inositol-1-4-5-trisphosphate and Diabetes-Mellitus--Type-2

Defects in pancreatic β-cell function and survival are key components in type 2 diabetes (T2D). An age-dependent deterioration in β-cell function has also been observed, but little is known about the molecular mechanisms behind this phenomenon. Our previous studies indicate that the regulation of cytoplasmic free Ca(2+) concentration ([Ca(2+)]i) may be critical and that this is dependent on the proper function of the mitochondria. The [Ca(2+)]i dynamics of the pancreatic β-cell are driven by an interplay between glucose-induced influx of extracellular Ca(2+) via voltage-dependent Ca(2+) channels and the inositol 1,4,5-trisphosphate (Ins(1,4,5)P3)-mediated liberation of Ca(2+) from intracellular stores. Our previous work has indicated a direct relationship between disruption of Ins(1,4,5)P3-mediated Ca(2+) regulation and loss of β-cell function, including disturbed [Ca(2+)]i dynamics and compromised insulin secretion. To investigate these processes in aging we used three mouse models, a premature aging mitochondrial mutator mouse, a mature aging phenotype (C57BL/6) and an aging-resistant phenotype (129). Our data suggest that age-dependent impairment in mitochondrial function leads to modest changes in [Ca(2+)]i dynamics in mouse β-cells, particularly in the pattern of [Ca(2+)]i oscillations. These changes are driven by modifications in both PLC/Ins(1,4,5)P3-mediated Ca(2+) mobilization from intracellular stores and decreased β-cell Ca(2+) influx over the plasma membrane. Our findings underscore an important concept, namely that even relatively small, time-dependent changes in β-cell signal-transduction result in compromised insulin release and in a diabetic phenotype.

Topics: Aging; Animals; Calcium; Calcium Signaling; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Humans; Inositol 1,4,5-Trisphosphate; Insulin-Secreting Cells; Mice

Excess consumption of carbohydrates, fat and calories leads to non-alcoholic fatty liver disease (NAFLD) and hepatic insulin resistance; these are major factors in the pathogenesis of type II diabetes. Hormones and catecholamines acting through G-protein coupled receptors (GPCRs) linked to phospholipase C (PLC) and increases in cytosolic Ca

Topics: Animals; Catecholamines; Diabetes Mellitus, Type 2; Diet, High-Fat; Glucagon; Hepatocytes; Inositol 1,4,5-Trisphosphate; Liver; Mice; Mice, Inbred C57BL; Non-alcoholic Fatty Liver Disease; Norepinephrine

The prevalence of acquired hyperlipidemia has increased due to sedentary life style and lipid-rich diet. In this work, a lipid-protein-protein interaction network (LPPIN) for acquired hyperlipidemia was prepared by incorporating differentially expressed genes in obese fatty liver as seed nodes, protein interactions from PathwayLinker, and lipid interactions from STITCH4.0. Cholesterol, diacylglycreol, phosphatidylinositol-bis-phosphate, and inositol triphosphate were identified as core lipids that influence the signaling pathways in the LPPIN. RACα serine/threonine-protein kinase (AKT1) was a highly essential central protein. The gastrin-CREB pathway was greatly enriched; all enriched pathways in the LPPIN showed crosstalk with the phosphatidylinositol-3-kinase-Akt pathway, correlating with the central role of AKT1 in the network. The disease clusters identified in the LPPIN were cardiovascular disease, cancer, Alzheimer's disease, and Type II diabetes. In this context, we note that the commercially approved drug targets for hyperlipidemia in each disease cluster may potentially be repurposed for treatment of the specific disease. We report here top 10 potential drug targets that may mediate progression from hyperlipidemia to the respective disease state. ToppGene Suite was employed to identify candidates followed by a) discarding high closeness centrality nodes, and b) selecting nodes with high bridging centrality. Three potential targets could be mapped to specific disease clusters in the LPPIN. Lipids associated with acquired hyperlipidemia and each disease cluster identified may be useful as prognostic fingerprints. These findings provide an integrative view of lipid-protein interactions leading to acquired hyperlipidemia and the associated diseases, and might prove useful in future translational pharmaceutical research.

