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M C Capogrossi

Publications and source records attributed to M C Capogrossi.

102 records · Page 6Linked to original sources
PubMed1987

Acidosis facilitates spontaneous sarcoplasmic reticulum Ca2+ release in rat myocardium.

Previous studies have shown that acidosis increases myoplasmic [Ca2+] (Cai). We have investigated whether this facilitates spontaneous sarcoplasmic reticulum (SR) Ca2+ release and its functional sequelae. In unstimulated rat papillary muscles, exposure to an acid solution (produced by increasing the [CO2] of the perfusate from 5 to 20%) caused a rapid increase in the mean tissue Cai, as measured by the photoprotein aequorin. This was paralleled by an increase in spontaneous microscopic tissue motion caused by localized Ca2+ myofilament interactions, as monitored in fluctuations in the intensity of laser light scattered by the muscle. In regularly stimulated muscles, acidosis increased the size of the Ca2+ transient associated with each contraction and caused the appearance of Cai oscillations in the diastolic period. In unstimulated single myocytes, acidosis depolarized the resting membrane potential by approximately 5 mV and enhanced the frequency of spontaneous contractile waves. The small sarcolemmal depolarization associated with each contractile wave increased and occasionally initiated spontaneous action potentials. In regularly stimulated myocytes, acidosis caused de novo spontaneous contractile waves between twitches; these waves were associated with a decrease in the amplitude of the subsequent stimulated twitch. Ryanodine (2 microM) abolished all evidence of spontaneous Ca2+ release during acidosis, markedly reduced the acidosis-induced increase in aequorin light, and reduced resting tension. We conclude that acidosis increases the likelihood for the occurrence of spontaneous SR Ca2+ release, which can cause spontaneous action potentials, increase resting tension, and negatively affect twitch tension.

Acidosis↗
PubMed1987

Synchronous occurrence of spontaneous localized calcium release from the sarcoplasmic reticulum generates action potentials in rat cardiac ventricular myocytes at normal resting membrane potential.

Under certain conditions, spontaneous release of Ca2+ from the sarcoplasmic reticulum occurs in resting mammalian myocardium. In single rat ventricular myocytes, such spontaneous Ca2+ release appears localized rather than homogeneous. When the increase in cytosolic Ca2+ is present in a single locus within a cell, it causes a small depolarization, which, at the normal resting potential, is subthreshold for generating an action potential. However, when spontaneous Ca2+ release occurs simultaneously at more than a single discrete locus, the resultant sarcolemmal depolarization is augmented to levels that can induce an action potential, even when this depolarization begins at the normal resting membrane potential. Thus, the synchronous occurrence of multifocal localized increases in cytosolic Ca2+ due to spontaneous Ca2+ release from the sarcoplasmic reticulum within ventricular myocytes is a mechanism for "abnormal automaticity."

Action Potentials↗
PubMed1986

Single adult rabbit and rat cardiac myocytes retain the Ca2+- and species-dependent systolic and diastolic contractile properties of intact muscle.

