake by cells diluted into a high K/low Na medium, where Ca uptake was primarily by Na/Ca exchange. The fast chelator 1,2-bis(o-aminophenoxy)ethane-N,N,-N',N'-tetraacetic acid (BAPTA) was 3-fold more effective than the slow chelator EGTA at preventing Ca efflux. BAPTA loading also caused an increase in sarcoplasmic reticulum (SR) Ca content. These results suggest that chelator loading had little effect on the rate of Ca influx by Ca channels or by Na/Ca exchange, and that the increased rate of 45Ca uptake seen with Quin-2 loading was caused by an inhibition of Ca efflux, either directly by chelation or by increased Ca uptake by the SR or by other intracellular organelles. This further suggests that most of the Ca entering the cell without chelator leaves again within the same beat, and that this may result from Ca efflux from a kinetically limited Ca pool in or around the diad cleft.

Aminoquinolines↗
PubMed1995

Procaine enhancement of the rapid cooling contracture and inhibition of the decay of potentiated state in rabbit papillary muscle.

We tested the possibility that inhibition of Ca2+ release from the sarcoplasmic reticulum decreases the transsarcolemmal Ca2+ efflux and results in an increased sarcoplasmic reticulum Ca2+ content. We used procaine to inhibit sarcoplasmic reticulum Ca2+ release in rabbit papillary muscles. Rapid cooling-induced contractures were used as an index of sarcoplasmic reticulum Ca2+ content. Decay of potentiated state was measured to study whether the transsarcolemmal Ca2+ efflux is decreased in the presence of procaine. Procaine (0.5, 1.0 mM) inhibited contractions elicited by electrical stimulation and enhanced the rapid cooling-induced contracture. The initial decay of potentiated state that occurs when the muscle was stimulated at a low frequency was slowed in the presence of procaine. These results suggest that procaine inhibition of Ca(2+)-induced release of sarcoplasmic reticulum Ca2+ decreases the transsarcolemmal Ca2+ efflux relative to the influx, resulting in an increased sarcoplasmic reticulum Ca2+ content.

Animals↗
PubMed1991

Effects of thiopental and halothane on spontaneous contractile activity induced in isolated ventricular muscles of the rabbit.

To see if the known properties of thiopental of reducing Ca2+ and K+ fluxes across the myocardial sarcolemma account for its arrhythmogenic action, we have evaluated the effect of the anesthetic on spontaneous contractile activity induced in isolated rabbit papillary muscles. Thiopental (20 mg/l) prolonged the duration of sustained automaticity induced by stimulation at 1-2 Hz in the presence of 1 mumol/l isoproterenol. Thiopental (10, 20 mg/l) shortened the delay before the onset of Ba(2+)-induced automaticity, which involves a decrease in a K+ current. The minimum concentration of Ba2+ required to induce automaticity was lowered by thiopental. Whether spontaneous activities were induced by high frequency stimulation in the presence of isoproterenol or by Ba2+, thiopental lowered the frequency of spontaneous beats. Thus, thiopental appears to have both arrhythmogenic and antiarrhythmic actions, and the former may be unmasked when catecholamines counteract the latter by increasing Ca2+ influx. Like thiopental, halothane (1.0%) decreased the frequency and force of Ba(2+)-induced automatic beats but, unlike thiopental, prolonged the delay before the onset of Ba(2+)-induced automaticity, indicating that halothane acts as a purely antiarrhythmic agent in this type of automaticity.

Animals↗
PubMed1990

Negative inotropic effect of ketamine in rabbit ventricular muscle.

The effect of ketamine on myocardial contractile force was examined in rabbit papillary muscles in order to determine the underlying mechanism of action of the anesthetic. Ketamine HCl (20 and 40 mg/L) inhibited rested-state contractions that are dependent on the transsarcolemmal influx of Ca2+ for activation and reduced the upstroke velocity of the slow action potential, which reflects Ca2+ influx through the slow Ca2+ channel. On the other hand, ketamine had a relatively small effect on potentiated-state contractions and no effect on rapid cooling induced contractures, both of which are activated by the release of Ca2+ stored in the sarcoplasmic reticulum. These results suggest that ketamine inhibition of transsarcolemmal Ca2+ influx plays a major role in the negative inotropic action of the anesthetic.

Animals↗
PubMed1989

Isoflurane and halothane inhibit tetanic contractions in rabbit myocardium in vitro.