Topics: Alzheimer Disease; Apolipoprotein B-100; Cardiovascular Diseases; Carrier Proteins; Cholesterol; Diabetes Mellitus, Type 2; Dietary Fats; Diglycerides; Disease Progression; Gene Expression Regulation; Humans; Hyperlipidemias; Hypolipidemic Agents; Inositol 1,4,5-Trisphosphate; Molecular Targeted Therapy; Phosphatidylinositol Phosphates; PPAR alpha; Proprotein Convertase 9; Protein Interaction Mapping; Proto-Oncogene Proteins c-akt; Signal Transduction

The present study was conducted to determine whether and how store-operated Ca(2+) entry (SOCE) in glomerular mesangial cells (MCs) was altered by high glucose (HG) and diabetes. Human MCs were treated with either normal glucose or HG for different time periods. Cyclopiazonic acid-induced SOCE was significantly greater in the MCs with 7-day HG treatment and the response was completely abolished by GSK-7975A, a selective inhibitor of store-operated Ca(2+) channels. Similarly, the inositol 1,4,5-trisphosphate-induced store-operated Ca(2+) currents were significantly enhanced in the MCs treated with HG for 7 days, and the enhanced response was abolished by both GSK-7975A and La(3+). In contrast, receptor-operated Ca(2+) entry in MCs was significantly reduced by HG treatment. Western blotting showed that HG increased the expression levels of STIM1 and Orai1 in cultured MCs. A significant HG effect occurred at a concentration as low as 10 mM, but required a minimum of 7 days. The HG effect in cultured MCs was recapitulated in renal glomeruli/cortex of both type I and II diabetic rats. Furthermore, quantitative real-time RT-PCR revealed that a 6-day HG treatment significantly increased the mRNA expression level of STIM1. However, the expressions of STIM2 and Orai1 transcripts were not affected by HG. Taken together, these results suggest that HG/diabetes enhanced SOCE in MCs by increasing STIM1/Orai1 protein expressions. HG upregulates STIM1 by promoting its transcription but increases Orai1 protein through a posttranscriptional mechanism.

Topics: Animals; Calcium Channel Agonists; Calcium Channel Blockers; Calcium Channels; Calcium Signaling; Cells, Cultured; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Glucose; Humans; Inositol 1,4,5-Trisphosphate; Ion Channel Gating; Male; Membrane Glycoproteins; Membrane Proteins; Mesangial Cells; Neoplasm Proteins; ORAI1 Protein; Rats; Rats, Sprague-Dawley; RNA Processing, Post-Transcriptional; RNA, Messenger; Stromal Interaction Molecule 1; Time Factors; Transcriptional Activation; Up-Regulation

Little is known about the molecular mechanisms underlying age-dependent deterioration in β-cell function. We now demonstrate that age-dependent impairment in insulin release, and thereby glucose homeostasis, is associated with subtle changes in Ca(2+) dynamics in mouse β-cells. We show that these changes are likely to be accounted for by impaired mitochondrial function and to involve phospholipase C/inositol 1,4,5-trisphosphate-mediated Ca(2+) mobilization from intracellular stores as well as decreased β-cell Ca(2+) influx over the plasma membrane. We use three mouse models, namely, a premature aging phenotype, a mature aging phenotype, and an aging-resistant phenotype. Premature aging is studied in a genetically modified mouse model with an age-dependent accumulation of mitochondrial DNA mutations. Mature aging is studied in the C57BL/6 mouse, whereas the 129 mouse represents a model that is more resistant to age-induced deterioration. Our data suggest that aging is associated with a progressive decline in β-cell mitochondrial function that negatively impacts on the fine tuning of Ca(2+) dynamics. This is conceptually important since it emphasizes that even relatively modest changes in β-cell signal transduction over time lead to compromised insulin release and a diabetic phenotype.