The systolic and diastolic properties of single myocytes and intact papillary muscles isolated from hearts of adult rats and rabbits were examined at 37 degrees C over a range of stimulation frequencies and bathing [Ca2+]o (Cao). In both rabbit myocytes and intact muscles bathed in 1 mM Cao, increasing the frequency of stimulation from 6 to 120 min-1 resulted in a positive staircase of twitch performance. During stimulation at 2 min-1, twitch performance also increased with increases in Cao up to 20 mM. In the absence of stimulation, both rabbit myocytes and muscles were completely quiescent in less than 15 mM Cao. Further increases in Cao caused the appearance of spontaneous asynchronous contractile waves in myocytes and in intact muscles caused scattered light intensity fluctuations (SLIF), which were previously demonstrated to be caused by Ca2+-dependent spontaneous contractile waves. In contrast to rabbit preparations, intact rat papillary muscles exhibited SLIF in 1.0 mM Cao. Two populations of rat myocytes were observed in 1 mM Cao: approximately 85% of unstimulated cells exhibited low-frequency (3-4 min-1) spontaneous contractile waves, whereas 15%, during a 1-min observation period, were quiescent. In a given Cao, the contractile wave frequency in myocytes and SLIF in intact muscles were constant for long periods of time. In both intact rat muscles and myocytes with spontaneous waves, in 1 mM Cao, increasing the frequency of stimulation from 6 to 120 min-1 resulted, on the average, in a 65% reduction in steady state twitch amplitude. Of the rat myocytes that did not manifest waves, some had a positive, some had a flat, and some had a negative staircase; the average steady state twitch amplitude of these cells during stimulation at 120 min-1 was 30% greater than that at 6 min-1. In contrast to rabbit preparations, twitch performance during stimulation at 2 min-1 saturated at 1.5 mM Cao in both intact rat muscles and in the myocytes with spontaneous waves. We conclude that the widely divergent, Ca2+-dependent systolic and diastolic properties of intact rat and rabbit cardiac muscle are retained with a high degree of fidelity in the majority of viable single myocytes isolated from the myocardium of these species, and that these myocytes are thus a valid model for studies of Ca2+-dependent excitation-contraction mechanisms in the heart.

Animals↗
PubMed1986

The interaction of electrically stimulated twitches and spontaneous contractile waves in single cardiac myocytes.

Spontaneous myofilament motion that propagates within cells as a contractile wave is a manifestation of localized Ca2+ release from sarcoplasmic reticulum (SR). At 37 degrees C, when bathing [Ca2+] (Cao) is 1.0 mM, rat myocytes exhibit contractile waves at rest and the interwave interval averages 9.1 +/- 1.5 s (n = 6). We determined whether there was an interaction between this type of SR Ca2+ release and that induced by electrical stimulation to cause a twitch, and whether such an interaction had functional significance. Progressive decreases in SR Ca2+ loading effected by graded concentrations of caffeine produced proportional decreases in the mechanical amplitude of the twitch and of the spontaneous contractile wave. Regular electrical stimulation in physiologic Cao abolished the waves and, after stimulation, waves did not reappear for a period of time (delay interval). Over a range of stimulation frequencies (6-72 min-1), the delay interval ranged from 11.4 +/- 3.6 to 12.4 +/- 1.7 s and was usually greater than the interwave interval at rest. The delay interval for a wave to occur after a twitch was reduced in the presence of increased Cao, glycosides, or catecholamines. When the interstimulus interval exceeded the delay interval, waves could appear between twitches and had a marked effect of shortening the duration of the action potential and decreasing the amplitude of the subsequent twitch. The magnitude of this effect varied inversely with time (up to 2 s) between the onset of the spontaneous diastolic wave and the subsequent stimulated twitch. A reduction of the interstimulus interval to less than the delay interval prevented the occurrence of diastolic waves. These results demonstrate the presence of an interaction between spontaneous and action potential-mediated Ca2+ release, which can be interpreted on the basis of a common Ca2+ pool and perhaps common release mechanisms. This interaction can explain many of the known effects of electrical stimulation on phenomena that are thought to result from spontaneous Ca2+ oscillations in intact tissue.

Animals↗
PubMed1986

Adenosine release from isolated rat adipocytes: influence of fat cell concentration and cell size.

The release of adenosine by isolated rat adipocytes into the incubation medium was studied in relation to fat cell size and concentration. Incubations were carried out for 60 min at 37 degrees C in Krebs-Ringer bicarbonate-albumin medium containing 6 mM glucose. 2'-Deoxycoformycin was added to inhibit endogenous adenosine deaminase activity (maximal suppression was achieved at 0.8 microM concentration of the inhibitor). The data show that (a) the amount of adenosine released into the medium was similar for the first and second 30-min incubation periods; (b) increasing adipocyte concentration markedly inhibited adenosine release; and (c) large fat cells (volume greater than 300 pl) released significantly more adenosine (per fat cell) into the medium than smaller fat cells (volume less than 180 pl) when incubated at concentrations of less than or equal to 350,000 cells/ml. Above this cell concentration, differences between adenosine release and cell size were not noted. Adenosine release by isolated rat adipocytes appears to be a precisely regulated process which is exquisitly sensitive to the number of fat cells in the incubation medium and, to a certain extent, to the adipocyte size.

