| 1. |
- Taneera, Jalal, et al.
(författare)
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A Systems Genetics Approach Identifies Genes and Pathways for Type 2 Diabetes in Human Islets
- 2012
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Ingår i: Cell Metabolism. - : Elsevier BV. - 1550-4131 .- 1932-7420. ; 16:1, s. 122-134
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Tidskriftsartikel (refereegranskat)abstract
- Close to 50 genetic loci have been associated with type 2 diabetes (T2D), but they explain only 15% of the heritability. In an attempt to identify additional T2D genes, we analyzed global gene expression in human islets from 63 donors. Using 48 genes located near T2D risk variants, we identified gene coexpression and protein-protein interaction networks that were strongly associated with islet insulin secretion and HbA(1c). We integrated our data to form a rank list of putative T2D genes, of which CHL1, LRFN2, RASGRP1, and PPM1K were validated in INS-1 cells to influence insulin secretion, whereas GPR120 affected apoptosis in islets. Expression variation of the top 20 genes explained 24% of the variance in HbA(1c) with no claim of the direction. The data present a global map of genes associated with islet dysfunction and demonstrate the value of systems genetics for the identification of genes potentially involved in T2D.
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| 2. |
- Eliasson, Lena, et al.
(författare)
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SUR1 Regulates PKA-independent cAMP-induced Granule Priming in Mouse Pancreatic B-cells.
- 2003
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Ingår i: Journal of General Physiology. - : Rockefeller University Press. - 0022-1295 .- 1540-7748. ; 121:3, s. 181-197
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Tidskriftsartikel (refereegranskat)abstract
- Measurements of membrane capacitance were applied to dissect the cellular mechanisms underlying PKA-dependent and -independent stimulation of insulin secretion by cyclic AMP. Whereas the PKA-independent (Rp-cAMPS–insensitive) component correlated with a rapid increase in membrane capacitance of ~80 fF that plateaued within ~200 ms, the PKA-dependent component became prominent during depolarizations >450 ms. The PKA-dependent and -independent components of cAMP-stimulated exocytosis differed with regard to cAMP concentration dependence; the Kd values were 6 and 29 µM for the PKA-dependent and -independent mechanisms, respectively. The ability of cAMP to elicit exocytosis independently of PKA activation was mimicked by the selective cAMP-GEFII agonist 8CPT-2Me-cAMP. Moreover, treatment of B-cells with antisense oligodeoxynucleotides against cAMP-GEFII resulted in partial (50%) suppression of PKA-independent exocytosis. Surprisingly, B-cells in islets isolated from SUR1-deficient mice (SUR1-/- mice) lacked the PKA-independent component of exocytosis. Measurements of insulin release in response to GLP-1 stimulation in isolated islets from SUR1-/- mice confirmed the complete loss of the PKA-independent component. This was not attributable to a reduced capacity of GLP-1 to elevate intracellular cAMP but instead associated with the inability of cAMP to stimulate influx of Cl- into the granules, a step important for granule priming. We conclude that the role of SUR1 in the B cell extends beyond being a subunit of the plasma membrane KATP-channel and that it also plays an unexpected but important role in the cAMP-dependent regulation of Ca2+-induced exocytosis.
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| 3. |
- Härndahl, Linda, et al.
(författare)
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Important role of phosphodiesterase 3B for the stimulatory action of cAMP on pancreatic beta -cell exocytosis and release of insulin.
- 2002
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 277:40, s. 37446-37455
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Tidskriftsartikel (refereegranskat)abstract
- Cyclic AMP potentiates glucose-stimulated insulin release and mediates the stimulatory effects of hormones such as glucagon-like peptide 1 (GLP-1) on pancreatic b-cells. By inhibition of cAMP-degrading phosphodiesterase (PDE) and, in particular, selective inhibition of PDE3 activity, stimulatory effects on insulin secretion have been observed. Molecular and functional information on b-cell PDE3 is, however, scarce. To provide such information, we have studied the specific effects of the PDE3B isoform by adenovirus-mediated overexpression. In rat islets and rat insulinoma cells, approximate 10-fold overexpression of PDE3B was accompanied by a 6-8-fold increase in membrane-associated PDE3B activity. The cAMP concentration was significantly lowered in transduced cells (INS-1(832/13), and insulin secretion in response to stimulation with high glucose (11.1 mM) was reduced by 40% (islets) and 50% (INS-1). Further, the ability of GLP-1 (100 nM) to augment glucose-stimulated insulin secretion was inhibited by approximately 30% (islets) and 70% (INS-1). Accordingly, when stimulating with cAMP, a substantial decrease (65%) in exocytotic capacity was demonstrated in patch-clamped single b-cells. In untransduced insulinoma cells, application of the PDE3-selective inhibitor OPC3911 (10 mM) was shown to increase glucose-stimulated insulin release as well as cAMP-enhanced exocytosis. The findings suggest a significant role of PDE3B as an important regulator of insulin secretory processes.
