scholarly journals Overexpression of the malate–aspartate NADH shuttle member Aralar1 in the clonal β-cell line BRIN-BD11 enhances amino-acid-stimulated insulin secretion and cell metabolism

2009 ◽  
Vol 117 (9) ◽  
pp. 321-330 ◽  
Author(s):  
Katrin Bender ◽  
Pierre Maechler ◽  
Neville H. McClenaghan ◽  
Peter R. Flatt ◽  
Philip Newsholme
Keyword(s):  
Insulin Secretion ◽  
Amino Acid ◽  
Cell Line ◽  
Recombinant Adenovirus ◽  
Cell Metabolism ◽  
Glutamate Release ◽  
Lactate Production ◽  
Control Site ◽  
Β Cell ◽  
Insulin Secreting Cells

In the present study, we have investigated the effects of the transduction with recombinant adenovirus AdCA-Aralar1 (aspartate–glutamate carrier 1) on the metabolism, function and secretory properties of the glucose- and amino-acid-responsive clonal insulin-secreting cell line BRIN-BD11. Aralar1 overexpression increased long-term (24 h) and acute (20 min) glucose- and amino-acid-stimulated insulin secretion, cellular glucose metabolism, L-alanine and L-glutamine consumption, cellular ATP and glutamate concentrations, and stimulated glutamate release. However, cellular triacylglycerol and glycogen contents were decreased as was lactate production. These findings indicate that increased malate–aspartate shuttle activity positively shifted β-cell metabolism, thereby increasing glycolysis capacity, stimulus–secretion coupling and, ultimately, enhancing insulin secretion. We conclude that Aralar1 is a key metabolic control site in insulin-secreting cells.

scholarly journals Pro-inflammatory cytokines increase glucose, alanine and triacylglycerol utilization but inhibit insulin secretion in a clonal pancreatic β-cell line

2007 ◽  
Vol 195 (1) ◽  
pp. 113-123 ◽  
Author(s):  
Aoife Kiely ◽  
Neville H McClenaghan ◽  
Peter R Flatt ◽  
Philip Newsholme
Keyword(s):  
Insulin Secretion ◽  
Inflammatory Cytokines ◽  
Prolonged Exposure ◽  
Cell Metabolism ◽  
Glutamate Release ◽  
Β Cell ◽  
Catabolic State ◽  
Stimulus Secretion Coupling ◽  
Factor Α ◽  
Pro Inflammatory Cytokines

We have investigated the effects of prolonged exposure (24 h) to pro-inflammatory cytokines on β-cell metabolism and insulin secretion using clonal BRIN-BD11 β cells. Addition of IL-1β, tumour necrosis factor-α and IFN-γ (at concentrations that did not induce apoptosis) inhibited chronic (24 h) and acute stimulated levels of insulin release (by 59 and 93% respectively), increased cellular glucose and alanine consumption, and also elevated lactate and glutamate release. However, ATP levels and cellular triacylglycerol were decreased while glutathione was increased. We conclude that sub-lethal concentrations of pro-inflammatory cytokines appear to shift β-cell metabolism away from a key role in energy generation and stimulus–secretion coupling and towards a catabolic state which may be related to cell defence.

scholarly journals l-Arginine is essential for pancreatic β-cell functional integrity, metabolism and defense from inflammatory challenge

2011 ◽  
Vol 211 (1) ◽  
pp. 87-97 ◽  
Author(s):  
Mauricio S Krause ◽  
Neville H McClenaghan ◽  
Peter R Flatt ◽  
Paulo I Homem de Bittencourt ◽  
Colin Murphy ◽  
...  
Keyword(s):  
Insulin Secretion ◽  
Amino Acid ◽  
Proinflammatory Cytokines ◽  
Glutamate Release ◽  
Glucose Consumption ◽  
Interferon Γ ◽  
Intermediary Metabolism ◽  
Β Cells ◽  
Β Cell ◽  
Functional Integrity

