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Biomedical subjects

M Fallert

Publications and source records attributed to M Fallert.

At least 19 recordsLinked to original sources

Kinetics of intracellular Ca2+ concentration changes and cell contraction of electrically stimulated cardiomyocytes as analysed by automated digital-imaging microscopy.

Enzymatically disaggregated, electrically stimulated cardiomyocytes from adult rats were examined by television-mediated vital microscopy for intracellular Ca2+ concentration and contractile activity. Using an inverted microscope in the epifluorescence mode, the Ca2+ signal was imaged with a low-light-level CCD camera and traced by means of the intracellular concentration of the fluorescent complex of Ca2+ with its indicator Fluo-3. Using the transmitted-light mode, cardiomyocytes that were not loaded were imaged with a conventional CCD camera with automatic gain control and traced by length measurements. Optical images of at least 40 cardiomyocytes per batch of cells from one heart were recorded in up to 20 microscopic fields of observation on videotape within 20 min. They were consecutively analysed by a personal computer installed with an image analysis card at a time-resolution of 20 ms, employing a discrete convolution operation, filtering and threshold setting for fluorescence measurements, and contour description and vectorial analysis for length measurements. Frames of fluorescent images were corrected for the halo effect caused by the increase in the Ca(2+)-dependent fluorescence signal after electrical stimulation. The cell contraction had to be measured in the transmission mode without Fluo-3 due to the inhibition caused by the intracellular Fluo-3. The following coefficients of variation (V) were determined: Vfluorescence < 0.033 and Vtransmission < 0.003 for the precision of measurement, and Vfluorescence < 0.05 and Vtransmission < 0.04 for the reproducibility. The system was validated with isoprenaline and ouabain as agents to modify the Ca(2+)-signal and the contraction. The response of cardiomyocytes of various rats to electrical stimulation, with respect to amplitude and its time point, had a V < 0.08 for both the Ca(2+)-signal and the contraction.

Animals↗

Repair of left ventricular aneurysm. Changes in ventricular mechanics, hemodynamics, and oxygen consumption.

Anteroapical left ventricular aneurysms were produced in 23 sheep by coronary arterial ligation. Plication of the aneurysm does not change stroke volume or cardiac output and does not significantly change left ventricular oxygen consumption from the preoperative value of 5.1 +/- 2.6 ml/100 gm per minute. Plication, however, does increase left ventricular end-systolic elastance from 3.2 +/- 0.9 to 4.4 +/- 1.5 mm Hg/mm (p = 0.005). In nine of these sheep the midsagittal plane of the left ventricle was imaged by means of an array of sonomicrometry crystals before and after plication of the aneurysm. Regional wall stresses at end-systole and end-diastole and changes in diastolic function were calculated for anterior and posterior ventricular walls in the border zone adjacent to the aneurysm and in more basilar myocardium remote from the infarct. Plication significantly reduced end-systolic wall stresses and systolic stress integrals in the posterior border zone and remote myocardium, but it did not significantly change anterior wall systolic stresses or stress integrals. Plication also decreased diastolic stretching of border zone myocardium. Plication of anteroapical left ventricular aneurysm produced a shorter, more spherical ventricle and removed the dyskinetic segments but altered deformation (strain) in both circumferential and longitudinal directions. The changes in ventricular wall geometry and deformation provide an explanation for the increased ventricular end-systolic elastance and unchanged stroke volume observed after aneurysm plication.

Animals↗

Influence of rubrospinal tract and the adjacent mesencephalic reticular formation on the activity of medullary respiratory neurons and the phrenic nerve discharge in the rabbit.

Suprapontine brain sites acting on the central respiratory system have been demonstrated to give rise to inspiratory as well as expiratory facilitatory effects. In the present study the inspiratory inhibitory effect which has been reported in the cat to be elicited consistently by electrical stimulation of the rubrospinal tract and the adjacent mesencephalic reticular formation was examined in the urethane-anaesthetized rabbit. Stimulation of these sites with single electrical shocks of moderate intensity induced a short latency (onset after 3.0 ms) transient (duration: 29 ms) inhibition of the phrenic nerve activity (PHR). Short volleys of stimuli applied in mid- to late-inspiration led to a premature off-switch of inspiration. The extracellularly recorded discharge activity of the different types of medullary respiration-related units (RRU) reflected these alterations, accordingly. Axonal connections of RRU with mesencephalic structures were evaluated. Examination of orthodromic responses of medullary RRU to stimulation of this pathway revealed that most bulbospinal inspiratory neurons (10 out of 13) were paucisynaptically inhibited after short latency (at least 1.2 ms). The conduction time from bulbospinal inspiratory neurons to the recording site of PHR was 1.6 ms. Thus, a disynaptic pathway--including bulbospinal inspiratory neurons--is suggested inducing inspiratory inhibition 3.0 ms after single shock midbrain stimulation. This inhibition results in disfacilitation of phrenic motoneurons. The fact that extensive electrolytic lesions of the pneumotaxic center in rostral pons did not abolish the observed inspiratory inhibitions excludes these structures from being involved.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of a calcium entry promoting 1,4,5,7-tetrahydro-furo[3,4-b] pyridine derivative on Na+-, Ca2+- and Mg2+-dependent bioelectrical activity of Purkinje fibres and papillary muscles.

