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Biomedical subjects

M J Berridge

Publications and source records attributed to M J Berridge.

17 recordsLinked to original sources

Relationship between phosphatidylinositol synthesis and recovery of 5-hydroxytryptamine-responsive Ca2+ flux in blowfly salivary glands.

Each salivary gland contains about 135 pmol of phosphatidylinositol. In glands prelabelled by incubation for 1 h with [32P]Pi or [3H]inositol there was a subsequent breakdown of 80% of the labelled phosphatidylinositol over a 2 h incubation period with 10 micrometer-5-hydroxytryptamine. However, there was no detectable decrease either in total phosphatidylinositol based on phosphorus analysis by chemical estimation or in the radioactivity of [32P]phosphatidylinositol in salivary glands of flies raised from the larval stage on diets containing[32P]Pi and whose phospholipids were uniformly labelled. These results suggest that the pool of phosphatidylinositol involved with Ca2+ gating is a small fraction of the total phosphatidylinositol content. Furthermore it is this small compartment that is preferentially radioactively labelled during short-term incubations with radioactively labelled precursors. In salivary glands incubated for 2 h with 10 micrometer-5-hydroxytryptamine there was a marked decrease in the flux of 45Ca2+ across the gland. After removal of the hormone, incubation of salivary glands for 1 h in the presence of 2mM-inositol, but not choline or ethanolamine, resulted in a recovery of hormone-responsive 45Ca2+ flux. Quantitative studies revealed that less than 9 pmol of phosphatidylinositol must be formed to fully restoret he 5-hydroxytryptamine-responsive 45Ca2+ flux.

Biological Transport

Relationship between hormonal activation of phosphatidylinositol hydrolysis, fluid secretion and calcium flux in the blowfly salivary gland.

The addition of 5-hydroxytryptamine to the isolated blowfly salivary gland stimulates fluid secretion, transepithelial calcium transport and the breakdown of 32P- or 3H-labelled phosphatidylinositol The breakdown of [32P]phosphatidylcholine and [32P]-phosphatidylethanolamine was not stimulated by 5-hydroxytryptamine. In salivary glands incubated with myo-[2-3H]inositol for 1--3 h, more than 95% of the label retained by the tissue was in the form of phosphatidylinositol. The addition of 5-hydroxytryptamine resulted in an increase in the accumulation of label in intracellular inositol 1:2-cyclic phosphate, inositol 1-phosphate and free inositol along with an increase in the release of [3H]inositol to the medium and saliva. The release of [3H]inositol to the medium served as a sensitive indicator of phosphatidylinositol breakdown. The release of [3H]inositol was not increased by cyclic AMP or the bivalent-cation ionophore A23187 under conditions in which salivary secretion was accelerated. The stimulation of fluid secretion by low concentrations of 5-hydroxytryptamine was potentiated by 3-isobutyl-1-methylxanthine, which had no effect on inositol release. The stimulation of fluid secretion by 5-hydroxytryptamine was greatly reduced in calcium-free buffer, but the breakdown of phosphatidylinositol continued at the same rate in the absence of calcium. These results support the hypothesis that breakdown of phosphatidylinositol by 5-hydroxytryptamine is involved in the gating of calcium.

Animals

Inhibition of phosphatidylinositol synthesis and the inactivation of calcium entry after prolonged exposure of the blowfly salivary gland to 5-hydroxytryptamine.

The incorporation of [32P]Pi into all salivary-gland phospholipids except phosphatidic acid was inhibited by 5-hydroxytryptamine. The accumulation of [32P]Pi into phosphatidic acid was actually enhanced by 5-hydroxytryptamine. There was an inhibition of labelled inositol incorporation into phosphatidylinositol by 5-hydroxytryptamine, which seems to be mediated by calcium because it was mimicked by the ionophore A23187, but was prevented if glands were stimulated with 5-hydroxytryptamine in the absence of external calcium. Inhibition of synthesis together with stimulation of breakdown will decrease the concentration of phosphatidylinositol, which could account for the inactivation of calcium transport observed at high 5-hydroxytryptamine concentrations. When salivary glands were stimulated with 1 micrometer-5-hydroxytryptamine, there was a rapid increase in the transfer of 45Ca2+ from the medium into the saliva, but with time this transport declined to a low value. If the glands were washed free of 5-hydroxytryptamine and incubated in the presence of 2mM-inositol for 1 h, the increase in calcium transport caused by 5-hydroxytryptamine was restored. There was little recovery in the absence of inositol. If glands were stimulated with 5-hydroxytryptamine in the absence of external calcium, a condition which prevents the inhibition of phosphatidylinositol synthesis, calcium transport in response to 5-hydroxytryptamine was greater than in glands preincubated with 5-hydroxytryptamine in the presence of calcium. The inactivation of calcium transport may result from a decrease in phosphatidylinositol concentration. These results support the hypothesis that the hydrolysis of phosphatidylinositol plays some role in either the opening or closing of calcium 'gates'.

Animals

A comparative survey of the function, mechanism and control of cellular oscillators.

This review attempts to survey in a uniform manner the available evidence concerning the generation and behaviour of several well-investigated cellular oscillators. Members of two broad classifications are contrasted: (i) cytoplasmic oscillations, where the periodic phenomena is generated by an instability pathway and (ii) membrane oscillators in which a membrane potential rhythm is generated at the membrane. Interactions between the cytoplasmic and membrane compartments are considered and the effects of these interactions on oscillatory behaviour is discussed. Because of their biological importance and the greater body of experimental results, particular attention is directed to a study of membrane potential oscillations. These systems can be approximately classified in two groups: (i) systems in which a periodic potential results from oscillatory changes in permeability and (ii) systems in which potential oscillations result from the periodic activity of an electrogenic pump. The examples considered include the glycolytic oscillator, oscillations in vein contraction in the slime mould Physarum polycephalum, rhythmic aggregation in Dictyostelium discoideum, neural oscillators, the periodic potential in Purkinje fibres and the sino-atrial node and rhythmic behaviour in smooth muscle. Questions considered include the generation of periodic activity, the modulation of the oscillation by drugs and other metabolic and membrane effectors and the question of the functional role of these oscillations.

