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Effects of age, diet and obesity on insulin secretion from isolated perfused rat pancreas: Response to glucose, arginine and glucagon-like peptide 1 (7-37)
Full text linkThe insulin secretory responses to glucose, arginine and glucagon-like peptide (GLP)-1-(7- 37 have been evaluated from the isolated perfused pancreas of rats with either acquired or genetic obesity, ie, a) fed ad libitum 14-mo old Sprague-Dawley rats as compared to age-matched animals subjected to two types of dietary restriction (every-other-day feeding, EOD, and 40% restriction? 40% DR), and b) 2.5-mo old genetically obese fa/fa rats as compared to the lean counterpart, In mature fed ad libitum rats, the glucose-stimulated insulin release from the perfused pancreas was increased 5-fold by addition of 0.1 nM GLP-1 (7-37), a subsequent challenge with high glucose resulted in an improvement of the first phase of insulin release, In 40% DR rats, a similar pattern of secretion was observed, with the difference of a lower response to arginine than in fed ad libitum animals, In EOD rats, the overall secretory performance of the perfused pancreas was approximately 50% lower than in the fed ad libitum group but probably adequate to the reduced weight of the animals, In genetically obese young rats, both the response to GLP-1 (7-37) anti the total insulin secretion were higher than in the lean controls. Interestingly, the maximal insulin outputs from the perfused pancreases were observed in both the groups of overweight animals, In conclusion no impairment in the secretory responsiveness of beta-cells occurs in obese animals, Conversely, at least within the age limits of the present study, the endocrine pancreas develops a compensatory ability to match the augmented insulin demand due to the over-weight. In the light of the observed great sensitivity of the isolated perfused pancreas to GLP-1 (7-37), changes in the responsiveness of beta-cells to incretins might be involved in the modulation of the endocrine pancreatic function of obese rats
STUDIES IN VIVO AND IN VITRO ON CHEMICALLY-INDUCED PRIMARY ISLET CELL TUMORS AND NON-TUMOR ENDOCRINE PANCREATIC TISSUE
Full text linkGLUCOSE STIMULATES INSULIN RELEASE FROM EXPERIMENTALLY INDUCED RAT INSULINOMAS IN VIVO AND IN VITRO
No full textTumor promoter phorbol myristate acetate inhibits Ca2+ influx through voltage-gated Ca2+ channels in two secretory cell lines, PC12 and RINm5F
No full textAbstract
Protein kinase C is known to be involved both in initiation and termination of cellular responses due to phosphoinositide breakdown. Here we report that in PC12 cells (a line of neurosecretory cells derived from a rat pheochromocytoma), pretreatment with nanomolar concentrations of phorbol myristate acetate, PMA, which is believed to specifically activate protein kinase C, inhibits the cytosolic-free Ca2+ concentration rise induced by depolarizing agents. In contrast, plasma membrane potential and 45Ca efflux from preloaded cells were unaffected by PMA pretreatment. Inhibition by PMA and diacylglycerol of the cytosolic-free Ca2+ concentration rise induced by depolarization was observed also in another cell line, the insulin secreting line RINm5F. These results raise the possibility that the voltage-gated Ca2+ channel is under inhibitory control by protein kinase C
Stimulated Ca2+ influx raises mitochondrial free Ca2+ to supramicromolar levels in a pancreatic beta-cell line. Possible role in glucose and agonist-induced insulin secretion.
No full textThe effects of stimulated Ca2+ influx on cytosolic ([Ca2+]c) or intramitochondrial free Ca2+ ([Ca2+]m) were examined in the new pancreatic beta-cell line, INS-1. [Ca2+]c was monitored by video imaging of single fura-2-loaded INS-1 cells, or in populations of cells transfected with non-targeted (cytosolic) aequorin. [Ca2+]m was measured after transfection with aequorin targeted to the mitochondria by fusion of the gene in frame with the signal peptide of cytochrome c oxidase subunit VIII. Two physiological stimuli of native beta-cells, glucose and ATP, raised [Ca2+]c in INS-1 cells largely by stimulating Ca2+ influx. Thus, glucose (20 mM) induced repetitive transient increases in [Ca2+]c (0.42 min-1, mean amplitude 229 nM above 102 nM basal). These transients were largely due to periodic stimulation of Ca2+ influx through voltage-sensitive Ca2+ channels, since they could be rapidly and reversibly blocked by chelation of external Ca2+, by addition of the hyperpolarizing agent diazoxide, or with the Ca2+ channel blocker SR 7037. ATP, by contrast, caused single transient [Ca2+]c increases, to about 300 nM above basal levels, which could be inhibited by > 90% upon external Ca2+ chelation. Challenge of aequorin-transfected cells with ATP increased [Ca2+]m to 4 microM or above, an effect blocked by EGTA. Furthermore, plasma membrane depolarization with high K+, used as a glucose surrogate to mimic, in a synchronized fashion, the influx-induced Ca2+ transients observed at the single-cell level, also increased [Ca2+]m to > 4 microM. Similar increases in [Ca2+]m were also measured in other aequorin-transfected insulin-secreting cells, RINm5F, during mobilization of internal Ca2+ with carbachol. In contrast, glucose-induced changes in [Ca2+]m were below the level of detection in INS-1 cell populations, consistent with the asynchrony of the [Ca2+]c transients induced by this nutrient at the single-cell level, and the consequent small average [Ca2+]c rise. These data are in line with the view that stimulated Ca2+ influx into excitable cells raises [Ca2+]m as efficiently as internal Ca2+ mobilization in nonexcitable cells. In the case of INS-1 and pancreatic beta-cells, this may be important both to enhance oxidative metabolism, hence fueling the secretory process, and also to maintain the production of metabolic signaling molecules
Voltage dependent calcium channels in adrenal glomerulosa cells and in insulin producing cells.
No full textMatrix alkalinization: a novel mitochondrial signal for sustained pancreatic beta-cell activation.
No full textSelective actions of mitochondrial fission/fusion genes on metabolism-secretion coupling in insulin releasing cells
No full textMitochondria form filamentous networks that undergo continuous fission/fusion. In the pancreatic beta-cells, mitochondria are essential for the transduction of signals linking nutrient metabolism to insulin granule exocytosis. Here we have studied mitochondrial networks in the insulinoma cell line INS-1E, primary rat and human beta-cells. We have further investigated the impact of mitochondrial fission/fusion on metabolism-secretion coupling in INS-1E cells. Overexpression of hFis1 caused dramatic mitochondrial fragmentation, whereas Mfn1 evoked hyperfusion and the aggregation of mitochondria. Cells overexpressing hFis1 or Mfn1 showed reduced mitochondrial volume, lowered cellular ATP levels, and as a consequence, impaired glucose-stimulated insulin secretion. Decreased mitochondrial ATP generation was partially compensated for by enhanced glycolysis as indicated by increased lactate production in these cells. Dominant-negative Mfn1 elicited mitochondrial shortening and fragmentation of INS-1E cell mitochondria, similar to hFis1. However, the mitochondrial volume, cytosolic ATP levels, and glucose-stimulated insulin secretion were little affected. We conclude that mitochondrial fragmentation per se does not impair metabolism-secretion coupling. Through their impact on mitochondrial bioenergetics and distribution, hFis1 and Mfn1 activities influence mitochondrial signal generation thereby insulin exocytosis
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