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B Himpens

Publications and source records attributed to B Himpens.

65 records · Page 4Linked to original sources
PubMed1990

Effects of substance P on [Ca2+]i and force in intact guinea pig ileal smooth muscle.

In intact sheets of the guinea pig ileal longitudinal muscle, loaded with fura-2, both substance P (SP)- and K(+)-induced contractile responses are preceded by cytoplasmic free Ca2+ ([Ca2+]i) variations. In response to SP (10(-10)-10(-7) M), [Ca2+]i and force increased concentration dependently. From 10(-9) M on, the response was biphasic: an initial Ca2+ spike and force transient were followed by a tonic component. The [Ca2+]i and force vs. log [SP] curves were sigmoidal for the initial phasic component, while a homologous receptor desensitization caused a reduced tonic component of the [Ca2+]i and contractile response at higher concentrations of SP. Both intracellular Ca2+ release and Ca2+ influx play a role in the effect of the peptide. In depolarizing solutions (140 mM K+) and in Ca2(+)-free medium (2 mM EGTA), SP induced a transient increase in [Ca2+]i and force. The Ca2+ stores used by SP and acetylcholine (ACh) overlap. [Ca2+]i and force fell to base-line levels when the extracellular Ca2+ was reduced from 1.2 to 0.2 mM during stimulation with SP. Verapamil reduced the tonic response. We also studied the relation between [Ca2+]i and force for the peak and steady-state values after stimulation with increasing concentrations of SP and K+. The phasic force response was linearly related to log [Ca2+]i. During the sustained response to K+, the Ca2+ sensitivity of the contractile/regulatory proteins was decreased, whereas no changes were observed during prolonged stimulation with the peptide.

Animals↗
PubMed1989

Cell calcium and its regulation in smooth muscle.

Two novel methods used to study smooth muscles-electron probe X-ray microanalysis and Ca2+-sensitive indicators (which are used for resolving, respectively, the spatial distribution and temporal distribution of calcium)-are briefly reviewed and the major findings obtained are summarized. In smooth muscle the sarcoplasmic reticulum is the major intracellular source of Ca2+; mitochondria do not play a significant role in the physiological regulation of [Ca2+]i. Under pathological conditions mitochondria can reversibly accumulate large amounts of calcium. Resting [Ca2+]i generally ranges from 80 to 200 nM, and is lower in phasic than in tonic smooth muscles. Removal of extracellular Ca2+ and Ca2+ entry blockers can reduce [Ca2+]i, but the effects of beta-adrenergic agents are variable. Increases in [Ca2+]i are triggered by electrical stimulation, depolarization with high K+, and excitatory agonists. Stretch, after a delay of several seconds, can cause an increase in [Ca2+]i in some smooth muscles. There is also a delay of approximately 200-400 ms between the initiation of the rise of Ca2+ and contraction that follows spontaneous action potentials or electrical stimulation. Agonist-induced Ca2+ release, a major mechanism of pharmacomechanical coupling, has been demonstrated in smooth muscles depolarized with high K; evidence suggests that it is mediated by G proteins that couple receptors to phospholipase C. Ca2+ release can be triggered directly in permeabilized smooth muscle with inositol 1,4,5-trisphosphate. Even though Ca2+ is the major physiological regulator of contraction, Ca2+ sensitivity of the regulatory-contractile apparatus differs in different (phasic and tonic) smooth muscles, and can be modulated in a given smooth muscle. The force [Ca2+]i ratio is higher during agonist-stimulated than during high K+-induced contractions, owing to agonist-induced increases in Ca2+ sensitivity mediated by G proteins. In some phasic smooth muscles (guinea pig ileum), the time course of the initial myosin light chain phosphorylation is extremely rapid and returns to basal levels while force remains elevated. In these smooth muscles there is also a marked decrease in the Ca2+ sensitivity of the regulatory-contractile apparatus during maintained depolarization in Ca2+-free or low Ca2+ solutions. It has been suggested that regulation of myosin light chain phosphatase plays a major role in the modulation of the Ca2+ sensitivity manifested as either potentiation or desensitization to [Ca2+]i.

Animals↗
PubMed1989

Desensitization to cytoplasmic Ca2+ and Ca2+ sensitivities of guinea-pig ileum and rabbit pulmonary artery smooth muscle.