Adenosine↗
PubMed1985

Frequency modulation and synchronization of spontaneous oscillations in cardiac cells.

Both intact mammalian cardiac muscle and single adult Ca2+-tolerant myocytes, under appropriate experimental conditions, exhibit periodic, spontaneous myofilament oscillations that originate locally within a cell and propagate longitudinally as contractile waves. We have used microscopic imaging techniques to study the effect of electrical stimulation on the oscillation characteristics in single rat and rabbit myocytes. Unstimulated rat cells bathed in Cao of 1-3 mM exhibited these oscillations. During stimulation at rates between 6 and 120 min-1, oscillations did not occur in the interval between stimulated contractions, and following termination of stimulation a transient suppression of the spontaneous oscillation frequency occurred. Conversely, with higher cell Ca2+ loading, achieved by increasing the [Ca2+]o or by addition of isoproterenol or ouabain, stimulation caused de novo oscillations in rabbit cells and increased the spontaneous oscillation frequency in rat cells to levels that resulted in their appearance between stimulated contractions. The tendency for myofilament motion to occur simultaneously at multiple foci was also increased by stimulation at high frequencies, and partial synchronization of these foci resulted in oscillations of an increased amplitude. The modulation of the spontaneous oscillation characteristics in single cells by stimulation may explain, in part, some effects of stimulation on Ca2+-dependent oscillatory phenomena in intact cardiac tissues.

Animals↗
PubMed1985

Direct observation of the "oxygen paradox" in single rat ventricular myocytes.

By phase contrast microscopy with video length tracking, we followed the sequence of morphological changes in individual isolated rat ventricular myocytes during anoxia followed by reoxygenation. Cells appeared normal during early anoxia. After a duration of anoxia T1, which varied from 17-47 minutes in different cells, each cell abruptly contracted an average of 33% in length to an inert rectangular form presumed to be a rigor state. Cells which were reoxygenated before the onset of rigor showed normal morphology and an unchanged extent of shortening on field stimulation, compared to control. Cells that were reoxygenated after a time in the rigor state, T2, either partially recovered to a shortened rectangular form capable of stimulated twitches or rounded up rapidly to a disordered hypercontracture form. The distribution of T1 was the same for cells which recovered and which hypercontracted. In contrast, the outcome of reoxygenation depended markedly on T2: all cells that were reoxygenated after less than 10 minutes of rigor recovered function, whereas all cells that spent more than 20 minutes in rigor hypercontracted when reoxygenated. The hypercontracture appears to be the cellular analog of the "oxygen paradox" in whole hearts. Its occurrence is reliably related to duration of rigor state but not to duration of hypoxia, because of marked cellular variability in the time of onset of rigor.

Adenosine Triphosphate↗
PubMed1985

Frequency, amplitude, and propagation velocity of spontaneous Ca++-dependent contractile waves in intact adult rat cardiac muscle and isolated myocytes.

