1. Volume 138, Issue 5, Pages 1976-1987.e5 (May 2010)
Dynamic Changes in Cytosolic and Mitochondrial ATP Levels in Pancreatic Acinar Cells 
Svetlana G. Voronina, Stephanie L. Barrow, Alec W.M. Simpson, Oleg V. Gerasimenko, Gabriela da Silva Xavier, Guy A. Rutter, Ole H. Petersen, Alexei V. Tepikin 
Gastroenterology 
Volume 138, Issue 5, Pages e5 (May 2010)
DOI: /j.gastro
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2. Figure 1
Effects of inhibitors of glycolysis and oxidative phosphorylation on the cytosolic adenosine triphosphate (ATP) level of pancreatic acinar cells. (A) Images show bioluminescence of pancreatic acinar cells. The images were acquired at time points indicated on the trace. Scale bar = 100 μm. Experiments were conducted with pyruvate (2 mM) and glucose (10 mM) present in the extracellular solution. (B) Images were acquired at time points indicated on the trace. Scale bar = 100 μm. Experiment was conducted with pyruvate (2 mM) present in the extracellular solution. Glucose was removed from the extracellular solution just before the beginning of the recording. (C) Experiment was conducted with pyruvate (2 mM) and glucose (10 mM) present in the extracellular solution. (D) Experiment was conducted with pyruvate (2 mM) present in the extracellular solution. Glucose was removed from the extracellular solution just before the beginning of the recording.
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3. Figure 2
Effects of cholecystokinin-8 (CCK) on cytosolic adenosine triphosphate (ATP) level. (A) Increase of bioluminescence in cells stimulated with 10 nM CCK. The left panel shows transmitted light image of isolated pancreatic acinar cells and clusters of pancreatic acinar cells. Central and right panels show images of bioluminescence recorded at time points indicated on the trace. Scale bar = 100 μm. (B) Following inhibition of mitochondrial ATP production with R/Olig, a high concentration of CCK triggers a further decrease in ATP level. Images of bioluminescence were recorded at time points indicated on the trace. Scale bar = 100 μm. (C) Increase of bioluminescence induced by a low (20 pM) concentration of CCK. (D) Following inhibition of mitochondrial ATP production with R/Olig a low concentration of CCK does not produce a further decrease in bioluminescence.
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4. Figure 3
Cholecystokinin-8 (CCK) increases the rate of cytosolic adenosine triphosphate (ATP) consumption. (A) Bioluminescence changes induced by CCK and subsequent inhibition of both oxidative phosphorylation and glycolysis. (B) Bioluminescence changes induced by inhibition of oxidative phosphorylation and glycolysis. (C) Decline of bioluminescence in experiments illustrated in (A) and (B) was approximated by an exponential function. The averaged time constants for the decline in the presence and absence of CCK are shown as bar graphs (±standard errors, number of experiments indicated on the bars).
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5. Figure 4
Thapsigargin (Tg) and ionomycin (Ion) induce an increase in cytosolic adenosine triphosphate (ATP). (A) Rise of bioluminescence induced by Tg. (B) Rise of bioluminescence induced by Ion.
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6. Figure 5
Effects of bombesin, acetylcholine (ACh), and caerulein on cytosolic adenosine triphosphate (ATP). (A) and (B) show effects of bombesin on cytosolic bioluminescence in cells with intact and inhibited (by rotenone [R]/oligomycin [Olig]) mitochondrial ATP production, respectively. (C) and (D) show effects of ACh on cytosolic bioluminescence in cells with intact and inhibited (by R/Olig) mitochondrial ATP production, respectively. (E) and (F) show effects of caerulein on cytosolic bioluminescence in cells with intact and inhibited (by R/Olig) mitochondrial ATP production, respectively.
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7. Figure 6
Effects of taurolithocholic acid 3-sulfate (TLC-S), palmitoleic acid ethyl ester (POAEE) and palmitoleic acid (POA) on cytosolic adenosine triphosphate (ATP). (A) and (B) show effects of TLC-S on cytosolic bioluminescence in cells with intact and inhibited (by rotenone [R]/oligomycin [Olig]) mitochondrial ATP production, respectively. (C) and (D) show effects of POAEE on cytosolic bioluminescence in cells with intact and inhibited (by R/Olig) mitochondrial ATP production, respectively. (E) Illustrates a decrease of bioluminescence induced by POA. (F) Following inhibition of mitochondrial ATP production by R/Olig, we have not observed further changes in bioluminescence upon application of POA.
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8. Figure 7
Effect of cholecystokinin-8 (CCK), palmitoleic acid (POA), palmitoleic acid ethyl ester (POAEE), and taurolithocholic acid 3-sulfate (TLC-S) on mitochondrial adenosine triphosphate (ATP) level in pancreatic acinar cells. Bioluminescence was recorded from cells transfected with mitochondrial-targeted luciferase (mLuc). (A) Effect of different concentrations of CCK on the bioluminescence. (Ai) Shows changes of bioluminescence induced by 20 pM CCK. (Aii) depicts changes of bioluminescence induced by 10 nM CCK. Note biphasic response of bioluminescence. At the end of each experiment, a combination of mitochondrial inhibitors rotenone (R)/oligomycin (Olig) was applied, note the abrupt and strong decrease in bioluminescence upon the application of R/Olig in all experiments shown on Figure 7. (B) Shows the effect of POA on bioluminescence. (C) Illustrates effect of POAEE on bioluminescence. (D) Effect of TLC-S on bioluminescence.
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9. Supplementary Figure 1
Pancreatic acinar cells retain polarized morphology, Ca2+ responses, mitochondrial membrane potential, and NAD(P)H responses following infection with replication deficient adenovirus. (A) Isolated doublet of pancreatic acinar cells following infection with a virus containing cytosolic luciferase (cLuc) and overnight incubation. Right panel shows transmitted light image, left panel shows confocal image of tetramethyl-rhodamine methyl ester (TMRM) fluorescence recorded from these cells. (B) Oscillatory [Ca2+]c response to 20 pM cholecystokinin-8 (CCK). In (B), (C), and (D), the fluorescence of Fluo 4 (F) was normalized to its fluorescence at the beginning of the experiment (F0). (C) [Ca2+]c response to 10 nM CCK. (D) Global long-lasting [Ca2+]c oscillations induced by 0.5 mM taurolithocholic acid 3-sulfate (TLC-S). (E) NAD(P)H response induced by 10 nM CCK. (F) NAD(P)H oscillations induced by TLC-S.
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10. Supplementary Figure 2
Effects of secretin, 8Br− adenosine 3′,5′-cyclic monophosphate (8Br-cAMP), VIP, and 8Br−guanosine 3′,5′-cyclic monophosphate (8Br-cGMP) on cytosolic adenosine triphosphate (ATP). Bioluminescence was recorded from cells transfected with cytosolic luciferase (cLuc). (A) Bioluminescence of pancreatic acinar cells before and after secretin application. (B) Bioluminescence of pancreatic acinar cells before and after 8Br-cAMP application. (C) VIP-induced decrease of bioluminescence of pancreatic acinar cells. (D) Bioluminescence of pancreatic acinar cells before and after 8Br-cGMP application.
Gastroenterology  , e5DOI: ( /j.gastro )
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