Dietary virgin olive oil enhances secretagogue-evoked calcium signaling in rat pancreatic acinar cells.

OBJECTIVE: We evaluated the long-term effects of a fat-enriched diet (virgin olive oil) on calcium mobilization and amylase secretion induced by cholecystokinin-octapeptide (CCK-8) in rat pancreatic acinar cells. Olive oil is a major component of the Mediterranean diet, and its role in human health is actively being debated. METHODS: Weaning male Wistar rats (21 d old) were assigned to one of two experimental groups and fed for 8 wk with a commercial chow (control group) or an experimental diet (olive group) containing 100 g/kg of virgin olive oil as dietary fat. Intracellular free calcium [Ca(2+)](i) levels were determined by loading the pancreatic cells with the fluorescent ratio-metric calcium indicator Fura-2 on an inverted fluorescent microscope. For measurement of amylase secretion, cells were incubated with the appropriate secretagogue for 30 min, and amylase activities in the supernatant were determined by the Phadebas blue starch method. Analysis of variance was used to test differences between groups. RESULTS: Compared with the control group, the CCK-8-induced increase in [Ca(2+)](i) occurred in cells from rats in the olive group (P < 0.05). This stimulatory effect of dietary virgin olive oil was observed in calcium oscillations and large [Ca(2+)](i) transients induced by low (20 pM/L) and high (10 nM/L) concentrations of CCK-8, respectively. In addition to the effects of dietary virgin olive oil on calcium mobilization, it increased (P < 0.05) amylase secretion in response to CCK-8. Olive oil treatment did not significantly alter resting [Ca(2+)](i) or amylase release values compared with the control group. Similar results were obtained when pancreatic acinar cells were stimulated with a high concentration of acetylcholine (10 microM/L). CONCLUSION: The present results demonstrate that a diet supplemented with virgin olive oil can modify pancreatic cell function as assessed by [Ca(2+)](i) mobilization and amylase release evoked by secretagogues in rat pancreatic acinar cells. A role for fatty acids in calcium signaling is suggested.

Effects of ageing on morphology, amylase release, cytosolic Ca2+ signals and acyl lipids in isolated rat parotid gland tissue.

Xerostomia (oral dryness sensation) is due to dryness of the oral cavity and it is more prevalent in the elderly. This study investigated the effect of ageing on parotid gland structure and function of control (2-6 months) and aged (12, 16-18 and 22-24 months) rats employing light microscopic, colorimetric, gas chromatographic and microspectrofluorimetric methods to investigate the morphological changes of the parotid glands, amylase release, endogenous lipid distribution and cytosolic free calcium levels, respectively. When compared to controls, age-related changes were apparent in glands obtained from rats aged 16-18 and 22-24 months, which included reduced acinar cell distribution, enlarged parotid ducts with fatty and connective tissue and mast cell infiltrations. Parotid acini from 12, 16-18 and 22-24-month-old glands showed significant (p < 0.05) age-related decreases in amylase release, compared to controls when challenged with acetylcholine (ACh). No change in basal calcium signals was observed in parotid acini from 2-6 to 16-18-month-old-animals. However, stimulation of 16-18-month-old parotid acini with 10(-5)M ACh resulted in a significant (p < 0.001) decrease in both peak and plateau phases of the cytosolic Ca2+ signal when compared to control. Gas chromatography of de novo and essential acyl lipids revealed no changes in the amount of either acyl lipid group in glands obtained from 2-6 to 22-24-month-old animals. Lipid analysis of phospholipid associated acyl chains showed a higher relative proportion of linoleic acid in older glands. The results reveal that ageing is associated with marked and distinct morphological changes including infiltrations of lipids and mast cells of the parotid gland and decreases in amylase release and cytosolic Ca2+ signals.

Evidence for secretion-like coupling involving pp60src in the activation and maintenance of store-mediated Ca2+ entry in mouse pancreatic acinar cells.

Store-mediated Ca2+ entry (SMCE) is one of the main pathways for Ca2+ influx in non-excitable cells. Recent studies favour a secretion-like coupling mechanism to explain SMCE, where Ca2+ entry is mediated by an interaction of the endoplasmic reticulum (ER) with the plasma membrane (PM) and is modulated by the actin cytoskeleton. To explore this possibility further we have now investigated the role of the actin cytoskeleton in the activation and maintenance of SMCE in pancreatic acinar cells, a more specialized secretory cell type which might be an ideal cellular model to investigate further the properties of the secretion-like coupling model. In these cells, the cytoskeletal disrupters cytochalasin D and latrunculin A inhibited both the activation and maintenance of SMCE. In addition, stabilization of a cortical actin barrier by jasplakinolide prevented the activation, but not the maintenance, of SMCE, suggesting that, as for secretion, the actin cytoskeleton plays a double role in SMCE as a negative modulator of the interaction between the ER and PM, but is also required for this mechanism, since the cytoskeleton disrupters impaired Ca2+ entry. Finally, depletion of the intracellular Ca2+ stores induces cytoskeletal association and activation of pp60(src), which is independent on Ca2+ entry. pp60(src) activation requires the integrity of the actin cytoskeleton and participates in the initial phase of the activation of SMCE in pancreatic acinar cells.