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| 4. |
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| 5. |
- Ivarsson, Rosita, et al.
(författare)
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Myosin 5a controls insulin granule recruitment during late-phase secretion.
- 2005
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Ingår i: Traffic: the International Journal of Intracellular Transport. - : Wiley. - 1398-9219. ; 6:11, s. 1027-1035
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Tidskriftsartikel (refereegranskat)abstract
- We have examined the importance of the actin-based molecular motor myosin 5a for insulin granule transport and insulin secretion. Expression of myosin 5a was downregulated in clonal INS-1E cells using RNAinterference. Stimulated hormone secretion was reduced by 46% and single-cell exocytosis, measured by capacitance recordings, was inhibited by 42% after silencing. Silencing of Slac-2c/MYRIP, which links insulin granules to myosin 5a, resulted in similar inhibition of single-cell exocytosis. Antibody inhibition of the myosin 5a-Slac-2c/MYRIP interaction significantly reduced the recruitment of insulin granules for release. The pool of releasable granules independent of myosin 5a activity was estimated to approximately 550 granules. Total internal reflection microscopy was then applied to directly investigate granule recruitment to the plasma membrane. Silencing of myosin 5a inhibited granule recruitment during late phase of insulin secretion. In conclusion, we propose a model where insulin granules are transported through the actin network via both myosin 5a-mediated transport and via passive diffusion, with the former playing the major role during stimulatory conditions.
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| 6. |
- Jing, Cuiping, et al.
(författare)
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?2NC: Redundant and Random Network Codingfor Robust H.264/SVC Transmission
- 2011
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Ingår i: 14th IEEE International Conference on Network-Based Information Systems (NBiS), 2011. - : IEEE Computer Society. - 9781457707896 - 9780769544588 ; , s. 634-639
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Konferensbidrag (refereegranskat)abstract
- In this paper we are interested in improving the performance of constructive network coding schemes for video transmission over packet lossy networks. A novel unequal packet loss protection scheme R2NC based on low-triangular global coding matrix with ladder-shaped partition will be presented, which combines redundant and random network coding for robust H.264/SVC video transmission. Firstly, the error-correcting capabilities of redundant network coding make our scheme resilient to loss. Secondly, the implementation of random network coding at the intermediate nodes with multiple input links can reduce the cost of network bandwidth, thus reducing the end to end delay for video transmission. Thirdly, the low-triangular global coding matrix with ladder-shaped partition is maintained throughout R2NC process to provide unequal erasure protection for H.264/SVC priority layers. The redundant network coding avoids the retransmission of lost packets and improves error correcting capabilities of lost packets. Based only on the knowledge of the loss rates on the output links, the source node and intermediate nodes can make decisions for redundant network coding and random network coding (i.e., how much redundancy to add at this node). However, the redundancy caused by redundant network coding makes the network load increases, in order to improve network throughput, we perform random network coding at the intermediate nodes. Our approach is grounded on the overall distortion of reconstructed video minimization by optimizing the amount of redundancy assigned to each layer. Experimental results are shown to demonstrate the significant improvement of H.264/SVC video reconstruction quality with R2NC over packet lossy networks.
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| 7. |
- Jing, Xingjun, et al.
(författare)
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CaV2.3 calcium channels control second-phase insulin release.
- 2005
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Ingår i: Journal of Clinical Investigation. - 0021-9738. ; 115:1, s. 146-154
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Tidskriftsartikel (refereegranskat)abstract
- Concerted activation of different voltage-gated Ca2+ channel isoforms may determine the kinetics of insulin release from pancreatic islets. Here we have elucidated the role of R-type CaV2.3 channels in that process. A 20% reduction in glucose-evoked insulin secretion was observed in CaV2.3-knockout (CaV2.3–/–) islets, close to the 17% inhibition by the R-type blocker SNX482 but much less than the 77% inhibition produced by the L-type Ca2+ channel antagonist isradipine. Dynamic insulin-release measurements revealed that genetic or pharmacological CaV2.3 ablation strongly suppressed second-phase secretion, whereas first-phase secretion was unaffected, a result also observed in vivo. Suppression of the second phase coincided with an 18% reduction in oscillatory Ca2+ signaling and a 25% reduction in granule recruitment after completion of the initial exocytotic burst in single CaV2.3–/– ß cells. CaV2.3 ablation also impaired glucose-mediated suppression of glucagon secretion in isolated islets (27% versus 58% in WT), an effect associated with coexpression of insulin and glucagon in a fraction of the islet cells in the CaV2.3–/– mouse. We propose a specific role for CaV2.3 Ca2+ channels in second-phase insulin release, that of mediating the Ca2+ entry needed for replenishment of the releasable pool of granules as well as islet cell differentiation.
