Intracellular Calcium Movements Research Articles

Effects of calcium on cholinergic-stimulated gastrin release in the rat

The effects of alterations in availability and access of extracellular media calcium on antral gastrin release were examined in the basal state and in response to cholinergic stimulation in rat antral organ culture experiments. In the presence of either divalent cationic chelator (EGTA) or calcium channel blocker (verapamil, nifedipine), carbachol-stimulated gastrin release was inhibited completely to values that were not significantly different from non-stimulated control. In the absence of added calcium chloride, carbachol stimulated gastrin release during the initial 30 min of culture but not at 60 and 120 min of culture. Inhibition by EGTA and verapamil of carbachol-stimulated gastrin release during the initial 30 min of culture suggests, but does not prove, that these agents may also affect intracellular availability and movement of calcium. Cholinergic stimulation of gastrin release demonstrated a concentration-dependent relationship with extracellular calcium: optimal culture media calcium concentration was 1 mM. In conclusion, these studies indicate that cholinergic stimulation of the gastrin cell requires availability of extracellular calcium.

Hemodynamic interferences between diltiazem and thiopental--experimental study in the pig

The clinical applications of calcium channel blockers are rapidly growing; as barbiturates interfere with intracellular calcium movements, the possibility of drug interaction at this level must be considered. We have studied the haemodynamic interaction of Thiopentone and Diltiazem in pigs. A loading dose of Diltiazem (0.15 mg X kg-1) was injected followed by a continuous administration of 0.07 mg X kg-1 X hour. From the fifth to the tenth minute, the reduced afterload was associated with a moderate depression of the contractility of the left ventricle. These values returned to the initial level within 30 minutes. Heart rate and cardiac output were not modified. The stability of these two parameters is thought to result from the complex interaction of both drugs on the sympathetic nervous system and the ventricular function.

Effects of acetylcholine, isoprenaline, phenylephrine and dibutyryl-cAMP on the contractility of isolated myocytes of the rat heart ventricles.

Single ventricular myocytes were isolated by enzymatic dissociation from the rat heart. The myocytes showed two types of spontaneous contractions: action potential dependent 'electric' contractions which occurred as short bursts, and slowly propagating contraction waves, i.e. phasic contractions. Electric contractions were normally found only in some of the myocytes, but in about 85% of the cells these could be induced by isoprenaline (ISO), phenylephrine (PHE), dibutyryl-cAMP (DB-cAMP) or acetylcholine (ACh). The inducing potency of the drugs was ACh greater than PHE greater than ISO greater than DB-cAMP. When electric contractions were blocked with verapamil, the frequency of phasic contractions was increased by ISO and DB-cAMP, whereas PHE and ACh had not this effect. In 45 min, ISO increased the frequency tenfold. High concentrations (10(-4) and 10(-5) M) of ISO lead to steady contracture of the myocytes in about 40% of the cells. The effect of DB-cAMP was weaker being only about 20% of that of ISO and it never caused steady contractures. The results suggest that alpha receptor activation by PHE does not affect the intracellular calcium movements but that ISO through beta receptor activation and DB-cAMP directly may increase the rate of calcium accumulation by sarcoplasmic reticulum (SR).

Verapamil and Collagen-Induced Platelet Reactions- Evidence for a Role for Intracellular Calcium in Platelet Activation

Calcium is considered to have an essential role in various platelet reactions. Using platelets preincubated with chlortetracycline, a fluorescent divalent cation indicator, and suspended in a calcium free medium, it was shown that collagen-induced intracellular calcium redistribution occurred before the platelet shape change, the release reaction and thromboxane B2 formation. Verapamil, at concentrations which affect intracellular calcium movements, inhibited intracellular calcium redistribution in platelets and the subsequent collagen-induced platelet reactions. Low concentrations of the ionophore A23187 overcame the inhibitory effect of verapamil. These experiments provide evidence that intracellular calcium mobilization is involved in the activation of platelets by collagen. Furthermore, calcium may be released from different cellular pools since platelet secretion, aggregation and thromboxane B2 formation were inhibited at lower concentrations of verapamil than was the platelet shape change.