4-(2-Difluoromethoxyphenyl)-2-methyl-5-oxo-1,4,5,7-tetrahydro- furo[3,4-b]pyridine-3-carboxylic acid ether ester (CGP 28 392), a dihydropyridine derivative with an annellated lactone ring, was examined in automatically discharging calf Purkinje fibres. Velocity of spontaneous depolarization during both the early and the late phase of the diastolic pause and the rate of rise of the action potential were increased, with the threshold potential at which the upstroke is generated being unaltered. These findings are discussed in terms of ionic mechanisms, including in particular the contribution of sodium ions during the late phase of the diastolic depolarization and the novel type of calcium channel which is supposed to contribute to pacemaker depolarization and action potential initiation. In general, the effects of equimolar concentrations of nifedipine were opposite to those of CGP 28 392. At 1 X 10(-5) mol/l, however, both CGP 28 392 and nifedipine accelerated the late phase of diastolic depolarization. In electrically stimulated, partially depolarized guinea-pig papillary muscles, CGP 28 392 also prolonged the slow action potential and markedly increased inotropism. In equimolar concentrations nifedipine shortened the slow action potential and inhibited contractility. The effects of CGP 28 392 are thus compatible with facilitation of calcium entry and--in Purkinje fibres--also with enhancement of fast sodium conductivity.

Action Potentials↗

Medullary respiratory-related neurons with axonal connections to rostral pons and their function in termination of inspiration.

In urethane-anaesthetized, paralyzed and artificially ventilated rabbits, medullary respiration-related neurons (RRU) were classified according to the phase relation of their burst discharge to phrenic nerve activity. Phase-bound inspiratory (I) or expiratory (E) neurons were discriminated from phase-spanning expiratory-inspiratory (EI) or inspiratory-expiratory (IE) units. Mechanisms of termination of inspiration by electrical stimulation of rostral pontine nuclei (Nc. parabrachialis medialis; Lc. coeruleus) were examined firstly to demonstrate whether RRU receive descending excitatory and inhibitory afferents as well as ascending efferents and secondly to analyse the time course of the neuronal pathways involved. Of 120 RRU, 38 neurons were demonstrated to receive pontine afferents. About 33% of all E neurons became orthodromically excited during rostral pons stimulation whereas 18.2% of all I cells became orthodromically inhibited. Some RRU were shown to project up to the rostral pons. 50% of these were of the phase-spanning IE type. The onset of inspiratory inhibition induced by rostral pons stimulation occurred 3.4 ms after the onset of single electrical pulse stimulation. Based on these results a neuronal model for a pontine mechanism terminating inspiration is proposed.

Animals↗

Bulbar respiratory neurons during artificial lung distension or collapse in spontaneously breathing rabbits.

Lung volume was altered setting the mean air pressure within a closed spirometer system to +5, +10, -5 or -10 cm H2O. In many respiratory modulated neurons (RMN) the duration of discharge changed in the same direction as the duration of the corresponding half cycle, but changes of duration of neuronal discharge were often larger. This was the case in many EI neurons for the duration of discharge during expiration, in many I units at high lung volume for the duration during inspiration and in many IE and E cells at low lung volume for the duration of discharge during expiration. The responses to inflation or collapse, however, differed in magnitude. At all lung volumes in part of the IE and E neurons and at low lung volume in part of the I units the duration of discharge during inspiration changed in opposite direction as the duration of inspiration. Neuronal desactivation at high lung volume or activation at low volume was termed alpha-type response; it occurred in many I neurons at high lung volume and for the inspiratory part of discharge of many EI units. Neuronal activation at high lung volume or desactivation at low volume was denoted as beta-type response; it was observed in many I neurons at low lung volume and in E, EI and IE cells for the expiratory part of discharge. In a large majority of all phase types of RMN, incidence of maximum spike density within the burst discharge was delayed in time when the corresponding half cycle was prolonged and vice versa; the shift of time incidence often exceeded the change of duration of inspiration or expiration.

Action Potentials↗

Inhibitory action of vagal nerve and spinal cord afferents on discharge pattern of bulbar respiratory modulated neurons.