Spontaneous contractile waves due to spontaneous calcium cycling by the sarcoplasmic reticulum occur in unstimulated bulk rat papillary muscle and single rat cardiac myocytes with intact sarcolemmal function. We used video analytic techniques to quantify the wave characteristics in both bulk muscle and myocytes; laser-light scattering techniques were also employed in muscle. In muscle bathed in physiological concentrations of calcium, the true periodicity of these waves was a fraction of 1 Hz and increased up to several hertz with increases in cell calcium. This was paralleled by an increase in the frequency of scattered laser light intensity fluctuations. In myocytes, a range of spontaneous contractile wave frequencies similar to that which occurred in the muscle was observed; it could be demonstrated that an increase in superfusate calcium concentrations (2-15 mM at 23 degrees C) increases the oscillation frequency but not amplitude. In both myocytes and muscle, low concentrations of caffeine (0.5 mM) and higher temperature increased the oscillation frequency but diminished their amplitude. However, the scattered light fluctuations did not change with temperature and decreased with caffeine. These results demonstrate that (1) the true frequency of spontaneous sarcoplasmic reticulum oscillations in the unstimulated rat muscle and single myocytes with intact sarcolemmal function is low, i.e., a fraction of a hertz; (2) with cell calcium loading, the oscillation frequency accelerates to those frequencies measured previously in the "calcium overload" state; (3) while scattered light fluctuations which sample myofilament motion are a sensitive, noninvasive method of detecting the oscillations in bulk muscle, they can be insensitive to the divergent changes in oscillation amplitude and frequency.

Animals↗
PubMed1985

Spontaneous myocardial calcium oscillations: overview with emphasis on ryanodine and caffeine.

All mammalian cardiac preparations exhibit the capacity for periodic spontaneous Ca2+ release from the sarcoplasmic reticulum (SR) (Ca2+ oscillations). The occurrence of such oscillations in unstimulated preparations and their periodicity depend on the species and the Ca2+ load on the cell. When the spontaneous frequency of these oscillations exceeds the rate of external simulation, they appear between stimulated contractions and impart a variable Ca2+-dependent component of diastolic tonus and a propensity for extrasystoles and arrhythmias to occur; these diastolic oscillations can also affect systolic function as well. Although enhancing the spontaneous frequency of Ca2+ release, caffeine depresses the oscillation amplitude, whereas ryanodine suppresses both frequency and amplitude. Detailed studies of oscillation characteristics and of the different effects of caffeine and ryanodine on them may provide an understanding of and may be useful for modeling SR Ca2+ uptake and release in intact preparations.

Alkaloids↗
PubMed1979

Suppression of food intake by adenosine and inosine.

The effect of adenosine and inosine on food consumption was tested by a 4 hr satiety assay in rats fasted overnight who then received injections subcutaneously, and also in ad libitum fed rats infused intravenously for 24 hr periods. It was found that adenosine, and to a lesser degree inosine, produced a significant suppression of food intake (86 and 69% reduction from control, respectively) during the 1st hr after subbcutaneous injection. This effect persisted for 4 hr, at the end of which both substances produced an average 37% reduction in food intake from control values. This effect of adenosine was also found in ad libitum-fed animals infused intravenously with adenosine for 24 hr (35% reduction from control). These studies report a previously undescribed effect of adenosine and add another substance to the list of agents shown to influence food intake regulation. Since adenosine is known to play a local regulatory role on adipose tissue metabolism and is generated and released from adipose tissue, the present findings may indicate an additional role for adenosine in providing a feedback regulatory signal between the adipose organ and the hypothalamic centers of food intake regulation.

Adenosine↗
PubMedDate not supplied

Optimum spectral windows to minimize quantum noise of ratiometric intracellular fluorescent probes.

When fluorescent indicators are used to measure intracellular ligands in single cells, the quality of the data is usually limited by quantum (shot) noise. For indicators which shift excitation or emission wavelengths upon ligand binding, a ratiometric method is usually employed. In choosing the spectral windows for excitation or collection of fluorescence, there is a trade-off between maximum sensitivity to ligand binding, and maximum collection of light. We show that there is a well-defined optimum choice of windows which minimizes the error caused by quantum noise in the estimated ligand concentration. An algorithm for determining these optimum windows is presented. As an example, we consider the measurement of intracellular calcium by indo-1 fluorescence emission ratio in cardiac myocytes. The optimum wavelength bands for collection of fluorescence are considerably wider than those commonly employed. The use of these windows in a pulsed-excitation time-resolved calcium measurement instrument resulted in improved signal to noise ratio of the calcium signal.

Calcium↗