Nutrient-induced intracellular calcium movement in rat pancreatic B cell.

Several insulin secretagogues increase 45Ca efflux from islets prelabeled with the tracer in the presence of glucose, an effect attributable to stimulation of 40Ca entry into islet cells. When the islets are prelabeled in the absence instead of presence of glucose, the 45Ca becomes more readily releasable, and the secretory response to nutrient or ionic secretagogues is decreased. Such a decrease is more marked in response to glucose than other secretagogues because the prior nutrient deprivation also impairs the metabolism of glucose in islet cells. In the islets prelabeled in the absence of glucose, both glyceraldehyde and 2-ketoisocaprote dramatically stimulated 45Ca efflux. This effect persists in the absence of extracellular Ca2+ or presence of the calmodulin-antagonist trifluoperazine. It may correspond to a nutrient-induced intracellular Ca2+ movement, the existence of which would be unmasked in islets prelabeled in the absence of glucose. This so-far-undetected Ca2+ movement may play a role in the secretory response to nutrient secretagogues.

External calcium and contractility in single giant muscle fibres

Isometric tension and intracellular calcium movements detected using the calcium sensitive photoprotein aequorin have been studied on single striated fibres. The decrease of tension and light response upon deprivation of external calcium would support the notion that calcium ions are involved in the transmission of the electrical potential change from the tubular system to the sarcoplasmic reticulum, by occupying sites on the external (extracellular) surface of tubular system in direct correspondence with the terminal sacs of the sarcoplasmic reticulum.

Immunocytochemical localization of the vitamin D-dependent calcium binding protein in chick duodenum.

The vitamin D-dependent calcium binding protein (CaBP) of chick duodenum has been localized by immunocytochemistry and by radioimmunoassay. Light microscopically, CaBP was seen to be present in the absorptive cells of the villi while in other cell types of the villi and the crypts, including goblet cells and endocrine cells, no CaBP was seen. At the electron microscopic level, CaBP was shown to be localized in the cytosol and the euchromatin of the nucleus but not in membrane-bounded cytoplasmic compartments. Quantitative evaluation of the immunocytochemical protein A-gold label showed that the terminal web and the cytosol of basal cellular regions were most highly labeled while the brush border was weakly labeled. The radioimmunoassay evaluation of intestinal subcellular fractions indicated that 96% of the homogenate CaBP is in the cytosol high-speed supernatant fraction. Collectively, these results support the hypothesis that the vitamin D-dependent intestinal CaBP may play a role in either regulation of intracellular calcium concentration or movement of calcium across the brush border membrane from the gut lumen.

Intracellular calcium: its release from granules during bursting activity in snail neurons.

Pentylenetetrazole induces a bursting activity accompanied by intracellular movement of calcium toward the cell membrane in the snail neuron. The calcium appears to originate from the dense lysosome-like granules in the cytoplasm. Pentylenetetrazole markedly depletes the granules of calcium and alters them ultrastructurally from dense granules to lamella type granules.

Regulation by magnesium of intracellular calcium movement in skinned muscle fibers.