Single vagal nerve stimuli delivered during burst discharge of respiratory modulated neurons entailed lengthening of an interspike interval in 37% of all units tested. Occasionally stimuli delivered to the spinal cord caused lengthening of interspike intervals in respiratory modulated neurons and in other bulbar units the discharge of which was not modulated with respiration. Vagal stimuli often entailed lengthening of the following interspike interval, while the interval during which stimuli were applied remained unaltered; latencies suggested that inhibitory pathways were polysynaptic. In expiratory-inspiratory (EI) and inspiratory (I) neurons, lengthenings of intervals roughly doubled those noted in inspiratory-expiratory (IE) and expiratory (E) units. Lengthening of intervals depended on stimulus incidence within the intervals and was greatest when stimuli were delivered during the late part of the intervals. In EI and I units, lengthening of intervals occurred mainly during the middle part of burst discharge, in IE and E neurons, however, towards the end of the burst. Conditioning of discharge of respiratory modulated neurons could be demonstrated.

Animals↗

Axonal projections and primary afferents of bulbar respiratory modulated and unmodulated neurons travelling with vagal nerve or spinal cord in the rabbit.

Respiratory modulated neurons of the rabbit were examined for efferent projections and afferent connections via the spinal cord or the vagal nerves. Variability of latency of responses proved to be related to latencies. When latency of evoked potentials is plotted versus variability of latency, monosynaptic responses can apparently be discriminated from oligo- or polysynaptic responses. Latency of antidromic responses depends from stimulus frequencies used. Results showed that only 12% of all respiratory modulated neurons have spinal descending axons conducting at 9-45 m per sec and 8% of the inspiratory group of neurons possess axons running along with the vagal nerves conducting at 20-37 m per sec. It is concluded that the respiratory network in the rabbit is essentially different from that in the cat.

Animals↗

Microelectrophoretic application of putative neurotransmitters onto various types of bulbar respiratory neurons.

Six putative neurotransmitters and the beta-receptor excitant isoproterenol were applied to bulbar respiratory neurons classified according to their burst discharge in the respiratory cycle and, for comparison, also to "unspecific" cells. With glycine inhibition occurred in I and prevailed in unspecific neurons. With GABA inhibition preponderated in I and E neurons. Glutamate excited E neurons. With dopamine inhibition prevailed in I and unspecific neurons. The majority of E neurons remained unaffected. With NE excitation occurred in EI and preponderated in I, IE, E and unspecific neurons, while some IE and E neurons were inhibited. The effects of isoproterenol did not allow any clear statement about receptor properties. In I units, however, activation was more frequent than inhibition. With 5-HT excitation prevailed in IE neurons. About half of the I cells remained unaffected and in the remainder inhibition preponderated over activation. This suggests the existence of two 5-HT-specific receptors in I and IE neurons. Comparison of the single effects revealed differences in the receptor properties of the various cell types. Results suggest that some cell types possess dopaminergic receptors and that these differ from NE-receptors, which have been found in all cell types. NE-receptor stimulation apparently can result in neuronal activation or inhibition.

Animals↗

Microelectrophoretic application of antagonists of putative neurotransmitters onto various types of bulbar respiratory neurons.

Seven antagonists of putative neurotransmitters were applied to bulbar respiratory neurons and, for comparison, also to unspecific cells. The antagonists exerted distinct effects when released alone, permitting to draw conclusions about receptor properties of the various cell types. With strychnine, specific antagonist of glycine, excitation prevailed in EI, I and E neurons. With bicuculline, specific antagonist of GABA, excitation preponderated in EI and E cells. About half of the unspecific neurons were activated and the remainder were unresponsive. GDEE (glutamatediethylester), antagonist of glutamate, excited part of the IE neurons and inhibited part of the E units, while the remainder of both types as well as 2 EI cells tested were not affected. With flupentixol, antagonist of dopamine, excitation prevailed in I neurons. About half of the IE and E units remained unaffected, while in the remainder E cells inhibition preponderated over excitation. With yohimbine, an alpha-adrenoceptor blocker, inhibition prevailed in E units. The two EI as well as the majority of the I neurons remained unaffected, with two cells of the latter type being activated. Propranolol, a beta-adrenoceptor blocker, inhibited about half of the E neurons, while the remainder as well as most IE and the 2 EI cells tested were not affected. Cyproheptadine, an antagonist of 5-HT, excited most E neurons. As concerns NE-receptors, those of the alpha-type might be involved in activation of part of the E cells only, whereas all other NE effects (inhibition or activation) are mediated by CNS-specific receptors different from the alpha- and beta-type. 5-HT effects apparently are mediated by two different receptor types.