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| 8. |
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| 9. |
- Jing, Xingjun
(författare)
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Ion channel control of phasic insulin secretion.
- 2007
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Glucose-stimulated insulin secretion exhibits a biphasic pattern. The mechanism underling biphasic insulin secretion is not fully understood, but consensus exists that an elevation in [Ca2+]i is required for both first- and second-phase insulin secretion. The molecular identity of the pancreatic β-cell L-type Ca2+ channel has not been established and it has variably been reported to be CaV1.2 (α1C) or CaV1.3 (α1D). Though the cellular background to the two phases of release remains unknown, it has been suggested to reflect the sequential release of distinct pools of granules, which vary with regard to release competence. This thesis investigated the role of different ion channels in insulin secretion. β-cell-selective ablation of the CaV1.2 gene(βCaV1.2-/- mouse) decreased the whole-cell Ca2+ current by only ~45%, but almost abolished first-phase insulin secretion and resulted in systemic glucose intolerance. High-resolution capacitance measurements of exocytosis in single β-cells revealed that the loss of first-phase insulin secretion in the βCaV1.2-/- mouse was associated with the disappearance of a rapid component of exocytosis reflecting fusion of secretory granules physically attached to the CaV1.2 channel. A 20% reduction in glucose-evoked insulin secretion was observed in CaV2.3-knockout (CaV2.3–/–) islets, close to the 17% inhibition by the R-type blocker SNX482. Genetic or pharmacological CaV2.3 ablation strongly suppressed second-phase secretion in vitro, as well as in vivo, whereas first-phase secretion was unaffected. Suppression of the second phase coincided with an 18% reduction in oscillatory Ca2+ signaling and a 25% reduction in granule recruitment after completion of the initial exocytotic burst in single CaV2.3–/– β-cells. Intracellular ClC-3 chloride channels have been implicated in the process of making insulin granules release-competent, a process referred to as priming. Analysis of insulin secretion in vivo and in vitro as well as capacitance measurements revealed that the secretory response of ClC-3 deficient β-cells was reduced, but not abolished. The presence of ClC-3 in insulin granules was detected in a high-purification fraction of LDCVs obtained by phogrin-GFP labelling. In conclusion: (1) CaV1.2 Ca2+ channels are required for first-phase insulin release and maintenance of systemic glucose tolerance. (2) CaV2.3 Ca2+ channels play an important role in second-phase insulin release. (3) ClC-3 chloride channels facilitate insulin secretion by enhancing properly acidification of insulin granules needed for granule priming.
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| 10. |
- Li, Dai-Qing, et al.
(författare)
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Suppression of sulfonylurea- and glucose-induced insulin secretion in vitro and in vivo in mice lacking the chloride transport protein ClC-3.
- 2009
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Ingår i: Cell metabolism. - : Elsevier BV. - 1932-7420 .- 1550-4131. ; 10:4, s. 309-15
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Tidskriftsartikel (refereegranskat)abstract
- Priming of insulin secretory granules for release requires intragranular acidification and depends on vesicular Cl(-)-fluxes, but the identity of the chloride transporter/ion channel involved is unknown. We tested the hypothesis that the chloride transport protein ClC-3 fulfills these actions in pancreatic beta cells. In ClC-3(-/-) mice, insulin secretion evoked by membrane depolarization (high extracellular K(+), sulfonylureas), or glucose was >60% reduced compared to WT animals. This effect was mirrored by a approximately 80% reduction in depolarization-evoked beta cell exocytosis (monitored as increases in cell capacitance) in single ClC-3(-/-) beta cells, as well as a 44% reduction in proton transport across the granule membrane. ClC-3 expression in the insulin granule was demonstrated by immunoblotting, immunostaining, and negative immuno-EM in a high-purification fraction of large dense-core vesicles (LDCVs) obtained by phogrin-EGFP labeling. The data establish the importance of granular Cl(-) fluxes in granule priming and provide direct evidence for the involvement of ClC-3 in the process.
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