The effect of Mg on Ca movement between the sarcoplasmic reticulum (SR) and myofilament space (MFS) was studied in skinned muscle fibers by using isometric force as an indicator of MFS Ca. In Ca-loaded fibers at 20 degrees C, the large force spike induced by Ca in 1 mM Mg (5 mM ATP) was strongly inhibited in 3 mM Mg, and force development was extremely slow. After a brief Ca stimulus in 1 mM Mg, relaxation in Ca-free solution was significantly faster in 3 mM Mg. These changes were due to altered Ca movements, since the effect of 3 mM Mg on steady force in CaEGTA solutions was small. Changes in Mg alone induced force transients apparently due to altered Ca movement. In relaxed fibers, decreasing the Mg to 0.25 mM caused phasic force development. In contracting fibers in Ca solutions, increasing the Mg caused a large transient relaxation. The effects of increased Mg were antagonized by 0.5 mM Cd, an inhibitor of the SR Ca transport system. The results indicate that active Ca uptake by the SR in situ is stimulated by Mg, and that it can affect local MFS [Ca++] in the presence of a substantial Ca source. These results provide evidence that an increased rate of Ca uptake in 3 mM Mg could account for inhibition of the large force spike associated with Ca-induced Ca release in skinned fibers.

Aequorin and Intracellular Calcium Movements in Skeletal Muscle Fibres (1)

(1975). Aequorin and Intracellular Calcium Movements in Skeletal Muscle Fibres (1) Archives Internationales de Physiologie et de Biochimie: Vol. 83, No. 1, pp. 15-18.

Dual action of chloroquine on frog's skeletal muscle contraction

Studies on the effect of chloroquine on frog's rectus abdominis muscle have been performed. Relatively low concentrations of chloroquine, 5 × 10 −5 and 2 × 10 −4 g/ml inhibited cholinesterase activity and potentiated acetylcholine (ACh)-induced contractions but antagonized carbachol and caffeine contractions, as well as ACh-induced contractions of eserinized muscle. High concentrations (5 × 10 −4 and 2 × 10 −3 g/ml) non-competitively antagonized contractions to ACh, carbachol, caffeine and potassium. It was suggested that the blocking action of chloroquine was due to its local anaesthetic property and interference with intracellular calcium movements.

Intracellular calcium movements in skinned muscle fibres.

1. Force development and (45)Ca movement between the internal membranes and the myofilament space were measured in segments of skinned muscle fibres.2. At 20 degrees C calcium ions in the myofilament space of calcium loaded fibres caused the internal membranes to release a substantial amount of calcium which produced a large force spike. Calcium produced a similar force spike in fibres containing only the physiological level of calcium when the temperature was lowered to about 6 degrees C.3. The force spike at 6 degrees C was inhibited when the magnesium concentration in the bathing solution was increased from 1 to 6 mM. The pattern of force development in preparations transferred directly from oil to a free calcium solution indicated that the normal concentration of Mg(2+) in the myofilament space of intact fibres is of the order of 10(-3)M.4. A possible role for the regenerative calcium release mechanism in the physiological activation of muscle fibres is suggested.

Intracellular calcium movements of frog skeletal muscle during recovery from tetanus.

Radioautographs of (45)Ca-labeled frog skeletal muscles have been prepared using freeze-dry and vapor fixation techniques to avoid displacement of the isotope during the preparation of the radioautographs. (45)Ca has been localized in resting muscles exposed to (45)Ca Ringer's for 5 min or 5 hr and in isotopically labeled muscles recovering from tetanic stimulation at room temperature or at 4 degrees C. In muscles soaked at rest for 5 min (45)Ca was present almost exclusively in the terminal cisternae. In all other muscles there were three sites at which the isotope was concentrated: (a) the terminal cisternae, (b) the intermediate cisternae and the longitudinal tubules, and (c) the A band portion of the myofibrils. The terminal cisternae were labeled more rapidly than the myofibrils, but both exchanges were accelerated by electrical stimulation. The amount of (45)Ca in the longitudinal tubules and the intermediate cisternae decreased with time after a tetanus as the amount in the terminal cisternae increased. It is proposed that electrical stimulation releases calcium from the terminal cisternae and that relaxation occurs from the binding of the released calcium by the longitudinal tubules and the intermediate cisternae. Complete recovery from mechanical activity involves the transport of this bound calcium into the reticulum and its subsequent binding by the terminal cisternae. Resting exchange of calcium occurs primarily between the terminal cisternae and the transverse tubules.