Alkaloids↗

Microelectrophoretic application of metabolic modifiers of the phosphofructokinase-hexosediphosphatase-system onto various types of bulbar respiratory neurons.

Three metabolic modifiers of the PFK-HDPase system were applied to bulbar respiratory neurons and, for comparison, also to unspecific cells. F-6-P did not alter the spike density of the majority of the respiratory and about half of the unspecific neurons. When responding, IE units were activated and unspecific cells were inhibited. Citrate did not alter the spike density of about half of all neurons tested. When responding, excitation prevailed in IE and E units, while unspecific cells were inhibited. AMP did not alter the spike density of the majority of the respiratory and about half of the unspecific cells. When responding, I neurons were activated, inhibition preponderated in IE units and both effects occurred in unspecific cells. The large number of zero effects suggests that the activity of only part of the cells is governed by the PFK-HDPase system.

Adenosine Monophosphate↗

Spatial distribution of various types of bulbar respiratory neurons in the rabbit.

In anesthetized rabbits, the burst activity of 277 single respiratory neurons was recorded extracellularly. The neurons were classified according to their spike incidence distribution within the respiratory cycle and to their response to lung distension or collapse (alpha or beta type). About one third of the neurons found in all animals were located at the level of the caudal end of the promontorium gliosum, widely scattered over the bulbar cross-section. More rostrally the units were located within a dorsal area neighbouring the tractus solitarius, more caudally within a ventral field surrounding the nucleus ambiguus. Most of the inspiratory neurons (the most frequently encountered type) were found within an antero-medial region of the medulla, extending down to about two thirds of its depth. Most of the inspiratory-expiratory units were found within an intermediate zone extending from antero-dorsal to postero-ventral. Most of the expiratory cells were found postero-ventrally. Most of the expiratory-inspiratory neurons (the rarest type) were again found within an antero-medial region at an intermediate depth. The alpha neurons were located in the rostral and the beta units in the caudal part of the medulla. The average proportion of respiratory neurons to non-respiratory units was estimated as about 1:36 and the total number of bulbar respiratory neurons as 500 on each side.

Animals↗

Patterns of bulbar respiratory neurons during and after artifical hyperventilation.

Single bulbar respiratory unit activity was recorded in urethane-anesthetized rabbits. During artificial ventilation, reflex contractions of the diaphragm were elicited by the forced deflations of the respirator. Out of 44 neurons, four I, one E and two EI units exhibited a shift of their discharge relative to the contractile phase of the diaphragm, compared to normal respiration. During hypocapnic apnea, some I, IE and E neurons became silent. Other cells belonging to all phase types, however, continued to fire tonically at a rate less than the peak rate in normal bursts. When rhythmic respiration resumed after termination of the apneic pause, five (I, IE and E) units exhibited a transient phase shift of their burst discharge compared to normal respiration. The results of simultaneous recordings of two neurons belonging to different phase types are compatible with the assumption that inspiratory units are periodically inhibited by inspiratory-expiratory cells.

Animals↗

Metabolic control of respiratory neuronal activity and the accompanying changes in breathing movements of the rabbit. 1. Mainpulation of inspiratory and expiratory-inspiratory neurons.

The property of the neuronal membrane to be permeable to metabolic modifiers of two regulatory enzymes has been utilized to manipulate the spike activity of inspiratory (I) and expiratory-inspiratory (EI) neurons of the bulbar respiratory centre. The neurons have been classified according to their response to lung distention or collapse (alpha- or beta-type) and to hyperventilation (tonic firing denoted by "+", cessation of activity by "-"). Using extracellular microelectrodes for single unit recording, the medulla oblongata was superfused with a metabolite-containing CSF. The various neuronal sub-types exhibited a differential activating or inhibitory response to one or several metabolic effectors. For example Ialpha+ units were activated by 5 mM glucose-6-phosphatase (G-6-P) and 3.5 mM 3-phosphoglycerate (3-PGA), which both inhibited Ibeta+ neurons, while 5 mM AMP inhibited Ialpha+ much more strongly than Ibeta+ cells. The spike density of Ialpha- and Ibeta- neurons was increased in the presence of 2.5 mM fructose-6-phosphate and 3.5--5 mM AMP, but became reduced by G-6-P. In contrast, 3 mM fructose-1,6-diphosphate and 5 mM 3-PGA activated the Ialpha- but inhibited the Ibeta- neurons. The EIbeta units were characteristically activated by 10 mM citrate, which inhibited all I-type neurons. Activations of the Ialpha and Ibeta neurons led to an accelerated respiratory rate and a higher tidal volume, while the opposite was true for EIbeta neurons. Intravenous injection of metabolites could not duplicate the striking effects under local applications.

Animals↗