Topics: Aging; Animals; Blood Glucose; Calcium; Diabetes Mellitus, Type 2; Electron Transport; Inositol 1,4,5-Trisphosphate; Insulin-Secreting Cells; Mice; Mice, Inbred C57BL; Mitochondria; Type C Phospholipases

This study examines the extent to which the antiapoptotic Bcl-2 proteins Bcl-2 and Bcl-x(L) contribute to diabetic Ca(2+) dysregulation and vessel contractility in vascular smooth muscle cells (VSMCs) through their interaction with inositol 1,4,5-trisphosphate receptor (InsP(3)R) intracellular Ca(2+) release channels. Measurements of intracellular ([Ca(2+)](i)) and sarcoplasmic reticulum ([Ca(2+)](SR)) calcium concentrations were made in primary cells isolated from diabetic (db/db) and nondiabetic (db/m) mice. In addition, [Ca(2+)](i) and constriction were recorded simultaneously in isolated intact arteries. Protein expression levels of Bcl-x(L) but not Bcl-2 were elevated in VSMCs isolated from db/db compared with db/m age-matched controls. In single cells, InsP(3)-evoked [Ca(2+)](i) signaling was enhanced in VSMCs from db/db mice compared with db/m. This was attributed to alterations in the intrinsic properties of the InsP(3)R itself because there were no differences between db/db and db/m in the steady-state [Ca(2+)](SR) or InsP(3)R expression levels. Moreover, in permeabilized cells the rate of InsP(3)R-dependent SR Ca(2+) release was increased in db/db compared with db/m VSMCs. The enhanced InsP(3)-dependent SR Ca(2+) release was attenuated by the Bcl-2 protein inhibitor ABT-737 only in diabetic cells. Application of ABT-737 similarly attenuated enhanced agonist-induced [Ca(2+)](i) signaling only in intact aortic and mesenteric db/db vessels. In contrast, ABT-737 had no effect on agonist-evoked contractility in either db/db or db/m vessels. Taken together, the data suggest that in type 2 diabetes the mechanism for [Ca(2+)](i) dysregulation in VSMCs involves Bcl-2 protein-dependent increases in InsP(3)R excitability and that dysregulated [Ca(2+)](i) signaling does not appear to contribute to increased vessel reactivity.

Topics: Animals; Aorta, Thoracic; bcl-X Protein; Biphenyl Compounds; Blood Glucose; Calcium; Calcium Signaling; Cells, Cultured; Diabetes Mellitus, Type 2; Diabetic Angiopathies; Disease Models, Animal; Dose-Response Relationship, Drug; Homeostasis; Inositol 1,4,5-Trisphosphate; Inositol 1,4,5-Trisphosphate Receptors; Male; Mice; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Nitrophenols; Piperazines; Proto-Oncogene Proteins c-bcl-2; Sarcoplasmic Reticulum; Sulfonamides; Time Factors; Up-Regulation; Vasoconstriction

In this study we have investigated muscarinic M₁, M₃ receptor kinetics and the functional role of IP3 and cGMP in the corpus striatum of both young and old diabetic and insulin-treated diabetic rats.. Radioreceptor binding assays was done in the corpus striatum using specific antagonists QNB and DAMP. IP3 and cGMP assay using [3H]IP3 and [3H]cGMP Biotrak assay system kits.. M₁ receptor increased and M₃ receptor decreased in control old rats when compared with young control rats. In young diabetic groups M₁ receptor increased and M₃ receptor decreased. Old diabetic groups showed reversed M₁ and M₃ receptors compared with their controls. IP3 and cGMP content increased in old control rats compared with young control rats. IP3 content increased in young diabetic rats and decreased in old diabetic rats. cGMP content was increased significantly in both young and old diabetic groups. Insulin treatment reversed these altered parameters near to control.. Our studies showed that M₁ and M₃ receptors, IP3 and cGMP were functionally regulated during diabetes as function of age, which will have immense clinical significance.

Topics: Age Factors; Animals; Blood Glucose; Body Weight; Carrier Proteins; Corpus Striatum; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Inositol 1,4,5-Trisphosphate; Insulin; Intracellular Signaling Peptides and Proteins; Male; Rats; Rats, Wistar; Receptor, Muscarinic M1; Receptor, Muscarinic M3; Streptozocin