1. The free cytoplasmic Ca2+ concentration [( Ca2+]i) was measured in the tonic rabbit pulmonary artery and the phasic ileum smooth muscle. 2. Force development and [Ca2+]i were determined during either cumulative or non-cumulative additions of [Ca2+]o to smooth muscles depolarized with 140 mM-K+ solutions. 3. The level to which [Ca2+]i declined in Ca2+-free, 140 mM-K+ solutions was significantly lower in the ileum (40 +/- 4 nM) than in pulmonary artery (77 +/- 5 nM) smooth muscle. 4. The level of [Ca2+]i reached during non-cumulative superfusion with 10 microM and 1 mM [Ca2+]o was higher in the pulmonary artery than in the ileum. 5. The force level reached for a given [Ca2+]i was also higher in the pulmonary artery than in the ileum. 6. During maintained depolarization there was a marked decrease in the sensitivity of ileum smooth muscle tension to [Ca2+]i. 7. We conclude that significant differences exist in the Ca2+ sensitivity of the regulatory/contractile apparatus among different smooth muscles; the lower sensitivity of depolarized ileum than pulmonary artery to [Ca2+]o is due to both differences in Ca2+ metabolism and in the Ca2+ sensitivity of the regulatory contractile system. We suggest that these two mechanisms also contribute to the decline in force during a phasic K+ contracture, and that desensitization to [Ca2+]i contributes to the decline of the K+ contracture in the ileum.

Animals↗
PubMed1988

Cytoplasmic free calcium, myosin light chain phosphorylation, and force in phasic and tonic smooth muscle.

The time course of [Ca2+]i, tension, and myosin light chain phosphorylation were determined during prolonged depolarization with high K+ in intact tonic (rabbit pulmonary artery) and phasic (longitudinal layer of guinea pig ileum) smooth muscles. [Ca2+]i was monitored with the 340 nm/380 nm signal ratio of the fluorescent indicator fura-2. The fluorescence ratio had a similar time course in both muscle types during depolarization with 109 mM [K+]o; after a transient peak, there was a decline to 70% of its peak value in tonic smooth muscle, and to 60% in phasic smooth muscle. Tension, however, continued to increase in the pulmonary artery, while in the ileum it declined in parallel with the [Ca2+]i. On changing [K+]o from 109 to 20 mM, tension and [Ca2+]i either remained unchanged or declined in parallel in the pulmonary artery. Phosphorylation of the 20-kD myosin light chain, measured during stimulation of muscle strips with 109 mM [K+]o in another set of experiments, increased from 3% to a peak of 50% in the intact pulmonary artery, and then declined to a steady state value of 23%. In the intact ileum, a very rapid, early transient phosphorylation (up to 50%) at 2-3 s was seen. This transient declined by 30 s to a value that was close to the resting level (7%), while tension remained at 55% of its peak force. A quick release during maintained stimulation induced no detectable change in the [Ca2+]i in either type of smooth muscle. We discuss the possibility that the slowly rising tonic tension in pulmonary artery could be due to cooperativity between phosphorylated and nonphosphorylated crossbridges.

Animals↗
PubMed1988

Free-calcium and force transients during depolarization and pharmacomechanical coupling in guinea-pig smooth muscle.

1. Fura2 was loaded by permeation and hydrolysis of the acetoxymethyl ester into smooth muscle cells of intact thin sheets of the longitudinal layer of the small intestine of the guinea-pig, to record Ca2+ transients during contraction. 2. Cytoplasmic Ca2+ ([Ca2+]i) was monitored by computing the ratio of the fluorescence signal excited at 340 and 380 nm wavelengths. The dye loading and the exposure to UV light required for the experiments had no significant effect on the contractile parameters observed. 3. Spontaneous, rhythmic increases in [Ca2+]i were often observed, preceding the onset of force. Removal of extracellular Ca2+ caused a very transient increase in [Ca2+]i accompanied by a phasic force transient; this was followed by a decline in [Ca2+]i and tension below control levels. Elevated Ca2+ from 1.2 to 15 mM also caused a fall in [Ca2+]i and a relaxation of basal tension. 4. Elevation of [K+]o increased [Ca2+]i. Graded concentrations of K+ caused graded changes in both fluorescence ratio and tension. 5. Carbachol evoked a transient increase in [Ca2+]i and contraction. Thereafter, in spite of the continued presence of the drug, both signals declined, presumably as the result of cholinergic desensitization. The initial phasic force response to carbachol was usually followed by an 'after-contraction', that was only occasionally accompanied by a similar (small) secondary rise in the fluorescence signal. 6. In depolarized smooth muscle, both in the presence and in the absence of extracellular Ca2+, carbachol induced a transient increase in [Ca2+]i, indicating that Ca2+ release from intracellular stores is a major mechanism of pharmacomechanical coupling. 7. In some preparations an applied stretch caused, after a few seconds, a rise in [Ca2+]i and force development.