Adrenal Cortex

Ion-selective electrode studies on the effects of 5-hydroxytryptamine on the intracellular level of potassium in an insect salivary gland.

Potassium-sensitive microelectrodes were used to measure the intracellular level of potassium in Calliphora salivary glands. During stimulation with 1 X 10(-8) M 5-HT, the potassium concentration increased from a resting level of 133.2 to 139.0 mM. When the external concentration of potassium was reduced below 2mM there was a rapid loss of potassium which could be partially reversed by 5-HT. During 5-HT stimulation, the addition of ouabain had no effect on a number of parameters including the rate of fluid secretion, the ionic composition of the saliva and the intracellular level of potassium. The possibility that potassium enters the cells passively is discussed.

Animals

Studies on the mechanism of fluid secretion by isolated salivary glands of Calliphora.

1. Potassium is the major cation in the secretion of the salivary glands of Calliphora and is necessary for full secretory rates. 2. Other ions (rubidium and sodium) can support secretion in the absence of potassium. 39. During stimulation with 5-HT a Nernst plot of the basal membrane potential has a slope of 53 mV for a tenfold change in external potassium concentration and the slope at rest deviates from this over the range I-20 mM external potassium. 4. Hyperpolarization of the basal membrane by 5-HT is abolished if the chloride in the bathing medium is replaced by isethionate. 5. The diuretic agent amiloride inhibits fluid secretion by a mechanism which may include a reduction in calcium entry in addition to its recognized effect on sodium permeability. 6. A model is proposed in which fluid secretion is driven by the active transport of potassium across the apical membrane with chloride following passively.

Amiloride

A freeze-fracture study of adult Calliphora salivary glands.

A freeze-fracture study of adult Calliphora salivary glands has revealed a high density (approx. 4500/mu2) of intramembraneous particles (80-110 A) in both the apical and basal membranes. Most of the particles were associated with the A face. The density of the stalked surface particles which coat the cytoplasmic surface of the apical membrane. The possible significance of these particles in ion transport is discussed.

Animals

Membrane permeability changes during stimulation of isolated salivary glands of Calliphora by 5-hydroxytryptamine.

1. The membrane resistance of isolated salivary glands was found to decrease in response to 5-HT. The change in resistance was calcium-dependent. 2. The resistance change of the apical membrane was found to be much greater than the change in resistance of the basal membrane. 3. Potential responses under current-clamped conditions showed that one part of the biphasic response to 5-HT (attributed to an increase in chloride permeability) could be reversed and the other part (attributed to an increase in a potassium pump) could not. 4. These observations have been incorporated into a model which, on evaluation, predicts all of the observed potential changes during the action of 5-HT. It suggests that the potential responses reflect changes in the internal chloride concentration produced by the calcium-dependent increases in chloride permeability.

Animals

Stimulus-secretion coupling in an insect salivary gland: cell activation by elevated potassium concentrations.

1. Fluid secretion by isolated salivary glands was stimulated by elevating the external potassium concentration. 2. The stimulatory effect of potassium was dependent on external calcium and was potentiated by a subthreshold dose of 5-hydroxytryptamine (5-HT). 3. During the action of 120 mM potassium there was a large calciumdependent decrease in transepithelial resistance similar to that produced with 5-HT at normal potassium concentrations. 4. These results on Calliphora salivary glands are compared with other cases where cells are activated by high potassium. In most cases, the effect of high potassium is dependent upon calcium, suggesting that the latter plays a primary role in cell activation.

5-Hydroxytryptophan

Control of cell division: a unifying hypothesis.

A constant feature of the initiation of cell division in a number of different cells is a rise in the intracellular level of calcium. The importance of cyclic nucleotides may depend on the way they interact with calcium. Cyclic AMP is apparently not an essential regulator of cell division but through its ability to modulate the intracellular level of calcium this cyclic nucleotide can exert profound effects on cell growth. In some systems (liver and salivary glands) cyclic AMP seems to augment the calcium signal whereas in others (lymphocytes and fibroblasts) it opposes calcium and can thus inhibit cell division. A rise in the level of calcium may be responsible for the parallel increase in cyclic GMP level which is usually associated with the stimulus to divide. An appealing feature of this calcium hypothesis is that it can account for the growth characteristics revealed by fibroblasts in tissue culture or embryonic cells during development. In both cases there is an initial phase of exponential growth during which I have proposed that the high level of calcium at mitosis persists into early G1 to provide the signal for the next division. In order to account for the sudden cessation of cell division at confluency, or at a specific stage during development, it is necessary to postulate that there is something different about the final mitosis which sets it apart from earlier mitoses. It is proposed that as the cells leave the last mitosis the level of calcium falls much more rapidly than it did during preceeding mitoses perhaps as a result of a more rapid rise in the level of cyclic AMP. This rapid rise in cyclic AMP level may have a dual function. Not only will it lower the level of calcium thus preventing further division, but it may also stimulate differentiation. Many of the embryonic cells which differentiate into specialized cells (lymphocytes, liver, salivary gland) retain the ability to divide if provided with appropriate stimuli. Although the nature of these stimuli vary considerably, they all seem to act by elevating the intracellular level of calcium.

Animals