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• W2110953604 endingPage "1820" @default.
• W2110953604 startingPage "1813" @default.
• W2110953604 abstract "The purpose of the present investigation was to establish a method for estimating intracellular Ca 2+ concentrations ([Ca 2+ ] i ) in isolated rat epitrochlearis muscles. Epitrochlearis muscles excised from 4-wk-old male Sprague-Dawley rats were loaded with a fluorescent Ca 2+ indicator, fura 2-AM, for 60–90 min at 35°C in oxygenated Krebs-Henseleit buffer. After fura 2 loading and subsequent 20-min incubation, the intensities of 500-nm fluorescence, induced by 340- and 380-nm excitation lights (F total 340 and F total 380), were measured. The fluorescences specific to fura-2 (F fura 2 340 and F fura 2 380) were calculated by subtracting the non-fura 2-specific component from F total 340 and F total 380, respectively. The ratio of F fura 2 340 to F fura 2 380 was calculated as R, and the change in the ratio from the baseline value (ΔR) was used as an index of the change in [Ca 2+ ] i . In resting muscle, ΔR was stable for 60 min. Incubation for 20 min with caffeine (3–10 mM) significantly increased ΔR in a concentration-dependent manner. Incubation with hypoxic Krebs-Henseleit buffer for 10–60 min significantly elevated ΔR, depending on the duration of the incubation. Incubation with 50 μM N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide for 20 min significantly elevated ΔR ( P < 0.05). No significant increases in ΔR were observed during incubation for 20 min with 2 mM 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside or with 2 mU/ml insulin. These results demonstrated that, by using the fura 2-AM fluorescence method, the changes in [Ca 2+ ] i can be monitored in the rat epitrochlearis muscle and suggest that the method can be utilized to observe quantitative information regarding [Ca 2+ ] i that may be involved in contraction- and hypoxia-stimulated glucose transport activity in skeletal muscle." @default.
• W2110953604 created "2016-06-24" @default.
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• W2110953604 date "2003-05-01" @default.
• W2110953604 modified "2023-10-15" @default.
• W2110953604 title "Changes in [Ca²⁺]_iinduced by several glucose transport-enhancing stimuli in rat epitrochlearis muscle" @default.
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• W2110953604 doi "https://doi.org/10.1152/japplphysiol.00780.2002" @default.
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