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New insights into cytosolic glucose levels during differentiation of 3T3-L1 fibroblasts into adipocytesKovačič, Petra Brina, nevrobiologinja ...Cytosolic glucose concentration reflects the balance between glucose entry across the plasma membrane and cytosolic glucose utilization. In adipocytes, glucose utilization is considered very rapid, ... meaning that every glucose molecule entering the cytoplasm is quickly phosphorylated. Thus, the cytosolicfree glucose concentration is considered to be negligible; however, it was never measured directly. In the present study, we monitored cytosolic glucose dynamics in 3T3-L1 fibroblasts and adipocytes by expressing a fluorescence resonance energy transfer (FRET)-based glucose nanosensor: fluorescent indicator protein FLIPglu-600mu. Specifically, we monitored cytosolic glucose responses by varying transmembrane glucose concentration gradient. The changes in cytosolic glucose concentration were detected in only56% of 3T3-L1 fibroblasts and in 14% of 3T3-L1 adipocytes. In adipocytes, the resting cytosolic glucose concentration was reduced in comparison with theone recorded in fibroblasts. Membrane permeabilization increased cytosolic glucose concentration in adipocytes, and glycolytic inhibitor iodoacetate failed to increase cytosolic glucose concentration, indicating low adipocyte permeability for glucose at rest. We also examined the effects of insulin and adrenaline. Insulin significantly increased cytosolic glucose concentration inadipocytes by a factor of 3.6; however, we recorded no effect on delta ratio(DeltaR) in fibroblasts. Adrenaline increased cytosolic glucose concentration in fibroblasts but not in adipocytes. However, in adipocytes in insulin-stimulated conditions, glucose clearance was significantly faster following adrenaline addition in comparison with controls (p < 0.001). Together, these results demonstrate that during differentiation, adipocytes develop more efficient mechanisms for maintaining low cytosolic glucose concentration, predominantly with reduced membrane permeability for glucose.Vir: The Journal of biological chemistry. - ISSN 0021-9258 (Vol. 286, no. 15, 2011, str. 13370-13381)Vrsta gradiva - članek, sestavni delLeto - 2011Jezik - angleškiCOBISS.SI-ID - 28263129
Avtor
Kovačič, Petra Brina, nevrobiologinja |
Chowdhury Haque, Helena |
Velebit Marković, Jelena |
Kreft, Marko |
Jensen, Jørgen, fizik |
Zorec, Robert, 1958-
Teme
Fibroblasts |
Adipocytes |
Cytosol |
Glucose |
Cell Membrane Permeability |
Fluorescent Dyes |
Epinephrine |
Escin |
Deoxyglucose |
Energy Metabolism |
Immunohistochemistry |
Microscopy, Fluorescence |
Cells, Cultured |
Adipociti |
Citosol |
Celična membrana, propustnost |
Glukoza |
Fibroblasti |
Escin |
Energetska presnova |
Imunohistokemija |
Deoksiglukoza |
Adrenalin |
Celične kulture |
Fluorescentna barvila |
Mikroskopija fluorescenčna
Vnos na polico
Trajna povezava
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Kovačič, Petra Brina, nevrobiologinja | 28638 |
Chowdhury Haque, Helena | 18548 |
Velebit Marković, Jelena | 32525 |
Kreft, Marko | 15666 |
Jensen, Jørgen, fizik | |
Zorec, Robert, 1958- | 03702 |
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