Animals↗
PubMed1987

Measurement by Quin2 of changes of the intracellular calcium concentration in strips of the rabbit ear artery and of the guinea-pig ileum.

Ca2+ transient and force development were investigated in smooth muscle strips of the rabbit ear artery and the longitudinal layer of the guinea-pig ileum by using the fluorescent indicator Quin2. Agonists only transiently increased the fluorescence intensity despite the enhanced contraction while excess potassium resulted in a maintained light signal. In Ca2+ free solutions the release by an agonist of Ca2+ from an intracellular store can be demonstrated. These observations illustrate the usefulness of the Ca2+ indicator Quin2 in the study of the excitation-contraction coupling in smooth muscle under various conditions.

Aminoquinolines↗
PubMed1986

Regulatory systems for the cytoplasmic calcium concentration in smooth muscle.

Changes of the intracellular Ca concentrations play a predominant though not unique role in the regulation of the force development of smooth muscle cells. Contraction is initiated by an influx of Ca2+ through the cell membrane or by the release of Ca2+ from intracellular storage sites. Ca influx occurs via voltage operated channels and receptor operated channels. The intracellular Ca release induced by agonists probably originates from the endoplasmic reticulum. The removal of Ca2+ from the cytoplasm occurs by extrusion across the plasmalemma and by reaccumulation in the endoplasmic reticulum. These active Ca2+ transport systems are catalysed by (Ca2+ +Mg2+) ATPases. Na-Ca exchange across the sarcolemma of smooth muscle is probably of minor importance since the (Ca2+ +Mg2+)ATPase activity of plasma membranes is higher than the activity of the Na+K+ ATPase, the ultimate energy source for Na+-dependent Ca2+ extrusion. The (Ca2+ +Mg2+)ATPase of the plasmalemma has a Mr of 130.000 and it is stimulated by calmodulin. It resembles the Ca2+ transport ATPase of erythrocyte membranes, including immunological cross-reactivity. The Ca2+ transport enzyme of the endoplasmic reticulum has a Mr of 100.000, is insensitive to calmodulin and resembles the Ca2+ pump of sarcoplasmic reticulum of skeletal muscle. However, antibodies against the Ca2+ pump of skeletal muscle do not cross react with the enzyme of smooth muscle. Subcellular fractionation of pig stomach smooth muscle indicates that in this tissue the large fraction of the (Ca2+ +Mg2+)ATPase activity is present in the plasma membrane while less activity is found in the endoplasmic reticulum.

Animals↗
PubMed1985

Excitation-contraction coupling in vascular smooth muscle cells and perivascular nerve stimulation.

Smooth muscle contraction depends largely on the increase of the cytoplasmic Ca2+ concentration. This change can be brought about by the opening of voltage-dependent Ca channels or receptor operated Ca channels. Although in some vascular smooth muscle cells the addition of noradrenaline does not appreciably change the membrane potential, it is observed that stimulation of the perivascular nerve fibres elicits excitatory junction potentials and action potentials. This difference between the action of exogenous and endogenous noradrenaline could be due to either the existence of intrajunctional gamma-receptors, which differ from the extrajunctional alpha-receptors or to the release from the nerve fibres of cotransmitters together with noradrenaline. It can be concluded that the clear distinction between electromechanical coupling and pharmacomechanical coupling only applies to some experimental conditions in vitro, but that the distinction cannot be used for conditions in vivo.

Adenosine Triphosphate↗
PubMedDate not supplied

Ca2+ homeostasis in vascular smooth muscle.

The free intracellular calcium concentration is an important link in the excitation-contraction coupling mechanism of vascular smooth muscle. In this review, some current topics about vascular smooth muscle as regards Ca2+ storage, Ca2+ release, Ca2+ extrusion and Ca2+ regulation are discussed. Particular attention is paid to Ca2+ mobilized from the sarcoplasmic reticulum, the physiologically important Ca2+ reservoir in vascular smooth muscle. This occurs through two Ca2+ release channels: the inositol 1,4,5-trisphosphate receptor and the ryanodine receptor; the characteristics, function and control of these two receptors are summarized. Emphasis is also placed on a role of the nucleus as a potential Ca2+ storage site.

Animals↗