Rabbit right ventricular papillary muscles were tetanized by rapid stimulation in the presence of 1 microM ryanodine, an inhibitor of sarcoplasmic reticular function. Tetanic contractions elicited in this manner increased in strength as extracellular calcium concentration [Ca+2]ext was raised from 0.5 to 5 mM, exhibited saturation behavior above [Ca+2]ext = 5 mM, and were blocked by nifedipine. Accompanying membrane potentials depolarized to +10 mV and repolarized to -60 mV between stimuli. These data suggest that rabbit myocardial tetany is supported in large part by extracellular calcium influx via slow (L-type) calcium channels, consistent with similar recent findings in the ferret. At [Ca+2]ext = 2.5 mM, isoflurane (0.6-2.3%, gas phase) and halothane (0.4-1.5%) inhibited the strength of tetanic contractions in dose-dependent fashion. At [Ca+2]ext = 20 mM neither isoflurane (1.2%) nor halothane (0.8%) inhibited tetanic contraction strength. These data demonstrate that isoflurane and halothane inhibit contractile activity that is dependent on transsarcolemmal calcium influx via pathways independent of the ryanodine-sensitive sarcoplasmic reticulum. The exact sites of inhibition (e.g., slow channel vs. intracellular transit vs. myofibrillar binding) are not identified, but inhibition by either anesthetic may be competitively reversed by high extracellular calcium concentrations.

Animals↗
PubMed1989

Effects of isoflurane and halothane on rapid cooling contractures in myocardial tissue.

The effects of isoflurane and halothane on the availability of Ca2+ stored in and released from the sarcoplasmic reticulum (SR) were studied in isolated rabbit papillary muscles by measuring the effects of the anesthetics on rapid cooling contractures, which are known to be activated by Ca2+ released from the SR. Isoflurane (0.3%) reduced the force of the rapid cooling contracture to 57% of control with a marked slowing of the average rate of contracture development and increased the force of the first contraction after rewarming to 133% of control. In contrast, 1.7% halothane, which reduced the rapid cooling contracture to 51% of control (comparable to the value in the presence of 0.3% isoflurane), had little effect on the rate of contracture development and strongly inhibited the first contraction after rewarming to 5% of control. Halothane, but not isoflurane, strongly inhibited postrest potentiated-state contractions, which are also known to be activated by Ca2+ released from the SR. These results suggest that 1) isoflurane inhibits rapid cooling-induced Ca2+ release from the SR without inhibiting Ca2+ release triggered by rapid depolarization as occurs in the potentiated-state contraction, and 2) halothane inhibits contractile activity dependent on Ca2+ released from the SR regardless of mechanism involved.

Animals↗
PubMed1984

The influence of serum potassium on the cerebral and cardiac toxicity of bupivacaine and lidocaine.

The influence of hyperkalemia on the central nervous system and cardiac toxicity of bupivacaine and lidocaine was studied in open-chested mechanically ventilated dogs. The seizure and cardiotoxic doses of intravenously administered lidocaine and bupivacaine were determined in two separate groups of normokalemic (2.7 +/- 0.05 SEM mEq/1) dogs. In the case of both anesthetics, the cardiotoxic dose was found to be approximately four times the seizure dose. Under conditions of hyperkalemia (5.4 +/- 0.08 SEM mEq/1), however, the cardiotoxic doses of both anesthetics were decreased significantly to approximately two times the seizure dose. Hyperkalemia did not change the seizure dose for either anesthetic. The cardiac to seizure dose ratio was decreased significantly for bupivacaine but not for lidocaine. The results of this study suggest that hyperkalemia enhances the cardiotoxic effects of both lidocaine and bupivacaine, with this enhancement being more pronounced in the case of bupivacaine.

Animals↗
PubMed1984

Hemodynamic effects of amrinone in dogs anesthetized with halothane: comparison with isoproterenol and dobutamine.

Baseline hemodynamic variables were established during morphine - N2O-O2 anesthesia in dogs following which N2O was discontinued and halothane 2% (end-tidal) in O2 was administered. Halothane caused significant decreases of 58, 55, and 66 percent, respectively, in cardiac output, mean arterial pressure, and left ventricular dP/dt and significantly increased pulmonary vascular resistance (161%) when compared to measurements obtained under morphine - N2O-O2 anesthesia. During halothane anesthesia, amrinone, isoproterenol, and dobutamine were administered by continuous intravenous infusion in doses which restored cardiac output to pre-halothane values. Left ventricular dP/dt was increased and pulmonary resistance was decreased significantly by the three positive inotropic agents. Isoproterenol and amrinone significantly decreased systemic vascular resistance 48 and 71 percent, respectively below the value for halothane alone. Dobutamine significantly increased mean arterial pressure. Heart rate and left ventricular end-diastolic pressure were not altered by any treatment. Ventricular arrhythmias were associated with the infusion of all three agents. It is concluded that amrinone, isoproterenol, and dobutamine were capable of restoring cardiac output which had been decreased by halothane. However, isoproterenol and amrinone caused marked decreases in systemic vascular resistance which precluded an increase in arterial pressure above the value encountered with halothane alone.

Aminopyridines↗