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- Berts, Alf, et al.
(författare)
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Oscillatory Ca2+ signaling in somatostatin-producing cells from the human pancreas
- 1997
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Ingår i: Metabolism. - 0026-0495 .- 1532-8600. ; 46:4, s. 366-369
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Tidskriftsartikel (refereegranskat)abstract
- Oscillatory Ca2+ signaling was studied in human somatostatin-releasing pancreatic δ cells identified by immunostaining. A ratiometric fura-2 technique was used for measuring cytoplasmic concentrations of Ca2+ and Sr2+ in δ cells exposed to the respective cation. Rhythmic activity in terms of slow (frequency, 0.1 to 0.4 per minute) oscillations from close to the basal level was seen in the presence of 3 to 20 mmol/L glucose during superfusion with medium containing 2.6 to 5 mmol/L Ca2+ or 5 mmol/L Sr2. These oscillations could be transformed into a sustained increase by decreasing extracellular Ca2+ or adding 1 mmol/L tolbutamide or 20 nmol/L glucagon. Addition of glucagon to a medium containing 20 mmol/L glucose resulted in the generation of short (< 30 seconds) transients, which disappeared upon exposure to 100 nmol/L of the intracellular Ca2+-adenosine triphosphatase (ATPase) inhibitor thapsigargin. When analyzing small aggregates of islet cells, it became evident that oscillatory activity in δ cells can be synchronous with that in adjacent non—δ cells. It is concluded that secretion of pancreatic somatostatin in man involves Ca2+ signaling similar to that regulating the pulsatile release of insulin.
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| 2. |
- Berts, Alf, et al.
(författare)
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Suppression of Ca2+ oscillations in glucagon-producing alfa2-cells by insulin/glucose and amino acids
- 1996
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Ingår i: Biochimica et Biophysica Acta. - 0006-3002 .- 1878-2434. ; 1310:2, s. 212-216
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Tidskriftsartikel (refereegranskat)abstract
- The cytoplasmic Ca2+ concentration ([Ca2+]i) was continuously monitored in single glucagon-producing α2-cells isolated from the mouse pancreas and later identified by immunostaining. Up to 60% of the α2-cells exhibited spontaneous [Ca2+]i oscillations (frequency 0.1–0.3/min) in a medium containing 3 mM glucose. In originating from a basal level of 60–100 nM, reaching peak values of 300–400 nM and promptly disappearing after blocking voltage-dependent Ca2+ channels with methoxyverapamil, the oscillations resembled those in insulin-releasing β-cells stimulated by glucose. The oscillatory activity was suppressed when combining elevation of glucose to 20 mM with the addition of 2–2000 ng/ml insulin. Whereas 10 mM of l-arginine or l-glycine transformed the oscillations into sustained elevation of [Ca2+];, there was no response to 1 mM tolbutamide or 0.1–1 mM γ-aminobutyric acid. The observations that α2-cells differ from islet cells secreting insulin and somatostatin in responding to adrenaline with mobilisation of intracellular calcium can be used for their rapid identification. It is suggested that the oscillations reflect periodic entry of Ca2+ due to variations of the membrane potential.
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| 4. |
- Bivehed, Erik, et al.
(författare)
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Visualizing DNA single- and double-strand breaks in the Flash comet assay by DNA polymerase-assisted end-labelling
- 2024
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Ingår i: Nucleic Acids Research. - : Oxford University Press. - 0305-1048 .- 1362-4962. ; 52:4
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Tidskriftsartikel (refereegranskat)abstract
- In the comet assay, tails are formed after single-cell gel electrophoresis if the cells have been exposed to genotoxic agents. These tails include a mixture of both DNA single-strand breaks (SSBs) and double-strand breaks (DSBs). However, these two types of strand breaks cannot be distinguished using comet assay protocols with conventional DNA stains. Since DSBs are more problematic for the cells, it would be useful if the SSBs and DSBs could be differentially identified in the same comet. In order to be able to distinguish between SSBs and DSBs, we designed a protocol for polymerase-assisted DNA damage analysis (PADDA) to be used in combination with the Flash comet protocol, or on fixed cells. By using DNA polymerase I to label SSBs and terminal deoxynucleotidyl transferase to label DSBs with fluorophore-labelled nucleotides. Herein, TK6-cells or HaCat cells were exposed to either hydrogen peroxide (H2O2), ionising radiation (X-rays) or DNA cutting enzymes, and then subjected to a comet protocol followed by PADDA. PADDA offers a wider detection range, unveiling previously undetected DNA strand breaks. Graphical Abstract
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| 9. |
- Gylfe, Erik, et al.
(författare)
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The Neurotransmitter ATP Triggers Ca2+ Responses Promoting Coordination of Pancreatic Islet Oscillations
- 2012
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Ingår i: Pancreas. - 0885-3177 .- 1536-4828. ; 41:2, s. 258-263
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Tidskriftsartikel (refereegranskat)abstract
- Objectives: Pulsatile insulin release into the portal vein is critically dependent on entrainment of the islets in the pancreas into a common oscillatory phase. Because the pulses reflect periodic variations of the cytoplasmic Ca2+ concentration ([Ca2+](i)), we studied whether the neurotransmitters adenosine triphosphate (ATP) and acetylcholine promote synchronization of [Ca2+](i) oscillations between islets lacking contact. Methods: Medium-sized and small mouse islets and cell aggregates were used for measuring [Ca2+](i) with the indicator fura-2. Results: Exposure to acetylcholine resulted in an initial [Ca2+](i) peak followed by disappearance of the [Ca2+](i) oscillations induced by 11-mmol/L glucose. The effect of ATP was often restricted to an elusive [Ca2+](i) peak. The incidence of distinct [Ca2+](i) responses to ATP increased under conditions (accelerated superfusion, small islets, or cell aggregates) intended to counteract purinoceptor desensitization owing to intercellular accumulation of ATP. Attempts to imitate neural activity by brief (15 seconds) exposure to ATP or acetylcholine resulted in temporary synchronization of the glucose-induced [Ca2+](i) oscillations between islets lacking contact. Conclusions: The data support the idea that purinergic signaling has a key role for coordinating the oscillatory activity of the islets in the pancreas, reinforcing previous arguments for the involvement of nonadrenergic, noncholinergic neurons.
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