In this work, our aim was to determine whetherl-arginine (a known insulinotropic amino acid) can promote a shift of β-cell intermediary metabolism favoring glutathione (GSH) and glutathione disulfide (GSSG) antioxidant responses, stimulus–secretion coupling and functional integrity. Clonal BRIN-BD11 β-cells and mouse islets were cultured for 24 h at variousl-arginine concentrations (0–1.15 mmol/l) in the absence or presence of a proinflammatory cytokine cocktail (interleukin 1β, tumour necrosis factor α and interferon γ). Cells were assessed for viability, insulin secretion, GSH, GSSG, glutamate, nitric oxide (NO), superoxide, urea, lactate and for the consumption of glucose and glutamine. Protein levels of NO synthase-2, AMP-activated protein kinase (AMPK) and the heat shock protein 72 (HSP72) were also evaluated. We found thatl-arginine at 1.15 mmol/l attenuated the loss of β-cell viability observed in the presence of proinflammatory cytokines.l-Arginine increased total cellular GSH and glutamate levels but reduced the GSSG/GSH ratio and glutamate release. The amino acid stimulated glucose consumption in the presence of cytokines while also stimulating AMPK phosphorylation and HSP72 expression. Proinflammatory cytokines reduced, by at least 50%, chronic (24 h) insulin secretion, an effect partially attenuated byl-arginine. Acute insulin secretion was robustly stimulated byl-arginine but this effect was abolished in the presence of cytokines. We conclude thatl-arginine can stimulate β-cell insulin secretion, antioxidant and protective responses, enabling increased functional integrity of β-cells and islets in the presence of proinflammatory cytokines. Glucose consumption and intermediary metabolism were increased byl-arginine. These results highlight the importance ofl-arginine availability for β-cells during inflammatory challenge.

scholarly journals Amino acid taste receptor regulates insulin secretion in pancreatic β-cell line MIN6 cells

2011 ◽  
Vol 16 (5) ◽  
pp. 608-616 ◽  
Author(s):  
Manami Oya ◽  
Hideyuki Suzuki ◽  
Yuichiro Watanabe ◽  
Moritoshi Sato ◽  
Takashi Tsuboi
Keyword(s):  
Insulin Secretion ◽  
Amino Acid ◽  
Cell Line ◽  
Taste Receptor ◽  
Pancreatic Β Cell ◽  
Β Cell ◽  
Min6 Cells

Volume-sensitive amino acid efflux from a pancreatic β-cell line

2000 ◽  
Vol 162 (1-2) ◽  
pp. 201-208 ◽  
Author(s):  
A.C.G. Grant ◽  
J. Thomson ◽  
V.A. Zammit ◽  
D.B. Shennan
Keyword(s):  
Amino Acid ◽  
Cell Line ◽  
Pancreatic Β Cell ◽  
Β Cell ◽  
Amino Acid Efflux

Time-resolved metabolomics analysis of β-cells implicates the pentose phosphate pathway in the control of insulin release

2013 ◽  
Vol 450 (3) ◽  
pp. 595-605 ◽  
Author(s):  
Peter Spégel ◽  
Vladimir V. Sharoyko ◽  
Isabel Goehring ◽  
Anders P. H. Danielsson ◽  
Siri Malmgren ◽  
...  
Keyword(s):  
Insulin Secretion ◽  
Pentose Phosphate Pathway ◽  
Metabolic Response ◽  
Cell Metabolism ◽  
Pentose Phosphate ◽  
Β Cells ◽  
Β Cell ◽  
Time Resolved ◽  
Atp Production ◽  
Rat Islets

Insulin secretion is coupled with changes in β-cell metabolism. To define this process, 195 putative metabolites, mitochondrial respiration, NADP+, NADPH and insulin secretion were measured within 15 min of stimulation of clonal INS-1 832/13 β-cells with glucose. Rapid responses in the major metabolic pathways of glucose occurred, involving several previously suggested metabolic coupling factors. The complexity of metabolite changes observed disagreed with the concept of one single metabolite controlling insulin secretion. The complex alterations in metabolite levels suggest that a coupling signal should reflect large parts of the β-cell metabolic response. This was fulfilled by the NADPH/NADP+ ratio, which was elevated (8-fold; P<0.01) at 6 min after glucose stimulation. The NADPH/NADP+ ratio paralleled an increase in ribose 5-phosphate (>2.5-fold; P<0.001). Inhibition of the pentose phosphate pathway by trans-dehydroepiandrosterone (DHEA) suppressed ribose 5-phosphate levels and production of reduced glutathione, as well as insulin secretion in INS-1 832/13 β-cells and rat islets without affecting ATP production. Metabolite profiling of rat islets confirmed the glucose-induced rise in ribose 5-phosphate, which was prevented by DHEA. These findings implicate the pentose phosphate pathway, and support a role for NADPH and glutathione, in β-cell stimulus-secretion coupling.

scholarly journals Investigation of Ultrasound-Mediated Intracellular Ca2+ Oscillations in HIT-T15 Pancreatic β-Cell Line

Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1129
Author(s):  
Chi Woo Yoon ◽  
Nan Sook Lee ◽  
Kweon Mo Koo ◽  
Sunho Moon ◽  
Kyosuk Goo ◽  
...  
Keyword(s):  
Insulin Secretion ◽  
Cell Line ◽  
Purinergic Signaling ◽  
Underlying Mechanism ◽  
Resting Cells ◽  
Pancreatic Β Cell ◽  
Pulsed Ultrasound ◽  
Β Cell ◽  
Secretion Pathways ◽  
Glucose Stimulated Insulin Secretion

In glucose-stimulated insulin secretion (GSIS) of pancreatic β-cells, the rise of free cytosolic Ca2+ concentration through voltage-gated calcium channels (VGCCs) triggers the exocytosis of insulin-containing granules. Recently, mechanically induced insulin secretion pathways were also reported, which utilize free cytosolic Ca2+ ions as a direct regulator of exocytosis. In this study, we aimed to investigate intracellular Ca2+ responses on the HIT-T15 pancreatic β-cell line upon low-intensity pulsed ultrasound (LIPUS) stimulation and found that ultrasound induces two distinct types of intracellular Ca2+ oscillation, fast-irregular and slow-periodic, from otherwise resting cells. Both Ca2+ patterns depend on the purinergic signaling activated by the rise of extracellular ATP or ADP concentration upon ultrasound stimulation, which facilitates the release through mechanosensitive hemichannels on the plasma membrane. Further study demonstrated that two subtypes of purinergic receptors, P2X and P2Y, are working in a competitive manner depending on the level of glucose in the cell media. The findings can serve as an essential groundwork providing an underlying mechanism for the development of a new therapeutic approach for diabetic conditions with further validation.

scholarly journals The biphasic effect of wortmannin on insulin secretion from pancreatic β cell line MIN6.

1994 ◽  
Vol 64 ◽  
pp. 112
Author(s):  
Takashi Sakurai ◽  
Shinji Hagiwara ◽  
Fumi Tashiro ◽  
Yoshiaki Hashimoto ◽  
Hiroshi Yano ◽  
...  
Keyword(s):  
Insulin Secretion ◽  
Cell Line ◽  
Pancreatic Β Cell ◽  
Β Cell ◽  
Biphasic Effect

Modulation of ATP-sensitive K+ channels by internal acidification in insulin-secreting cells

1994 ◽  
Vol 267 (4) ◽  
pp. C1036-C1044 ◽  
Author(s):  
Z. Fan ◽  
Y. Tokuyama ◽  
J. C. Makielski
Keyword(s):  
Insulin Secretion ◽  
Cell Line ◽  
Single Channel ◽  
Inhibitory Effect ◽  
Channel Activity ◽  
Patch Clamp Technique ◽  
Open Probability ◽  
Activation Effect ◽  
Insulin Secreting Cells ◽  
Single Channel Currents

The effect of intracellular acidification (low pHi) on open probability of the ATP-sensitive K+ (KATP) channel was examined in insulin-secretion cells using an inside-out configuration of the patch-clamp technique. In an insulin-secreting cell line beta-TC3, KATP single-channel currents (IKATP) were readily recorded in the absence of internal ATP. ATP (50 microM and 0.5 mM) dramatically decreased the channel activity. A step decrease of intracellular pH (pHi) from 7.4 to 6.7 or 6.3 in the presence of ATP gradually increased the channel activity. In addition, low pHi in the presence of ATP could partially restore channel activity lost in a process called "rundown." Kinetic analysis revealed a change in channel gating at low pHi with ATP. The bursting durations of IKATP at pHi 6.3 in the presence of ATP were significantly longer than those at pHi 7.4 in the absence of ATP. These results suggest that the increased channel activity at low pHi might have resulted from a mechanism involving an alteration of channel conformation. We also observed an inhibitory effect of low pHi on channel activity. However, the inhibitory effect was much more apparent at pHi 5.7 and was only partially reversible. The activation effect of low pHi on IKATP in the presence of ATP was also observed in acutely isolated rat islet cells and in another insulin-secretion cell line RINm5F, although the effect was weaker and was variable among experiments. We conclude that, as in frog skeletal muscle and cardiac muscle, an increase in channel activity at low pHi is one of the mechanisms underlying proton modulation of IKATP in insulin-secreting cells.

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